Migratory potential of transplantable neural tumor cell lines

Experimentally induced primitive neuroectodermal tumor (PNET) cell lines were transplanted into neonatal and adult rat brain and examined neuropathologically for their tumorigenic potential. Both cell lines showed a striking migratory behavior in both neonatal and adult brain. Migration of tumor cells was found in host brain parenchyma, along white matter tracts and associated with CSF pathways. These neural tumor cell lines provide a valuable tool for the development of strategies against strongly migrating neural tumors. Received: 22 June 1998 / Revised: 5 November 1998, 30 November 1998 / Accepted: 30 November 1998     

Selective polyunsaturated fatty acids enrichment in phospholipids from neuronal-derived cell lines

Most studies aimed at exploring the molecular and cellular properties of plasma membranes in neural tissues make use of cell lines. However, cell membrane lipid composition of cell lines is notably different from that of brain tissues where they presumably derive from. Using septal-derived SN56 cells and hippocampal-derived HT22 cells, we demonstrated that cell lines exhibit lower contents of saturated (18:0) and long polyunsaturated fatty acids (PUFA; 20:4n-6 and especially 22:6n-3), as well as higher monounsaturated fatty acid contents (mainly 18:1n-9), compared to mouse brain. Also, cell lines exhibited higher contents of sterol esters and lower contents of cholesterol and phospholipids, especially phosphatidylethanolamine and phosphatidylserine. We have also evaluated the effects of different (n-3/n-6) PUFA enrichments on fatty acid and phospholipid contents in these cell lines. Our results show that enrichment of culture medium with 22:6n-3 and 20:4n-6 in a 70/30 proportion during 48 h, using fat-free bovine serum albumin as vehicle, successfully readjusted fatty acid profiles in cell line-polar lipids to values found in natural nerve cells. Interestingly, no differences in cell survival were observed upon enrichment. The generalization of these methodologies would allow a more feasible adaptation of cellular models to the study of in vivo nerve physiology.     

Antioxidant activity of X-34 in synaptosomal and neuronal systems

Inhibiting aggregation and deposition of amyloid beta-peptide (Abeta) in brain is a therapeutic strategy for Alzheimer's disease (AD). A Congo-red-like molecule, X-34, is reported to bind to Abeta deposits. Oxidative stress associated with Abeta is hypothesized to be critical for the neurotoxic properties of this peptide. The present study was undertaken to test the hypothesis that X-34, with its salicylate groups, would act as an antioxidant. When challenged by hydroxyl or peroxyl free radicals or Abeta(1-42), oxidative stress and neurotoxicity occurred in neural systems as assessed by several indices. However, pretreatment of synaptosomes and primary neuronal cell culture with X-34 greatly ameliorated lipid peroxidation induced by these free radicals and Abeta(1-42). Protein oxidation was not prevented by X-34. These results are discussed in terms of potential therapeutic use of X-34 and related compounds in AD.     

A system, using neural cell lines, to characterize HSV-1 vectors containing genes which affect neuronal physiology, or neuronal promoters.

Among the potential uses of defective herpes simplex virus (HSV-1) vectors are to study neuronal physiology, neuronal gene regulation, and to perform gene therapy of neuronal diseases. The prototype HSV-1 vector, pHSVlac, stably expresses Escherichia coli beta-galactosidase from the HSV-1 immediate early (IE) 4/5 promoter in cultured rat peripheral and CNS neurons, and in neurons in the adult rat brain. The LacZ gene and the IE 4/5 promoter in pHSVlac can be replaced with genes which affect neuronal physiology or cellular promoters, respectively. A system is required to characterize these HSV-1 vectors; cultured neurons, a mixture of different kinds of neurons and glia, cannot be used. In contrast, neural cell lines represent a homogenous population of neural cells available in virtually unlimited quantities. A system, using neural cell lines, to characterize HSV-1 vectors carrying other genes or promoters is now reported: First, 4 assays are described to detect HSV-1 vector DNA, RNA transcribed from the vector, and to quantitate beta-galactosidase expression. Second, 8 cell lines derived from rodents, primates, and humans were infected with pHSVlac virus and shown to express beta-galactosidase. The cell lines tested included adrenergic and cholinergic mouse neuroblastoma cells, rat pheochromocytoma cells, rodent pituicytes, and human neuroblastoma cells. Infection of these cell lines should prove useful for characterizing HSV-1 vectors with molecular and biochemical assays. Third, differentiated rat pheochromocytoma and mouse neuroblastoma cells, which resemble neurons, were infected with pHSVlac virus and shown to stably express beta-galactosidase. Infection of these cells should be useful for determining the effect of various HSV-1 vectors on neuronal physiology. Thus, HSV-1 vectors containing various genes or promoters can be characterized using the system described in this study.     

