I am a neurobiologist working on the pathogenesis of Down syndrome and Alzheimer’s disease. All adults with Down syndrome will develop Alzheimer disease neuropathology by age 40 years. We have been involved in identifying neuronal networks that undergo significant degeneration in mouse models of Down syndrome and Alzheimer’s disease. Currently, my group is working on the role of norepinephrinergic system in failed contextual learning in the Ts65Dn mouse model of Down syndrome. We are hoping that improving this system would restore cognitive function in children and reduce the severity of Alzheimer pathology in adults with Down syndrome. In a double-blind placebo-controlled clinical trial, we are currently testing whether improved beta2 adrenergic signaling can improve cognitive function in individuals with mild to moderate dementia of Alzheimer type.
Deleterious Effects of Chronic Folate Deficiency in the Ts65Dn Mouse Model of Down Syndrome
Susan Helm, Morgnan Blayney, Taylor Whited, Mahjabin Nootozi, Sen Lin, Semira Kern, David Green, and Ahmad Salehi
Folate is an important B vitamin naturally found in the human diet and plays a critical role in methylation of nucleic acids. Indeed, abnormalities in this major epigenetic mechanism play a pivotal role in the pathogenesis of cognitive deficit and intellectual disability in humans. The most common cause of cognitive dysfunction in children is Down syndrome (DS). Since folate deficiency is very common among the pediatric population, we questioned whether chronic folate deficiency (CFD) exacerbates cognitive dysfunction in a mouse model of DS. To test this, adult Ts65Dn mice and their disomic littermates were chronically fed a diet free of folic acid while preventing endogenous production of folate in the digestive tract for a period of 8 weeks. Our results show that the Ts65Dn mouse model of DS was significantly more vulnerable to CFD in terms of plasma homocysteine and N5-methyltetrahydrofolate (5-MTHF) levels. Importantly, these changes were linked to degenerative alterations in hippocampal dendritic morphology and impaired nest building behavior in Ts65Dn mice. Based on our results, a rigorous examination of folate intake and its metabolism in individuals with DS is warranted.
Books Series [Book]
Common Pathogenic Mechanisms between Down Syndrome and Alzheimer’s Disease: Steps toward Therapy Book Series: Recent Advances in Alzheimer Research
Edited by Ahmad Salehi, Cristy Phillips, and Michael Rafii
GABAergic Hyperinnervation of Dentate Granule Cells in the Ts65Dn Mouse Model of Down Syndrome: Exploring the Role of App
Fatemeh.S Mojabi, Atoossa Fahimi, Shahrzad Moghadam, Sarah Moghadam, M.Windy McNerney, Ravikumar Ponnusamy, Alexander Kelschevnikov, William C Mobley, and Ahmad Salehi
It has been suggested that increased GABAergic innervation
in the hippocampus plays a significant role in cognitive dysfunction
in Down syndrome (DS). Bolstering this notion, are studies linking hyperinnervation
of the dentate gyrus (DG) by GABAergic terminals to failure
in LTP induction in the Ts65Dn mouse model of DS. Here, we used
extensive morphometrical methods to assess the status of GABAergic
interneurons in the DG of young and old Ts65Dn mice and their 2N controls.
We detected an age-dependent increase in GABAergic innervation
of dentate granule cells (DGCs) in Ts65Dn mice. The primary source of
GABAergic terminals to DGCs somata is basket cells (BCs). For this reason,
we assessed the status of these cells and found a significant increase
in the number of BCs in Ts65Dn mice compared with controls. Then we
aimed to identify the gene/s whose overexpression could be linked to
increased number of BCs in Ts65Dn and found that deleting the third
copy of App gene in Ts65Dn mice led to normalization of the number of
BCs in these mice. Our data suggest that App overexpression plays a
major role in the pathophysiology of GABAergic hyperinnervation of the
DG in Ts65Dn mice
Brain Structure and Function
Physical exercise induces structural alterations in the hippocampal astrocytes: exploring the role of BDNF-TrkB signaling
Atoossa Fahimi, Mehmet Akif Bkhtir, Sarah Moghadam, Fatemeh Mojabi, Krithika Sumanth, M. Windy McNerney, Ravikumar Ponnusamy, and Ahmad Salehi
While it has been known that physical activity can improve cognitive function and protect against neurodegeneration, the underlying mechanisms for these protective effects are yet to be fully elucidated. There is a large body of evidence indicating that physical exercise improves neurogenesis and maintenance of neurons. Yet, its possible effects on glial cells remain poorly understood. Here, we tested whether physical exercise in mice alters the expression of trophic factor-related genes and the status of astrocytes in the dentate gyrus of the hippocampus. In addition to a significant increase in Bdnf mRNA and protein levels, we found that 4 weeks of treadmill and running wheel exercise in mice, led to (1) a significant increase in synaptic load in the dentate gyrus, (2) alterations in astrocytic morphology, and (3) orientation of astrocytic projections towards dentate granule cells. Importantly, these changes were possibly linked to increased TrkB receptor levels in astrocytes. Our study suggests that astrocytes actively respond and could indeed mediate the positive effects of physical exercise on the central nervous system and potentially counter degenerative processes during aging and neurodegenerative disorders.
Curr Alzheimer Res
Curr Alzheimer Res (cont.)
Noradrenergic System in Down Syndrome and Alzheimer’s Disease A Target for Therapy.
C Phillips, A Fahimi, FS Das, D Mojabi, R Ponnusamy, and A Salehi
Locus coeruleus (LC) neurons in the brainstem send extensive noradrenergic (NE)-ergic terminals to the majority of brain regions, particularly those involved in cognitive function. Both Alzheimer's disease (AD) and Down syndrome (DS) are characterized by similar pathology including significant LC degeneration and dysfunction of the NE-ergic system. Extensive loss of NE-ergic terminals has been linked to alterations in brain regions vital for cognition, mood, and executive function. While the mechanisms by which NE-ergic abnormalities contribute to cognitive dysfunction are not fully understood, emergent evidence suggests that rescue of NE-ergic system can attenuate neuropathology and cognitive decline in both AD and DS. Therapeutic strategies to enhance NE neurotransmission have undergone limited testing. Among those deployed to date are NE reuptake inhibitors, presynaptic α-adrenergic receptor antagonists, NE prodrugs, and β-adrenergic agonists. Here we examine alterations in the NE-ergic system in AD and DS and suggest that NE-ergic system rescue is a plausible treatment strategy for targeting cognitive decline in both disorders.
A Special Regenerative Rehabilitation and Genomics Letter: Is There a “Hope” Molecule?
and Cristy Phillips
Further validation to support clinical translation of [(18)F]FTC-146 for imaging sigma-1 receptors.
B Shen, ML James, L Andrews, C Lau, S Chen, M Palner, Z Miao, NC Arksey, AJ Shuhendler, S Scatliffe, K Kaneshige, SM Parsons, CR McCurdy, A Salehi, SS Gambhir, and FT Chin
This study aims to further evaluate the specificity and selectivity of [(18)F]FTC-146 and obtain additional data to support its clinical translation. The binding of [(19)F]FTC-146 to vesicular acetylcholine transporter (VAChT) was evaluated using [(3)H]vesamicol and PC12(A123.7) cells in an in vitro binding assay. The uptake and kinetics of [(18)F]FTC-146 in S1R-knockout mice (S1R-KO) compared to wild-type (WT) littermates was assessed using dynamic positron emission tomography (PET) imaging. Ex vivo autoradiography and histology were conducted using a separate cohort of S1R-KO/WT mice, and radiation dosimetry was calculated from WT mouse data (extrapolated for human dosing). Toxicity studies in Sprague-Dawley rats were performed with a dose equivalent to 250× the anticipated clinical dose of [(19)F]FTC-146 mass. VAChT binding assay results verified that [(19)F]FTC-146 displays negligible affinity for VAChT (K i = 450 ± 80 nM) compared to S1R. PET images demonstrated significantly higher tracer uptake in WT vs. S1R-KO brain (4.57 ± 1.07 vs. 1.34 ± 0.4 %ID/g at 20-25 min, n = 4, p < 0.05). In S1R-KO mice, it was shown that rapid brain uptake and clearance 10 min post-injection, which are consistent with previous S1R-blocking studies in mice. Three- to fourfold higher tracer uptake was observed in WT relative to S1R-KO mouse brains by ex vivo autoradiography. S1R staining coincided well with the autoradiographic data in all examined brain regions (r (2) = 0.85-0.95). Biodistribution results further demonstrated high [(18)F]FTC-146 accumulation in WT relative to KO mouse brain and provided quantitative information concerning tracer uptake in S1R-rich organs (e.g., heart, lung, pancreas) for WT mice vs. age-matched …
EJNMMI Res. (cont.)
S1R-KO mice. The maximum allowed dose per scan in humans as extrapolated from mouse dosimetry was 33.19 mCi (1228.03 MBq). No significant toxicity was observed even at a 250X dose of the maximum carrier mass [(19)F]FTC-146 expected to be injected for human studies.Together, these data indicate that [(18)F]FTC-146 binds specifically to S1Rs and is a highly promising radiotracer ready for clinical translation to investigate S1R-related diseases.
Increased Incidence of Intermittent Hypoxemia in the Ts65Dn Mouse Model of Down Syndrome.
