共查询到20条相似文献,搜索用时 49 毫秒
1.
Susana Beatriz Etcheverry Evelina Gloria Ferrer Luciana Naso Josefina Rivadeneira Victoria Salinas Patricia Ana María Williams 《Journal of biological inorganic chemistry》2008,13(3):435-447
Vanadium compounds are known for a variety of pharmacological properties. Many of them display antitumoral and osteogenic
effects in several cell lines. Free radicals induce the development of tumoral processes. Natural polyphenols such as flavonoids
have antioxidant properties since they scavenge different free radicals. For these reasons it is interesting to investigate
the effects of a new complex generated between the vanadyl(IV) cation and the flavonoid hesperidin. The complex has been synthesized
and characterized by physicochemical methods. Spectroscopic analysis revealed a 1:1 stoichiometry of ligand:VO and coordination
by deprotonated cis-hydroxyl groups to the disaccharide moiety of the ligand. The complex improves the superoxide dismutase (SOD)-like activity
of the ligand, but the scavenging of other radicals tested does not change upon complexation. When tested on two tumoral cell
lines in culture (one of them derived from a rat osteosarcoma UMR106 and the other from human colon adenocarcinoma Caco-2),
the complex enhanced the antiproliferative effects of the free ligand, and this effect correlated with the morphological alterations
toward apoptosis. Also, on the osteoblastic cell line the complex stimulated cell proliferation and collagen type I production
at low concentrations. At higher doses the complex behaved as a cytotoxic compound for the osteoblasts. 相似文献
2.
Cecilia Gotti Emanuele Sher Donatella Cabrini Gianpietro Bondiolotti Enzo Wanke Enzo Mancinelli Francesco Clementi 《Differentiation; research in biological diversity》1987,34(2):144-155
The differentiation of human nerve cells was investigated using a cell model comprising human neuroblastoma (IMR32) cells that were induced to differentiate by the addition of 5-bromo-2'-deoxyuridine (BrdU) or N6-O2-dibutyryl cyclic adenosine 3'-5' monophosphate (Bt2cAMP). As parameters of differentiation, we studied neurite outgrowth, cholinergic receptors, voltage-activated ion channels, tyrosine hydroxylase activity, and neurotransmitter content. BrdU induced marked morphological differentiation, as indicated by the number and length of neurites, as well as an increase in the number of alpha-bungarotoxin binding sites, muscarinic receptors, and voltage-dependent Na channels. In addition, BrdU induced an increase in tyrosine hydroxylase activity as well as in serotonin, dopamine, and noradrenaline content. Bt2cAMP had a less dramatic effect on the morphological appearance of the cells, induced the expression of alpha-bungarotoxin binding sites (but not of muscarinic receptors), and produced a marked increase in the serotonin and noradrenaline content. Not only the number but also the functional properties of nicotinic and muscarinic receptors were differently affected by the two drugs. We conclude that Bt2cAMP and BrdU induce a different pattern of differentiation in the same cells, and that the expression of specific neuronal markers can be modulated to yield functionally different neurons. 相似文献
3.
T Fukushige M Nagoshi Y Hachitanda T Ueno Y Zaizen S Suita M Tsuneyoshi Y Kaneko A Nakagawara 《Virchows Archiv. B, Cell pathology including molecular pathology》1992,62(3):159-166
A new human cell line, termed Muraoka, has been established from the recurrent tumor of a case of congenital primitive neuroectodermal tumor (PNET) arising at the temporofacial region of a male infant. The microscopic findings of this cell line were epithelioid, and the xenografted tumor in a nude mouse consisted of the malignant epithelioid cells. Immunohistochemically, the cells were positive for neuron-specific enolase, S-100 protein, carcinoembryonic antigen, cytokeratin, epithelial membrane antigen, and glial fibrillary acidic protein. These findings were quite similar to those of the epithelioid cells in the original tumor and of the xenografted tumor cells. Neither chromosomal abnormalities nor N-myc amplification were observed. Morphological differentiation after treatment with N6-2'-O-dibutyryladenosine 3':5'-cyclic monophosphate (Bt2-cAMP), all-trans-retinoic acid (RA), prostaglandin E1 (PGE1), and 5-bromo-2'-deoxyuridine (BrdU) showed two different results. Bt2-cAMP and PGE1 induced neuronal differentiation with the extension of neurites, whereas RA and BrdU predominantly induced Schwannian differentiation (flat cells). In these respects, the cell line Muraoka seems to be useful for studying characteristics of PNET as well as for developing the new treatments against such tumors. 相似文献
4.
