Number Of Cholinergic Neurons Research Articles

Effect of Basic Fibroblast Growth Factor on Hippocampal Cholinergic Neurons in a Rodent Model of Ischaemic Encephalopathy

Ischaemic encephalopathy (IE) is a debilitating condition resulting from stroke that can lead to impaired learning and memory related to damage of cholinergic neurons in the hippocampus. The present study used an animal model of IE to test the hypothesis that treatment with basic fibroblast growth factor (bFGF) can reduce cognitive symptoms of IE by increasing the number of cholinergic neurons in the CA region of the hippocampus. The animal model of IE was created surgically by double ligation of the bilateral common carotid arteries. Three groups of Sprague-Dawley rats: sham, IE and IE with bFGF treatment group, were measured for changes in learning and memory using the Morris water maze test. Microscopic and immunohistochemical techniques were used to identify cells that bind bFGF and cholinergic neurons. IE rats treated with bFGF had better scores in the Morris water maze test than the untreated IE group, indicating improved learning and memory in the treated group. Microscopy showed that bFGF crossed the blood-brain barrier, was taken up by neurons in the hippocampus, and that the number of cholinergic neurons in the treated group was significantly increased. These results may provide an experimental basis for the treatment of IE by subcutaneous injection of bFGF.

Hypercholesterolemia in rats impairs the cholinergic system and leads to memory deficits

Alzheimer's disease (AD) is a chronic brain disorder characterized by cognitive impairment, cholinergic dysfunction, inflammation, tau and beta-amyloid pathology and vascular damage. Recent studies have shown, that high cholesterol levels are linked to the pathology of AD. The aim of our present work was to study the effects of hypercholesterolemia in adult rats. Five months after 5% cholesterol-enriched diet plasma cholesterol levels and total weight were significantly enhanced compared to controls. Spatial memory was studied in an 8-arm radial maze and cholesterol-treated rats showed an impaired learning and long-term memory. Hypercholesterolemia significantly reduced the number of cholinergic neurons in the basal nucleus of Meynert and decreased acetylcholine levels in the cortex. Nerve growth factor was only slightly enhanced in the cortex of cholesterol-treated animals. Levels of amyloid precursor protein, beta-amyloid(1–42), as well as tau and phospho-tau 181 were significantly enhanced in the cortex of cholesterol-fed rats. Hypercholesterolemia markedly increased several cerebral inflammatory markers and enhanced microglial CD11b-like immunoreactivity. Vascular density, stained by RECA-1 was not changed. However, cholesterol induced cortical microbleedings illustrated by intensive anti-rat IgG-positive spots in the cortex. In conclusion, our data demonstrate that hypercholesterolemia in rats caused memory impairment, cholinergic dysfunction, inflammation, enhanced cortical beta-amyloid and tau and microbleedings, all indications, which resemble an AD-like pathology.

Calpain inhibition prevents amyloid-β-induced neurodegeneration and associated behavioral dysfunction in rats

Amyloid-β (Aβ) is toxic to neurons and such toxicity is – at least in part – mediated via the NMDA receptor. Calpain, a calcium dependent cystein protease, is part of the NMDA receptor-induced neurodegeneration pathway, and we previously reported that inhibition of calpain prevents excitotoxic lesions of the cholinergic nucleus basalis magnocellularis of Meynert. The present study reveals that inhibition of calpain is also neuroprotective in an in vivo model of Aβ oligomer-induced neurodegeneration in rats. Aβ-induced lesions of the nucleus basalis induced a significant decrease in the number of cholinergic neurons and their projecting fibers, as determined by analysis of choline-acetyltransferase in the nucleus basalis magnocellularis and cortical mantle of the lesioned animals. Treatment with the calpain inhibitor A-705253 significantly attenuated cholinergic neurodegeneration in a dose-dependent manner. Calpain inhibition also significantly diminished the accompanying neuroinflammatory response, as determined by immunohistochemical analysis of microglia activation. Administration of β-amyloid markedly impaired performance in the novel object recognition test. Treatment with the calpain inhibitor, A-705253, dose-dependently prevented this behavioral deficit. In order to determine whether pre-treatment with the calpain inhibitor is necessary to exhibit its full protective effect on neurons we induced Aβ toxicity in primary neuronal cultures and administered A-705253 at various time points before and after Aβ oligomer application. Although the protective effect was higher when A-705253 was applied before induction of Aβ toxicity, calpain inhibition was still beneficial when applied up to 1 h post-treatment. We conclude that inhibition of calpains may represent a valuable strategy for the prevention of Aβ oligomer-induced neuronal decline and associated cognitive deterioration.

