Suganuma T, Muramatsu H, Muramatsu T, Ihida K, Kawano J, Murata F
Department of Anatomy, Faculty of Medicine, Kagoshima University, Japan.
J Histochem Cytochem. 1991 Mar;39(3):299-309. doi: 10.1177/39.3.1899684.
We prepared a monoclonal antibody (MAb) against N-acetylglucosaminide beta 1----4 galactosyltransferase purified from F9 embryonal carcinoma cells. The MAb recognized the protein portion of the enzyme, since it inhibited galactosyltransferase activity, reacted with the enzyme both from F9 cells and from bovine milk, and did not exhibit anti-carbohydrate activity. Using this MAb, we studied the subcellular localization of the enzyme by immunoelectron microscopy. Intense staining was observed in trans-Golgi stacks within testicular interstitial cells and mucous neck cells, confirming the specificity of the immunological reaction. Cell surface galactosyltransferase was detected in the following regions: cultured cells such as F9 embryonal carcinoma cells, testicular interstitial cells, seminiferous tubule epithelial cells, Sertoli cells, the head of the epididymal sperm, epididymal epithelial cells, and apical surfaces of epithelial cells in the fundic gland and of intestinal goblet cells. The use of Triton X-100 intensified the cell surface immunoreactivity, and in certain cases the mode of distribution of the cell surface enzyme was different from that described in previous reports. In addition, nuclear envelopes of cultured cells were distinctly stained. The possible significance of the latter finding is discussed in relation to recent advances in nuclear localization of glycoproteins.
我们制备了一种针对从F9胚胎癌细胞中纯化的N-乙酰葡糖胺β1----4半乳糖基转移酶的单克隆抗体（MAb）。该单克隆抗体识别该酶的蛋白质部分，因为它抑制半乳糖基转移酶活性，与来自F9细胞和牛乳中的酶都发生反应，且不表现出抗碳水化合物活性。利用这种单克隆抗体，我们通过免疫电子显微镜研究了该酶的亚细胞定位。在睾丸间质细胞和黏液颈细胞的反式高尔基体堆叠中观察到强烈染色，证实了免疫反应的特异性。在以下区域检测到细胞表面半乳糖基转移酶：培养细胞，如F9胚胎癌细胞、睾丸间质细胞、生精小管上皮细胞、支持细胞、附睾精子头部、附睾上皮细胞，以及胃底腺上皮细胞和肠杯状细胞的顶端表面。使用Triton X-100增强了细胞表面免疫反应性，并且在某些情况下，细胞表面酶的分布模式与先前报道的不同。此外，培养细胞的核膜被明显染色。结合糖蛋白核定位的最新进展，讨论了后一发现的可能意义。