Monoamine oxidase A regulates neural differentiation of murine embryonic stem cells

Monoamine oxidase (MAO) A is the major metabolizing enzyme of serotonin (5-hydroxytryptamine, 5-HT) which regulates early brain development. In this study, wild-type (WT) and MAO A^neo embryonic stem (ES) cell lines were established from the inner cell mass of murine blastocysts and their characteristics during ES and differentiating stages were studied. Our results show that the differentiation to neural cells in MAO A^neo ES cells was reduced compared to WT, suggesting MAO A played a regulatory role in stem cells neural differentiation.     

Membrane protein and glycoprotein composition of beef brain synaptic vesicles

Synaptic vesicles were isolated from adult bovine cortical gray matter by differential centrifugation and membrane filtration of a hypoosmotically lysed crude mitochondrial fraction. Vesicle preparations were analyzed for purity by electron microscopy and enzyme assays. Polyacrylamide gel electrophoresis of SDS-solubilized and 2-mercaptoethanol-reduced vesicle membrane proteins revealed 4 major proteins with molecular weights ranging from 17,000 to 60,000, and about 10 minor proteins with molecular weights up to 170,000. The protein profile of the Triton X-100-extracted vesicle membranes was less complex, with 1 major protein and 5 minor bands. The major protein of the Triton extract was identified as a glycoprotein with a molecular weight of 45,000. Two additional minor PAS-positive bands were seen, with molecular weights of 78,000 and 95,000.     

Differentiation and heterogeneity in T-antigen immortalized precursor cell lines from mouse cerebellum.

Recently, various techniques have been developed to transfer oncogenes into brain cells in order to generate immortalized neural cell lines. It is of interest to establish how well such cell lines reflect their cellular origin. Here we report the characterization of sixteen cell lines from mouse cerebellum and, as a control, six cell lines from skin. Lines were established by immortalizing postnatal primary cell cultures with a retrovirus carrying a modified temperature-sensitive variant of SV40 large T antigen. The cell lines reflect many properties of the cell type from which they were derived. All of the sixteen cerebellar lines expressed one or more markers of the neural precursor cells, namely, nestin and epitopes for NG2 and A2B5. In contrast, none of the six skin lines expressed neural precursor markers. Both types of cell lines expressed vimentin and fibronectin. Differentiation occurred in some of the cerebellar lines and was enhanced in defined medium. A small percentage of cerebellar cells, usually less than 5%, was positive for a marker of differentiation, e.g., glial fibrillary acidic protein (GFAP), galactocerebroside (GalC), or L1. Expression of GFAP colocalized with that of nestin at varying levels of intensity, indicating a gradual replacement of nestin by GFAP in the cytoskeleton. Both the cells positive for precursor markers and those positive for differentiation markers tended to be located in clusters, suggesting that stochastic processes or cell-cell interactions are important for the determination of the fate of cells within a clonal cell line in vitro. The degree of differentiation seemed to correlate with a shift from serum-containing to defined medium, but not with a shift from the permissive to the nonpermissive temperature for T antigen expression. The immortalization approach described here thus allows the establishment of cell lines which are "captured" in the precursor state of the developing mouse neuroepithelium.     