Narcoleptic orexin receptor knockout mice express enhanced cholinergic properties in laterodorsal tegmental neurons

Pharmacological studies of narcoleptic canines indicate that exaggerated pontine cholinergic transmission promotes cataplexy. As disruption of orexin (hypocretin) signaling is a primary defect in narcolepsy with cataplexy, we investigated whether markers of cholinergic synaptic transmission might be altered in mice constitutively lacking orexin receptors (double receptor knockout; DKO). mRNA for Choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT) and the high-affinity choline transporter (CHT1) but not acetylcholinesterase (AChE) was significantly higher in samples from DKO than wild-type (WT) mice. This was region-specific; levels were elevated in samples from the laterodorsal tegmental nucleus (LDT) and the fifth motor nucleus (Mo5) but not in whole brainstem samples. Consistent with region-specific changes, we were unable to detect significant differences in Western blots for ChAT and CHT1 in isolates from brainstem, thalamus and cortex or in ChAT enzymatic activity in the pons. However, using ChAT immunocytochemistry, we found that while the number of cholinergic neurons in the LDT and Mo5 were not different, the intensity of somatic ChAT immunostaining was significantly greater in the LDT, but not Mo5, from DKO than from WT mice. We also found that ChAT activity was significantly reduced in cortical samples from DKO compared with WT mice. Collectively, these findings suggest that the orexins can regulate neurotransmitter expression and that the constitutive absence of orexin signaling results in an up-regulation of the machinery necessary for cholinergic neurotransmission in a mesopontine population of neurons that have been associated with both normal rapid eye movement sleep and cataplexy.

Neuroinflammation not associated with cholinergic degeneration in aged-impaired brain

Degeneration of the cholinergic neurons in the basal forebrain and elevation of inflammatory markers are well-established hallmarks of Alzheimer's disease; however, the interplay of these processes in normal aging is not extensively studied. Consequently, we conducted a neuroanatomical investigation to quantify cholinergic neurons and activated microglia in the medial septum/vertical diagonal band (MS/VDB) of young (6 months) and aged (28 months) Fisher 344 × Brown Norway F(1) rats. Aged rats in this study were impaired relative to the young animals in spatial learning ability as assessed in the Morris water maze. Stereological analysis revealed no difference between aged and young rats in the total numbers of cholinergic neurons, demonstrating that loss of cholinergic neurons is not a necessary condition to observe impaired spatial learning in aged rats. In this same region, the total number of activated microglia was substantially greater in aged rats relative to young rats. Jointly, these data demonstrate that aging is characterized by an increase in the basal inflammatory state within the MS/VDB, but this inflammation is not associated with cholinergic neuron death.

Lesion and stimulation of the ventral tegmental area increases cholinergic activity in the rat brain.

Our previous study indicated that unilateral lesion of the ventral tegmental area (VTA) facilitates contralateral VTA stimulation-induced feeding or exploration. The present study was aimed to determine the possible role of the central cholinergic systems in this effect. Immunohistochemistry for choline acetyltransferase (ChAT) was used to measure the number of active cholinergic neurons in their major groups (Ch1-Ch6) and in striatal regions in rats subjected to unilateral electrocoagulation and contralateral VTA electrical stimulation (L/S group) in comparison to the unilaterally stimulated (S), unilaterally lesioned (L) and sham (Sh) groups. The study showed that unilateral VTA lesion increased (as compared to Sh group) the number of ChAT+ neurons in the Ch1-Ch3 and unilateral VTA stimulation increased the number in the Ch1 and the ventral pallidum only. The most sensitive to these changes in the mesolimbic system were cholinergic structures providing hippocampal afferentation. Surprisingly, there was no significant increase in the number of ChAT+ neurons in the L/S group. The obtained results did not confirm any evident influence of the cholinergic systems on the VTA lesion-induced facilitation of the behavioral response evoked by contralateral VTA stimulation.

