Cheng-Chew S B, Leung P Y
Department of Physiology, Faculty of Medicine, Chinese University of Hong Kong, Shatin, N.T., Hong Kong.
Histochem Cell Biol. 1996 Jun;105(6):443-52. doi: 10.1007/BF01457657.
Using an ultrastructural postembedding immunogold technique, we demonstrated vasoactive intestinal polypeptide (VIP)- and calcitonin gene-related peptide (CGRP)-like immunoreactivity in the Merkel cell dense-cored granules of skin and sinus hair follicles of adult cat and dog. The VIP-like substance was located in cat Merkel cells while both VIP- and CGRP-like substances were colocalised in dog Merkel cells. In cat Merkel cells, the magnitude of labelling of VIP was qualitatively higher than in dog Merkel cells. In the dog Merkel cell, CGRP appeared as the most abundant peptide. Dense-cored granules were labelled for these peptides. In addition, mast cells encountered in the dermal region of dog skin were also found to be immunolabelled by VIP antiserum. The immunoreaction was found to be confined to the secretory granules of the cells. Furthermore, all non-myelinated nerve plexuses encountered in the dermal region of the skin and the sinus hair follicles of the various mammalian species studied were immunolabelled by CGRP antiserum. The specific location was again restricted to the dense-cored granules present in these nerves. As VIP and CGRP have potent vasodilatory effects, our observations suggest that Merkel cells may play a separate or synergistic role in regulatory functions of the skin neuroendocrine cell, exerting their influence by paracrine, endocrine and neurocrine pathways, or a combination of these. Different methodologies of double labelling with different sizes of gold particles are also discussed.
采用超微结构包埋后免疫金技术,我们在成年猫和狗的皮肤及鼻窦毛囊的默克尔细胞致密核心颗粒中证实了血管活性肠肽(VIP)和降钙素基因相关肽(CGRP)样免疫反应性。猫默克尔细胞中存在VIP样物质,而狗默克尔细胞中VIP样和CGRP样物质共定位。在猫默克尔细胞中,VIP标记的强度在质量上高于狗默克尔细胞。在狗默克尔细胞中,CGRP是最丰富的肽。致密核心颗粒被这些肽标记。此外,在狗皮肤真皮区域发现的肥大细胞也被VIP抗血清免疫标记。免疫反应局限于细胞的分泌颗粒。此外,在所研究的各种哺乳动物物种的皮肤真皮区域和鼻窦毛囊中遇到的所有无髓神经丛都被CGRP抗血清免疫标记。特定位置再次局限于这些神经中存在的致密核心颗粒。由于VIP和CGRP具有强大的血管舒张作用,我们的观察结果表明,默克尔细胞可能在皮肤神经内分泌细胞的调节功能中发挥单独或协同作用,通过旁分泌、内分泌和神经分泌途径或这些途径的组合发挥其影响。还讨论了用不同大小金颗粒进行双重标记的不同方法。