Merchenthaler I, Maderdrut J L, Weber E, Petrusz P
Department of Anatomy, University Medical School, Pécs, Hungary.
Brain Res. 1988 Jun 14;452(1-2):87-96. doi: 10.1016/0006-8993(88)90012-1.
Double-staining in either vibratome or paraffin sections using contrasting chromogens revealed an alpha-melanocyte-stimulating hormone (alpha-MSH)-containing cell group in the arcuate nucleus, a metorphamide-containing cell group in the paraventricular hypothalamus, and an extensive group of magnocellular perikarya in the zona incerta (ZI) and the lateral hypothalamus (LH) that appeared to contain both antigens. Staining of adjacent paraffin sections also suggested that most (and perhaps all) of the magnocellular perikarya in the ZI and LH that contained metorphamide-like immunoreactivity also contained alpha-MSH-like immunoreactivity. Metorphamide-like immunoreactivity in the ZI and the LH was abolished by absorption of the antiserum with metorphamide but was unaffected by absorption with alpha-MSH. alpha-MSH-like immunoreactivity in the ZI and LH was abolished by absorption of the antiserum with alpha-MSH but was unaffected by absorption with metorphamide. Antisera directed against [Met5]-enkephalin (Met-ENK), [Met5]-enkephalin-Arg6,Gly7,Leu8 (ENK-8), [Met5]-enkephalin-Arg6,Phe7 (ENK-7), neuropeptide Y, and FMRF-amide did not stain magnocellular perikarya in the ZI and LH. Pretreatment of paraffin sections with trypsin resulted in the appearance of [Met5]-enkephalin-Arg6-like immunoreactivity in the ZI and LH. Pretreatment of paraffin sections with trypsin did not reveal any occult Met-ENK-, ENK-7- or ENK-8-like immunoreactivity in either the ZI or the LH. These observations indicate that magnocellular neurons in the ZI and LH contain both a metorphamide-like and an alpha-MSH-like peptide but do not express either the preproenkephalin or the prepro-opiomelanocortin48 gene.
在振动切片机切片或石蜡切片中使用对比色原进行双重染色，结果显示在弓状核中有一组含α-促黑素细胞激素（α-MSH）的细胞群，室旁下丘脑中有一组含甲硫啡肽的细胞群，以及在未定带（ZI）和外侧下丘脑（LH）中有一大群大细胞神经元胞体，这些细胞似乎同时含有这两种抗原。相邻石蜡切片的染色还表明，ZI和LH中大多数（可能全部）含有甲硫啡肽样免疫反应性的大细胞神经元胞体也含有α-MSH样免疫反应性。用甲硫啡肽吸收抗血清后，ZI和LH中的甲硫啡肽样免疫反应性消失，但用α-MSH吸收则无影响。用α-MSH吸收抗血清后，ZI和LH中的α-MSH样免疫反应性消失，但用甲硫啡肽吸收则无影响。针对[Met5]-脑啡肽（Met-ENK）、[Met5]-脑啡肽-Arg6,Gly7,Leu8（ENK-8）、[Met5]-脑啡肽-Arg6,Phe7（ENK-7）、神经肽Y和FMRF酰胺的抗血清未对ZI和LH中的大细胞神经元胞体进行染色。用胰蛋白酶预处理石蜡切片后，ZI和LH中出现了[Met5]-脑啡肽-Arg6样免疫反应性。用胰蛋白酶预处理石蜡切片后，在ZI或LH中均未发现任何隐匿的Met-ENK-、ENK-7-或ENK-8样免疫反应性。这些观察结果表明，ZI和LH中的大细胞神经元同时含有甲硫啡肽样和α-MSH样肽，但不表达前脑啡肽原或前阿黑皮素原48基因。