Devsmita Das, Brian Medina, Mehmet Baktir, F Mojabi, Atoossa Fahimi, Ravikumar Ponnusamy, and Ahmad Salehi
In addition to nervous system, cardiovascular and respiratory systems are primarily affected in Down syndrome (DS). The Ts65Dn mouse model is widely used to recapitulate cognitive dysfunction in DS. While these mice consistently show failure in learning and memory along with functional and structural abnormalities in the hippocampal region, the underlying mechanisms behind cognitive dysfunction remain to be fully elucidated. Convergent evidence implicates chronic episodes of hypoxemia in cognitive dysfunction in people with DS. Using an infra-red detection system to assess oxygen saturation in free-moving mice, we assessed blood oxygenation in both adolescent and adult Ts65Dn mice and found a significant increase in the incidence of hypoxemia in both groups. Notably, the severity of hypoxemia increased during the dark cycle, suggesting a link between hypoxemia and increased motor activity. Postmortem analysis showed significant increase in the expression of mitochondrial Cox4i2, the terminal enzyme of the mitochondrial respiratory chain and oxygen response element. Altogether these data suggest early and chronic occurrence of hypoxemia in the Ts65Dn mouse model of DS, which can contribute to cognitive dysfunction in these mice.
Increased cortical synaptic activation of TrkB and downstream signaling markers in a mouse model of Down Syndrome.
R Nosheny, PV Belichenko, BL Busse, AM Weissmiller, V Dang, A Fahimi, A Salehi, SJ Smith, and WC Mobley
Down Syndrome (DS), trisomy 21, is characterized by synaptic abnormalities and cognitive deficits throughout the lifespan and with development of Alzheimer's disease (AD) neuropathology and progressive cognitive decline in adults. Synaptic abnormalities are also present in the Ts65Dn mouse model of DS, but which synapses are affected and the mechanisms underlying synaptic dysfunction are unknown. Here we show marked increases in the levels and activation status of TrkB and associated signaling proteins in cortical synapses in Ts65Dn mice. Proteomic analysis at the single synapse level of resolution using array tomography (AT) uncovered increased colocalization of activated TrkB with signaling endosome related proteins, and demonstrated increased TrkB signaling. The extent of increases in TrkB signaling differed in each of the cortical layers examined and with respect to the type of synapse, with the most marked increases seen in inhibitory synapses. These findings are evidence of markedly abnormal TrkB-mediated signaling in synapses. They raise the possibility that dysregulated TrkB signaling contributes to synaptic dysfunction and cognitive deficits in DS.
Journal of visualized experiments (JoVE)
Assessment of Dendritic Arborization in the Dentate Gyrus of the Hippocampal Region in Mice
Devsmita Das, Cristy Phillips, Bill Lin, Fatemeh Mojabi, Van Dang, Mehmet Baktir, Ravikumar Ponnusamy, and Ahmad Salehi
We describe two methods for visualization and quantification of dendritic arborization in the hippocampus of mouse models: real-time and extended depth of field imaging. While the former method allows sophisticated topographical tracing and quantification of the extent of branching, the latter allows speedy visualization of the dendritic tree.
The Link Between Physical Activity and Cognitive Dysfunction in Alzheimer Disease.
Cristy Phillips, Mehmet Akif Baktir, Devsmita Das, Bill Lin, and Ahmad Salehi
Alzheimer's disease (AD) is a chronic, neurodegenerative disorder that adversely affects neurons in the brain, ultimately resulting in loss of memory and language, behavioural disturbances, and dependence on caregivers. The strongest risk factor for AD is aging, a risk that doubles every five years after the age of 65 years.1 Increasing population, longevity, and economic prosperity have contributed to concern of a dementia epidemic in the aging population. Currently, 26 million individuals are affected by AD worldwide, a number that is expected to approximate 106 million by the year 2050, provoking serious clinical, social, ethical, and economical problems.
Neurobiol Learn Mem
Nest building is impaired in the Ts65Dn mouse model of Down syndrome and rescued by blocking 5HT2a receptors
Craig Heller, Ahmad Salehi, Bayarsaikhan Chuluun, Devsmita Das, Bill Lin, Sara Moghadam, Craig Garner, and Damien Colas
The Role of NMDA Receptors in the Pathophysiology and Treatment of Mood Disorders
Mehdi Ghasemi, Cristy Phillips, Ludwig Trillo, Zuri De Miguel, Devsmita Das, and Ahmad Salehi
Mood disorders such as major depressive disorder and bipolar disorder are chronic and recurrent illnesses that cause significant disability and affect approximately 350 million people worldwide. Currently available biogenic amine treatments provide relief for many and yet fail to ameliorate symptoms for others, highlighting the need to diversify the search for new therapeutic strategies. Here we present recent evidence implicating the role of N-methyl-D-aspartate receptor (NMDAR) signaling in the pathophysiology of mood disorders. The possible role of NMDARs in mood disorders has been supported by evidence demonstrating that: (i) both BPD and MDD are characterized by altered levels of central excitatory neurotransmitters; (ii) NMDAR expression, distribution, and function are atypical in patients with mood disorders; (iii) NMDAR modulators show positive therapeutic effects in BPD and MDD patients; and (iv) conventional antidepressants/mood stabilizers can modulate NMDAR function. Taken together, this evidence suggests the NMDAR system holds considerable promise as a therapeutic target for developing next generation drugs that may provide more rapid onset relief of symptoms. Identifying the subcircuits involved in mood and elucidating the role of NMDARs subtypes in specific brain circuits would constitute an important step toward the development of more effective therapies with fewer side effects.
Frontiers in Cellular Neuroscience
Neuroprotective Effects of Physical Activity on the Brain A Closer Look at Trophic Factor Signaling
Cristy Phillips, Mehmet A Bakhtir, Malathi Siravatsan, and Ahmad Salehi
While the relationship between increased physical activity and cognitive ability has been conjectured for centuries, only recently have the mechanisms underlying this relationship began to emerge. Convergent evidence suggests that physical activity offers an affordable and effective method to improve cognitive function in all ages, particularly the elderly who are most vulnerable to neurodegenerative disorders. In addition to improving cardiac and immune function, physical activity alters trophic factor signaling and, in turn, neuronal function and structure in areas critical for cognition. Sustained exercise plays a role in modulating anti-inflammatory effects and may play a role in preserving cognitive function in aging and neuropathological conditions. Moreover, recent evidence suggests that myokines released by exercising muscles affect the expression of brain-derived neurotrophic factor synthesis in the dentate gyrus of the hippocampus, a finding that could lead to the identification of new and therapeutically important mediating factors. Given the growing numbers of individuals with cognitive impairment in the US population, a better understanding of how these factors work in aggregate to contribute to cognition is imperative, and constitutes an important first step toward developing non-pharmacological therapeutic strategies to improve cognition in vulnerable populations.
Prog Neuropsychopharmacol Biol Psychiatry
Prog Neuropsychopharmacol Biol Psychiatry (cont.)
Neurotransmitter-based strategies for the treatment of cognitive dysfunction in Down syndrome.
Devsmita Das, Cristy Phillips, Wayne Hsieh, Krithika Sumanth, Van Dang, and Ahmad Salehi
Down syndrome (DS) is a multisystem disorder affecting the cardiovascular, respiratory, gastrointestinal, neurological, hematopoietic, and musculoskeletal systems and is characterized by significant cognitive disability and a possible common pathogenic mechanism with Alzheimer's disease. During the last decade, numerous studies have supported the notion that the triplication of specific genes on human chromosome 21 plays a significant role in cognitive dysfunction in DS. Here we reviewed studies in trisomic mouse models and humans, including children and adults with DS. In order to identify groups of genes that contribute to cognitive disability in DS, multiple mouse models of DS with segmental trisomy have been generated. Over-expression of these particular genes in DS can lead to dysfunction of several neurotransmitter systems. Therapeutic strategies for DS have either focused on normalizing the expression of triplicated genes with important role in DS or restoring the function of these systems. Indeed, our extensive review of studies on the pathogenesis of DS suggests that one plausible strategy for the treatment of cognitive dysfunction is to target the cholinergic, serotonergic, GABA-ergic, glutamatergic, and norepinephrinergic system. However, a fundamental strategy for treatment of cognitive dysfunction in DS would include reducing to normal levels the expression of specific triplicated genes in affected systems before the onset of neurodegeneration.
Neuroscience & Biobehavioral Reviews
Neuroscience & Biobehavioral Reviews (cont.)
Ascending monoaminergic systems alterations in Alzheimer’s disease. Translating basic science into clinical care
Ludwig Trllo, Devsmita Das, Wayne Hsieh, Brian Medina, Bill Lin, Van Dang, Martha Millan Sanchez, Zurine DeMiguel, J Wesson Ashford, and Ahmad Salehi
Extensive neuropathological studies have established a compelling link between abnormalities in structure and function of subcortical monoaminergic (MA-ergic) systems and the pathophysiology of Alzheimer's disease (AD). The main cell populations of these systems including the locus coeruleus, the raphe nuclei, and the tuberomamillary nucleus undergo significant degeneration in AD, thereby depriving the hippocampal and cortical neurons from their critical modulatory influence. These studies have been complemented by genome wide association studies linking polymorphisms in key genes involved in the MA-ergic systems and particular behavioral abnormalities in AD. Importantly, several recent studies have shown that improvement of the MA-ergic systems can both restore cognitive function and reduce AD-related pathology in animal models of neurodegeneration. This review aims to explore the link between abnormalities in the MA-ergic systems and AD symptomatology as well as the therapeutic strategies targeting these systems. Furthermore, we will examine possible mechanisms behind basic vulnerability of MA-ergic neurons in AD.
