Effect of subventricular zone irradiation on prognosis of patients with glioblastoma / 中华放射肿瘤学杂志

Objective:To investigate the role of subventricular zone (SVZ) irradiation in the prognosis of patients with glioblastoma (GBM), and to analyze the factors affecting the prognosis of patients with GBM.Methods:Clinical data of 52 patients with GBM treated in the Affiliated Cancer Hospital of Nanjing Medical University from 2017 to 2020 were analyzed retrospectively. According to the median dose of ipsilateral or contralateral SVZ, the patients were divided into the high-dose group and low-dose group. The prognostic differences between two groups were compared and the prognostic factors were analyzed.Results:The median progression-free survival (PFS) was 17.1 months (95% CI:12.4-30.7)and the median overall survival (OS) was 38.3 months (95% CI:20.4-44.5). Univariate analysis showed that whether the tumor invading SVZ ( P = 0.039), the degree of resection ( P = 0.009) and MGMT promoter methylation status ( P = 0.039) were the influencing factors of PFS. Age ( P = 0.018), Kanofsky performance score ( P = 0.043), whether the tumor invading SVZ ( P = 0.038), degree of resection ( P = 0.020) and MGMT promoter methylation status ( P = 0.019) were the influencing factors of OS. The analysis of SVZ dose as a continuous variable showed that SVZ dose was the influencing factor of PFS ( P < 0.05) rather than OS ( P ≥ 0.05). Whether the tumor invading SVZ or not, there was no significant difference in survival between the high-dose and low-dose groups. Multivariate analysis showed that whether the tumor invading SVZ and MGMT promoter methylation were the independent prognostic factor for PFS (both P < 0.05), and OS (both P < 0.05). The SVZ dose related variables were not statistically significant in multivariate analysis. Conclusions:Patients with tumors directly invading SVZ achieve worse survival. Increasing the ipsilateral or contralateral SVZ dose does not improve patient survival. Whether SVZ irradiation affects the survival of patients still needs to be further confirmed by prospective randomized clinical studies.

Penetrative needling of scalp-points improves neurological function by promoting stem cell proli-feration and activating Shh/Gil1 signaling in intracerebral hemorrhage rats / 针刺研究

OBJECTIVE: To observe the effect of penetrative needling from "Baihui" (GV20) to "Qubin" (GB7) on neural stem cell proliferation and sonic hedgehog (Shh) signaling in subventricular zone (SVZ) in intracerebral hemorrhage (ICH) rats so as to explore its mechanisms underlying improvement of ischemic injury of brain. METHODS: Male SD rats were randomly divided into blank control, model, acupuncture and agonist (Purmorphamine, an activator of Shh signaling pathway) groups (n＝18 in each group, 6 for H．E. stain, 6 for examining neuronal cell proliferation, and 6 for immunohistochemistry). The ICH model was established by injecting autogenous blood (50 µL) into the right caudate nucleus. The neurological defect was scored with refe-rence to Bederson's method. Penetrative needling from GV20 to GB7 was performed by manipulating the needle for 6 min (repeated 3 times in 30 min), once daily for 7 days. Intraperitoneal injection of Purmorphamine (1 mg/mL, 1 mg/kg) was performed, once daily for 7 days. Histopathological changes of the hemorrhagic penumbra region were observed under microscope after H．E. stain, the newborn neural stem cell proliferation (BrdU＋/Nestin＋ double labeled cells) in the SVZ was observed by immunofluorescence after intraperitoneal injection of BrdU (50 mg/kg), and the expression of Shh and glioma-associated hemolog-1 (Gli1) detected by immunohistochemistry. RESULTS: After modeling, the neurological score and expression levels of Shh and Gil1 proteins were significantly increased in the model group relevant to the blank control group (P0.05). Outcomes of H．E. stain showed obvious edema, disordered arrangement of cells, infiltration of inflammatory cells and red blood cells with glial cell hyperplasia around the hematoma area in the model group, which was relatively milder in both acupuncture and agonist groups such as in basic disappearance of edema and inflammatory reaction. CONCLUSION: Penetrative needling from GV20 to GB7 can obviously improve neurological function in ICH rats, which is related to its effects in activating Shh/Gil1 signaling and in further promoting neural stem cell proliferation in the SVZ region.