Trichloroethylene induces dopaminergic neurodegeneration in Fisher 344 rats

Trichloroethylene, a chlorinated solvent widely used as a degreasing agent, is a common environmental contaminant. Emerging evidence suggests that chronic exposure to trichloroethylene may contribute to the development of Parkinson's disease. The purpose of this study was to determine if selective loss of nigrostriatal dopaminergic neurons could be reproduced by systemic exposure of adult Fisher 344 rats to trichloroethylene. In our experiments, oral administration of trichloroethylene induced a significant loss of dopaminergic neurons in the substantia nigra pars compacta in a dose-dependent manner, whereas the number of both cholinergic and GABAergic neurons were not decreased in the striatum. There was a robust decline in striatal levels of 3, 4-dihydroxyphenylacetic acid without a significant depletion of striatal dopamine. Rats treated with trichloroethylene showed defects in rotarod behavior test. We also found a significantly reduced mitochondrial complex I activity with elevated oxidative stress markers and activated microglia in the nigral area. In addition, we observed intracellular alpha-synuclein accumulation in the dorsal motor nucleus of the vagus nerve, with some in nigral neurons, but little in neurons of cerebral cortex. Overall, our animal model exhibits some important features of Parkinsonism, and further supports that trichloroethylene may be an environmental risk factors for Parkinson's disease.

Effects of NT-4 gene modified fibroblasts transplanted into AD rats

It is well known that fibroblasts can act as a cell vector to express functional protein, like neurotrophin-4 (NT-4). The present study evaluated the effect of NT-4 gene modified fibroblasts grafted into the hippocampus of AD rat model. AD rats were reproduced by bilateral transection of the hippocampal fimbria-fornix. The transplanted fibroblasts steadily expressed NT-4 proteins at least 2 months after transplantation. This correlated with a significant rescue in the number of cholinergic neurons in the host hippocampus. Morris water maze tests demonstrated significant improvements in learning and memory, especially in rats receiving NT-4-modified fibroblasts. The present results showed that NT-4 gene modified fibroblasts could provide a long-term and steady expression of NT-4, and it significantly improved the behavior of AD rats. These findings should have important clinical applications in providing a long-term NT-4 Secretion for the treatment of AD.

Accumulation of reelin‐positive plaques is accompanied by a decline in basal forebrain projection neurons during normal aging

Besides its critical role during neurodevelopment, the extracellular glycoprotein reelin is also a pivotal regulator of adult synaptic function and plasticity, and altered reelin-mediated signalling has been suggested to contribute to neuronal dysfunction associated with Alzheimer's disease. We have recently discovered, in aged rodents and non-human primates, a pronounced decline in reelin-positive interneurons and concomitant accumulation of reelin in extracellular amyloid-like deposits, both being associated with episodic-like memory impairments. Here, we report that these age-related neuropathological changes in hippocampus, entorhinal and piriform cortices of aged wild-type mice are accompanied by abnormal axonal varicosities and altered expression profiles of calcium-binding proteins in plaque-dense areas, as well as a significant reduction in the number of parvalbumin-positive gamma-aminobutyric acid (GABA)ergic projection neurons in basal forebrain areas, including medial septum (MS), ventral and horizontal diagonal Band of Broca (VDB/HDB) and substantia innominata (SI), compared with young subjects. In addition, a significant reduction in the number of choline acetyltransferase-positive cholinergic projection neurons was evident in the HDB/SI area but not in the MS of aged compared with young wild-type mice. No reelin-deposits were found in these basal forebrain regions. Our findings suggest that the elevated reelin plaque load in the projection areas of afferent subcortical GABAergic and cholinergic neurons including hippocampus, entorhinal and piriform cortices affects the axonal integrity and survival of these neurons, potentially contributing to the cognitive impairments observed in aged wild-type mice.

The pro-apoptotic substance thapsigargin selectively stimulates re-growth of brain capillaries.

