Immunocytochemical localization and development of multiple kinds of neuropeptides and neuroendocrine proteins in the chick ultimobranchial gland
Immunocytochemical localization and development of multiple kinds of neuropeptides and neuroendocrine proteins in the chick ultimobranchial gland
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鸡上鳃腺多种神经肽和神经内分泌蛋白的免疫细胞化学定位和发育
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发表时间：
1991
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通讯作者：
Y. Kameda
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作者：
Y. Kameda
后鳃腺是由C细胞群组成的内分泌器官。在鸡中，腺体由对神经丝免疫反应的神经纤维丰富供应。免疫细胞化学染色发现，鸡后鳃腺C细胞除降钙素外，还对多种神经肽和神经内分泌蛋白呈免疫反应性，降钙素基因相关肽（CGRP）、生长抑素、神经降压素、嗜铬粒蛋白A和酪氨酸羟化酶。此外，还检测到脑啡肽免疫反应细胞，其显示出长的胞质突起和大的胞体，与C细胞特征不同。用计算机辅助图像分析系统测定后鳃腺单位面积上这些细胞的密度：降钙素细胞42.9 ± 10.0%、降钙素基因相关肽细胞26.9 ± 5.6%、神经降压素细胞8.6 ± 6.9%、生长抑素细胞3.1 ± 1.4%、嗜铬粒蛋白A细胞11.8 ± 1.8%、酪氨酸羟化酶细胞10.0 ± 5.2%、酪氨酸羟化酶细胞10.0 ± 5.2%。脑啡肽细胞2.9 ± 1.3%。鸡后鳃腺内也有密集分布的肽能神经纤维。大量P物质免疫反应阳性的曲张纤维分布在C细胞群和血管附近。脑啡肽免疫反应性纤维在C细胞簇周围也很突出。甘丙肽、血管活性肠肽（VIP）和酪氨酸羟化酶免疫反应性纤维仅分布在血管周围。随后，这些神经肽，神经内分泌蛋白，肽能神经支配的个体发育在鸡在不同的发育阶段进行了研究。在10天大的胚胎中，几乎所有的C细胞中都存在降钙素的弱至中等强度的免疫反应性。生长抑素，CGRP，酪氨酸羟化酶的免疫反应性开始出现在这个年龄。在孵育12天时，在后鳃腺的实质中首次检测到P物质免疫反应性纤维。还观察到大量脑啡肽免疫反应性纤维和细胞。在孵育14天时，生长抑素和脑啡肽免疫反应细胞达到最大数量;单位面积生长抑素和脑啡肽免疫反应细胞的密度分别为21.2 ± 3.2%和12.9 ± 3.1%。在这个年龄，P物质免疫反应纤维在整个腺体中变得众多。此后，降钙素-，CGRP-，酪氨酸羟化酶-免疫反应细胞的数量随着胚胎年龄的增加而逐渐增加，而生长抑素-和脑啡肽-免疫反应细胞开始减少。嗜铬粒蛋白A和神经降压素免疫反应性细胞分别在孵育16天和18天开始出现。甘丙肽、VIP和酪氨酸羟化酶免疫反应纤维在胚胎期不明显。
The ultimobranchial gland is an endocrine organ consisting of C cell groups. In chickens, the glands are richly supplied by nerve fibers immunoreactive for neurofilaments. It was found by immunocytochemical staining that C cells of chick ultimobranchial glands showed immunoreactivities for multiple kinds of neuropeptides and neuroendocrine proteins in addition to calcitonin, i.e., calcitonin gene‐related peptide (CGRP), somatostatin, neurotensin, chromogranin A, and tyrosine hydroxylase. Furthermore, enkephalin‐immunoreactive cells that showed long cytoplasmic processes and large cell bodies, being distinct from the C cell feature, were detected. The densities of these cells per unit area of ultimobranchial gland were assessed using computer‐assisted image analysis system; calcitonin cells were 42.9 ± 10.0%; CGRP cells 26.9 ± 5.6%; neurotensin cells 8.6 ± 6.9%; somatostatin cells 3.1 ± 1.4%, chromogranin A cells 11.8 ± 1.8%; tyrosine hydroxylase cells 10.0 ± 5.2%; enkephalin cells 2.9 ± 1.3%. Dense distributions of peptidergic nerve fibers were also detected in chick ultimobranchial glands. Numerous varicose fibers immunoreactive for substance P were distributed in the close vicinity to C cell clusters and blood vessels. Enkephalin‐immunoreactive fibers were also prominent around C cell clusters. Galanin‐, vasoactive intestinal peptide (VIP)‐, and tyrosine hydroxylase‐immunoreactive fibers were distributed around blood vessels only. Subsequently, the ontogeny of these neuropeptides, neuroendocrine proteins, and peptidergic innervations was examined in chickens at various developmental stages. In 10‐day‐old embryos, weak to moderately intense immunoreactivity for calcitonin was already present in almost all C cells. Immunoreactivities for somatostatin, CGRP, and tyrosine hydroxylase began to appear at this age. At 12 days of incubation, substance P‐immunoreactive fibers were first detected in the parenchyma of ultimobranchial glands. Considerable numbers of enkephalin‐immunoreactive fibers and cells were also observed. At 14 days of incubation, the largest populations of somatostatin‐ and enkephalin‐immunoreactive cells were attained; the densities of somatostatin‐ and enkephalin‐immunoreactive cells per unit area were 21.2 ± 3.2% and 12.9 ± 3.1%, respectively. Substance P‐immunoreactive fibers became numerous throughout the gland at this age. Thereafter, calcitonin‐, CGRP‐, tyrosine hydroxylase‐immunoreactive cells progressively increased in number with embryonic age, whereas somatostatin‐ and enkephalin‐immunoreactive cells started to decrease. Chromogranin A‐ and neurotensin‐immurioreactive cells began to appear at 16 days and 18 days of incubation, respectively. Galanin‐, VIP‐, and tyrosine hydroxylase‐immunoreactive fibers were inconspicuous during embryonic life.