A reignited debate over the cell(s) of origin for glioblastoma and its clinical implications / 医学前沿

Glioblastoma (GBM) is the most common and lethal primary neoplasm in the central nervous system. Despite intensive treatment, the prognosis for patients with GBM remains poor, with a median survival of 14-16 months. 90% of GBMs are primary GBMs that are full-blown at diagnosis without evidences of a pre-existing less-malignant precursor lesion. Therefore, identification of the cell(s) of origin for GBM-the normal cell or cell type that acquires the initial GBM-promoting genetic hit(s)-is the key to the understanding of the disease etiology and the development of novel therapies. Neural stem cells and oligodendrocyte precursor cells are the two major candidates for the cell(s) of origin for GBM. Latest data from human samples have reignited the longstanding debate over which cells are the clinically more relevant origin for GBMs. By critically analyzing evidences for or against the candidacy of each cell type, we highlight the most recent progress and debate in the field, explore the clinical implications, and propose future directions toward early diagnosis and preventive treatment of GBMs.

Effect of subventricular zone invasion on prognosis of glioma patients / 中国肿瘤临床

This work was supported by Science and Technology Program of Health and Family Planning Commission of Jiangxi Province (No.20161106) Abstract Objective: To investigate the prognostic value of subventricular zone (SVZ) invasion in glioma patients. Methods: The clini-cal data of 175 patients with glioma diagnosed based on pathology in Jiangxi Province Cancer hospital between January 2010 and July 2015 were analyzed retrospectively. There were 59 cases of World Health Organization (WHO) gradeⅡ, 59 cases of WHO gradeⅢ, and 57 cases of WHO gradeⅣat the first diagnosis. There were 75 cases of SVZ invasion (SVZ+) and 100 cases of SVZ non-invasion (SVZ-) according to preoperative magnetic resonance imaging. The survival outcomes of both cohorts were compared using the Log-rank test. The correlation between the recurrence pattern and SVZ involvement was analyzed using Chi-square tests. Results: The me-dian follow-up time was 63 months. The 5-year overall survival (OS) and progression-free survival (PFS) rates were 42.2% and 37.5%, respectively. These were 20.9% and 15.3% in the SVZ+group, compared with 57.1% and 44.1% in the SVZ-group, respectively (P<0.001 and P<0.001, respectively). The SVZ+group had fewer cases of total resection, larger lesions (maximum diameter greater than 5.0 cm), and more cases of gradeⅣ(P<0.001, P<0.001, and P=0.018, respectively). There were 89 cases of recurrence. The total recur-rence rate was 62.7% in the SVZ+group, compared with 42.0% in the SVZ-group (P=0.007); the distant recurrence rates were 21.3% and 7.0% (P=0.004), respectively. Conclusions: SVZ invasion is a poor prognostic factor for OS and PFS in gliomas, which is positively correlated with a low total resection rate, large lesions, and gradeⅣlesions, and increases the probability of total recurrence and dis-tant recurrence.

Effects of donepezil on the expression of neurotrophic related factors in subventricular zone of cerebral infarction mice / 中华行为医学与脑科学杂志