Neuronal differentiation is accompanied by NSP-C expression 总被引:5,自引:0,他引:5
Jurgen Hens Ronny Nuydens Hugo Geerts Nicole H. M. Senden Wim J. M. Van de Ven Anton J. M. Roebroek Helgi J. K. van de Velde Frans C. S. Ramaekers J. L. V. Broers 《Cell and tissue research》1998,292(2):229-237
Neuroendocrine-specific protein (NSP) reticulons are expressed in neural and neuroendocrine tissues and cell cultures derived therefrom, while most other cell types lack NSP-reticulons. Three major subtypes have been identified so far, designated NSP-A, NSP-B, and NSP-C. We have investigated the correlation between the degree of neuronal differentiation, determined by morphological and biochemical criteria, and NSP-reticulon subtype expression. For this purpose, several human neuroblastoma cell lines, exhibiting different degrees of neuronal differentiation, were examined immuno(cyto) chemically. It became obvious that the expression of NSP-C, as detected by immunofluorescence microscopy and Western blotting, is most prominent in cell lines with a high degree of neuronal differentiation, such as LA-N-5. Such highly differentiated cells also express other neural and neuroendocrine markers, such as neural cell adhesion molecule (NCAM), neurofilament proteins, synaptophysin, and chromogranin. NSP-A was observed in all cell lines to a different extent. However, no clear correlation was observed with the degree of neuronal differentiation as defined by other neuronal and neuroendocrine markers or morphology. NSP-B could not be detected. The induction of neuronal differentiation with nerve growth factor, dbcAMP, and retinoic acid in the rat pheochromocytoma cell line PC12 and the human teratocarcinoma cell line hNT2, respectively, induced the expression of NSP-A and NSP-C in these cell lines parallel to the induction of neurofilament protein expression. It is concluded that NSP-C expression, in particular, is strongly correlated with neuronal differentiation. 相似文献
5.
The extracellular matrix (ECM) plays an important role in cell differentiation and apoptosis. Collagen is the major component of ECM. Here, an ESR signal of the hydroxyl radicals (.OH) generated via Fe2+ -mediated Fenton reaction was found to be significantly inhibited by type I collagen. Further study showed that type I collagen also inhibited cell apoptosis induced by.OH, as evidenced by morphological criteria (DAPI and annexin V staining) and quantitive assays for apoptotic cells (MTT and flow-cytometric assay for subG1 cells). By addition of type I collagen in HeLa cells, the lipid peroxidation caused by.OH was inhibited and the cellular GSH was protected. In comparison with type I collagen, BSA and the denatured collagen, gelatin, lacked such antioxidative and antiapoptotic effects. Together, the results suggest that type I collagen can uniquely prevent.OH-mediated apoptosis by scavenging free radicals. 相似文献
6.