Distribution of phosphate-independent MAP2 epitopes revealed with monoclonal antibodies in microwave-denatured human nervous system tissues

In contrast with results obtained in experimental animals, antibodies to microtubule associated protein-2 (MAP2) preferentially label abnormal structures in human nervous system tissue samples, but the normal sites at which MAP2 is expressed are not well-defined. To determine the distribution of MAP2 in the human central (CNS) and peripheral (PNS) nervous systems, we prepared monoclonal antibodies (MAbs) specific to MAP2, and compared the localization of this MAP in postmortem bovine and human tissues as well as in several human neural cell lines that express either neurofilament (NF) or glial filament (GF) proteins. Eight MAbs specific for phosphate-independent epitopes in bovine and human MAP2 were obtained, and those that performed well in tissues produced immunoreactivity confined to the somatodendritic domain of neurons in bovine and human CNS and PNS tissues. Other neural cells (e.g. astrocytes) did not express MAP2 immunoreactivity using these MAbs. Postmortem delays of less than 24 h prior to tissue denaturation did not affect the distribution of MAP2 immunoreactivity. However, microwave denaturation of these tissues preserved MAP2 immunoreactivity better than fixation with Bouin's solution or formalin. Microwave treatment also improved the immunoreactivity of several MAbs for NF and GF proteins. Finally, MAP2 was not detected in human neural cell lines that express NF (2) or GF (1) proteins. We conclude that microwave denaturation provides an effective means to preserve the immunoreactivity of normal human neuronal cytoskeletal proteins, and that this method of tissue denaturation allows the normal distribution of MAP2 to be defined in postmortem samples of human CNS and PNS tissues.     

Identification of KLRC2 as a candidate marker for brain tumor-initiating cells

ABSTRACT

Objective: Brain tumor-initiating cells are characterized by their features of self-renewal, multi-lineage differentiation, and tumorigenicity. We analyzed the gene expression of brain tumor-initiating cells to identify their novel cellular markers.

Methods: We performed cDNA microarray, in silico expressed sequence tags (ESTs), RT-PCR, and q-PCR analyses.

Results: We identified 10 genes that were more highly expressed in brain tumor-initiating cells than in neural stem cells. In addition, we identified 10 other genes that were more highly expressed in brain tumor-initiating cells than in glioma cell line cells from the cDNA microarray analysis. Using the EST database, we looked to see if the 20 genes were expressed more highly in gliomas, compared with normal adult brains. Among the 20 genes, five (KLRC2, HOXB2, KCNJ2, KLRC1, and COL20A1) were expressed more than twice in glioma samples, compared with normal adult brains, and, therefore, were referred for further evaluation. RT-PCR was conducted using cDNA samples obtained from neural stem cells, normal brain tissue, fetal brain tissue, glioma cell lines, and glioma tumor-initiating cell lines. KLRC2, a transmembrane activating receptor in natural killer cells, was expressed more highly in glioma-initiating cells than in neural stem cell lines or normal adult brain tissue. The q-PCR analysis revealed that expression of KLRC2 was significantly higher in brain tumor-initiating cells compared to normal brain controls.

Conclusion: KLRC2 could be a novel cellular marker for brain tumor-initiating cells.     

Neurotrophic effects of transferrin on embryonic chick brain and neural retina cell cultures