Objective To investigate the effect of donepezil on subventricular zone ( SVZ) neuro-genesis related neurotrophic factors after cerebral infarction. Methods Mice were randomly assigned into three groups: vehicle-treated sham group (Sham+vehicle,n=18),vehicle-treated middle cerebral artery oc-clusion (MCAO) group (MCAO + vehicle,n=30) and donepezil-treated MCAO group (MCAO+donepezil, n=30). Middle cerebral artery occlusion( MCAO) was induced by thread-occlusion method. Nissl staining was used to measure the infarct volume and the modified neurological severity score(mNSS) was used to as-sess neurologic function and brain water content was detected to assess brain edema degree. Proliferative cells and neuroblasts were labeled with 5-bromodeoxyuridine ( BrdU) and doublecortin ( DCX). The SVZ BrdU+/DCX+cells were detected by immunofluorescence. The expression of glial cell line-derived neurotro-phic factor (GDNF),brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) were detec-ted by Western blot. Results The infarct volume of MCAO + donepezil group ((13. 33±4. 55)%) was sig-nificantly lower than that of MCAO + vehicle group ((31. 33±3. 93)%,t=7. 34,P<0. 05). The neurologic deficits were significantly ameliorated after donepezil treatment,and the brain water content of MCAO + done-pezil group ((71. 82±10. 18)%)was significantly less than that of MCAO + vehicle group ((85. 93± 7. 54)%,F=13. 480,P<0. 05). All differences were statistically significant (P<0. 05). The area of BrdU+/DCX+cells within SVZ of MCAO + vehicle group ((6. 16±1. 79)%) was significantly larger than that of sham + vehicle group ((2. 25±1. 09)%),and was fewer than that of MCAO+donepezil group ((16. 19± 2. 16)%,F=102. 756,P<0. 05). MCAO significantly promoted the expression of GDNF,BDNF and NGF within SVZ compared with sham operation,and donepezil increased these protein levels(F=15. 114,27. 121, 27. 398,P<0. 05). Conclusion Donepezil regulates neurogenesis via increasesing the expression of GDNF, BDNF and NGF within SVZ after cerebral infarction.

Effect of donepezil on the proliferation of neurons in subventricular zone and the mechanism of Src signaling pathway in mice with cerebral ischemia / 中华行为医学与脑科学杂志

Objective To investigate the role of Src signaling pathway in neurogenesis promoted by choline acetyltransferase (CHAT) + neurons in the subventricular zone (SVZ) after ischemic stroke.Methods The eighty-four mice were randomly assigned into four groups:sham-operated mice treated with vehicle (Sham+vehicle,n=18),middle cerebral artery occlusion (MCAO)-operated mice treated with vehicle (MCAO+vehicle,n=22),MCAO mice treated with donepezil (MCAO+donepezil,n=21),MCAO mice treated with donepezil and Src inhibitor KX2-391 (MCAO+donepezil+KX2-391,n=23).Mice were subjected to the temporary MCAO model of ischemic stroke.Modified neurological severity score (mNSS) was used to assess neurologic function of the mice.Proliferative cells were labeled with Ki67,and neuroblasts with doublecortin (DCX).The expression of Ki67+/DCX+ in the SVZ was detected by immunofluorescence.The expression of Ki67,phospho-epidermal growth factor receptor (p-EGFR),p-Raf,Src and p-Akt in the SVZ were quantified by Western blot.Results MCAO+donepezil+KX2-391 group showed worse performance in the mNSS test than MCAO+donepezil group (P＜0.05).Ten days after MCAO,the number of Ki67+/DCX+ cells in the SVZ of MCAO+donepezil group was 125.33± 13.71/area,which was 71.67± 18.35/area in MCAO+ donepezil+KX2-391 group (P＜0.05).What's more,the expression of proteins Ki67,p-EGFR,p-Raf,Src and p-Akt in mice of MCAO+donepezil group was markedly increased,which was (1.39±0.23),(1.42±0.19),(0.88±0.13),(1.14±0.19),(1.04±0.18) and it was decreased in MCAO+donepezil+KX2-391 group,which was 0.84±0.26,0.94±0.26,0.73±0.15,0.71±0.18,0.81±0.19(P＜0.05).Conclusion CHAT+ neurons in SVZ may promote neurogenesis after stroke via Src-epidermal growth factor receptor (EGFR) signaling pathway.

Research advances in subventricular zone irradiation in glioblastoma / 中华放射肿瘤学杂志

Glioblastoma multiforme (GBM,WHO grade IV) contains some glioma stem cells which have unique self-renewal capacity and multilineage potency.There are numerous neural stem cells in the subventricular zone (SVZ) of adult human brain;it may also act as a storehouse of glioma stem cells that can promote the development and recurrence of a tumor.GBM involving SVZ is prone to early recurrence and intracranial metastasis after resection,so irradiation of the SVZ potentially influences the survival of GBM patients.This review provides a summary of related experimental and clinical studies,and discusses the value of irradiation of the SVZ in GBM patients and the direction of future research.

