IMMUNOCYTOCHEMIICAL AND MORPHOMETRICAL STUDY OF THE NEURONAL PRECURSOR CELLS DERIVED FROM THE SUBVENTRICULAR ZONE OF THE POSTNATAL RAT FOREBRAIN IN VITRO
- VernacularTitle:离体培养的生后大鼠脑室下层神经元前体细胞免疫细胞化学和形态学研究
- Author:
Dianshuai GAO
;
Fengzhen ZHANG
;
Deguang WANG
;
Changfu ZHOU
- Publication Type:Journal Article
- Keywords:
neuronal precursor cells;
subventricular zone;
cell culture;
rat
- From:
Chinese Journal of Neuroanatomy
2000;16(3):261-264
- CountryChina
- Language:Chinese
-
Abstract:
In order to investigate the chemical and morphornetrical properties of the neuronal precursor cells derived from thesubventricular zone(SVZ) of the postnatal rat forebrain in vitro. The cell-type specific antibodies were used for the immunocy-tochemical staining ,and the morphometric parameters which were the mean soma diameter and the ellipticity index (i. e. , thesmallest soma diameter divided by the largest soma diameter) of every SVZ-derived cell were measured for identifying the pheno-types of the SVZ cells in vitro. The experiment animals were SD rats (weights: 100～ 150 g), the SVZ cells derived from thepostnatal rats were cultured on poly-D-lysine-coated 24-well glass chamber slides in the Neurobasal Medium supplemented withB27 in 5% CO2 at 37 C. The following results were obtained.. At 1 day in vitro, almost all SVZ cells (〉90%) from the postna-tal rat forebrain expressed Tujl, an antibody that recognizes neuron-specific tubulin. Likewise, the preponderance of the SVZcells expressed the polysialylated neural cell adhesion molecule (PSA-N-CAM) ; The majority of the SVZ Tujl-positive cells cul-tured were the cells that had oval-shaped bodies with two short, unbranched processes protruded from every two poles, theirmean soma diameter were 8.42±1.03μm and their ellipticity index were 0.57±0.12. Meanwhile, there were approximately20% of the SVZ cells in culture that were sphere-shaped cells with mean soma diameter 7.20±l.04 μm , and it might be observed that these cells connected with one another. As the time in culture went on, these sphere-shaped SVZ-derived cells alsotransformed to oval-shaped ones as described above, but it could be observed that the cells were still connected in the processesof them. By 3 and 5 days in culture, the SVZ cells had larger cell somas (average diameter 9. 07±1.07 μm), and often consider-ably longer processes but still with few branches. Immunocytochemical staining revealed that the majority of the SVZ cells in cul-ture remained Tujl-positive, PSA-N-CAM-positive. By 7 days in culture, the Tujl-positive cells in culture showed remarkablemorphological changes, and possessed typical neuronal phenotypes, which had more larger cell somas (average diameter 12.8 ±1.13 μm), and had more longer, branched processes. Our results showed that the SVZ in the postnatal SD rats contained theneuronal precursor cells which were PSA-N-CAM-positive and could differentiate into new neurons in vitro.