Thapsigargin is a pro-apoptotic chemical, which has been shown to be useful to study cell death of cholinergic or dopaminergic neurons, or cells, which degenerate in Alzheimer's disease or Parkinson's disease, respectively. The aim of the present work was to study the effects of thapsigargin in the well established organotypic brain co-slice model composed of the basal nucleus of Meynert (nBM), ventral mesencephalon (vMes), dorsal striatum (dStr) and parietal cortex (Ctx). Cholinergic acetyltransferase-positive neurons in the nBM and dStr and dopaminergic tyrosine hydroxylase-positive neurons in the vMes survived, when cultured for 4 weeks with nerve growth factor and glial cell line-derived neurotrophic factor. Nerve fibers of cholinergic nBM neurons grew into the cortex and dopaminergic nerve fibers sprouted into dopamine D2 receptor-positive dStr. The whole co-slice contained a dense laminin-positive capillary network. Treatment of co-cultures with 3 microM thapsigargin for 24 hr significantly decreased the number of cholinergic neurons and dopaminergic neurons. This cell death displayed apoptotic DAPI-positive malformed nuclei and enhanced TUNEL-positive cells. Thapsigargin selectively stimulated the laminin-positive capillary growth between the nBM and Ctx. In conclusion, the induced cell death of cholinergic and dopaminergic neurons may be accompanied by enhanced angiogenic activity.

Age-related changes in number and expression patterns of cholinergic and GABAergic basal forebrain neurons in spatially characterized young and aged F344 rats

Age-related changes in number and expression patterns of cholinergic and GABAergic basal forebrain neurons in spatially characterized young and aged F344 rats

Rotenone induces cell death of cholinergic neurons in an organotypic co-culture brain slice model.

In Alzheimer and Parkinson's disease cell death of cholinergic and dopaminergic neurons are characteristic hallmarks, respectively. It is well established that rotenone, an inhibitor of complex I of the electron transport chain, induces cell death of dopaminergic neurons, however, not much is known on the effects of rotenone on cholinergic neurons. The aim of the present study was to evaluate the effects of rotenone on cholinergic neurons in an organotypic in vitro brain co-slice model. When co-cultures were treated with 10 μM rotenone for 24 h a significantly decreased number of cholinergic neurons was found in the basal nucleus of Meynert but not in the dorsal striatum. This cell death exhibited apoptotic DAPI-positive malformed nuclei and enhanced TUNEL-positive cells. In summary, inhibition of complex I of the electron transport chain may play a role in neurodegeneration of cholinergic neurons.

Enhanced spatial memory and hippocampal long‐term potentiation in p75 neurotrophin receptor knockout mice

Previous reports have described increases in the size and number of cholinergic neurons in the basal forebrain in p75 neurotrophin receptor (p75(NTR)) knockout mice. In an earlier study, we also found improved spatial memory in these mice, raising the possibility that p75(NTR) regulates hippocampal function by its effects on the cholinergic basal forebrain. We therefore investigated hippocampal long-term potentiation in p75(NTR) knockout mice that shared the same genetic background as control 129/Sv mice. We also investigated heterozygous mice, carrying just one functional p75(NTR) allele. The p75(NTR) knockout mice had enhanced long-term potentiation in the Schafer collateral fiber synapses of the hippocampus. Heterozygous mice had an intermediate level, greater than controls but less than knockout mice. Hippocampal choline acetyltransferase activity was also markedly elevated in p75(NTR) knockout mice, with a smaller increase in heterozygous mice. In the Barnes maze, p75(NTR) knockout mice displayed markedly superior learning to controls, and this was evident over the three age brackets tested. At each age, the performance of heterozygous mice was intermediate to the other groups. In the open field test, p75(NTR) knockout mice exhibited greater stress-related behavioral responses, including freezing, than did control animals. There were no differences between the three groups in a test of olfactory function. The dose-dependent effects of p75(NTR) gene copy number on hippocampal plasticity and spatial memory indicate that p75(NTR) has profound effects on hippocampal function. Bearing in mind that p75(NTR) is very sparsely expressed in the adult hippocampus and has a potent effect on hippocampal choline acetyltransferase activity, the effects of p75(NTR) on hippocampal function are likely to be mediated indirectly, by its actions on basal forebrain cholinergic neurons.

