Kumamoto K, Matsuura T, Amagai T, Kawata M
Department of Anatomy, Meiji College of Oriental Medicine, Kyoto, Japan.
Cell Tissue Res. 1995 Jul;281(1):1-10. doi: 10.1007/BF00307953.
Oxytocin-like and vasopressin-like immunoreactive cells, and the cells expressing mRNAs for these peptides in the spleen of the C57BL/6 mouse were studied by immunohistochemistry, immuno-electron microscopy and in situ hybridization. Immunoreactive cells were distributed mainly in the splenic cord and marginal zone, whereas there were few in the lymphocyte-packed periarteriolar-lymphoid sheath, lymphoid follicle and germinal center. More numerous vasopressin-positive cells were seen in the splenic cord. The colocalization of oxytocin-like and vasopressin-like immunoreactivity in the same cells was identified by the investigation of mirror sections. By the pre-embedding immuno-electron-microscopic method using antisera against oxytocin and vasopressin, immunopositive reaction products were localized in the matrix around the specific granules, small clear vesicles and mitochondrial membrane of the eosinophils. No immunoreactivity to these peptides was found within the specific granules of the eosinophils. In situ hybridization with synthetic oligonucleotide probes labeled with 32P revealed the presence of mRNAs for oxytocin and vasopressin in the cells of the spleen, the distribution of the mRNAs for these peptides being the same as that of immunopositive cells. These observations suggest that eosinophils synthesize both oxytocin and vasopressin and store them in the matrix. Possible differences in the mechanism of synthesis and storage of these peptides between peripheral eosinophils and hypothalamic neurons are discussed.
通过免疫组织化学、免疫电子显微镜和原位杂交技术,对C57BL/6小鼠脾脏中类催产素和类血管加压素免疫反应性细胞以及表达这些肽mRNA的细胞进行了研究。免疫反应性细胞主要分布在脾索和边缘区,而在富含淋巴细胞的动脉周围淋巴鞘、淋巴滤泡和生发中心则很少。在脾索中可见较多的血管加压素阳性细胞。通过对镜像切片的研究,确定了同一细胞中类催产素和类血管加压素免疫反应性的共定位。采用抗催产素和抗血管加压素抗血清的包埋前免疫电子显微镜方法,免疫阳性反应产物定位于嗜酸性粒细胞特异性颗粒周围的基质、小透明囊泡和线粒体膜上。在嗜酸性粒细胞的特异性颗粒内未发现对这些肽的免疫反应性。用32P标记的合成寡核苷酸探针进行原位杂交,结果显示脾脏细胞中存在催产素和血管加压素的mRNA,这些肽的mRNA分布与免疫阳性细胞的分布相同。这些观察结果表明,嗜酸性粒细胞可合成催产素和血管加压素并将其储存于基质中。文中还讨论了外周嗜酸性粒细胞与下丘脑神经元在这些肽的合成和储存机制上可能存在的差异