Kent C, Coupland R E
Department of Human Morphology, University of Nottingham Medical School, Queen's Medical Centre, U.K.
J Anat. 1989 Oct;166:213-25.
The localisation of chromogranins A and B, met-enkephalin-arg6-gly7-leu8 (met-enk 8) and protein gene product 9.5 (PGP 9.5) in the adrenal medulla and extra-adrenal chromaffin tissue has been studied in the developing rat by immunogold-silver staining. In the adult rat adrenal the cytoplasm of all medullary chromaffin cells showed a positive response with chromogranin A and B; in each case occasional groups of cells with a low reactivity that may have been NA cells were seen. Chromogranin A was first detected in adrenal medullary and extra-adrenal chromaffin cells at 18 days of gestation whilst chromogranin B was not detected in animals younger than 7 days. In 15 days old animals the adrenal medullary response to A and B was of the same intensity as that seen in the adult. Less than 1% of adult medullary chromaffin cells were responsive to met-enk 8 staining and medullary cells were unreactive in the fetus, with only extra-adrenal chromaffin tissue responding prenatally. During the first postnatal week immunoreactive cells appeared in the adrenal medulla in considerably greater proportions than in the adult gland. In contrast, positively stained nerve terminals associated with chromaffin cells and abundant in the adult adrenal were not detected during the first week of life. Immunoreactive nerve terminals were first seen early in the second week of life at a time when positive chromaffin cells were becoming less common. PGP 9.5 was located in all chromaffin cells of the adult adrenal and was readily detected in chromaffin cells in the adrenal and in extra-adrenal locations of the earliest stage examined (E16). Our findings suggest that the ontogenesis of the chromogranin-like immunostaining reflects the maturation of chromaffin granules and the PGP 9.5 immunostaining detected a protein common to cells of neuronal origin and expressed at an early stage of differentiation. The reciprocal relationship between the presence of enkephalins in chromaffin cells and in their presynaptic terminals merits further investigation.
通过免疫金银染色法,对发育中的大鼠肾上腺髓质和肾上腺外嗜铬组织中嗜铬粒蛋白A和B、甲硫氨酸脑啡肽-精氨酸6-甘氨酸7-亮氨酸8(met-enk 8)和蛋白基因产物9.5(PGP 9.5)的定位进行了研究。在成年大鼠肾上腺中,所有髓质嗜铬细胞的细胞质对嗜铬粒蛋白A和B均呈阳性反应;在每种情况下,偶尔可见反应性较低的细胞群,可能是去甲肾上腺素能(NA)细胞。妊娠18天时,首次在肾上腺髓质和肾上腺外嗜铬细胞中检测到嗜铬粒蛋白A,而在7日龄以下的动物中未检测到嗜铬粒蛋白B。在15日龄动物中,肾上腺髓质对A和B的反应强度与成年动物相同。成年髓质嗜铬细胞中对met-enk 8染色有反应的细胞不到1%,胎儿期髓质细胞无反应,只有肾上腺外嗜铬组织在出生前有反应。在出生后的第一周内,肾上腺髓质中出现免疫反应性细胞的比例远高于成年腺体。相比之下,在出生后的第一周内未检测到与嗜铬细胞相关且在成年肾上腺中丰富的阳性神经末梢。免疫反应性神经末梢最早在出生后第二周早期出现,此时阳性嗜铬细胞变得不那么常见。PGP 9.5位于成年肾上腺的所有嗜铬细胞中,并且在最早检查阶段(E16)的肾上腺和肾上腺外部位的嗜铬细胞中很容易检测到。我们的研究结果表明,嗜铬粒蛋白样免疫染色的个体发生反映了嗜铬颗粒的成熟,并且PGP 9.5免疫染色检测到一种神经元起源细胞共有的蛋白质,该蛋白质在分化早期表达。嗜铬细胞及其突触前终末中脑啡肽的存在之间的相互关系值得进一步研究。