Angiogenesis promotion in peri-infarction region by ChAT-positive neurons in subventricular zone after ischemic stroke / 实用医学杂志

Objective To observe the activities of ChAT + neurons in subventricular zone (SVZ) after ischemic stroke and their effects on angiogenesis in peri-infarction region and related signaling pathways. Methods C57BL/6 mice were randomly assigned into sham group,middle cerebral artery occlusion (MCAO) group and atropine group. Ischemic models were made by permanent coagulation of the distal middle cerebral artery. The expression of ChAT,AChE in SVZ and VEGF,VEGFR2,pERK in peripheral regions of ischemic injury was evaluated by Western blotting and immunofluorescence. 5-bromodeoxyuridine(BrdU)/CD31 double-labeled cells were also tested by immunofluorescence. Results At 14 d after the surgery,the ratio of ChAT/AChE in SVZ increased after stroke(P < 0.05). Compared with those in Sham group,the levels of VEGF,VEGFR2 and pERK were higher in MCAO group(P<0.05)and VEGFR2-positive and BrdU/CD31-positive cells increased significantly. However,lower expression of VEGF,VEGFR2 and pERK and less VEGFR2-positive and BrdU/CD31-positive cells were found in atropine group when compared with that in MCAO group. Conclusions The activities of ChAT +neurons in SVZ are enhanced after ischemic injury and they can promote angiogenesis in peripheral region of ischemic injury via upregulating VEGF-VEGFR2 signaling pathway and improving the brain function restoration.

Culture, characterization and differentiation of neural precursors from the central nervous system of guinea pigs (Cavia porcellus Linnaeus, 1758) / Cultura, caracterização e diferenciação de precursores neurais do sistema nervoso central de porquinho-da-índia (Cavia porcellus Linnaeus, 1758)

Potentially neurogenic areas were initially identified by incorporation of bromodeoxyuridine (BrdU) in cells underlying the subventricular zone (SVZ) of the lateral ventricles wall, hippocampus and olfactory bulbs of newborn guinea pigs. Neural precursors from the SVZ were cultured in suspension, generating neurospheres (NSFs), which, upon dissociation were able to generate new NSFs. Upon culture in the absence of growth factors, cells dissociated from NSFs displayed evidence for neural differentiation, giving rise to cells from neural lineage. Flow cytometry analysis for of NSFs-derived cells after differentiation revealed approximately 13.3% nestin positive, 5.5% Beta-III-tubulin positive, 9% GFAP positive and 7.8% mGalC positive. Functional assays by measurement of calcium influx upon gamma butiric amino acid (GABA) and glutamate stimuli, revealed stimulation in differentiated cells, an indicator of neuronal differentiation. The ability of guinea pig SVZ cells to originate functional neurons in vitro is promising for research and towards a future use of neural stem cells in the therapy of neurological disorders.(AU)

Áreas potencialmente neurogênicas foram identificadas por incorporação de bromodeoxiuridina (BrdU) na zona subventricular (SVZ) dos ventrículos laterais, hipocampo e bulbos olfatórios de cobaias neonatos. Precursores neurais provenientes da SVZ foram cultivados em suspensão, resultando na geração de neuroesferas (NSFs), que quando dissociadas foram capazes de proliferar e gerar novas NSFs. Quando cultivadas na ausência de fatores de crescimento, as células provenientes de NSFs dissociadas apresentaram evidências de diferenciação neuronal, dando origem a células da linhagem neural. Citometria de fluxo em células das NSFs após a diferenciação revelou aproximadamente 13,3% positivas para nestina, 5,5% positivas para Beta-III-tubulina, 9% positivas para GFAP e 7,8% positivas para mGalC. Testes de funcionalidade pela mensuração de influxo de cálcio após estímulo com ácido gama amino butírico (GABA) e glutamato revelaram a estimulação de células diferenciadas, um indicador de função neuronal. A capacidade de células da SVZ de fetos de cobaias originarem células neurais funcionais in vitro é promissora para a pesquisa e eventual uso terapêutico de células tronco em disordens do sistema nervoso.(AU)