Structural and functional cardiac cholinergic deficits in adult neurturin knockout mice

Previous work provided indirect evidence that the neurotrophic factor neurturin (NRTN) is required for normal cholinergic innervation of the heart. This study used nrtn knockout (KO) and wild-type (WT) mice to determine the effect of nrtn deletion on cardiac cholinergic innervation and function in the adult heart. Immunohistochemistry, confocal microscopy, and quantitative image analysis were used to directly evaluate intrinsic cardiac neuronal development. Atrial acetylcholine (ACh) levels were determined as an indirect index of cholinergic innervation. Cholinergic function was evaluated by measuring negative chronotropic responses to right vagal nerve stimulation in anaesthetized mice and responses of isolated atria to muscarinic agonists. KO hearts contained only 35% the normal number of cholinergic neurons, and the residual cholinergic neurons were 15% smaller than in WT. Cholinergic nerve density at the sinoatrial node was reduced by 87% in KOs, but noradrenergic nerve density was unaffected. Atrial ACh levels were substantially lower in KO mice (0.013 +/- 0.004 vs. 0.050 +/- 0.011 pmol/microg protein; P < 0.02) as expected from cholinergic neuron and nerve fibre deficits. Maximum bradycardia evoked by vagal stimulation was reduced in KO mice (38 +/- 6% vs. 69 +/- 3% decrease at 20 Hz; P < 0.001), and chronotropic responses took longer to develop and fade. In contrast to these deficits, isolated atria from KO mice had normal post-junctional sensitivity to carbachol and bethanechol. These findings demonstrate that NRTN is essential for normal cardiac cholinergic innervation and cholinergic control of heart rate. The presence of residual cardiac cholinergic neurons and vagal bradycardia in KO mice suggests that additional neurotrophic factors may influence this system.

Effects of two years of conjugated equine estrogens on cholinergic neurons in young and middle-aged ovariectomized monkeys

The effect of estrogen on the number and size of cholinergic neurons in the basal forebrain was examined in surgically menopausal young and middle-aged cynomolgus monkeys. Young and middle-aged female monkeys were ovariectomized and treated with conjugated equine estrogens (Premarin) at doses that are equivalent to those currently prescribed to postmenopausal women. In the medial septum/diagonal band (MS/DB), no effect of treatment with Premarin was observed in the cholinergic neurons in either ovariectomized young or middle-aged monkeys. However, the number and size of cholinergic neurons in the MS/DB of middle-aged monkeys was greater than that in the young monkeys. In the nucleus basalis of Meynert (NBM) of middle-aged monkeys, the number of cholinergic neurons in the intermediate region (Ch4i) was greater in Premarin-treated monkeys as compared to controls and numbers of neurons in this region were greater at higher levels of estrogen. No effects of estrogen were observed in other NBM regions in the middle-aged monkeys and the size of cholinergic neurons was unaffected by Premarin. These findings suggest that treatment with Premarin has selective beneficial effects on cholinergic neurons in the basal forebrain but that these effects are both age and region specific.

Effects of engrafted neural stem cells in Alzheimer's disease rats

Cell therapy is thought to have a central role in restorative therapy, which aims to restore the function of the damaged nervous system. Neural stem cells (NSCs) can differentiate into neurons, astrocytes and oligodendrocytes. The purpose of this study was to evaluate the therapeutic effects of transplanting NSCs into rats which have the animal model of Alzheimer's disease (AD). NSCs from the hippocampus and NSCs-derived glial cells labeled with 5′-Bromo-2′-deoxyuridine (BrdU) were transplanted into two groups of transected rat basal forebrain. Nestin staining, glial fibrillary acidic protein (GFAP) staining and double-labeling immunofluorescence were used to detect the engrafted cells in the basal forebrain. Immunohistochemical detection of p75NGFR showed that the number of cholinergic neurons of the NSCs-transplanted group was significant higher than that of the glia-transplanted group in medial septum (MS) and vertical diagonal branch (VDB) (P<0.05). Learning and memory abilities were also measured by Y-maze test. The results indicate that transplanted NSCs can differentiate into cholinergic neurons, which may play an important role in the therapeutic effects of transplanted NSCs.

Sex differences in water maze performance and cortical neurotrophin levels of LHX7 null mutant mice

Mice lacking both alleles of the LIM-homeobox gene Lhx7 display dramatically reduced number of forebrain cholinergic neurons. Given the fact that sex differences are consistently observed in forebrain cholinergic function, in the present study we investigated whether the absence of LHX7 differentially affects water maze performance in the two sexes. Herein we demonstrate that LHX7 null mutants display a sex-dependent impairment in water maze, with females appearing more affected than males. Moreover, neurotrophin assessment revealed a compensatory increase of brain-derived neurotrophic factor and neurotrophin 3 in the neocortex of both male and female mutants and an increase of nerve growth factor levels only in the females. Nevertheless, the compensatory increase of cortical neurotrophin levels did not restore cognitive abilities of Lhx7 homozygous mutants. Finally, our analysis revealed that cortical neurotrophin levels correlate negatively with water maze proficiency, indicating that there is an optimal neurotrophin level for successful cognitive performance.