The viability and differentiation promoting effects of various transferrins [iron-saturated (holo) and iron-depleted (apo) human and chick ovo (conalbumin)-transferrins, and bovine apo-transferrin] were studied, using serum-free, flat-sedimented cell cultures of embryonic chick brain and neural retina. The effects of transferrin (Tf) on the cell cultures depended on the type of Tf used and the parameter measured.Significant differences between brain and neural retina cultures in the effects of apo-ovoTf and iron [supplemented as ammonium-iron (III) citrate] were detected. Maximal levels of mitochondrial activity were observed in the presence of 2 mg/l apo-ovoTf in neural retina cell cultures. In brain cell cultures, 40 mg ovoTf/l were needed to achieve maximal levels. In brain, but not in neural, retina cell cultures ovoTf and optimal concentrations of Fe³⁺ exhibited similar effects on biochemical parameters of cell function and differentiation. Although, in the absence of ovoTf, neuronal outgrowth on areas not covered by glial cells was inhibited in both cell cultures, the differences were more prominent in neural retina cell cultures. Our data strongly suggest that Tf plays a key role in processes not connected directly with its iron transport capability.     

人脑胶质瘤干细胞增殖和分化过程中NOTCH-1信号通路的调控

背景：研究发现NOTCH-1信号通路在神经干细胞或神经前体细胞的自我更新、增殖及分化中起重要作用。 目的：探讨NOTCH-1信号通路在人脑胶质瘤干细胞增殖和分化过程中的调控作用。 设计、时间及地点：开放性实验,于2005-01/2007-03在解放军第三军医大学新桥医院完成。 材料：人脑胶质瘤组织、正常成人脑组织由解放军第三军医大学新桥医院神经外科提供。U251胶质瘤细胞株由解放军第三军医大学新桥医院神经外科吕胜青副教授惠赠;CHG-5胶质瘤细胞株由解放军第三军医大学西南医院病理研究所卞修武教授、姚晓红博士惠赠。 方法：取人脑胶质瘤组织、正常成人脑组织、U251及CHG-5胶质瘤细胞株,采用免疫磁珠法分选获得CD133+脑胶质瘤干细胞,加入DMEM/F12无血清培养基进行增殖培养,形成细胞克隆后,加入含体积分数为10%胎牛血清的培养液,2 h后行抗CD133和抗巢蛋白免疫荧光双标染色。 主要观察指标：人脑胶质瘤干细胞的生长和鉴定,采用WST-8法、免疫组化实验、流式细胞仪、免疫荧光双标实验检测NOTCH-1信号通路蛋白的表达。 结果：在无血清培养基中,细胞呈悬浮生长,培养24~48 h可见单个细胞开始分裂生长,形成肿瘤球,将肿瘤球转入含胎牛血清的培养基后,4 h周边细胞伸出突起并逐渐分化,24 h后肿瘤球迁移出的细胞增多,形成单细胞层。肿瘤球能同时表达干细胞标志物CD133和巢蛋白,CD133+脑胶质瘤干细胞核浆比例达2/3~3/4,突起少,胞浆中线粒体等细胞器较少,核糖体丰富,未见胶质丝等分化结构,符合干细胞超微结构特点。NOTCH-1蛋白在人脑胶质瘤组织中的表达明显强于正常成人脑组织（P < 0.01）,在CD133+ U251及CHG-5胶质瘤细胞中有很强的表达,在GFAP+和MAP2+ U251及CHG-5胶质瘤细胞中的表达强弱不等,在MBP+ U251及CHG-5胶质瘤细胞中呈弱表达或不表达。在脑胶质瘤干细胞增殖过程中能检测到较强的NOTCH-1信号通路关键基因NOTCH-1,CBF-1,HES-1 mRNA及NOTCH-1,HES-1蛋白表达;而在细胞分化时NOTCH-1信号通路关键基因NOTCH-1,CBF-1,HES-1 mRNA和HES-1蛋白表达逐渐减弱。 结论：NOTCH-1信号通路的关键蛋白分子NOTCH-1在人脑胶质瘤组织和胶质瘤细胞株中均有表达,而在正常成人脑组织中仅微弱表达。NOTCH-1信号通路关键基因NOTCH-1和HES-1的表达强弱可能参与了胶质瘤干细胞增殖和分化的调控。     

Selective expression of JNK isoforms and stress-specific JNK activity in different neural cell lines