Acteoside promotes proliferation of neural stem cells from adult mice by activating PI3 K/AKT pathway / 中国药理学通报

Aim Toclarifytheeffectofacteosideon proliferation of neural stem cells (NSCs ) from adult mice,as well as the involved signaling pathway.Meth-ods NSCswereisolatedfromthesubventricularzone (SVZ)of adult C57BL/6 mice,then identified by im-munofluorescence staining with Nestin,the marker of NSCs.NSCs were exposed to acteoside (5,10,20,40μmol·L-1 )in absence of mitogen(EGF/bFGF)for 24 h.We employed CCK8 assay to detect NSCs viability and BrdU staining to identify NSCs proliferation.We performed Western blot to quantify the expression level ofp-AktinducedbyacteosideonNSCs.Results With-out mitogen,acteoside increased NSCs proliferation by activating p-Akt,which can be blocked by LY294002, the inhibitor of PI3K/AKT signaling pathway.Conclu-sion ActeosidepromotestheproliferationofNSCsfrom adult mice by activating PI3K/AKT pathway.

Donepezil promotes neurogenesis in the subventricular zone：an experimental study of cerebral ischemia in mice / 实用医学杂志

Objective To investigate the influence of donepezil on neurogenesis in the subventricular zone (SVZ) under the cerebral ischemic conditions. Methods Sixty mice were randomly assigned to a sham +vehicle group, a middle cerebral artery occlusion(MCAO) + vehicle group and a MCAO + donepezil group, 20 in each group. A model of MCAO was established. Neural stem cells/progenitor cells were labeled by Mash1, and neuroblasts were labeled by doublecortin (DCX). The expressions of DCX in the SVZ cells of each group were detected by immunofluorescence. The expressions of Mash1 and DCX in the SVZ cells of each group were quantified by Western blot. Results 10 days after MCAO was undergone, DCX-positive cells were seen in the SVZ of each group, and the expressions of DCX and Mash1 in MCAO + vehicle group were markedly increased as compared with the sham + vehicle group (P < 0.05), and that in MCAO + donepezil group were significantly increased as compared with the other two groups (P < 0.05). Conclusion As a kind of acetylcholinesterase inhibitors, donepezil can promote neurogenesis in SVZ under the cerebral ischemic conditions.

The effects of functional electrical stimulation on motor function and on the proliferation, migration and differentiation of neural stem cells in the subventricular zone / 中华物理医学与康复杂志

Objective To investigate the effects of functional electrical stimulation (FES) on motor function and on the expression, proliferation, migration and differentiation of endogenous neural stem cells in the subventricular zone (SVZ) after cerebral ischemia.Methods Middle cerebral artery occlusion (MCAO) was used to induce a model of cerebral ischemia in 108 rats using the modified Zea-Longa method of intraluminal filament occlusion.They were then randomly divided into an FES group, a placebo stimulation group and a control group with 36 cases in each.Superficial FES electrodes were pasted on the paralyzed forelimbs of the rats in the first two groups, though FES treatment was administered only to the FES group beginning on the 3rd day after the MCAO operation.The stimulation was designed to produce extension of the wrist and digits of the paralyzed forelimb.Before, and after 1,3, 7 and 14 days of the treatment, the neurological deficit was evaluated using modified neurological severity scoring (mNSS).BrdU +/GFAP+, BrdU+/DCX+ and BrdU+/NeuN + cells in the SVZ were detected using immunofluorescence technique.Results After 7 and 14 days of treatment, the average motor function of the rats in the FES group had improved significantly when compared with the averages of the other two groups.Compared with the other two groups, the average number of BrdU +/GFAP+ positive cells in the ischemic SVZ was also significantly greater in the FES group after 7 and 14 days of treatment.After 14 days, BrdU +/Dcx + positive cells in the FES group had also increased significantly more,but only a few BrdU +/NeuN + cells had appeared in any of the three groups.Conclusion FES can improve motor function after cerebral ischemia, and promote proliferation and differentiation of neural stem cells in the SVZ.

