Mao J N, Burnside J, Li L, Tang J, Davolos C, Cogburn L A
Department of Animal and Food Sciences, College of Agriculture and Natural Resources, Delaware Agricultural Experiment Station, University of Delaware, Newark 19717-1303, USA.
Endocrinology. 1999 Mar;140(3):1165-74. doi: 10.1210/endo.140.3.6603.
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.
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。