Biological Psychiatry (cont.)
Formoterol, a Long-Acting β2 Adrenergic Agonist, Improves Cognitive Function and Promotes Dendritic Complexity in a Mouse Model of Down Syndrome
Van Dang, Kara Martin, Devsmita Das, Bill Lin, Priyanka Naik, Devan Patel, Rachel Nosheney, John Ashford, and Ahmad Salehi
Down syndrome is associated with significant failure in cognitive function. Our previous investigation revealed age-dependent degeneration of locus coeruleus, a major player in contextual learning, in the Ts65Dn mouse model of Down syndrome. We studied whether drugs already available for use in humans can be used to improve cognitive function in these mice.
We studied the status of β adrenergic signaling in the dentate gyrus of the Ts65Dn mouse model of Down syndrome. Furthermore, we used fear conditioning to study learning and memory in these mice. Postmortem analyses included the analysis of synaptic density, dendritic arborization, and neurogenesis.
We found significant atrophy of dentate gyrus and failure of β adrenergic signaling in the hippocampus of Ts65Dn mice. Our behavioral analyses revealed that formoterol, a long-acting β2 adrenergic receptor agonist, caused significant improvement in the cognitive function in Ts65Dn mice. Postmortem analyses revealed that the use of formoterol was associated with a significant improvement in the synaptic density and increased complexity of newly born dentate granule neurons in the hippocampus of Ts65Dn mice.
Our data suggest that targeting β2 adrenergic receptors is an effective strategy for restoring synaptic plasticity and cognitive function in these mice. Considering its widespread use in humans and positive effects on cognition in Ts65Dn mice, formoterol or similar β2 adrenergic receptor agonists with ability to cross the blood brain barrier might be attractive candidates for clinical trials to improve cognitive function in individuals with Down syndrome.
Progress in Brain Research. Down Syndrome: From Understanding the Neurobiology to Therapy
Progress in Brain Research. Down Syndrome: From Understanding the Neurobiology to Therapy (cont.)
Discoveries in Down syndrome: moving basic science to clinical care
Alexander Kleschevnikov, Pavel Belichenko, Ahmad Salehi, and Chenbiao Wu
This review describes recent discoveries in neurobiology of Down syndrome (DS) achieved with use of mouse genetic models and provides an overview of experimental approaches aimed at development of pharmacological restoration of cognitive function in people with this developmental disorder. Changes in structure and function of synaptic connections within the hippocampal formation of DS model mice, as well as alterations in innervations of the hippocampus by noradrenergic and cholinergic neuromodulatory systems, provided important clues for potential pharmacological treatments of cognitive disabilities in DS. Possible molecular and cellular mechanisms underlying this genetic disorder have been addressed. We discuss novel mechanisms engaging misprocessing of amyloid precursor protein (App) and other proteins, through their affect on axonal transport and endosomal dysfunction, to "Alzheimer-type" neurodegenerative processes that affect cognition later in life. In conclusion, a number of therapeutic strategies have been defined that may restore cognitive function in mouse models of DS. In the juvenile and young animals, these strategists focus on restoration of synaptic plasticity, rate of adult neurogenesis, and functions of the neuromodulatory subcortical systems. Later in life, the major focus is on recuperation of misprocessed App and related proteins. It is hoped that the identification of an increasing number of potential targets for pharmacotherapy of cognitive deficits in DS will add to the momentum for creating and completing clinical trials.
NoteDown Syndrome: From Understanding the Neurobiology to Therapy is part of the series Progress in Brain Research.
Biological Psychiatry (cont.)
Neurobiological Elements of Cognitive Dysfunction in Down Syndrome: Exploring the Role of APP
Martha Millan Sanchez, Sietske Heyn, Devsmita Das, Sarah Moghadam, Kara Martin, and Ahmad Salehi
Down syndrome (DS) is the most common cause of cognitive dysfunction in children. Additionally, most adults with DS will eventually show both clinical and neuropathologic hallmarks of Alzheimer's disease (AD). The hippocampal formation constitutes the primary target for degeneration in both AD and DS. Over the past few years, we have studied the molecular mechanisms behind degeneration of this region and its major inputs in mouse models of DS. Our investigation has suggested that the loss of hippocampal inputs, particularly cholinergic and noradrenergic terminals, leads to de-afferentation of this region in the Ts65Dn mouse model of DS. Interestingly, we were able to link the overexpression of amyloid precursor protein (App) gene to degeneration of cholinergic and noradrenergic neurons in DS mouse models. We examined the underlying mechanisms of degeneration of multiple systems with extensive projections to the hippocampus in DS and its mouse models and the role of App overexpression in neurodegeneration. Understanding mechanisms behind hippocampal dysfunction has helped us to test several therapeutic strategies successfully in mouse models of DS. Here we review these strategies and mechanisms and discuss ways to translate our findings into possible interventions in humans.
BDNF polymorphism predicts the rate of decline in skilled task performance and hippocampal volume in healthy individuals
Martha Millan Sanchez, Devsmita Das, Joy Taylor, Art Noda, Jerrome Yesavage, and Ahmad Salehi
Numerous studies have indicated a link between the presence of polymorphism in brain-derived neurotrophic factor (BDNF) and cognitive and affective disorders. However, only a few have studied these effects longitudinally along with structural changes in the brain. This study was carried out to investigate whether valine-to-methionine substitution at position 66 (val66met) of pro-BDNF could be linked to alterations in the rate of decline in skilled task performance and structural changes in hippocampal volume. Participants consisted of 144 healthy Caucasian pilots (aged 40–69 years) who completed a minimum of 3 consecutive annual visits. Standardized flight simulator score (SFSS) was measured as a reliable and quantifiable indicator for skilled task performance. In addition, a subset of these individuals was assessed for hippocampal volume alterations using magnetic resonance imaging. We found that val66met substitution in BDNF correlated longitudinally with the rate of decline in SFSS. Structurally, age-dependent hippocampal volume changes were also significantly altered by this substitution. Our study suggests that val66met polymorphism in BDNF can be linked to the rate of decline in skilled task performance. Furthermore, this polymorphism could be used as a predictor of the effects of age on the structure of the hippocampus in healthy individuals. Such results have implications for understanding possible disabilities in older adults performing skilled tasks who are at a higher risk for cognitive and affective disorders.
Journal of Alzheimer's Disease
Hippocampal Network Alterations in Alzheimer’s Disease and Down Syndrome: From Structure to Therapy
Martha Millan Sanchez, Sarah Moghadam, Priyanka Naik, Kara Martin, and Ahmad Salehi
Hippocampal structural and functional alterations in Alzheimer's disease (AD), detected by advanced imaging methods, have been linked to significant abnormalities in multiple internal and external networks in this critical brain region. Uncovering the temporal and anatomical pattern of these network alterations would provide important clues into understanding the pathophysiology of AD and suggest new therapeutic strategies for this multi-system and prevalent disorder. Over the last decade, we have focused on studying brain structures that provide major projections to the hippocampus (HC) and the pattern of de-afferentation of this area in mouse models of AD and a related neurodegenerative disorder, i.e. Down syndrome (DS). Our studies have revealed that major inputs into the hippocampal structure undergo significant age-dependent alterations. Studying locus coeruleus (LC), the sole source of noradrenergic terminals for the HC, it has been shown that these neurons show significant age-dependent degeneration in both mouse models of DS and AD. Furthermore, increasing noradrenergic signaling was able to restore cognitive function by improving synaptic plasticity, and possibly promoting microglia recruitment, and amyloid β (Aβ) clearance in transgenic (tg) mouse models of AD. Here, we re-examine the effects of alterations in major inputs to the hippocampal region and their structural and functional consequences in mouse models of neurodegenerative disorders. We will conclude that improving the function of major hippocampal inputs could lead to a significant improvement in cognitive function in both AD and DS.
Science Translational Medicine
Science Translational Medicine (cont.)
Restoration of Norepinephrine-Modulated Contextual Memory in a Mouse Model of Down Syndrome
Ahmad Salehi, Mehrdad Faizi, Damien Colas, Janice Valletta, Jessania Laguna, Ryoko Takimoto-Kimura, Alexander Kleschevnikov, Steve Wagner, Paul Aisen, Mehrdad Shamloo, and William Mobley
Down syndrome (trisomy 21) is the most common cause of mental retardation in children and leads to marked deficits in contextual learning and memory. In rodents, these tasks require the hippocampus and are mediated by several inputs, particularly those originating in the locus coeruleus. These afferents mainly use norepinephrine as a transmitter. To explore the basis for contextual learning defects in Down syndrome, we examined the Ts65Dn mouse model. These mice, which have three copies of a fragment of mouse chromosome 16, exhibited significant deficits in contextual learning together with dysfunction and degeneration of locus coeruleus neurons. However, the postsynaptic targets of innervation remained responsive to noradrenergic receptor agonists. Indeed, despite advanced locus coeruleus degeneration, we were able to reverse contextual learning failure by using a prodrug for norepinephrine called L-threo-3,4-dihydroxyphenylserine, or xamoterol, a β1-adrenergic receptor partial agonist. Moreover, an increased gene dosage of App, …
Science Translational Medicine (cont.)
in the context of Down syndrome, was necessary for locus coeruleus degeneration. Our findings raise the possibility that restoring norepinephrine-mediated neurotransmission could reverse cognitive dysfunction in Down syndrome.