The effects of repetitive transcranial magnetic stimulation on proliferation and differentiation of neural stem cells / 대한해부학회지

Repetitive transcranial magnetic stimulation (rTMS) is a new method for treating many neurological conditions; however, the exact therapeutic mechanisms behind rTMS-induced plasticity are still unknown. Neural stem and progenitor cells (NS/PCs) are active players in brain regeneration and plasticity but their behavior in the context of rTMS therapy needs further elucidation. We aimed to evaluate the effects of rTMS on proliferation and differentiation of NS/PCs in the subventricular zone (SVZ) of adult mouse brain. Adult male mice (n=30) were divided into rTMS (1-Hz and 30-Hz) and sham groups and treated for 7 or 14 consecutive days. Harvested NS/PCs from the SVZ were cultured in the neurosphere assay for 8 days and the number and size of the resulting neurospheres as well as their in vitro differentiation capacity were evaluated. After one week of rTMS treatment at 1-Hz and 30-Hz compared with sham stimulation, the mean neurosphere forming frequency per brain was not different while this measure significantly increased after two weeks (P<0.05). The mean neurosphere diameter in 1-Hz treatment paradigm was significantly larger compared with sham stimulation at both 1 and 2 weeks. In contrast, 30-Hz treatment paradigm resulted in significantly larger neurospheres only after 2 weeks. Importantly, rTMS treatment at both frequencies increased neuronal differentiation of the harvested NS/PCs. Furthermore, one week in vitro rTMS treatment of NS/PCs with both 1-Hz and 30-Hz increased NS/PCs proliferation and neuronal differentiation. It is concluded that both 1-Hz and 30-Hz rTMS treatment increase NS/PCs proliferation and neuronal differentiation.

Polarized and Stage-Dependent Distribution of Immunoreactivity for Novel PDZ-Binding Protein Preso1 in Adult Neurogenic Regions

BACKGROUND: Adult neural stem cells have the potential for self-renewal and differentiation into multiple cell lineages via symmetric or asymmetric cell division. Preso1 is a recently identified protein involved in the formation of dendritic spines and the promotion of axonal growth in developing neurons. Preso1 can also bind to cell polarity proteins, suggesting a potential role for Preso1 in asymmetric cell division. METHODS: To investigate the distribution of Preso1, we performed immunohistochemistry with adult mouse brain slice. Also, polarized distribution of Preso1 was assessed by immunocytochemistry in cultured neural stem cells. RESULTS: Immunoreactivity for Preso1 (Preso1-IR) was strong in the rostral migratory stream and subventricular zone, where proliferating transit-amplifying cells and neuroblasts are prevalent. In cultured neural stem cells, Preso1-IR was unequally distributed in the cell cytosol. We also observed the distribution of Preso1 in the subgranular zone of the hippocampal dentate gyrus, another neurogenic region in the adult brain. Interestingly, Preso1-IR was transiently observed in the nuclei of doublecortin-expressing neuroblasts immediately after asymmetric cell division. CONCLUSION: Our study demonstrated that Preso1 is asymmetrically distributed in the cytosol and nuclei of neural stem/progenitor cells in the adult brain, and may play a significant role in cell differentiation via association with cell polarity machinery.

Effects of olfactory bulb lesion on neural stem cells proliferation and expression of NMDA-receptor subunit 2B in the subventricular zone of rats / 中华行为医学与脑科学杂志