Youichirou Ninomiya Richard Adams Gillian M. Morriss-Kay Kazuhiro Eto 《Journal of cellular physiology》1997,172(1):25-35
Apoptotic cell death was observed during aggregate culture of the mouse embryonal carcinoma cell line P19 exposed to all-trans retinoic acid (tRA). This finding was confirmed by genomic DNA agarose gel electrophoresis and transmission electron microscopy. Apoptosis was associated with P19 cell neuronal differentiation; alternative causes of cell death, i.e., cavitation-related, cytotoxicity of tRA, or spontaneous cell death were excluded. Analysis by flow cytometry revealed that the apoptosis was likely to occur in multiplying cells that underwent to reentering into S phase. We therefore examined 5-bromo-2′-deoxyuridine (BrdU) incorporation and proliferating cell nuclear antigen (PCNA) expression and localization in the aggregates by immunofluorescent staining. Although the P19 cells in the aggregates exposed to tRA incorporated BrdU at an equivalent level to those not exposed to tRA, the cells showed diminished PCNA expression and nuclear accumulation. We propose that P19 apoptosis during neuronal differentiation is a model system in which programmed cell death occurs simultaneously with cell division leading to differentiation. J. Cell. Physiol. 172:25–35, 1997. © 1997 Wiley-Liss, Inc. 相似文献
7.
Trophic molecules are key regulators of survival, growth and differentiation of neural cells. Neuronal cell type Neuro-2a is a good model to study development and molecules modulating this process, and retinoic acid (RA) and neurotrophins (NGF, BDNF, NT-3 and NT-4) have been shown to be active in this modulation. The purpose of the present study was the functional analysis of these trophic molecules in our short-term bioassay of Neuro-2a cells, an immortalised murine neuroblastoma cell line. Through cell counting, image process and arithmetic combination of digital parameters of treated and untreated cultures, we show that RA inhibits growth and induces morphological neuronal phenotype of treated cells. Through DNA labelling with BrdU we also show that NGF, BDNF, and NT-3 increase survival and proliferation of cells, grown in serum-deprived media. From these results we conclude that neurotrophins have manifest trophic effects on cells improving survival, growth and proliferation and we also confirm the growth arrest and differentiation properties of RA on Neuro-2a cells. 相似文献
8.
Kanski J Aksenova M Stoyanova A Butterfield DA 《The Journal of nutritional biochemistry》2002,13(5):273-281
In this study, free radical scavenging abilities of ferulic acid in relation to its structural characteristics were evaluated in solution, cultured neurons, and synaptosomal systems exposed to hydroxyl and peroxyl radicals. Cultured neuronal cells exposed to the peroxyl radical initiator AAPH die in a dose-response manner and show elevated levels of protein carbonyls. The presence of ferulic acid or similar phenolic compounds, however, greatly reduces free radical damage in neuronal cell systems without causing cell death by themselves. In addition, synaptosomal membrane systems exposed to oxidative stress by hydroxyl and peroxyl radical generators show elevated levels of oxidation as indexed by protein oxidation, lipid peroxidation, and ROS measurement. Ferulic acid greatly attenuates these changes, and its effects are far more potent than those obtained for vanillic, coumaric, and cinnamic acid treatments. Moreover, ferulic acid protects against free radical mediated changes in conformation of synaptosomal membrane proteins as monitored by EPR spin labeling techniques. The results presented in this study suggest the importance of naturally occurring antioxidants such as ferulic acid in therapeutic intervention methodology against neurodegenerative disorders such as Alzheimer's disease in which oxidative stress is implicated. 相似文献
9.
The influence of 5-bromo-2'-deoxyuridine (BrdU) on rat embryo development and neurogenesis was investigated using a rat conceptus culture system during organogenesis (pregnancy days 10-13). The embryos and visceral yolk sacs of conceptuses cultured with BrdU were examined for overall growth, morphological anomalies, incorporation of radiolabeled BrdU into DNA, and neurotransmitter enzyme activities in embryos. In addition, neural tubes from cultured whole embryos were isolated and mechanically dissociated into fragments and cultured again to assess neural cell differentiation into neuron-like cells. BrdU was found to incorporate differentially into embryonic and visceral yolk sac DNA with simultaneous stage-specific retardation and anomalous organogenesis in proportion to the increasing concentrations used. Neural tube differentiation of cultured embryos was markedly altered, and there were morphologically distinct neural anomalies. The neurite outgrowth from neuroblast cells (type 1) of explanted spinal neural tube fragments from BrdU-treated embryos was markedly reduced in length and number compared to those from similar areas of embryos grown without BrdU. In contrast, BrdU at the same doses did not affect differentiation of a number of neural tissue-related enzymes. These results indicate that BrdU incorporation into DNA of primordial embryonic cells significantly affects neurogenesis and differentiation of neurites from neuroblasts, which is a specific neural cytodifferentiation characteristic of neuronal cells. 相似文献
10.