Cell Stem Cell
Cell Stem Cell (cont.)
Neuroprotection of Host Cells by Human Central Nervous System Stem Cells in a Mouse Model of Infantile Neuronal Ceroid Lipofuscinosis
Stanley Tamaki, Yakop Jacobs, Monika Dohse, Alexandra Capela, Jonathan Cooper, Michael Teitsma, Donping He, Robert Tushinski, Pavel Belichenko, Ahmad Salehi, William Mobley, Tred Gage, Stephen Huhn, Ann Tsukamoto, Irving Weissma, and Nobuko Uchida
On the Cover
Neurodegeneration of purkinje cells. In a mouse model of neuronal ceroid lipofuscinosis (NCL), a lysosomal storage disorder, accumulation of lipofuscin granules is followed by progressive neuronal cell death. Purkinje neurons in the cerebellum are severely affected as illustrated by loss of cell bodies and dendritic arbors. Massive lipofuscin deposits remain in the purkinje cell layer in the degenerating brain. The work by Tamaki et al. (pages 310–319) demonstrates that transplantation of human neural stem cells leads to neuroprotection of host cells in the NCL mouse brain. The cover image shows a section of cerebellum from a mouse with disease symptoms. Cover design by Monika Dohse.
Infantile neuronal ceroid lipofuscinosis (INCL) is a fatal neurodegenerative disease caused by a deficiency in the lysosomal enzyme palmitoyl protein thioesterase-1 (PPT1). Ppt1 knockout mice display hallmarks of INCL and mimic the human pathology: accumulation of lipofuscin, degeneration of CNS neurons, and a shortened life span. Purified non-genetically modified human CNS stem cells, grown as neurospheres (hCNS-SCns), were transplanted into the brains of immunodeficient Ppt1(-/)(-) mice where they engrafted robustly, migrated extensively, and produced sufficient levels of PPT1 to alter host neuropathology. Grafted mice displayed reduced autofluorescent lipofuscin, significant neuroprotection …
Cell Stem Cell (cont.)
of host hippocampal and cortical neurons, and delayed loss of motor coordination. Early intervention with cellular transplants of hCNS-SCns into the brains of INCL patients may supply a continuous and long-lasting source of the missing PPT1 and provide some therapeutic benefit through protection of endogenous neurons. These data provide the experimental basis for human clinical trials with these banked hCNS-SCns.
The Journal of Neuroscience
The “Down syndrome critical region” is sufficient in the mouse model to confer behavioral, neurophysiological, and synaptic phenotypes characteristic of Down syndrome
Nadia Belichenko, Pavel Belichenko, Alexander Kleschevnikov, Ahmad Salehi, Roger Reeves, and William Mobley
Down syndrome (DS) can be modeled in mice segmentally trisomic for mouse chromosome 16. Ts65Dn and Ts1Cje mouse models have been used to study DS neurobiological phenotypes including changes in cognitive ability, induction of long-term potentiation (LTP) in the fascia dentata (FD), the density and size of dendritic spines, and the structure of synapses. To explore the genetic basis for these phenotypes, we examined Ts1Rhr mice that are trisomic for a small subset of the genes triplicated in Ts65Dn and Ts1Cje mice. The 33 trisomic genes in Ts1Rhr represent a "DS critical region" that was once predicted to be sufficient to produce most DS phenotypes. We discovered significant alterations in an open field test, a novel object recognition test and in a T-maze task. As in Ts65Dn and Ts1Cje mice, LTP in FD of Ts1Rhr could be induced only after blocking GABAA-dependent inhibitory neurotransmission. In addition, widespread enlargement of dendritic spines and decreased density of spines in FD were preserved in Ts1Rhr. Twenty of 48 phenotypes showed significant differences between Ts1Rhr and 2N controls. We conclude that important neurobiological phenotypes characteristic of DS are conserved in Ts1Rhr mice. The data support the view that biologically significant trisomic phenotypes occur because of dosage effects of genes in the Ts1Rhr trisomic segment and that increased dosage is sufficient to produce these changes. The stage is now set for studies to decipher the gene(s) that play a conspicuous role in creating these phenotypes.
The Journal of Comparative Neurology
Excitatory-inhibitory relationship in the fascia dentata in the Ts65Dn mouse model of Down syndrome
Pavel Belichenko, Alexander Kleschevnikov, E. Masilah, Chengbioa Wu, Ryoko Takimoto-Kitamura, Ahmad Salehi, and William Mobley
Down syndrome (DS) is a neurological disorder causing impaired learning and memory. Partial trisomy 16 mice (Ts65Dn) are a genetic model for DS. Previously, we demonstrated widespread alterations of pre- and postsynaptic elements and physiological abnormalities in Ts65Dn mice. The average diameter of presynaptic boutons and spines in the neocortex and hippocampus was enlarged. Failed induction of long-term potentiation (LTP) due to excessive inhibition was observed. In this paper we investigate the morphological substrate for excessive inhibition in Ts65Dn. We used electron microscopy (EM) to characterize synapses, confocal microscopy to analyze colocalization of the general marker for synaptic vesicle protein with specific protein markers for inhibitory and excitatory synapses, and densitometry to characterize the distribution of the receptor and several proteins essential for synaptic clustering of neurotransmitter receptors. EM analysis of synapses in the Ts65Dn vs. 2N showed that synaptic opposition lengths were significantly greater for symmetric synapses (approximately 18%), but not for asymmetric ones. Overall, a significant increase in colocalization coefficients of glutamic acid decarboxylase (GAD)65/p38 immunoreactivity (IR) (approximately 27%) and vesicular GABA transporter (VGAT)/p38 IR (approximately 41%) was found, but not in vesicular glutamate transporter 1 (VGLUT1)/p38 IR. A significant overall decrease of IR in the hippocampus of Ts65Dn mice compared with 2N mice for glutamate receptor 2 (GluR2; approximately 13%) and anti-gamma-aminobutyric acid (GABAA) receptor β2/3 subunit (approximately 20%) was also found. The study of proteins essential for synaptic clustering of receptors revealed a significant increase in puncta size for neuroligin 2 (approximately 13%) …
The Journal of Comparative Neurology (cont.)
and GABAA receptor-associated protein (GABARAP; approximately 13%), but not for neuroligin 1 and gephyrin. The results demonstrate a significant alteration of inhibitory synapses in the fascia dentata of Ts65Dn mice.
Intracellular Traffic and Neurodegenerative Disorders [Book]
Axonal Transport of Neurotrophic Signals: An Achilles’ Heel for Neurodegeneration?
Edited by Ahmad Salehi, Chengbioa Wu, Ke Zhan, and William Mobley
Wordpress The most effective treatments for neurodegenerative disorders, including Alzheimer's disease, will come from studies of the pathogenesis of age-related cognitive failure and understanding of the underlying mechanisms. Given the marked similarities in pathological and clinical phenotypes between Alzheimer's disease and Down syndrome, studies of the pathogenesis of one can be expected to complement and support those in the other. Alzheimer's disease and Down syndrome are characterized by dysfunction and loss of several biochemically and anatomically defined neuronal populations. The pathological involvement of hippocampus, in particular, is an early feature of both disorders, as is the degeneration of neurons whose axons innervate this region. Long, thin and poorly myelinated axons project from a number of subcortical and brain stem nuclei to modulate hippocampally mediated cognitive functions. In studies on mouse models of Down's syndrome, we uncovered evidence for the involvement of a particular neuronal population heavily innervating the hippocampus. In an extensive series of experiments, we found evidence that failed retrograde transport of nerve growth factor signaling in cholinergic neurons of the basal forebrain is linked to their vulnerability and that these changes are caused by increased gene dose and overexpression of the gene for amyloid precursor protein. These findings raise the possibility that intracellular trafficking defects created by changes in amyloid precursor protein expression or processing make an important contribution to pathogenesis and set the stage for studies to explore the molecular mechanisms of degeneration of cholinergic neurons and to define new therapeutic targets for these neurons. An important unanswered question is whether or not similar mechanisms operate within other vulnerable populations, …
Intracellular Traffic and Neurodegenerative Disorders (cont.)
innervating hippocampus to cause de-afferentation and dysfunction of this critical brain region.
