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围产期大鼠胰腺中促甲状腺激素释放激素前体肽的鉴定、表征及定位

Identification, characterization and localization of thyrotropin-releasing hormone precursor peptides in perinatal rat pancreas.

作者信息

Wu P, Jackson I M

机构信息

Division of Endocrinology, Brown University, Rhode Island Hospital, Providence 02903.

出版信息

Regul Pept. 1988 Sep;22(4):347-60. doi: 10.1016/0167-0115(88)90111-5.

Abstract

The sequence of rat hypothalamic prepro TRH, deduced from its complementary DNA, contains five TRH progenitor sequences and six cryptic sequences separated by paired basic amino acid residues. We have utilised antisera against two synthetic peptides corresponding to sequences within proTRH, [Tyr53] preproTRH (53-74), part of the amino terminal leader sequence of proTRH and [Cys 74,83] preproTRH-(75-82), representing a TRH progenitor sequence flanked by cysteine residues (pCC10) in radioimmunoassays (RIA) to identify and chromatographically characterize proTRH derived peptides in extracts of rat perinatal pancreas and to localize these peptides immunohistochemically. Two forms of immunoreactive pYT22 (ipYT22) were observed, similar in size to ipYT22 seen in extracts of adult rat brain. By RIA immunoreactive pCC10 was detectable in neonatal but not fetal pancreas. However, immunohistochemical double staining of both fetal and neonatal rat pancreas colocalized both ipYT22 and ipCC10 with immunoreactive insulin in the B-cell of the developing Islets of Langerhans. These findings indicate that the B-cell of the perinatal pancreas synthesizes TRH from a prohormone encoded by a mRNA similar to that present in adult rat hypothalamus.

摘要

从大鼠下丘脑前促甲状腺激素释放激素(TRH)的互补DNA推导得出的序列,包含五个TRH前体序列和六个隐蔽序列,这些序列由成对的碱性氨基酸残基分隔。我们利用针对两种与促TRH内序列相对应的合成肽的抗血清,即[酪氨酸53]前促TRH(53 - 74),它是促TRH氨基末端前导序列的一部分,以及[半胱氨酸74,83]前促TRH -(75 - 82),它代表一个两侧有半胱氨酸残基的TRH前体序列(pCC10),在放射免疫分析(RIA)中鉴定并通过色谱法表征大鼠围产期胰腺提取物中源自促TRH的肽,并通过免疫组织化学对这些肽进行定位。观察到两种形式的免疫反应性pYT22(ipYT22),其大小与成年大鼠脑提取物中所见的ipYT22相似。通过RIA,在新生胰腺中可检测到免疫反应性pCC10,但在胎儿胰腺中未检测到。然而,对胎儿和新生大鼠胰腺进行的免疫组织化学双重染色显示,在发育中的胰岛B细胞中,ipYT22和ipCC10均与免疫反应性胰岛素共定位。这些发现表明,围产期胰腺的B细胞从一种由与成年大鼠下丘脑存在的mRNA相似的前体激素编码的前体合成TRH。

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