Induction of erythroid differentiation in Friend leukemia cells by bromodeoxyuridine 总被引:1,自引:0,他引:1
Treatment of Friend leukemia cells with BrdU, the thymidine analog which interferes with DMSO induced differentiation in these cells as well as the expression of differentiated character in many other cell systems, is capable of inducing erythroid differentiation. Globin mRNA, as assayed by hybridization to globin cDNA, increases 2.5- to 30-fold after appropriate treatment with BrdU. This effect was observed with several different subclones of three independent Friend tumor cell lines. After BrdU treatment, globin mRNA content may reach up to 10-20% of the levels in DMSO induced cultures. The induction of erythroid differentiation is also apparent when accumulated heme content or the appearance of benzidine positive cells is monitored. One Friend cell line (745) we examined was not induced by BrdU although it incorporated an amount of BrdU into its DNA comparable to that incorporated by the other cell lines. In addition, BrdU did interfere with DMSO induction in this cell line. These results suggest that two different mechanisms may be operative in regulating erythroid differentiation in Friend leukemia cells. While BrdU interferes with the mechanism activated by DMSO treatment, this analog could independently activate an alternative mechanism. 相似文献
11.
Through free radical-mediated peroxidation, cyclooxygenase (COX) can metabolize dihomo-γ-linolenic acid (DGLA) and arachidonic acid (AA) to form well-known bioactive metabolites, namely, the 1-series of prostaglandins (PGs1) and the 2-series of prostaglandins (PGs2), respectively. Unlike PGs2, which are generally viewed as proinflammatory and procarcinogenic PGs, PGs1 may possess anti-inflammatory and anti-cancer activity. Previous studies using ovine COX along with spin trapping and the LC/ESR/MS technique have shown that certain exclusive free radicals are generated from different free radical reactions in DGLA and AA peroxidation. However, it has been unclear whether the differences were associated with the contrasting bioactivity of DGLA vs AA. The aim of this study was to refine the LC/MS and spin trapping technique to make it possible for the association between free radicals and cancer cell growth to be directly tested. Using a colon cancer cell line, HCA-7 colony 29, and LC/MS along with a solid-phase extraction, we were able to characterize the reduced forms of radical adducts (hydroxylamines) as the free radicals generated from cellular COX-catalyzed peroxidation. For the first time, free radicals formed in the COX-catalyzed peroxidation of AA vs DGLA and their association with cancer cell growth were assessed (cell proliferation via MTS and cell cycle distribution via propidium iodide staining) in the same experimental setting. The exclusive free radicals formed from the COX-catalyzed peroxidation of AA and DGLA were shown to be correlated with the cell growth response. Our results indicate that free radicals generated from the distinct radical reactions in COX-catalyzed peroxidation may represent the novel metabolites of AA and DGLA that correspond to their contrasting bioactivity. 相似文献
12.
The principal aim of the current study was to identify the initial cell targets of hydroxyl free radicals. Our recent report showed that proteins were oxidized before lipids in U937 cells exposed to peroxyl radicals. Extending this finding, we investigated whether a similar oxidation sequence occurs in other lines of cells, whether hydroxyl radicals can also initiate cell protein oxidation, and whether DNA fragmentation is an early event in radical-induced cell damage. Mouse myeloma Sp2/0-Ag14 and U937 cells were exposed to hydroxyl radicals generated in solution by gamma irradiation and the formation of protein peroxides measured by a ferric-xylenol orange assay. No lipid peroxidation or DNA damage was evident by the time of significant formation of protein peroxides. DNA fragmentation was detectable after prolonged incubation at 37 degrees C and was characteristic of enzymatic action rather than of random scission by the radicals. Yields of protein hydroperoxides in the irradiated cells were independent of composition of the medium, suggesting that only the radicals produced within the cells or immediately near the cell surface were effective in oxidizing the cell proteins. The results are consistent with the hypothesis that proteins are major initial targets of free radicals in cells and suggest that treatments leading to the prevention of protein oxidation or to harmless reduction of protein peroxides is likely to result in alleviation of radical-induced biological damage. 相似文献
13.