The Journal of Comparative Neurology
Synaptic and cognitive abnormalities in mouse models of Down syndrome: exploring genotype-phenotype relationship genotype-phenotype relationships
Pavel Belichenko, Alexander Kleschevnikov, Ahmad Salehi, Charles Epstein, and William Mobley
Down syndrome (DS) is caused by trisomy of human chromosome 21. Because Ts65Dn and Ts1Cje mice are segmentally trisomic for a region of mouse chromosome 16, they genetically model DS and are used to study pathogenic mechanisms. Previously, we provided evidence for changes in both the structure and function of pre- and postsynaptic elements in the Ts65Dn mouse. Striking changes were evident in the size of the dendritic spines and in the ability to induce long-term potentiation (LTP) in the fascia dentata (FD). To explore the genetic basis for these changes, we examined Ts1Cje mice, which are trisomic for a completely overlapping but smaller segment of mouse chromosome 16. As in the Ts65Dn mouse, there was a regionally selective decrease in the density of dendritic spines (approximately 12%), an increase in the size of spine heads (approximately 26%), a decrease in the length of spine necks (approximately 26%), and reorganization of inhibitory inputs with a relative decrease in inputs to dendrite shafts and spine heads and a significant increase to the necks of spines (6.4%). Thus, all of the Ts65Dn phenotypes were present, but they were significantly less severe. In contrast, and just as was the case for the Ts65Dn mouse, LTP could not be induced unless the selective gamma-aminobutyric acid (GABAA) receptor antagonist picrotoxin was applied. Therefore, there was conservation of important synaptic phenotypes in the Ts1Cje mice. The analysis of data from this and earlier studies points to genotype-phenotype linkages in DS whose complexity ranges from relatively simple to quite complex.
Mental Retardation and Developmental Disabilities Research Reviews
Using mouse models to explore genotype-phenotype relationship in Down syndrome
Ahmad Salehi, Mehrdad Faizi, Pavel Belichenko, and William Mobley
Down Syndrome (DS) caused by trisomy 21 is characterized by a variety of phenotypes and involves multiple organs. Sequencing of human chromosome 21 (HSA21) and subsequently of its orthologues on mouse chromosome 16 have created an unprecedented opportunity to explore the complex relationship between various DS phenotypes and the extra copy of approximately 300 genes on HSA21. Advances in genetics together with the ability to generate genetically well-defined mouse models have been instrumental in understanding the relationships between genotype and phenotype in DS. Indeed, elucidation of these relationships will play an important role in understanding the pathophysiological basis of this disorder and helping to develop therapeutic interventions. A successful example of using such a strategy is our recent studies exploring the relationship between failed nerve growth factor (NGF) transport and amyloid precursor protein (App) overexpression. We found that increased dosage of the gene for App is linked to failed NGF signaling and cholinergic neurodegeneration in a mouse model of DS. Herein, we discuss several mouse models of DS and explore the emergence of exciting new insights into genotype-phenotype relationships, particularly those related to nervous system abnormalities. An important conclusion is that uncovering these relationships is enhanced by working from carefully defined phenotypes to the genes responsible.
Pharmacological Mechanisms in Alzheimer's Therapeutics [Book]
Cholinergic Neurodegeneration in Alzheimer’s Disease: Basis for Nerve Growth Factor Therapy?
Edited by Ahmad Salehi, Alexander Kleschevnikov, and William Mobley
Neurotrophins play an important role in the survival, differentiation, and maintenance of neurons selectively involved in a number of disorders of the nervous system. Nerve growth factor (NGF) plays a vital role for basal forebrain cholinergic neurons (BFCNs), including the maintenance of the cholinergic phenotype in adults. Recognition of this role has suggested the use of NGF to ameliorate the loss of these neurons in Alzheimer’s disease (AD). While clinical studies directed at supplying NGF to patients continue to be pursued, fundamental questions remain as to the relationship between selective vulnerability of cholinergic neurons and the actions of NGF. In this chapter, we review the structure and function of the basal forebrain cholinergic system, its role in higher cognitive functions, and the importance of NGF actions on these cells. Studies that link changes in NGF signaling to the degeneration of BFCNs are then discussed, as are current approaches to NGF-related treatments. Finally, on the basis of recent findings in mouse models of AD and Down syndrome, we suggest that impaired retrograde axonal transport of NGF plays a significant role in pathogenesis. This insight may guide future studies of pathogenesis and innovative treatment for BFCNs and, perhaps, other neurons affected in AD.
Increased App expression in a mouse model of Down’s syndrome disrupts NGF transport and causes cholinergic neuron degeneration
Ahmad Salehi, Jean-Dominique Delcroix, Pavel Belichenko, Ke Zhan, Chenbioa Wu, Janice Valleta, Ryoko Takimoto-Kimura, Alexander Kleschevnikov, Kumar Sambamurti, Peter Chung, Weiming Xia, Angela Villar, William Campbell, Laura Kulnane, Randy Nixon, Bruce Lamb, Charles Epstein, Gorazd Stokin, Larry Goldstein, and William Mobley
Down Syndrome (DS) caused by trisomy 21 is characterized by a variety of phenotypes and involves multiple organs. Sequencing of human chromosome 21 (HSA21) and subsequently of its orthologues on mouse chromosome 16 have created an unprecedented opportunity to explore the complex relationship between various DS phenotypes and the extra copy of 300 genes on HSA21. Advances in genetics together with the ability to generate genetically well-defined mouse models have been instrumental in understanding the relationships between genotype and phenotype in DS. Indeed, elucidation of these relationships will play an important role in understanding the pathophysiological basis of this disorder and helping to develop therapeutic interventions. A successful example of using such a strategy is our recent studies exploring the relationship between failed nerve growth factor (NGF) transport and amyloid precursor protein (App) overexpression. We found that increased dosage of the gene for App is linked to failed NGF signaling and cholinergic neurodegeneration in a mouse model of DS. …
Herein, we discuss several mouse models of DS and explore the emergence of exciting new insights into genotype-phenotype relationships, particularly those related to nervous system abnormalities. An important conclusion is that uncovering these relationships is enhanced by working from carefully defined phenotypes to the genes responsible.
NGF and Related Molecules in Health and Disease [Book]
Trafficking the NGF signal: implications for normal and degenerating neurons
Edited by Jean-Dominique Delcroix, Janice Valleta, Chengbioa Wu, Charles Howe, Chun-Fai Lai, John Cooper, Pavel Belichenko, Ahmad Salehi, and William Mobley
Nerve growth factor (NGF) activates TrkA to trigger signaling events that promote the survival, differentiation and maintenance of neurons. The mechanism(s) that controls the retrograde transport of the NGF signal from axon terminals to neuron cell bodies is not known. The ‘signaling endosome’ hypothesis stipulates that NGF, TrkA and signaling proteins are retrogradely transported on endocytic vesicles. Here, we provide evidence for the existence of signaling endosomes. Following NGF treatment, clathrin-coated vesicles (CCVs) contain NGF bound to TrkA together with activated signaling proteins of the Ras/pErk1/2 pathway. NGF signals from isolated CCVs through the Erk1/2 pathway. Early endosomes appear to represent a second type of signaling endosomes. We found that NGF induced a sustained activation of Rap1, a small monomeric GTP-binding protein of the Ras family, and that this activation occurred in early endosomes that contain key elements of Rap1/pErk1/2 pathway. We discuss the possibility that the failure of retrograde NGF signaling in a mouse model of Down syndrome (Ts65Dn) may be due to the failure to retrograde transport signaling endosomes. It is important to define further the significance of signaling endosomes in the biology of both normal and degenerating neurons.
NoteNGF and Related Molecules in Health and Disease is part of the series Progress in Brain Research.
The Journal Comparative Neurology
Synaptic structural abnormalities in the Ts65Dn mouse model of Down Syndrome
Pavel Belichenko, Alexander Kleschevnikov, Ahmad Salehi, Charles Epstein, and William Mobley
The Ts65Dn mouse is a genetic model for Down syndrome. Although this mouse shows abnormalities in cognitive function that implicate hippocampus as well as marked deficits in hippocampal long-term potentiation, the structure of the hippocampus has been little studied. We characterized synaptic structure in Ts65Dn and control (2N) mice, studying the hippocampus (fascia dentata, CA1) as well as the motor and somatosensory cortex, entorhinal cortex, and medial septum. Confocal microscopy was used to examine immunostained presynaptic boutons and to detail the structure of dendrites after Lucifer yellow microinjection. Both presynaptic and postsynaptic elements were significantly enlarged in Ts65Dn in all regions examined. The changes were detected at the youngest age examined (postnatal day 21) and in adults. In studies detailing the changes in fascia dentata and motor cortex, the enlargement of spines affected the entire population, resulting in the presence of spines whose volume was greatly increased. Electron microscopy confirmed that boutons and spines were enlarged and demonstrated abnormalities in the internal membranes of both. In addition, spine density was decreased on the dendrites of dentate granule cells, and there was reorganization of inhibitory inputs, with a relative decrease in inputs to dendrite shafts and an increase in inputs to the necks of spines. Taken together, the findings document widespread abnormalities of synaptic structure that recapitulate important features seen in Down syndrome. They establish the Ts65Dn mouse as a model for abnormal synapse structure and function in Down syndrome and point to the importance of studies to elucidate the mechanisms responsible for synapse enlargement.
