Distribution of the α1 to α6 chains of type IV collagen in bovine follicles
Distribution of the α1 to α6 chains of type IV collagen in bovine follicles
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DOI：
10.1095/biolreprod59.6.1334
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发表时间：
1998-12-01
影响因子：
3.6
通讯作者：
Rodgers, RJ
中科院分区：
文献类型：
--
作者：
Rodgers, HF;Irvine, CM;Rodgers, RJ
在卵泡发育过程中，上皮样颗粒细胞的增殖和分化状态发生变化，液体在卵泡基底层的运动使腔形成。IV型胶原是许多基底层的重要组成部分，每个分子由三条α链组成；然而，已经确定了六种不同的IV型胶原链，尚不清楚这些链中哪一种存在于卵泡基底层，以及在卵泡发育过程中基底层的IV型胶原组成是否发生了变化。因此，我们使用针对非保守非胶原结构域的抗体对牛卵巢中的6条链中的每一条进行了免疫定位。此外，用胶原酶消化解剖的卵泡以释放NC结构域，然后用标准的Western免疫印迹方法检测NC1结构域。几乎所有原始滤泡和前腔滤泡的滤泡基底层所有IV型胶原α链都呈阳性，IV型胶原和因子viii相关抗原的共定位允许区分滤泡基底层和内皮基底层。IV型胶原α 1、α 2、α 3、α 4和α 5链仅在部分腔泡（22 / 17、21 / 20、18 / 15、28 / 14、23 / 12）中存在，且染色强度低于腔前卵泡，α 1和α 2链的染色在与识别的基底膜无关的区域弥散分布在整个卵泡中。免疫染色的特异性α1和α2链IV型胶原蛋白被西方免疫印迹证实,作为基板的检测到大约一半的窦的卵泡检查,IV型胶原蛋白α4也与3 beta-hydroxysteroid dehydrogenase-immunopositive细胞外壁interna, IV型胶原蛋白α6中检测出的只有一个基板检查16窦的毛囊。因此，在卵泡发育过程中，滤泡基底膜的组成发生变化，所有IV型胶原链的免疫染色水平降低，随着卵泡大小的增加，IV型胶原α 6的免疫反应性丧失。此外，随着卵泡膜的发育，细胞外基质中出现了α 1和α 2的免疫反应性。
During follicular development the proliferative and differentiated state of the epithelioid granulosa cells changes, and the movement of fluid across the follicular basal lamina enables the formation of an antrum. Type IV collagen is an important component of many basal laminae, Each molecule is composed of three alpha chains; however, six different type IV collagen chains have been identified, It is not known which of these chains are present in the follicular basal lamina and whether the type IV collagen composition of the basal lamina changes during follicular development. Therefore, we immunolocalized each of the six chains in bovine ovaries using antibodies directed to the nonconserved non-collagenous (NC) domains. Additionally, dissected follicles were digested with collagenase to release the NC domains, and the NC1 domains were then detected by standard Western immunoblot, methods. The follicular basal lamina of almost all primordial and preantral follicles was positive for all type IV collagen alpha chains, Colocalization of type IV collagen and factor VIII-related antigen allowed for discrimination between the follicular and endothelial basal laminae. Type IV collagen alpha 1, alpha 2, alpha 3, alpha 4, and alpha 5 chains were present within the follicular basal lamina of only a proportion of antral follicles (17 of 22, 20 of 21, 15 of 18, 14 of 28, and 12 of 23, respectively), and staining was less intense than in the preantral follicles, Staining for the alpha 1 and alpha 2 chains was diffusely distributed throughout the theca in regions not associated with recognized basal laminae. The specificity of this immunostaining for alpha 1 and alpha 2 chains of type IV collagen was confirmed by Western immunoblots, As well as being detected in the basal lamina of approximately half of the antral follicles examined, type IV collagen alpha 4 also colocalized with 3 beta-hydroxysteroid dehydrogenase-immunopositive cells in the theca interna, Type IV collagen alpha 6 was detected in the basal lamina of only one of the 16 antral follicles examined. Thus, the follicular basal lamina changes in composition during follicular development, with immunostaining levels being reduced for all type IV collagen chains and immunoreactivity for IV collagen alpha 6 being lost as follicle size increases. Additionally, immunoreactivity for alpha 1 and alpha 2 appears in the extracellular matrix of the theca as it develops.