Lehner B Sandner B Marschallinger J Lehner C Furtner T Couillard-Despres S Rivera FJ Brockhoff G Bauer HC Weidner N Aigner L 《Cell and tissue research》2011,345(3):313-328
5-Bromo-2'-deoxyuridin (BrdU) is frequently used in anaylsis of neural stem cell biology, in particular to label and to fate-map dividing cells. However, up to now, only a few studies have addressed the question as to whether BrdU labeling per se affects the cells to be investigated. Here, we focused on the potential impact of BrdU on neurosphere cultures derived from the adult rat brain and on proliferation of progenitors in vivo. In vitro, neurospheres were pulsed for 48?h with BrdU, and cell proliferation, cell cycle, differentiation, survival and adhesion properties were subsequently analyzed. BrdU inhibited the expansion of neural progenitors as assessed by MTS assay and increased the fraction of cells in the G0/G1-phase of the cell cycle. Moreover, BrdU increased cell death and dose-dependently induced adherence of NPCs. Cell adherence was accompanied by a reduced amount of active matrix-metalloproteinase-2 (MMP-2). Furthermore, BrdU repressed neuronal and oligodendroglial differentiation, whereas astroglial fate was not affected. In contrast to the in vitro situation, BrdU apparently did not influence endogenous proliferation of NPCs or neurogenesis in concentrations that are typically used for labeling of neural progenitors in vivo. Our results reveal so far uncharacterized effects of BrdU on adult NPCs. We conclude that, because of its ubiquitous use in stem cell biology, any potential effect of BrdU of NPCs has to be scrutinized prior to interpretation of data. 相似文献
14.
Takahiko Fukushige Makoto Nagoshi Yoichi Hachitanda Takaki Ueno Yoshio Zaizen Sachiyo Suita Masazumi Tsuneyoshi Yasuhiko Kaneko Akira Nakagawara 《Virchows Archiv. B, Cell pathology including molecular pathology》1992,62(1):159-166
A new human cell line, termedMuraoka, has been established from the recurrent tumor of a case of congenital primitive neuroectodermal tumor (PNET) arising at
the temporofacial region of a male infant. The microscopic findings of this cell line were epithelioid, and the xenografted
tumor in a nude mouse consisted of the malignant epithelioid cells. Immunohistochemically, the cells were positive for neuron-specific
enolase, S-100 protein, carcinoembryonic antigen, cytokeratin, epithelial membrane antigen, and glial fibrillary acidic protein.
These findings were quite smiliar to those of the epithelioid cells in the original tumor and of the xenografted tumor cells.
Neither chromosomal abnormalities nor N-myc amplification were observed. Morphological differentiation after treatment with
N6-2′-Odibutyryladenosine 3′:5′-cyclic monophosphate (Bt2cAMP), all-trans-retinoic acid (RA), prostaglandin E1 (PGE1, and 5-bromo-2′-deoxyuridine (BrdU) showed two different results. Bt2-cAMP and PGE1 induced neuronal differentiation with the extension of neurites, whereas RA and BrdU predominantly induced Schwannian differentiation
(flat cells). In these respects, the cell lineMuraoka seems to be useful for studying characteristics of PNET as well as for developing the new treatments against such tumors. 相似文献
15.