Journal of Neural Transmission
Alzheimer’s disease and NGF signaling
Ahmad Salehi, Jean-Dominique Delcroix, and Dick Swaab
Age-related degeneration of basal forebrain cholinergic neurons (BFCNs) occurs early and contributes significantly to cognitive decline in Alzheimer's disease (AD). Proper function and morphology of BFCNs depends on the supply of nerve growth factor (NGF) from the cortex and the hippocampus. A large number of experiments have shown that decreased supply of NGF at the level of BFCN cell bodies leads to loss of neuronal markers and shrinkage, mimicking what is observed in AD. The delivery of sufficient amounts of NGF signal to BFCN cell bodies depends on the effective participation of several factors including sufficient synthesis and release of NGF, adequate synthesis and expression of NGF receptors by BFCNs, normal signaling and retrograde transport of NGF-receptor complex, and finally effective induction of gene expression by NGF. In the past few years it has become clear that decreased amounts of NGF at the level of BFCN cell bodies is largely due to failed retrograde transport rather than decreased synthesis, binding or expression of NGF receptors in the BFCN terminals. We will discuss in vivo evidence supporting decreased retrograde transport of NGF in a mouse model with BFCN degeneration, and will attempt to match these findings with our studies in postmortem human AD brain. We will speculate about the possible mechanisms of failed NGF retrograde transport and its relationship to AD pathology.
Trends in Neurosciences
Traffic at the intersection of neurotrophic factor signaling and neurodegeneration
Ahmad Salehi, Jean-Dominique Delcroix, and William Mobley
Advances in understanding the biology of neurotrophic factors and their signaling pathways have provided important insights into the normal growth, differentiation and maintenance of neurons. Stimulated by neuropathological observations and genetic discoveries, studies in cell and animal models of neurodegenerative disorders have begun to clarify pathogenetic mechanisms. We examine the intersection of these research themes and identify several potential mechanisms for linking failed neurotrophic factor signaling to neurodegeneration. Studies of nerve growth factor signaling in a mouse model of Down syndrome encourage the views that neuronal dysfunction and atrophy might be linked to failed neurotrophic support and that additional studies focused on this possibility would enhance our understanding of the mechanisms of neurodegenerative disorders and their treatment.
Proceedings of the National Academy of Sciences
Failed retrograde transport of NGF in a mouse model of Down’s syndrome: reversal of cholinergic neurodegenerative phenotypes following NGF infusion
John Cooper, Ahmad Salehi, Jean-Dominique Delcroix, Charles Howe, Pavel Belichenko, Jane Chua-Couzens, Joshua Kilbridge, Elaine Carlson, Charles Epstein, and William Mobley
Age-related degeneration of basal forebrain cholinergic neurons (BFCNs) contributes to cognitive decline in Alzheimer's disease and Down's syndrome. With aging, the partial trisomy 16 (Ts65Dn) mouse model of Down's syndrome exhibited reductions in BFCN size and number and regressive changes in the hippocampal terminal fields of these neurons with respect to diploid controls. The changes were associated with significantly impaired retrograde transport of nerve growth factor (NGF) from the hippocampus to the basal forebrain. Intracerebroventricular NGF infusion reversed well established abnormalities in BFCN size and number and restored the deficit in cholinergic innervation. The findings are evidence that even BFCNs chronically deprived of endogenous NGF respond to an intervention that compensates for defective retrograde transport. We suggest that age-related cholinergic neurodegeneration may be a treatable disorder of failed retrograde NGF signaling.
Neurobiology of Aging
Molecular misreading: a new type of transcript mutation expressed during aging
Fred van Leeuwen, David Fischer, Darlene Kamel, Jacqueline Slujis, Marc Sonnemans, Rob Benne, Dick Swaab, Ahmad Salehi, and Elly Hol
Dinucleotide deletions (e.g. ΔGA, ΔGU) are created by molecular misreading in or adjacent to GAGAG motifs of neuronal mRNAs. As a result, the reading frame shifts to the +1 frame, and so-called "+1 proteins" are subsequently synthesized. +1 Proteins have a wild-type N-terminus, but an aberrant C-terminus downstream from the site of the dinucleotide deletion. Molecular misreading was discovered in the rat vasopressin gene associated with diabetes insipidus and subsequently in human genes linked to Alzheimer's disease (AD), e.g. β amyloid precursor protein (βAPP) and ubiquitin-B (UBB). Furthermore, βAPP(+1) and UBB(+1) proteins accumulate in the neuropathological hallmarks (i.e. in the tangles, neuritic plaques, and neuropil threads) of AD. As these +1 proteins were also found in elderly nondemented controls, but not in younger ones (<51 years), molecular misreading in nondividing cells might act as a factor that only becomes manifest at an advanced age. Frameshift mutations (UBB(+1)) and pretangle staining (Alz-50 and MC1) seem to occur independently of each other during early stages of AD. We recently detected +1 proteins, not only in proliferating cells present in non-neuronal tissues such as the liver and epididymis, but also in neuroblastoma cell lines. These observations suggest that molecular misreading is a general source of transcript errors that are involved in cellular derangements in various age-related pathologies.
Sex- and age-related P75 neurotrophin receptor expression in the human supraoptic nucleus
Tatjana Ishunina, Ahmad Salehi, and Dick Swaab
The human supraoptic nucleus (SON) is the main production site of plasma vasopressin. Previously, using the Golgi apparatus and cell size as measures for neuronal metabolic activity, an activation of vasopressinergic neurons was found during ageing in the human SON in women but not in men. We hypothesized that the low-affinity neurotrophin receptor p75 (p75(NTR)) might be involved in the mechanism of activation of vasopressin neurons in postmenopausal women, since this receptor was found to be expressed in the SON neurons of aged individuals, and because p75(NTR) expression was shown to be suppressed by estrogens. Therefore, we investigated whether p75(NTR) immunoreactivity in the SON neurons was age- and sex-dependent. For this purpose, we studied paraffin sections of the SON in 32 postmortem brains of control patients ranging in age from 29 to 94 years with an anti-p75(NTR) antibody and determined the area of p75(NTR) immunoreactivity per neuron using an image analysis system. To study whether the p75(NTR) might also participate in the activation of SON neurons, we related Golgi apparatus size to the area of p75(NTR) immunoreactivity per cell in the same patients. We found that the area of p75(NTR) immunoreactivity per cell correlated indeed significantly with age and with Golgi apparatus size only in women but not in men. Therefore, our results suggest that p75(NTR) is involved in postmenopausal activation of vasopressinergic neurons in the human SON.
P75 neurotrophin receptor in the nucleus basalis of meynert in relation to age, sex, and Alzheimer’s disease
Ahmad Salehi, Marta Ocampo, Joost Verhaagen, and Dick Swaab
In a previous study we showed that the staining of tyrosine kinase receptors (trks), which are high-affinity neurotrophin receptors (NTRs), is strongly diminished in the nucleus basalis of Meynert (NBM) of Alzheimer's disease (AD) patients, which may explain the lack of effect of NGF therapy in AD patients so far. Since the literature regarding the expression of low-affinity NTRs was rather controversial, the aim of the present study was to examine (i) possible changes in the staining of low-affinity NTRs, i.e., p75 in the human NBM, an area that is severely affected in AD; and (ii) alterations of these receptors in relation to risk factors for AD, e. g., age, sex, and menopause. Brain material of 31 controls and 30 AD patients was obtained at autopsy, embedded in paraffin, and stained immunocytochemically. Using an image analysis system, we quantified p75 immunoreactivity in both cell bodies and fibers at the level of the NBM. Our results showed a significant diminishment of p75 immunoreactivity in both cell bodies and fibers of NBM neurons in AD. We did not find any relationship between age or sex and the expression of p75 receptor in cell bodies. However, there was a clearly positive relationship between age and fiber staining in AD patients which suggests the occurrence of a p75 transport disorder as an early event in the process of AD. These observations and the earlier reported decreased staining of trk receptors show that degeneration of NBM neurons in AD is associated with a decreased neurotrophin responsiveness of NBM neurons in AD and that therapeutic strategies should be directed toward upregulation of receptors or facilitation of transport before an effect of neurotrophins in AD may be expected.
Journal of Neuroendocrinology
Activity of vasopressinergic neurons of the human supraoptic nucleus is age- and sex-dependent
Tatjana Ishunina, Ahmad Salehi, Michel Hofman, and Dick Swaab
In the human hypothalamus, arginine-vasopressin (AVP) is produced for a major part by the neurones of the supraoptic nucleus (SON). Since plasma AVP levels in men were reported to be higher than those of women and we did not find a sex difference in the neurone number, a higher vasopressinergic neurone activity was supposed to be present in the SON of men. Therefore we studied the size of the Golgi-apparatus (GA), which has been demonstrated previously to be a sensitive parameter for protein synthetic ability of neurones, in 15 men and 17 women ranging in age from 29 to 94 years. A polyclonal antibody against immunoaffinity purified MG-160, a sialoglycoprotein of the medial cisternae of the GA was applied on paraffin-embedded sections containing the dorsolateral SON (dl-SON) from which 90-95% of neurones are vasopressinergic. SON areas that contain oxytocin (OT) cells were excluded on the basis of adjacent sections stained with a monoclonal antibody against OT. By means of an image analysis system the size of the GA and the cellular profile area were determined in dl-SON neurones with a nucleolus. Our results showed indeed an age-dependent sex difference in the size of the GA that appeared to be twice as large in young men (≤ 50 years old) than in young women of the same age. The size of the GA increased with age in women but not in men. In addition, the mean cell profile area, another measure for neuronal activity, was significantly larger in young men than in young women and was in old women larger than in young women. In conclusion, these data show the presence of a sex-dependent age-difference in the activity of vasopressinergic neurones in dl-SON which may relate to differences in AVP and sex hormone levels and kidney AVP receptors.