Stimulation of choline acetyltransferase by C3d, a neural cell adhesion molecule ligand

Septal cholinergic neurons project to the hippocampus and release acetylcholine, a neurotransmitter involved in learning and memory. The enzyme choline acetyltransferase (ChAT) is responsible for synthesizing acetylcholine. Promoting ChAT activity and acetylcholine release can lead to new treatments for neurodegenerative diseases with cholinergic deficits, such as Alzheimer's disease. We present evidence that the synthetic molecule C3d, which is a peptide mimetic of the neural cell adhesion molecule (NCAM), promotes ChAT activity in cultures of rat embryonic septal neurons. Our data demonstrate that ChAT activity triggered by C3d is dependent on the fibroblast growth factor receptor (FGFR) and the mitogen-activated protein kinase (MAPK) pathway. C3d did not affect the number of cholinergic neurons in culture, indicating that NCAM homophilic binding enhances ChAT activity, without affecting cholinergic cell survival. In conclusion, the NCAM mimetic peptide C3d promotes ChAT activity in septal neurons through FGFR and MAPK. These findings are relevant to the design of new strategies aimed at stimulating cholinergic function and improving cognition in disorders such as Alzheimer's disease.

Involvement of angiotensin converting enzyme in cerebral hypoperfusion induced anterograde memory impairment and cholinergic dysfunction in rats

Forebrain cholinergic dysfunction is the hallmark of vascular dementia (VaD) and Alzheimer's dementia (AD) induced by cerebral hypoperfusion during aging. The aim of the present study is to evaluate the role of angiotensin converting enzyme (ACE) in cerebral hypoperfusion-induced dementia and cholinergic dysfunction. Chronic cerebral hypoperfusion (CHP) was induced by permanent bilateral common carotid artery (2VO) occlusion in rats. Chronic cerebral hypoperfusion resulted in anterograde memory impairment revealed from Morris water maze (MWM) and passive avoidance step through tasks (PA), which was significantly attenuated by ACE inhibitor, captopril. Cerebral hypoperfusion down-regulated the relative expression of cholinergic muscarinic receptor (ChM-1r) and choline acetyltransferase (ChAT) as well as up-regulated the angiotensin II type-1 receptor (AT-1) expression in hippocampus of vehicle treated CHP group on the 54th day post-hypoperfusion. The diminished number of presynaptic cholinergic neurons and the pyramidal neurons were evident from ChAT-immunofluorescence and the hematoxylin and eosin (H&E) staining studies respectively in hippocampal Cornu ammonis1 (CA1); region of vehicle-treated hypoperfused animals. Further the lipid peroxidation level was also found to be elevated in the hippocampus of the vehicle-treated group. Our results demonstrated that continuous captopril treatment (50 mg/kg, i.p. twice daily) for 15 days mitigated the hypoperfusion-induced cholinergic hypofunction and neurodegeneration in hippocampus. The present study robustly reveals that the angiotensinergic system plays a pivotal role in progression of neuronal death and memory dysfunctions during cerebral hypoperfusion.

BDNF improves the effects of neural stem cells on the rat model of Alzheimer's disease with unilateral lesion of fimbria-fornix

In this study, neural stem cells (NSCs) were obtained from the hippocampus using the serum-free culturing. NSCs labeled with 5'-bromo-2'-deoxyuridine (BrdU) were transplanted into transected rat basal forebrain followed by the injection of brain-derived neurotrophic factor (BDNF) into the lateral ventricle. Nestin staining and double-labeling immunohistochemistry were used to detect cell survival and neuronal differentiation of the BrdU labeled cells in the basal forebrain and it was observed that labeled NSCs differentiated into neurons and astrocytes in the basal forebrain. Immunohistochemical detection of p75(NGFR) indicated that the number of cholinergic neurons of the combination groups treated by NSCs, BDNF, and NSCs groups had more significant improvement than that of the injured groups in medial septum (MS) and vertical diagonal branch (VDB). Learning and memory abilities were also measured by Y-maze test and the results support that BDNF can enhance the treatment effects of NSCs transplanted into brain lesion model.