Selective attachment of neural cells to specific substrates including Cell-Tak, a new cellular adhesive 总被引:2,自引:0,他引:2
M F Notter 《Experimental cell research》1988,177(2):237-246
Adherence of embryonic hypothalamic cells and a homogeneous neuronal cell line was assessed on various substrates and compared to attachment to the new cellular and tissue adhesive, Cell-Tak. Cell-Tak provided the most advantageous surface with 100% of fetal brain cells attaching in 5 h. Attachment of hypothalamic cells to compounds such as poly-D-lysine or collagen within this time was increased by 45 and 25%, respectively, over tissue-culture plastic. All cells of the clonal cell line N2AB-1 attached to Cell-Tak in the presence or absence of fetal calf serum and were found to be resistant to trypsin removal. Conditioned medium from these cells enhanced attachment of N2AB-1 twofold when compared to adherence to tissue-culture plastic. Striking morphological changes were seen in N2AB-1 after culturing on Cell-Tak for 2 days. Thirty percent of the population extended long neurites when grown on Cell-Tak with serum. Without serum, 30 to 50% of the cells extended very broad neurites often branched at the end, which were morphological changes not seen on plastic surfaces. These findings indicate that Cell-Tak is an optimal adhesive for primary neural cell culture and maintenance. Moreover, this adhesive protein appears to induce neuritogenesis and cellular differentiation in a neuronal cell line. 相似文献
16.
ESR spin trapping technique was used to detect and analyze free radical formation. When 6-hydroxydomine (6-OHDA) was incubated alone or in the presence of a free radical generating system (H2O2 and FeSO4), hydroxyl free radicals were observed in a concentration-dependent manner. Glutathione was found to be the most effective scavenger of the ESR signal when compared with vitamin E or Mannitol. The addition of ethanol resulted in the formation of the pure hydroxyethyl free radicals. The amount of hydroxyethyl free radicals in the system was dependent upon the concentration of ethanol and the formation of hydroxyethyl free radicals correlated well with the extent of lipid peroxidation and the loss of enzymic activity of the membrane-bound (Na+, K+)-ATPase. We suggest that in the biological system ethanol may potentiate the neurotoxicity of 6-OHDA with the formation of hydroxyethyl free radicals, which are longer-lived and far more damaging to membranes that the hydroxyl radicals. These data lead us to further hypothesize that the neuronal degeneration caused by 6-OHDA and other compounds that generate free radicals could be potentiated in the presence of ethanol. 相似文献
17.
When diaziquone was irradiated with 500 nm visible light, hydroxyl free radicals as well as the diaziquone semiquinone were produced. The diaziquone semiquinone is a stable free radical that exhibits a characteristic 5-line electron spin resonance (ESR) spectrum. Since hydroxyl free radicals are short lived, and not observable by conventional ESR, the nitrone spin trap 5,5-dimethyl-1-pyrroline-1-oxide (DMPO) was used to convert hydroxyl radicals into longer lived ESR detectable spin adducts. The formation of hydroxyl radicals was further confirmed by investigating reactions in which hydroxyl radical scavangers, sodium formate and dimethylsulfoxide, compete with the spin traps DMPO or POBN (alpha-(4-Pyridyl-1-oxide)-N- tert-butylnitrone) for hydroxyl free radicals. The products of these scavenging reactions were also trapped with DMPO or POBN. If drug free radicals and hydroxyl free radicals are important in the activity of quinone-containing antitumor agents, AZQ may have a potential in photoirradiation therapy or photodynamic therapy. 相似文献
18.
