Majzoub J A, Rich A, van Boom J, Habener J F
J Biol Chem. 1983 Dec 10;258(23):14061-4.
Vasopressin and oxytocin are nonapeptide hormones that regulate water metabolism and lactation, respectively. To study the regulation of the vasopressin and oxytocin genes at the mRNA level, we constructed a series of synthetic oligonucleotides, from 8 to 15 bases in length, for use in filter-blot hybridization assays (Northern blots) of hypothalamic mRNA levels and for primed synthesis of cDNAs from which we determined the nucleotide sequences of the 5' regions of the vasopressin and oxytocin mRNAs. A 20-fold increase occurred in the amounts of the two mRNAs present in the hypothalami of rats drinking 2% saline for three weeks. In addition, the sequence analyses of the cDNAs provided the complete amino acid sequences of the NH2-terminal signal peptides of the rat vasopressin and oxytocin precursors. Thus, synthetic oligonucleotides consisting of as few as eight nucleotides can be used to prime reverse transcription of specific cDNAs from hypothalamic RNA, and pentadecanucleotide hybridization probes readily detect changes in levels of vasopressin and oxytocin mRNAs in response to osmotic stress.
血管加压素和催产素是分别调节水代谢和泌乳的九肽激素。为了在mRNA水平研究血管加压素和催产素基因的调控，我们构建了一系列长度为8至15个碱基的合成寡核苷酸，用于下丘脑mRNA水平的滤膜杂交分析（Northern印迹）以及用于从其确定血管加压素和催产素mRNA 5'区域核苷酸序列的cDNA的引物合成。饮用2%盐水三周的大鼠下丘脑内这两种mRNA的量增加了20倍。此外，cDNA的序列分析提供了大鼠血管加压素和催产素前体NH2末端信号肽的完整氨基酸序列。因此，由少至八个核苷酸组成的合成寡核苷酸可用于引发下丘脑RNA中特定cDNA的逆转录，并且十五聚体杂交探针能够轻松检测血管加压素和催产素mRNA水平对渗透压应激的变化。
