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家鸡垂体和卵巢中β-聚糖的信使核糖核酸及蛋白质表达分析

Messenger RNA and protein expression analysis of betaglycan in the pituitary and ovary of the domestic hen.

作者信息

Sweeney Sheila A, Johnson Patricia A

机构信息

Department of Animal Science, Cornell University, Ithaca, New York 14853, USA.

出版信息

Biol Reprod. 2005 Jan;72(1):172-8. doi: 10.1095/biolreprod.104.030171. Epub 2004 Sep 22.

Abstract

Betaglycan was originally characterized as the type III receptor for TGFbeta, yet recent research has indicated that betaglycan can serve as an accessory receptor for inhibin. To understand better the action of inhibin in avian follicular development, we have investigated the expression of betaglycan in the pituitary gland and ovary of the hen. In experiments 1 and 2, betaglycan mRNA was detected at 6 kilobases (kb) by Northern blot analysis (n = 5) in chicken pituitary, granulosa, and theca layers and whole ovary. Expression of betaglycan was greatest in the pituitary gland in experiment 1 and greater in the granulosa layer of small yellow follicles (SYF) compared with the granulosa layer of larger follicles. In experiment 2, betaglycan mRNA was more abundantly expressed in the theca layer compared with the granulosa layer for all follicle sizes, although there was no significant difference in betaglycan expression in the theca layer among follicle sizes. In experiment 3, immunohistochemical analysis revealed betaglycan protein in the anterior pituitary as well as in the ovary (n = 4) and SYF (n = 4). Colocalization studies revealed a high abundance of cells within the anterior pituitary expressing both betaglycan and FSH (n = 4). Betaglycan protein was found in the granulosa layer; however, markedly enhanced staining was observed in the theca layer of ovarian follicles. Our results provide evidence for expression of betaglycan mRNA and protein colocalization with FSH in the anterior pituitary, consistent with known inhibin effects. Ovarian localization of betaglycan, particularly in the theca layer, suggests a paracrine role for inhibin in the hen.

摘要

β聚糖最初被鉴定为转化生长因子β(TGFβ)的Ⅲ型受体,但最近的研究表明,β聚糖可作为抑制素的辅助受体。为了更好地理解抑制素在禽类卵泡发育中的作用,我们研究了母鸡垂体和卵巢中β聚糖的表达。在实验1和实验2中,通过Northern印迹分析(n = 5)在鸡垂体、颗粒层、卵泡膜层和整个卵巢中检测到6千碱基(kb)的β聚糖mRNA。实验1中β聚糖在垂体中的表达最高,与较大卵泡的颗粒层相比,小黄卵泡(SYF)颗粒层中的表达更高。在实验2中,对于所有卵泡大小,卵泡膜层中β聚糖mRNA的表达均比颗粒层更丰富,尽管卵泡膜层中β聚糖的表达在不同卵泡大小之间没有显著差异。在实验3中,免疫组织化学分析显示垂体前叶以及卵巢(n = 4)和SYF(n = 4)中存在β聚糖蛋白。共定位研究显示垂体前叶中有大量细胞同时表达β聚糖和促卵泡激素(FSH)(n = 4)。在颗粒层中发现了β聚糖蛋白;然而,在卵巢卵泡的卵泡膜层中观察到染色明显增强。我们的结果为垂体前叶中β聚糖mRNA的表达以及与FSH的蛋白共定位提供了证据,这与已知的抑制素作用一致。β聚糖在卵巢中的定位,特别是在卵泡膜层中的定位,表明抑制素在母鸡中具有旁分泌作用。

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