The function of c-Jun N-terminal kinases (JNKs) in the nervous system is poorly understood and the majority of the data has been gained in neuronal and non-neuronal cell lines. Thus, it is not clear to which extent the expression pattern and the degree of activation of the three JNK isoforms in different cell lines are representative for their activation in the adult brain. In the present study, the expression of JNK isoforms and the activity of JNK1 were determined following UV irradiation and exposure to H(2)O(2) and TNFalpha in three neural cell lines, rat PC12, murine Neuro2A and human SHSY5Y. These cell lines differ in their expression of JNK isoforms: PC12 cells express JNK1 and JNK2, whereas Neuro2A and SHSY5Y cells displays the expression of JNK1, JNK2 and JNK3. JNK3 was not inducible following stress and differentiation in PC12 cells. The stimulation paradigms evoked different degree of cell death: UV irradiation resulted in death of around 50% in all three cell lines; exposure to 200 microM H(2)O(2) for 6 h resulted in the death of 43% Neuro2A cells and 31% PC12 cells, SHSY5Y cells are less sensitive to H(2)O(2) since only 5 mM H(2)O(2) killed 59% of SHSY5Y cells after 6 h. Exposure to 50 ng/ml TNFalpha did not induce cell death in SHSY5Y, Neuro2A and naive PC12 cells. Although differentiated PC12 cells exhibit a similar activation of JNK1 compared to naive PC12 cells after exposure to TNFalpha, 42% of differentiated PC12 cells died after 24 h. H(2)O(2) that evoked only a moderate JNK1 activity in Neuro2A and PC12 cells induced only a moderate cell death. In contrast, SHSY5Y cells exhibit a much stronger JNK1 activation accompanied with a higher degree in cell death after exposure to H(2)O(2). JNK1 activity induced by UV irradiation, however, could not be correlated with the extend of cell death. These data clearly demonstrate that expression and activation of JNK depends on the neuronal cell type and the applied stress paradigms, and that JNK activity is not simply linked to cell death.     

An oligodendrocyte precursor cell line from rat optic nerve

We have established permanent cell lines from the optic nerve of the rat with a temperature sensitive immortalizing oncogene (Simian Virus 40 large T-antigen carrying both the tsA58 and U19 mutations). The oncogene was transduced into primary cultures via a replication deficient retrovirus, and infected cells were selected with the antibiotic G418. A clonal cell line (tsU19-5) displayed some properties of oligodendrocyte precursors: it proliferated, bound the monoclonal antibody A2B5 (which recognizes minor ganglioside species), and expressed the intermediate filament vimentin and the enzyme 2',3'-cyclic-nucleotide 3'-phosphodiesterase (CNP) at 33 degrees C (the permissive temperature for the oncogene). At 39 degrees C (the non-permissive temperature), some cells had the potential to differentiate further, and expressed several oligodendrocyte specific components: galactocerebroside, myelin basic protein, proteolipid protein and CNP. These results suggest that conditional oncogenes can establish neural precursor cell lines which are still capable of differentiation in vitro.     

Differential expression of glial fibrillary acidic protein in human glioma cell lines

Summary We have obtained a cDNA fragment to human glial fibrillary acidic protein (GFAP) by immunoscreening a gt11 human brain cDNA library with antibody to bovine GFAP. The highly homologous nucleotide sequence of this clone with that of the mouse GFAP enabled the identification of this cDNA as one encoding GFAP. As this cDNA hybridized with a single major RNA species in Northern blots of RNA from human and mouse brain tissues and gave one or two bands in Southern blots of human genomic DNA, it was considered to be specific for GFAP. Using this cDNA as a probe we investigated the levels of GFAP expression in ten human glioma cell lines. A 3.5-kb GFAP mRNA was detected in five of the ten glioma cell lines, one of which was U-251 MG cell line and the other four were clones derived from the same tumor (CL1, 2, 3, and 4). There was a difference in the amount of GFAP mRNA among U-251 MG and the four clonal cell lines. Quantitative evaluation of this difference by RNA dot blot analysis revealed that the amount of GFAP mRNA expressed in CL3 was about 1/5 and in CL4 about 1/10 the amount expressed in U-251 MG, CL1, and CL2. Semiquantitative Western blot analysis showed that GFAP levels corresponded to the GFAP mRNA levels in these cell lines. By Southern blot analysis of genomic DNA the GFAP gene was similarly detected in all of these cell lines regardless of the level of GFAP expression. Thus, by using a cDNa to human GFAP we have demonstrated the presence of clonal cell lines from human glioma showing different levels of GFAP expression, which may provide a useful basis for further investigations on the regulation of GFAP gene expression in glial cells.     

