Shimosegawa T, Uchida T, Kobayashi S, Imura H, Mochizuki T, Yanaihara C, Yanaihara N
Nihon Naibunpi Gakkai Zasshi. 1984 Jun 20;60(6):779-87. doi: 10.1507/endocrine1927.60.6_779.
The occurrence and localization of Met-enkephalin-Arg-Gly-Leu (Met-Enk-Arg-Gly-Leu)-like immunoreactivity in the canine pancreas were studied by immunocytochemistry. For light microscopy, Bouin fixed and paraffin embedded sections were immunostained by the PAP method. For electron microscopy, ultrathin sections of 4% paraformaldehyde-fixed and Araldite embedded materials were immunostained by a protein A-colloidal gold method. The results obtained were as follows: In light microscopy, Met-Enk-Arg-Gly-Leu-like immunoreactivity was localized in nerve fibers of autonomic ganglion, in beaded nerve terminals around blood vessels of the exocrine parenchyma and pancreatic islet, in eighty-five out of one hundred and seventy-four ganglion cell somas of intra- and interlobular ganglia, and in ganglion cell somas and nerve fibers of neuro-insular complexes. In electron microscopy, colloidal gold particles representing Met-Enk-Arg-Gly-Leu-like immunoreactivity were concentrated on secretory type granules of the ganglion cell somas and on large cored synaptic vesicles of the nerve terminals around blood capillaries. All the positive immunoreactions disappeared with preincubation of the anti-Met-Enk-Arg-Gly-Leu serum (dilution, 1:4,000) with Met-Enk-Arg-Gly-Leu (10 micrograms/ml). Immunostaining was not affected by preincubation of the antiserum with Met-enkephalin (10 micrograms/ml), Leu-enkephalin (10 micrograms/ml) and Met-enkephalin-Arg-Phe (20 micrograms/ml). Out of the three precursors for a variety of opioid peptides throughout the body, i.e. preproenkephalin A, preproopiomelanocortin and preproenkephalin B, only preproenkephalin A contains Met-Enk-Arg-Gly-Leu.(ABSTRACT TRUNCATED AT 250 WORDS)
通过免疫细胞化学方法研究了甲硫氨酸脑啡肽-精氨酸-甘氨酸-亮氨酸(Met-Enk-Arg-Gly-Leu)样免疫反应性在犬胰腺中的发生及定位。光学显微镜检查时,用波因氏固定、石蜡包埋的切片采用过氧化物酶抗过氧化物酶(PAP)法进行免疫染色。电子显微镜检查时,用4%多聚甲醛固定、环氧树脂包埋材料的超薄切片采用蛋白A-胶体金法进行免疫染色。结果如下:在光学显微镜下,Met-Enk-Arg-Gly-Leu样免疫反应性定位于自主神经节的神经纤维、外分泌实质和胰岛血管周围的串珠状神经末梢、174个小叶内和小叶间神经节中的85个神经节细胞胞体以及神经胰岛复合体的神经节细胞胞体和神经纤维。在电子显微镜下,代表Met-Enk-Arg-Gly-Leu样免疫反应性的胶体金颗粒集中在神经节细胞胞体的分泌型颗粒和毛细血管周围神经末梢的大核心突触小泡上。用Met-Enk-Arg-Gly-Leu(10微克/毫升)对抗Met-Enk-Arg-Gly-Leu血清(稀释度1:4000)进行预孵育后,所有阳性免疫反应消失。用甲硫氨酸脑啡肽(10微克/毫升)、亮氨酸脑啡肽(10微克/毫升)和甲硫氨酸脑啡肽-精氨酸-苯丙氨酸(20微克/毫升)对抗血清进行预孵育不影响免疫染色。在全身多种阿片肽的三种前体中,即前脑啡肽原A、前阿黑皮素原和前脑啡肽原B,只有前脑啡肽原A含有Met-Enk-Arg-Gly-Leu。(摘要截短于250字)