Journal of Neural Transmission
Diminished neuronal metabolic activity in Alzheimer’s disease
Ahmad Salehi, and Dick Swaab
An increasing number of studies have appeared in the literature suggesting that Alzheimer's disease (AD) is a hypometabolic brain disorder. Decreased metabolism in AD has been revealed by a variety of in vivo and postmortem methods and techniques including positron emission tomography and glucose metabolism. We used the size of the Golgi apparatus (GA) and cell profile area as indicators of neuronal activity in postmortem material. Using an antibody against MG-160, a sialoglycoprotein of the medial cisternae of the GA, we were able to visualize and quantify the GA area. In a series of experiments, we tried to relate neuronal metabolism to different hallmarks of AD, i.e. plaques and tangles, and also to genetic risk factors for AD like age and (apolipoprotein E) ApoE polymorphism. Our results showed that in AD there is indeed a clear reduction in brain metabolism in several severely affected brain regions including the nucleus basalis of Meynert (NBM), the CA1 area of the hippocampus and the hypothalamic tuberomamillary nucleus. However, the reduction in neuronal activity did not seem to be caused by the presence of neuropathological hallmarks of AD, i.e. plaques and tangles. There was, however, a clear relationship between the presence of ApoE ε4 alleles and a decrease in GA size. Our data suggest that decreased neuronal activity and neuropathological hallmarks of AD, such as plaques and tangles, are basically independent phenomena. Moreover, ApoE ε4 may participate in the pathogenesis of AD by decreasing neuronal metabolism. The main implication of these findings is that therapeutic strategies in AD should be focussed on reactivation of neuronal metabolism.
Neurobiology of Aging
A sex difference and no effect of ApoE type on the amount of cytoskeletal alterations in the nucleus basalis of Meynert in Alzheimer’s disease age- and sex-dependent
Ahmad Salehi, Victoria Martinez, and Dick Swaab
In the nucleus basalis of Meynert (NBM) we studied the presence of early cytoskeletal alterations as shown by the antibody Alz-50 in ApoE-typed patients. Using an image analysis system, the area covered by Alz-50 staining and the percentage of neurons stained by Alz-50 were determined. There were no significant differences in the area covered by Alz-50 or in the proportion of Alz-50-stained neurons in the nucleus basalis of Meynert of Alzheimer’s disease (AD) patients with one or two ApoE ε4 alleles as compared with those without any ApoE ε4 allele. However, there was a significant sex difference in Alz-50 staining: female Alzheimer’s disease patients showed more severe early cytoskeletal alterations than males. We also found a significant relationship between the number of Alz-50-stained neurons and the severity of dementia.
Journal of Alzheimer's Disease
Activity of Hippocampal CA1 Neurons in Alzheimer’s Disease is not Affected by the Presence of Adjacent Neuritic Plaques in Alzheimer’s disease
Ahmad Salehi, Chris Pool, Monique Mulder, Rifka Ravid, Nikolas Gonatas, and Dick Swaab
Alzheimer's disease (AD) is neuropathologically characterized by neuritic plaques (NPs) and neurofibrillary tangles and functionally by a decreased metabolic rate of neurons. Our previous studies showed that in brain areas which are extensively affected by plaques and tangles, i.e. the CA1 area of the hippocampus and the hypothalamic tuberomamillary nucleus, neuronal protein synthetic ability is significantly lower in AD patients than in controls. However, the presence of tangles as shown by Bodian staining appeared not to be directly related to decreased protein synthetic ability of neurons. In order to study to what extent the metabolic function of neurons might be affected by the other neuropathological hallmark of AD, i.e. NPs, which are presumed to contain neurotoxic compounds, we studied eight severely demented AD patients matched for the ApoE genotype (ApoE 3/3). Using an image analysis system, the size of the neuronal Golgi apparatus (GA) and of the cell profile area was measured as a parameter for protein synthetic activity in the CA1 area of these patients. NPs were stained by Bodian, and subsequently the distance of each neuron with an immunostained GA to the nearest NP was measured. Our results showed that neither NP density nor the distance between NPs and neurons correlated with the protein synthetic ability of neurons as judged by the size of the GA. Based on these results we suggest that in AD the presence of NPs and decreased neuronal protein synthetic ability are basically two independent phenomena.
Proceedings of the National Academy of Sciences
Aggravated decrease in the activity of nucleus basalis neurons in Alzheimer’s disease is apolipoprotein E-type dependent
Ahmad Salehi, Elisabeth Dubelaar, Monique Mulder, and Dick Swaab
As reported before, the metabolic activity of nucleus basalis neurons is reduced significantly in Alzheimer patients. Because the apolipoprotein E (ApoE) ε4 genotype is a major risk factor for Alzheimer's disease (AD), we determined whether the decrease in metabolic activity in nucleus basalis neurons in AD is ApoE-type dependent. The size of the Golgi apparatus (GA) was determined as a measure of neuronal metabolic activity in 30 controls and 41 AD patients with a known ApoE genotype by using an image analysis system in the nucleus basalis of Meynert. A polyclonal antibody directed against MG-160, a sialoglycoprotein of the GA, was used to visualize this organelle. There was a very strong reduction in the size of the GA in the nucleus basalis of AD patients. Furthermore, a strong and significant extra reduction in the size of the GA was found in the nucleus basalis neurons of AD patients with either one or two ApoE ε4 alleles compared with Alzheimer patients without ApoE ε4 alleles. Our data show that the decreased activity of nucleus basalis neurons in AD is ApoE ε4 dependent and suggest that ApoE ε4 participates in the pathogenesis of AD by decreasing neuronal metabolism.
Alzheimer Disease & Associated Disorders
Limited effect of neuritic plaques on neuronal density in the hippocampal CA1 area of Alzheimer patients
Ahmad Salehi, Jan Bakker, Monique Mulder, and Dick Swaab
Neurofibrillary tangles (NFTs) and neuritic plaques (NPs) are the classic neuropathological hallmarks of Alzheimer disease (AD). It is generally assumed that the pathogenic process of AD could start by local neurotoxicity induced by the β-amyloid core of plaques, followed by the appearance of NFTs and eventually cell death. To determine whether or not local neurotoxicity around NPs is indeed a major pathogenetic mechanism, we used an image analysis system to measure the neuronal density around Bodian-stained NPs in the hippocampal CA1 area of eight AD patients. Neuronal density, as measured within two arbitrary concentric circles around NPs with a radius of 74 and 123.5 μm, respectively, was on average 19% and 16% lower than the density in similar control circles without NPs in the same section. Furthermore, neuronal density around NPs was inversely related to their size. To investigate the impact of such a local reduction in cell density around NPs on the entire CA1 area, we also determined the proportion of the CA1 covered by the NPs and the arbitrary concentric circles around them. This appeared to be 16.3% of the total CA1 area, which means that the negative effect of NPs on the cell density can only explain 2.6% of cell death in the entire CA area. In conclusion, this study suggests that although NPs have a local negative effect on neighboring neurons, their contribution to the strong decrease in CA1 cell numbers is limited.
Neuronal degeneration and regeneration:from basic mechanisms to prospects for therapy [Book]
Neurotrophin receptors in Alzheimer’s disease
Edited by Ahmad Salehi, Joost Verhaagen, and Dick Swaab
Note Ahmad Salehi was also one of the editors of Neuronal degeneration and regeneration:from basic mechanisms to prospects for therapy. The book is part of the series Progress in Brain Research.
Frameshift Mutants of β Amyloid Precursor Protein and Ubiquitin-B in Alzheimer’s and Down Patients
Fred van Leeuwen, Dominique de Kleijn, Helma van den Hurk, Andrea Neubauer, Marc Sonnemans, Jacqueline Slujis, Soner Köccü, Ravindra Ramdjielal, Ahmad Salehi, Gerard Martens, Frank Grosveld, John Burbach, and Elly Hol
Alzheimer's is characterized by short-term memory loss and plaques and tangles in the cerebral cortex. Aberrant ubiquitin protein, likely resulting from an RNA mutation, is detectable in plaques (vague spots) and tangles (flame shapes, ~40 µm long) in the hippocampus (a region involved in short-term memory) from an Alzheimer's patient. This mutation mechanism is likely a factor in nonfamilial Alzheimer's and other neurodegenerative pathologies.
The cerebral cortex of Alzheimer's and Down syndrome patients is characterized by the presence of protein deposits in neurofibrillary tangles, neuritic plaques, and neuropil threads. These structures were shown to contain forms of beta amyloid precursor protein and ubiquitin-B that are aberrant (+1 proteins) in the carboxyl terminus. The +1 proteins were not found in young control patients, whereas the presence of ubiquitin-B+1 in elderly control patients may indicate early stages of neurodegeneration. The two species of +1 proteins displayed cellular colocalization, suggesting a common origin, operating at the transcriptional level or by posttranscriptional editing of RNA. This type of transcript mutation is likely an important factor in the widely occurring nonfamilial early- and late-onset forms of Alzheimer's disease.