Folic acid deficiency inhibits neural rosette formation and neuronal differentiation from rhesus monkey embryonic stem cells

Evidence from epidemiological studies has proved that periconceptional use of folic acid (FA) can significantly reduce the risk of neural tube defects (NTDs). However, it is hard to explore when and how FA plays roles in neurogenesis and brain development in vivo, especially in human or other nonhuman primate systems. Primate embryonic stem cell (ESC) lines are ideal models for studying cell differentiation and organogenesis in vitro. In the present study, the roles of FA in neural differentiation were assessed in a rhesus monkey ESC system in vitro. The results showed no significant difference in the expression of neural precursor markers, such as nestin, Sox-1, or Pax-6, among neural progenitors obtained from different FA concentrations or with the FA antagonist methotrexate (MTX). However, FA depletion decreased cell proliferation and affected embryoid body (EB) and neural rosette formation, as well as neuronal but not neuroglia differentiation. Our data imply that the ESC system is a suitable model for further exploring the mechanism of how FA works in prevention of NTDs in primates.     

Models for the in vitro assessment of neurotoxicity in the nervous system in relation to xenobiotic and neurotrophic factor-mediated events.

We have been investigating the use of three culture types for both screening and mechanistic neurotoxicology in vitro. These are the neuroblastoma cell lines (IMR32 - human; C-1300 - mouse), primary mixed monolayer cultures of the rat and chick embryonic midbrain ('micromass' systems) and organotypic whole rat brain reaggregate cultures. The performance of these models for neurotoxicity resting has been investigated with ethylcholine mustard aziridinium (ECMA), vincristine, aluminium, glutamate receptor antagonists, MPTP, and 'hypothyroidism'. From a 'screening' viewpoint, in vitro exposure through a tiered testing system (ranging from simple cytotoxicological parameters in the neural cell lines to neurotransmitter measurements in the organotypic cultures) may permit detection of CNS neurotoxicity and delineation of possible mechanisms. The type of developmental neurotoxicological information gained is highlighted in the cases of aluminum and the glutamate receptor antagonists. High concentrations of aluminum caused significant neural cell death in differentiated neuroblastoma cell lines after approximately two weeks exposure in vitro. In contrast, cell death was detected in the developing midbrain cultures as early as 24 - 48 hr. Studies in whole brain reaggregates suggest that cholinotoxicity may occur in a similar time-frame and is consistent with some of aluminium's effects in vivo. Preliminary experiments have shown that exposure of immature developing midbrain rat primary cultured neurones to the glutamate receptor antagonists, AP3 and MK-801 induces neural cell death which may relate to control of NGF by glutamate cells. Developing neural culture systems may prove useful for testing agents which cause neurotoxicity through disturbances of neurotrophic function.     

