Roles of NG2 Glia in Cerebral Small Vessel Disease / 神经科学通报·英文版

Cerebral small vessel disease (CSVD) is one of the most prevalent pathologic processes affecting 5% of people over 50 years of age and contributing to 45% of dementia cases. Increasing evidence has demonstrated the pathological roles of chronic hypoperfusion, impaired cerebral vascular reactivity, and leakage of the blood-brain barrier in CSVD. However, the pathogenesis of CSVD remains elusive thus far, and no radical treatment has been developed. NG2 glia, also known as oligodendrocyte precursor cells, are the fourth type of glial cell in addition to astrocytes, microglia, and oligodendrocytes in the mammalian central nervous system. Many novel functions for NG2 glia in physiological and pathological states have recently been revealed. In this review, we discuss the role of NG2 glia in CSVD and the underlying mechanisms.

Method of transfection of STOP gene lentiviral vector into oligodendrocyte precursor cells of BTBR mouse model of autism / 解剖学报

Objective To investigate the effect of microtubule binding protein STOP on myelin formation of oligodendrocyte in BTBR mice spectrum disorder in vitro, a highly purified primary culture method of oligodendrocyte precursor cells from cerebral cortex of BTBR mice was established. Establishment of a highly efficient transfection method for overexpression of STOP gene in oligodendrocyte precursor cells of BTBR mice cerebral cortex using lentiviral vector. Methods BTBR mice were used as experimental objects, 6-10 suckling mice were taken each time, repeat 3 times independently. The single cell suspension was prepared by trypsin digestion, and the primary oligodendrocyte precursor cells were obtained by immunomagnetic bead cell sorting method . After 5 days of culture, the cell purity was identified by oligodendrocyte precursor cell marker staining. The primary cultured oligodendrocyte precursor cells were transfected with STOP gene vector constructed in the early stage of the project group. 72-96 hours after transfection, the fluorescence staining of oligodendrocyte precursor cells was observed under fluorescence microscope, and the transfection rate and cell survival rate were calculated. Results The oligodendrocyte precursor cells of BTBR mice extracted by immunomagnetic beads sorting method basically adhered to the wall completely after 48 hours, and the cells had strong ability of proliferation. On the fifth da)' of culture, the purity of the cells was more than 95% identified by immunofluorescence. A lentivirus transfection method for primary oligodendrocyte precursor cells of BTBR mice with high transfection efficiency was established. The fluorescence expression of the cells was obvious after being photographed by high connotation microscope, the lentivirus transfection rate of oligodendrocyte precursor cells was increased to 60%-70%. Conclusion The primaiy oligodendrocyte precursor cells of BTBR mouse cerebral cortex with high purity were successfull)' isolated and cultured. A method for lentivirus infection of primaiy oligodendrocyte precursor cells in the cerebral cortex of BTBR mice is successfully established.

Progress in Methodology for Oligodendrocyte Culture in Vitro (review) / 中国康复理论与实践

It is important to establish a simple method of culture in vitro to obtain purity oligodendrocyte. This paper introduced the re-searches about co-culture in vitro with rat cortical gliocyte, proliferation and differentiation of oligodendrocyte progenitor, differentiation of oligodendrocyte and identification of cell surface antigens at different stages of development.

Culture and isolation of oligodendrocyte precursor cells from neonatal rat cerebral cortices / 中国矫形外科杂志

[Objective]To observe the growth pattern of oligodendrocyte precursor cells(OPCs) in the primary culture from neonatal rat cerebral cortices and study the methods of cell culture and isolation in order to obtain purified OPCs for the experiments of cell transplantation.[Method]The mixed glial cells from the cerebral cortices of 48-hour-old Sprague-Dawley(SD) rats were cultured in vitro.Until 9-10 days in vitro OPCs were isolated and purified by the shaking process and differential adhesion,and then continued OPCs culture in the defined medium.The growth pattern of OPCs in vitro was investigated by contrast phase microscopy and OPC s were further identified with the immunocytochemical techniques.[Result]The distinct stratification of OPCs and astrocytes developed around 9-10 days in primary culture.At this point,the OPCs scattered on the top of the monolayer astroctyes and the soma of most OPCs typically appeared oval or round with two or three processes.The purity of the isolated OPCs reached 95% and these OPCs further developed into the mature oligdendrocytes which were immunoreactive to the specific antigen of oligodendrocyte lineage cells,Oligo2.[Conclusion]OPCs separated from the cerebral cortices of neonatal SD rats can be maintained as immature precursor cells in culture,and OPCs are able to be purified by shaking and differential adhesion under the condition of appropriate cell stratification.

Postnatal development and perinatal electrophysiological characteristics of oligodendrocyte precursor cell in rats / 第三军医大学学报

Objective To observe the postnatal development and perinatal electrophysiological characteristics of oligodendrocyte precursor cell (OPC) in rats. Methods Immunohistochemistry and Western blot were applied to determine the expression of NG2 OPC in cerebral cortex and hippocampus at various developmental stages of SD rats. Electrophysiological characteristics of OPC were also recorded in slices of 7-day rats. Results The majority of hippocampal and cerebral OPC exhibited stellate shape,a small cell body with a few processus. Total population of the NG2 immunopositive OPC was numerous at P7d in cerebral cortex and hippocampus. OPC expressed in adult rats with slightly more quantity. Moreover,OPC in hippocampus of P7d rats typically exhibited small inward sodium current and weak active responses,whereas only outward potassium current and inactive responses were recorded in white matter OPC of P7d rats. Conclusion Total population of OPC and relative optical density of NG2 are the highest in P7d rats at the postnatal developmental stages. OPC in cerebrum and hippocampus of P7d rats displays electrophysiological heterogeneity.

Morphological localization of NG2 positive cells in the brain of adult rats / 第三军医大学学报

Objective To observe the localization of NG2 positive cells and morphological character in the brain of adult rats. Methods Immunohistochemical method was applied to determine the expression of NG2 positive cells in the cerebrum cortex, hippocampus, dentate gyrus, thalamencephalon and hypothalamus of adult rats. Image analysis program Image Pro Plus 5.0 was used to count the positive cells and for statistic analysis. Results NG2 positive cells were strongly expressed in multiple brain regions of adult rats, of which strongest signals were centralized in gray and white matter of cerebral cortex, hippocampus, dentate gyrus, thalamic subventricular zone and hypothalamic periventricular region. The NG2 positive cells were seen with abundant process arborization which bifurcated two or more times. The soma of NG2 positive cell displays a star-like morphology with different shapes in the gray and white matter of cerebrum cortex. Conclusion NG2 positive cells are numerous in adult rat brain and display the special glial with a stellate morphology.