Connections, Cognition and Alzheimer's Disease [Book]
Reduced neuronal activity is one of the major hallmarks of Alzheimer’s disease
Edited by Dick Swaab, Paul Lucassen, J. van de Nes, Rifka Ravid, and Ahmad Salehi
Neuropeptides vasopressin (AVP), oxytocin (OXT) and corticotropin-releasing hormone (CRH) in the human hypothalamus: activity changes in aging, Alzheimer’s disease and depression
Paul Lucassen, F. Tilders, Ahmad Salehi, and Dick Swaab
Co-localization of high-affinity neurotrophin receptors in nucleus basalis of Meynert neurons and their differential reduction in Alzheimer’s disease
Ahmad Salehi, Joost Verhaagen, Paul Dijkhuizen, and Dick Swaab
It has been suggested that degeneration of neurons in Alzheimer's disease is the result of diminished trophic support. However, so far no evidence has been forwarded that neuronal degeneration in Alzheimer's disease is causally related to insufficient production of neurotrophins. The present study deals with (i) the expression and co-localization of tyrosine kinase receptors (trks) in the human nucleus basalis of Meynert and (ii) alterations of these receptors in Alzheimer's disease in the nucleus basalis of Meynert, an area severely affected in Alzheimer's disease. The expression of trkA, trkB and trkC in the nucleus basalis of Meynert of control and Alzheimer's disease brains was studied using three polyclonal antibodies specifically recognizing the extracellular domain of trkA, trkB and trkC. Brain material of eight controls and seven Alzheimer's disease patients was obtained at autopsy, embedded in paraffin and stained immunocytochemically. Using an image analysis system, we determined the proportion of trk neurons expressing the different trk receptors in controls and Alzheimer's disease patients. In control brains, trkA, trkB and trkC were differentially expressed in numerous nucleus basalis of Meynert neurons. The highest proportion of neurons was found to express trkB (75%), followed by trkC (58%) and trkA (54%). Furthermore, using consecutive sections, a clear co-localization of trk receptors was observed in the same neurons. The highest degree of co-localization was observed between trkA and trkB. In Alzheimer's disease patients, the number of immunoreactive neurons and the staining intensity of individual neurons was reduced dramatically. Reduction in the proportion of neurons expressing trkA was 69%, in trkB 47% and in trkC 49%, which indicated a differential reduction in the amount of trk receptors …
in Alzheimer's disease. These observations indicate that nucleus basalis of Meynert neurons can be supported by more than one neurotrophin and that the degeneration of these neurons in Alzheimer's disease is associated with a decreased expression of trk receptors, suggesting a decreased neurotrophin responsiveness of nucleus basalis of Meynert neurons in Alzheimer's disease.
Journal of Neuropathology & Experimental Neurology
Decreased activity of hippocampal neurons in Alzheimer’s disease is not related to the presence of neurofibrillary tangles
Ahmad Salehi, Rifka Ravid, Nikolas Gonatas, and Dick Swaab
Numerous studies have established the key role of the Golgi apparatus (GA) in post-translational processing, transport and targeting of proteins destined for secretion, lysosomes and plasma membranes. Moreover, several studies performed in our laboratories have shown that the size of the immunocytochemically detected neuronal GA is a reliable index of neuronal activity in aging, Alzheimer's disease (AD) and amyotrophic lateral sclerosis. It has been suggested that in AD there is decreased neuronal activity, e.g. in terms of glucose metabolism and protein synthetic capability. To further explore the hypothesis of decreased neuronal activity in AD, in this study the size of the GA was measured in pyramidal neurons of the CAI area of the hippocampus of non-demented controls and AD patients. The size of the GA was measured separately in neurons with and without neurofibrillary tangles (NFT). Moreover, in order to establish a correlation between the density of NFT and the size of the GA, the density of extraneuronal NFT was determined around each neuron and related to the size of its GA. The results, quantified by image analysis, indicate that there is a significant reduction in GA size in the neurons of the CAI area of the hippocampus of AD patients. However, there was no significant relationship between the size of the GA and the presence or absence of intracellular NFT. In addition, there was no correlation between the density of extracellular NFT and GA size of adjacent neurons. These findings are consistent with the conclusion that in AD there is evidence of decreased protein processing and secretion in the affected neurons of the CAI area of the hippocampus. However, we failed to detect a relationship between intracellular or extracellular NFT and neuronal protein synthetic ability. …
Journal of Neuropathology & Experimental Neurology (cont.)
These results justify the hypothesis that in AD a primary lesion is hypoactivity of neurons that is not directly linked with the development of intracellular or extracellular NFT.
Decreased protein synthetic activity of the hypothalamic tuberomamillary nucleus in Alzheimer’s disease as suggested by smaller Golgi apparatus.
Ahmad Salehi, S. Heyn, Nikolas Gonatas, and Dick Swaab
The nucleus tuberalis lateralis (NTL) and tuberomamillary nucleus (TM), which are located close together in the tuberal region of the human hypothalamus, are differentially affected by Alzheimer's disease (AD), In the AD, the NTL shows only early cytoskeletal alterations, i.e. pre-tangle stages, while the TM is characterised by advanced Alzheimer's changes, e.g. neurofibrillary degeneration, senile plaques and amyloid deposition. Earlier we showed that the early cytoskeletal alterations in the NTL are not accompanied by changes in protein synthetic activity. The present study was carried out in order to measure the protein synthetic activity of the neighbouring area, the TM, which is severely affected by advanced Alzheimer changes. A polyclonal antibody against MG-160, a conserved membrane sialoglycoprotein of the Golgi apparatus, was used to stain this organelle and using an image analysis system, the size of the Golgi apparatus was measured as an index for synthetic and secretory activity in 15 Alzheimer patients and 21 controls. A significant decrease in the size of the Golgi apparatus was found in the TM neurons in AD, although the cell profile area remained unchanged. These data suggest that the protein synthetic and secretory activity of TM neurons is indeed decreased in AD.
Early cytoskeletal changes as shown by Alz-50 are not accompanied by decreased neuronal activity
Ahmad Salehi, J. van de Nes, Michel Hofman, Nikolas Gonatas, and Dick Swaab
The nucleus tuberalis lateralis (NTL) is located in the basolateral part of the hypothalamus and is only present as a well-delineated nucleus in human and higher primates. In Alzheimer's disease (AD), NTL neurons show strong early cytoskeletal alterations, as revealed by the antibody Alz-50, but practically no senile plaques or neurofibrillary tangles. To study whether the activity of NTL neurons decreases when cytoskeletal changes appear, i.e., during aging and in AD, we applied a polyclonal antibody raised against the medial cisternae of the Golgi apparatus (GA). The size of the GA and the cell profile of NTL neurons, two established parameters for neuronal activity, were measured by an image analysis system. No significant change in the size of the profiles of the GA or of the neurons was observed in this nucleus during aging or AD. Earlier studies have shown that there is no decrease in cell number in the NTL in AD. We conclude that in the NTL an early hallmark of AD, i.e., cytoskeletal changes as stained by Alz-50, does not correlate with decreased neuronal activity, as reflected by the size of the GA, nor with a decrease in cell number. In addition, we found that the very early occurring and abundant presence of lipofuscin in NTL neurons does not go together with decreased neuronal activity.
Journal of Neuroendocrinology
Activation of vasopressin neurons in aging and Alzheimer’s disease
Paul Lucassen, Ahmad Salehi, Chris Pool, Nikolas Gonatas, and Dick Swaab
The supraoptic (SON) and paraventricular nuclei (PVN) of the human hypothalamus are production sites of vasopressin (AVP) and oxytocin (OXT). Although the hypothalamus is affected in Alzheimer's disease (AD), previous work has not only shown that in these two nuclei no neurons are lost, neither during aging nor in AD, but that the number of AVP-expressing neurons and their nucleolar size had even increased with age. These observations indicated that the peptide synthesis of the AVP neurons was activated in the oldest age-groups. Recently published, qualitative observations, using the area of the Golgi Apparatus (GA) as a sensitive parameter for neurosecretory activity, confirmed the activation of SON and PVN neurons with age in human; however, in this report the neurons were not identified according to their neuropeptide content. In the present quantitative study we determined whether the AVP neurons were indeed activated as a result of the aging process in controls and AD patients. We applied a polyclonal antiserum directed against the medial cisternae of the GA on formalin-fixed, paraffin-embedded tissue sections taken from the dorsolateral SON (dl-SON) of 10 controls and 10 AD patients, and performed our measurements in this area that is known to be predominantly occupied (90–95%) by AVP neurons. In addition, the sparse OXT cells present in the area of study, were excluded from the measurements on the basis of alternative sections stained for OXT. In the dl-SON, the area occupied by the GA and the cellular profile area per patient were quantified by means of image analysis. The results show a significant increase in GA area with age in controls and in AD, demonstrating an activation of the AVP neurons in the dl-SON of the human hypothalamus in these two conditions. No changes were observed in the cellular profile areas with age, neither in the controls nor in AD, …
Journal of Neuroendocrinology (cont.)
suggesting that the GA area is a much more sensitive parameter for monitoring activity changes in post-mortem material than neuronal size. It is proposed that this activation of AVP cells with age, which has been suggested to be a compensatory response to the age-related loss of AVP receptors in the kidney, might be the basis of the stability of these neurons in aging and AD.
Neuronal atrophy, not cell death, is the main hallmark of Alzheimer’s disease.
Dick Swaab, MA Hofman, Paul J Lucassen, Ahmad Salehi, and Harry B Uylings