Polysialic acid regulates the clustering, migration, and neuronal differentiation of progenitor cells in the adult hippocampus 总被引:1,自引:0,他引:1
Burgess A Wainwright SR Shihabuddin LS Rutishauser U Seki T Aubert I 《Developmental neurobiology》2008,68(14):1580-1590
Newborn cells of the adult dentate gyrus in the hippocampus are characterized by their abundant expression of polysialic acid (PSA), a carbohydrate attached to the neural cell adhesion molecule (NCAM). PSA+ newborn cells of the dentate gyrus form clusters with proliferating neural progenitor cells, migrate away from these clusters, and terminally differentiate. To identify the roles of PSA in the development of adult progenitors of the dentate gyrus, we injected endoneuraminidase N (endoN) into the hippocampus of adult rats to specifically cleave PSA from NCAM. Two days later, we administered the mitotic marker, 5-bromo-2'-deoxyuridine (BrdU). Three days after BrdU injection, BrdU+ cells were found inside and outside the clusters of newborn cells. In endoN-treated animals, the total number of BrdU+ cells was not changed but significantly more BrdU+ cells were present within clusters, suggesting that PSA normally facilitates the migration of progenitors away from the clusters. Seven days post-BrdU injection, endoN-treated animals had significantly more BrdU+ cells which were also positive for the mature neuronal nuclear marker NeuN compared with controls, indicating that the loss of PSA from progenitor cells increases neuronal differentiation. This report is the first demonstration that PSA is involved in controlling the spatio-temporal neuronal maturation of adult hippocampal progenitors in the normal brain. In vitro, the removal of PSA from adult-derived neural progenitors significantly enhanced neuronal differentiation, strengthening our in vivo findings and indicating that PSA removal on isolated progenitor cells, apart from a complex in vivo environment, induces neuronal maturation. 相似文献
19.
目的:建立NTera2/CloneD1细胞向神经元分化的模型,检测神经元限制性沉默因子(NRSF)经分化培养基诱导后表达的变化。方法:收集正常培养的NTera2/CloneD1细胞及经全反式维甲酸(RA)、阿糖胞苷(AraC)、尿苷分阶段诱导共28 d的细胞,显微镜下观察诱导前后细胞的形态学变化;免疫荧光法检测NTera2/CloneD1细胞诱导前后干性标志Nestin、Sox2和成熟神经元特异性标志NF-200、β-tubulinⅢ的表达情况;应用RT-PCR和免疫荧光法对NRSF进行mRNA和蛋白水平的检测。结果:显微镜下观察到正常培养的NTera2/CloneD1细胞呈克隆样生长,经分化培养基诱导后的NTera2/CloneD1细胞表现出典型的神经元样细胞形态。免疫荧光检测表明,未诱导的NTera2/CloneD1细胞表达神经干细胞的标志Sox2、Nestin,不表达成熟神经元特异性蛋白NF-200、β-tubulinⅢ;而经RA等诱导分化的细胞则不表达Sox2、Nestin,表达NF-200、β-tubulinⅢ。RT-PCR和免疫荧光检测显示,NRSF在诱导分化后的NTera2/CloneD1细胞中的表达量显著降低。结论:建立了NTera2/CloneD1细胞向神经元分化的模型,NRSF在诱导后的NTera2/CloneD1细胞中表达量显著下调,提示NTera2/CloneD1细胞在诱导过程中可能通过下调NRSF,使受到NRSF负性调控的神经元特异性蛋白启动表达并上调,进而实现NTera2/CloneD1细胞向神经元的定向分化。 相似文献
20.
The terminal differentiation of neurons occurs as precisely timed waves, with specific neuronal types differentiating in defined sequences. The precision of neuronal differentiation in the central nervous system offers an unusual opportunity to study terminal differentiation in vivo. The p34cdc2 kinase complex and the anti-oncogenes p53 and RB are central in the regulatory network that controls cell proliferation. We found high levels of expression of CDC2 mRNA and protein in proliferating neuronal precursor cells. The expression of both CDC2 and cyclin A was dramatically downregulated upon terminal differentiation of neurons in vivo and in a neuronal precursor cell line, ST15A. p53 mRNA expression was also downregulated but to a lesser extent; RB mRNA levels were unchanged during neuronal differentiation. Immunohistochemistry showed that p34cdc2 was expressed not only in the neuronal precursors of the cerebellar external granule layer but also in the early differentiating granule neurons. The expression of p34cdc2 in early neurons suggests a function for this enzyme in the events that occur soon after proliferation ceases. On the basis of the results reported here and other recent findings, we propose a model in which terminal differentiation is achieved by a switch in the neuronal precursors from p34cdc2-based proliferation to a differentiated state controlled by p34cdc2-related kinases. 相似文献