Wang Fang-Yu, Watanabe Masahito, Zhu Ren-Min, Maemura Kentaro
Department of Gastroenterology, Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, Jiangsu Province, China.
World J Gastroenterol. 2004 Dec 15;10(24):3608-11. doi: 10.3748/wjg.v10.i24.3608.
To investigate the expression between gamma-aminobutyric acid (GABA) and glutamate decarboxylase and its relation with differentiation and maturation of jejunal epithelial cells in rat jejunum.
Immunohistochemical expression of GABA and glutamate decarboxylase (GAD, including two isoforms, GAD65 and GAD67) was investigated in rat jejunum. Meanwhile, double staining was performed with GAD65 immunohistochemistry, followed by lectin histochemistry of fluorescent wheat germ agglutinin. Furthermore, evaluation of cell kinetics in jejunum was conducted by (3)H-thymidine autoradiography and immunohistochemistry using a monoclonal antibody to proliferating cell nuclear antigen (PCNA).
The cells showing positive immunoreactivity GABA and GAD65 were mainly distributed in the villi in rat jejunum, while jejunal epithelial cells were negative for GAD67. Positive GABA or GAD65 staining was mainly located in the cytoplasm and along the brush border of epithelial cells in the middle and upper portions. In addition, a few GABA and GAD65 strongly positive cells were scattered in the upper two thirds of jejunal villi. Double staining showed that GAD65 immunoreactivity was not found in goblet cells. (3)H-thymidine-labeled nuclei were found in the lower and middle portions of jejunal crypts, which was consistent with PCNA staining. Therefore, GABA and GAD65 were expressed in a maturation or functional zone.
The characteristic expression of GABA and GAD suggests that GABA might be involved in regulation of differentiation and maturation of epithelial cells in rat jejunum.
研究大鼠空肠中γ-氨基丁酸(GABA)与谷氨酸脱羧酶的表达及其与空肠上皮细胞分化和成熟的关系。
研究大鼠空肠中GABA和谷氨酸脱羧酶(GAD,包括两种同工型,GAD65和GAD67)的免疫组织化学表达。同时,进行GAD65免疫组织化学双重染色,随后进行荧光麦胚凝集素的凝集素组织化学。此外,通过³H-胸腺嘧啶核苷放射自显影术和使用抗增殖细胞核抗原(PCNA)单克隆抗体的免疫组织化学对空肠中的细胞动力学进行评估。
显示GABA和GAD65免疫反应阳性的细胞主要分布在大鼠空肠的绒毛中,而空肠上皮细胞GAD67呈阴性。GABA或GAD65阳性染色主要位于上皮细胞中上部的细胞质和沿刷状缘处。此外,少数GABA和GAD65强阳性细胞散在于空肠绒毛的上三分之二处。双重染色显示杯状细胞中未发现GAD65免疫反应性。³H-胸腺嘧啶核苷标记的细胞核见于空肠隐窝的中下部,这与PCNA染色一致。因此,GABA和GAD65在成熟或功能区表达。
GABA和GAD的特征性表达表明GABA可能参与大鼠空肠上皮细胞分化和成熟的调节。