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鸡胚羽毛发育过程中皮肤糖胺聚糖的组织化学定位。

Histochemical localization of skin glycosaminoglycans during feather development in the chick embryo.

作者信息

Jahoda Colin A B, Mauger Annick, Sengel Philippe

机构信息

Unité Associée au CNRS no 682, Morphogenèse expérimentale, Département de Biologie, Université de Grenoble 1, B.P. 68, 38402, Saint Martin d'Hères, France.

出版信息

Rouxs Arch Dev Biol. 1987 Jul;196(5):303-315. doi: 10.1007/BF00395954.

Abstract

The distribution of glycosaminoglycans (GAGs) was studied in embryonic chick skin, using alcian blue staining with critical electrolyte concentration and glycanase treatment, immunofluorescence and transmission electron microscopy. Light microscopy revealed an uneven distribution of sulphated and non-sulphated GAGs at all stages of feather development. Along the dermal-epidermal junction and throughout the depth of the dermis, staining was stronger inside the feathers than in the interplumar skin. With increasing MgCl concentration, the decrease in stain intensity along the dermal-epidermal junction was stronger in interplumar skin than inside feather structures, indicating that sulphated GAGs are more abundant within feathers than in interplumar skin. The same differential sensitivity to electrolyte concentration was noted in the dermis, except at the feather placode stage, when labelling inside the dermal condensation was virtually wiped out at 0.6 M MgCl and higher concentrations, whereas it persisted in the surrounding dermis up to 0.8 M MgCl, indicating that the dermal condensation contains a larger amount of hyaluronate than non-feather-forming dermis. Enzyme treatment of sections with Streptomyces hyaluronidase as compared with those treated with chondroitinase ABC corroborated these findings. Immunofluorescent detection of heparan sulphate proteoglycan revealed the presence of the antigen along the dermal-epidermal junction at all stages of feather development, with peaks of brightness in discrete spots of feather structures. Electron microscopy revealed the presence of ruthenium red and tannic acid positive material in the dermal-epidermal junctional zone and inside the dermis. The density of marked granules was somewhat higher in intraplumar than in interplumar regions. These observations demonstrate that certain sulphated and non-sulphated GAGs are distributed in a microheterogeneous manner, which appears to be related to the morphogenetic events of feather development. They are discussed in view of the possible role these components might play in dermal-epidermal interactions. They strengthen the notion, already gained from previous studies on the localization of interstitial collagens and fibronectin, that extracellular matrix components play an important structural and informative role in organogenesis.

摘要

利用临界电解质浓度的阿尔新蓝染色、聚糖酶处理、免疫荧光和透射电子显微镜技术,研究了胚胎期鸡皮肤中糖胺聚糖(GAGs)的分布情况。光学显微镜显示,在羽毛发育的各个阶段,硫酸化和非硫酸化GAGs的分布不均匀。沿着真皮-表皮交界处以及整个真皮深度,羽毛内部的染色比羽间皮肤更强。随着MgCl浓度的增加,羽间皮肤中沿真皮-表皮交界处染色强度的降低比羽毛结构内部更强,这表明羽毛内部的硫酸化GAGs比羽间皮肤中更丰富。在真皮中也观察到了对电解质浓度的相同差异敏感性,但在羽基板阶段除外,此时在0.6M MgCl及更高浓度下,真皮凝聚物内部的标记几乎消失,而在周围真皮中,直至0.8M MgCl时标记仍持续存在,这表明真皮凝聚物中含有的透明质酸比非羽毛形成的真皮更多。与用软骨素酶ABC处理的切片相比,用链霉菌透明质酸酶处理切片的酶处理结果证实了这些发现。硫酸乙酰肝素蛋白聚糖的免疫荧光检测显示,在羽毛发育的各个阶段,抗原均沿真皮-表皮交界处存在,在羽毛结构的离散斑点处亮度峰值出现。电子显微镜显示,在真皮-表皮交界区和真皮内部存在钌红和单宁酸阳性物质。羽内区域标记颗粒的密度略高于羽间区域。这些观察结果表明,某些硫酸化和非硫酸化GAGs以微异质性方式分布,这似乎与羽毛发育的形态发生事件有关。鉴于这些成分可能在真皮-表皮相互作用中发挥的作用,对其进行了讨论。它们强化了从先前关于间质胶原和纤连蛋白定位的研究中已经获得的观念,即细胞外基质成分在器官发生中发挥重要的结构和信息作用。

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