Objective To investigate the effects of olfactory bulb(OB) lesion on neural stem cells proliferation and expression of NMDA receptor subunit 2B in subventricular zone(SVZ) of rats.Methods Sixty adult female SD rats were randomly divided into normal group,saline group and OB lesion group.OB lesion was induced by N-methyl-D-aspartic acid (NMDA) injection.Each group was respectively divided into four time points including 3 d,7 d,14 d and 28 d.Immunohistochemistry staining was used to detect the number of Nestin,Ki67 and NR2B-positive cells in the SVZ.Results (1) Nestin positive cells in the SVZ were shown at the different time of three groups.Seven days after OB lesion,IOD value of nestin-positive cells began to increase((29601± 1788)/0.01 mm2,P＜0.05),reached the maximum at 14 d((49800±3701)/0.01 mm2,P＜0.05) and still sustained a high level at 28 day((27600±3209)/0.01 mm2,P＜0.05).(2)Ki67 positive cells in the SVZ were shown at the different time of three groups.The number of Ki67-positive cells was increased significantly at 7 d,14 d and 28 d after OB lesion compared to normal group and saline group (P＜0.05).(3)NR2B immune expression in the SVZ was shown at the different time of three groups.The NR2B-positive cells increased at 3 d after OB lesion(58.80±2.95,P＜0.05),reached the maximum at 14 d(68.40±4.04,P＜0.05).At 28 d of OB lesion,the number of positive cells was reduced,but still sustained a high level(62.20±3.56,P＜0.05).(4)The positive cells of NR2B and Ki67 were highly positive correlation at different time after OB lesion(r=0.968,P＜0.05).Conclusions OB lesion can stimulate neural stem cell proliferation and increases the expression of NR2B.The increased mode of NR2B is in accordance with the schedule of the neural stem cells increase induced by OB lesion.Therefore,it indicates that the NMDA receptor subunit 2B may be involved in neural stem cell proliferation.

Identification and culture of neural stem cells isolated from adult rat subventricular zone following fluid percussion brain injury / 药物分析学报

Objective To analyze proliferation and differentiation of glial fibrillary acid protein (GFAP)- and nestin-positive (GFAP+/nestin+) cells isolated from the subventricular zone following fluid percussion brain injury to determine whether GFAP+/nestin+ cells exhibit characteristics of neural stem cells. Methods Male Sprague-Dawley rats, aged 12 weeks and weighing 200-250 g, were randomly and evenly assigned to normal control group and model group. In the model group, a rat model of fluid percussion brain injury was established. Five days later, subventricular zone tissue was resected from each group and made into single cell suspension. After serum-free neural stem cell medium culture and subsequent serum-induced differentiation, cell type, proliferation and differentiation capacities were determined by immunofluorescence staining and flow cytometry. Results At 3-7 days after fluid percussion brain injury, nestin+/GFAP+ cells in the single cell suspension from the model group significantly outnumbered those from the normal control group (P<0.01). In the model group, an increased number of small neurospheres with smooth cell edge and bulged center formed after primary culture, and were clearly visible with the increase of culture time and medium replacement. After several passages, many clonal spheres were obtained, suggesting strong self-proliferatiing capacity. Neurospheres from the model group differentiated into astrocytes, neurons and oligodendrocytes. Conclusion GFAP+/nestin+ cells isolated from the adult rat subventricular zone after fluid percussion brain injury are thought to be neural stem cells because of their self-renewal and multi-differentiation capacities.

Neurogenesis in the brain / 西安交通大学学报(医学版)

Neurogenesis is sustained throughout adulthood in the mammalian brain due to the proliferation and differentiation of adult neural progenitor cells found in the subventricular zone of the lateral ventricles and subgranular zone of the dentate gyrus. This review covers recent findings that elucidate different aspects of regulation of neurogenesis, including proliferation, migration and differentiation into mature neurons and functional integration into the existing neural circuits. Furthermore, this review also discusses the effects of pathological conditions on adult neurogenesis in both rodent models and human patients as well as some of the potential problems or limitations in neurogenesis research, which may shed some light on developing novel research strategies for replacement treatment of neurological disorders.

Identification and culture of neural stem cells isolated from adult rat subventricular zone following fluid percussion brain injury / 西安交通大学学报·英文版

