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性成熟鸡睾丸中与细胞内结构域相对应的独特截短催乳素受体转录本的表征。

Characterization of unique truncated prolactin receptor transcripts, corresponding to the intracellular domain, in the testis of the sexually mature chicken.

作者信息

Mao J N, Burnside J, Li L, Tang J, Davolos C, Cogburn L A

机构信息

Department of Animal and Food Sciences, College of Agriculture and Natural Resources, Delaware Agricultural Experiment Station, University of Delaware, Newark 19717-1303, USA.

出版信息

Endocrinology. 1999 Mar;140(3):1165-74. doi: 10.1210/endo.140.3.6603.

Abstract

We have examined expression of the chicken PRL receptor (cPRLR) gene in different tissues of the chicken by Northern blot analysis. Most tissues examined (ovary, testis, oviduct, kidney, and fat) possess a prominent full-length (4.6-kb) cPRLR transcript. A larger (11.7-kb) transcript is also detected in ovary, oviduct, testis, and kidney after longer exposure. A unique pattern of cPRLR expression was found in the testis of sexually mature chickens, which have an unusually high abundance of three small transcripts (1.2, 1.7, and 2 kb) in addition to the 4.6-kb transcript found in other tissues. Three domain-specific complementary DNA (cDNA) probes were constructed that correspond to the first and second ligand-binding regions in the extracellular domain and the transmembrane-intracellular domain. With these probes, Northern blot analysis of polyadenylated RNA prepared from the testes of a mature (22-week-old) chicken indicates that the highly abundant (1.2- and 1.7-kb) and less abundant (2.0-kb) cPRLR transcripts in testis hybridize only to the intracellular domain probe. Two types of truncated testis-specific cPRLR transcripts were identified using 5'-RACE (rapid amplification of cDNA ends) analysis of polyadenylated RNA from the testis of a 22-week-old chicken. The predominant truncated cDNA sequence contains the highly conserved box 1 motif [(+)box 1 cDNA] and diverges (at nucleotide 1396) from that of the cPRLR cDNA, just downstream of the transmembrane domain. The other truncated cDNA lacks the box 1 motif [(-)box 1 cDNA], which is replaced by 39 bases that could encode a hydrophobic N-terminus with a putative 5'-untranslated region of 131 bases. Young chickens predominately express the full-length cPRLR messenger RNA (4.6 kb) in the testis. At the onset of sexual maturity, there is a dramatic increase in abundance of the testis-specific (+)box 1 transcript, whereas expression of the full-length cPRLR is depressed. The presence of truncated [(+) or (-)box 1] cPRLR transcripts in the sexually mature chicken testis suggests a complex mechanism of PRL action on gonadal function.

摘要

我们通过Northern印迹分析检测了鸡催乳素受体(cPRLR)基因在鸡不同组织中的表达情况。大多数检测的组织(卵巢、睾丸、输卵管、肾脏和脂肪)都有一条明显的全长(4.6 kb)cPRLR转录本。长时间曝光后,在卵巢、输卵管、睾丸和肾脏中也检测到一条更大的(11.7 kb)转录本。在性成熟鸡的睾丸中发现了一种独特的cPRLR表达模式,除了在其他组织中发现的4.6 kb转录本外,该睾丸中还有三种异常丰富的小转录本(1.2、1.7和2 kb)。构建了三种与细胞外结构域的第一和第二配体结合区域以及跨膜-细胞内结构域相对应的结构域特异性互补DNA(cDNA)探针。使用这些探针,对一只成熟(22周龄)鸡睾丸中制备的聚腺苷酸化RNA进行Northern印迹分析表明,睾丸中高度丰富的(1.2和1.7 kb)以及不太丰富的(2.0 kb)cPRLR转录本仅与细胞内结构域探针杂交。使用5'-RACE(cDNA末端快速扩增)分析法对一只22周龄鸡睾丸中的聚腺苷酸化RNA进行分析,鉴定出两种类型的截短型睾丸特异性cPRLR转录本。主要的截短型cDNA序列包含高度保守的框1基序[(+)框1 cDNA],并在跨膜结构域下游(核苷酸1396处)与cPRLR cDNA序列不同。另一种截短型cDNA缺乏框1基序[(-)框1 cDNA],取而代之的是39个碱基,这些碱基可编码一个具有131个碱基推定5'-非翻译区的疏水N末端。幼鸡在睾丸中主要表达全长cPRLR信使RNA(4.6 kb)。在性成熟开始时,睾丸特异性(+)框1转录本的丰度急剧增加,而全长cPRLR的表达则受到抑制。性成熟鸡睾丸中截短型[(+)或(-)框1]cPRLR转录本的存在提示了催乳素对性腺功能作用的复杂机制。

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