Functional Recovery in Hemiparkinsonian Primates Transplanted with Polymer-Encapsulated PC12 Cells

Cross-species neural grafting of cell lines immunoisolated by a permselective polymer membrane represents a promising source of neuroactive molecules for the treatment of neurodegenerative diseases. Utilization of a cell line of xenogeneic origin is advantageous since the transplanted cells will be rejected by the host immune system in case of breakage of the immunoisolating envelope. Polymer-encapsulated PC12 cells, a dopaminergic cell line derived from a rat pheochromocytoma, were transplanted in five Macaca fascicularis monkeys which had been previously rendered hemiparkinsonian by a unilateral carotid injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Well-preserved, tyrosine hydroxylase positive encapsulated PC12 cells were observed in the lesioned striatum for up to 5 months after implantation. Four out of five monkeys which received polymer-encapsulated PC12 cells showed significant behavioral improvement, whereas three monkeys implanted with either encapsulated bovine chromaffin cells or empty polymer envelopes showed no amelioration.     

Chemokines influence the migration and fate of neural precursor cells from the young adult and middle-aged rat subventricular zone

We have previously demonstrated a role for the chemokines MCP-1, MIP-1α and GRO-α in directing subventricular zone (SVZ)-derived neural precursor cell migration towards the site of cell death in the adult rodent brain. However the influence of chemokines such as MCP-1, MIP-1α and GRO-α on the differentiation of adult neural precursor cells has not previously been investigated. Further, as the majority of neurological disorders and injuries occur during ageing, it is important to investigate the effect of chemokines on adult neural precursor cell cultures obtained from the ageing brain. This study therefore examined the effect of MCP-1, MIP-1α and GRO-α on SVZ-derived neural precursor cell differentiation in vitro, and assessed whether precursor cells from the middle-aged rat brain (13 months old) follow the same migratory and differential profile as neural precursor cells obtained from the young adult rat brain (2 months old). We observed that each of the chemokines examined generated differing effects in regards to neuronal or glial differentiation. Further, both MIP-1α and GRO-α increased total cell number, suggesting an effect on precursor cell proliferation and/or survival. In agreement with cultures obtained from young adult brains, SVZ-derived neural precursor cells cultured from the middle-aged brain exhibited chemotactic migration in response to a concentration gradient. These results indicate that the chemokines MCP-1, MIP-1α and GRO-α can influence both the migration and fate choice of SVZ-derived neural precursor cells, as well as promoting cell viability. While a response to each of these chemokines is maintained in the middle-aged brain, a distinct age-related alteration in differential fate can be identified.     

Human immunodeficiency virus type 1 Tat modulates proliferation and differentiation of human neural precursor cells: implication in NeuroAIDS

Human immunodeficiency virus type 1 (HIV-1) and viral proteins affect neuronal survival and neuron-glial cell interactions, which culminate in neurological disorders. HIV-1 infects regions of neurogenesis in human adult and pediatric brain. However, little is known about the effect of HIV-1 or viral proteins on the properties of human neural precursor cells (hNPCs), particularly neurogenesis, hence a detailed investigation on these lines is warranted. Human neural precursor cells were cultured in presence and absence of HIV-1B transactivating protein Tat to investigate if HIV-1 viral protein alters the properties of human neural precursor cells. Cellular and molecular approaches were adopted to study the effect of HIV-1B transactivating protein Tat on proliferation and differentiation potential of human fetal brain-derived NPCs. Cell proliferation assays such as BrdU and Ki67 staining and pathway-specific cDNA and protein arrays were used in the study. Data reveal that HIV-1B Tat protein severely affects proliferation of hNPCs, as evident by lower incorporation of BrdU and Ki67 staining as well as neurosphere assay. HIV-1 Tat substantially attenuated neurogenesis, as evident by the smaller numbers of Tuj-1- and doublecortin-positive cells differentiated from hNPCs, without affecting their viability. These data suggest that HIV-1 Tat alters the properties of human neural precursor cells via attenuation of the cell cycle regulatory unit cyclin D1 and the mitogen-activated protein kinase (MAPK) pathway, particularly extracellular signal-related kinase 1/2 (ERK1/2). The study provides new insights into cellular and molecular mechanisms that may modulate human neural precursor cell properties in HIV/AIDS (acquired immunodeficiency syndrome) individuals. Validation with autopsy brain samples is necessary to further substantiate these important observations.