Objective: To analyze proliferation and differentiation of glial fibrillary acid protein (GFAP)- and nestin-positive (GFAP+/nestin+) cells isolated from the subventricular zone following fluid percussion brain injury to determine whether GFAP+/nestin+ cells exhibit characteristics of neural stem cells. Methods: Male Sprague-Dawley rats, aged 12 weeks and weighing 200-250 g, were randomly and evenly assigned to normal control group and model group. In the model group, a rat model of fluid percussion brain injury was established. Five days later, subventricular zone tissue was resected from each group and made into single cell suspension. After serum-free neural stem cell medium culture and subsequent serum-induced differentiation, cell type, proliferation and differentiation capacities were determined by immunofluorescence staining and flow cytometry. Results: At 3-7 days after fluid percussion brain injury, nestin+/GFAP+ cells in the single cell suspension from the model group significantly outnumbered those from the normal control group (P < 0.01). In the model group, an increased number of small neurospheres with smooth cell edge and bulged center formed after primary culture, and were clearly visible with the increase of culture time and medium replacement. After several passages, many clonal spheres were obtained, suggesting strong self-proliferatiing capacity. Neurospheres from the model group differentiated into astrocytes, neurons and oligodendrocytes. Conclusion: GFAP+/nestin+ cells isolated from the adult rat subventricular zone after fluid percussion brain injury are thought to be neural stem cells because of their self-renewal and multi-differentiation capacities.

Induction of Striatal Neurogenesis by Voluntary Exercise and Environmental Enrichment

OBJECTIVE: To investigate that neurogenesis in the subventricular zone (SVZ), which is already known as neurogenic area where neural stem/progenitor cells persist, and the striatum, which is non-neurogenic area, might be induced by voluntary exercise (VEx) or environmental enrichment (EE), and compare the extent of the neurogenesis with untreated controls. METHOD: Total 12 C57BL/6 mice, 2~3 months old, were recruited as follows; voluntary wheel runner, EE and control. For 2 weeks, VEx group was housed in rat cage (48x26 cm) with 2 running wheels with 3~4 animals/cage, and EE group was housed in the living condition of huge cage (86x76 cm), social interaction (13~14 mice/cage) and objects such as toys, tunnels and running wheel, whereas control group was placed in the standard cage (30x18 cm). RESULTS: VEx and EE tended to increase the densities of mitotic marker BrdU+ cells in SVZ and striatum. They also exhibited more BrdU+ cells (/mm3) into the striatum, even though they did not show statistical significance. Moreover, EE group showed significant increment of the newly generated neurons coexpressed with BrdU+ and betaIII-tubulin+ (/mm3) in SVZ and striatum as compared to those of controls. CONCLUSION: Voluntary physical exercise and EE induced cell proliferation and neurogenesis in both SVZ and striatum. Characteristically, EE could significantly induce neurogenesis in striatum, non-neurogenic area as well as SVZ, typical neurogenic area. Therefore, this strategy might be used to activate neural regeneration in various central nervous system diseases.

Erythropoietin Attenuates Brain Injury, Subventricular Zone Expansion, and Sensorimotor Deficits in Hypoxic-Ischemic Neonatal Rats

The aim of this study was to investigate the effect of erythropoietin (EPO) on histological brain injury, subventricular zone (SVZ) expansion, and sensorimotor function deficits induced by hypoxia-ischemia (HI) in newborn rat pups. Seven-day-old male rat pups were divided into six groups: normoxia control, normoxia EPO, hypoxia control, hypoxia EPO, HI control, and HI EPO group. Sham surgery or HI was performed in all animals. HI was induced by ligation of the right common carotid artery followed by 90 min of hypoxia with 8% oxygen. Recombinant human EPO 3 U/g or saline was administered intraperitoneally, immediately, at 24- and 48-hr after insult. At two weeks after insult, animals were challenged with cylinder-rearing test for evaluating forelimb asymmetry to determine sensorimotor function. All animals were then sacrificed for volumetric analysis of the cerebral hemispheres and the SVZ. The saline-treated HI rats showed marked asymmetry by preferential use of the non-impaired, ipsilateral paw in the cylinder-rearing test. Volumetric analysis of brains revealed significantly decreased preserved ipsilateral hemispheric volume and increased ipsilateral SVZ volume compared with the sham-operated animals. Treatment of EPO significantly improved forelimb asymmetry and preserved ipsilateral hemispheric volume along with decreased expansion of ipsilateral SVZ following HI compared to the saline-treated HI rats. These results support the use of EPO as a candidate drug for treatment of neonatal hypoxic-ischemic brain damage.