Mesotocin gene expression in the diencephalon of domestic fowl: Cloning and sequencing of the MT cDNA and distribution of MT gene expressing neurons in the chicken hypothalamus
Mesotocin gene expression in the diencephalon of domestic fowl: Cloning and sequencing of the MT cDNA and distribution of MT gene expressing neurons in the chicken hypothalamus
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DOI：
10.1046/j.1365-2826.1997.00643.x
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发表时间：
1997-10-01
影响因子：
3.2
通讯作者：
Grossmann, R
中科院分区：
文献类型：
--
作者：
Barth, SW;Bathgate, RAD;Grossmann, R
本研究对鸡下丘脑中催产素（mesotocin，MT）基因的结构和表达进行了研究。采用锚定和巢式RT-PCR结合环形RACE-PCR的方法，获得了鸡MT cDNA全长序列，该cDNA及其衍生的氨基酸序列符合催产素样基因家族的结构。然而，与大多数哺乳动物物种不同，MT和AVT cDNA序列之间似乎没有频繁的基因转换。Southern杂交检测到1.2kb的特异性杂交信号，表明鸡基因组中只有一个基因拷贝。北方杂交检测到0.6kb的特异性杂交信号。原位杂交组织化学检测到MT-mRNA主要定位于小细胞，室旁核的大细胞和室周亚群，并且当与在相同区域中含有精氨酸-血管催产素（AVT）-mRNA的神经元的分布相比时，在外侧亚群中表达MT基因的神经元少得多。只有少数和分散的神经元表达MT基因被发现在腹侧和外部亚组的视上核，其中许多神经元含有AVT转录本，在连续的部分证明。终纹床核和延髓背侧核均未发现MT-mRNA的特异性标记。用免疫细胞化学方法检测AVT和原位杂交方法检测MT-mRNA的表达。在室旁核中发现一些神经元同时含有AVT和MT基因产物，而在视上核中则没有。
This study focuses on the structure and expression of the mesotocin (MT) gene in the chicken hypothalamus. Using an anchored and nested RT-PCR strategy, combined with circular RACE-PCR, the full length sequence of the chicken MT cDNA was obtained, The cDNA and derived amino acid sequences conformed to the structure of the oxytocin-like gene family. However, unlike most mammalian species, there does not appear to be frequent gene conversion between the MT and AVT cDNA sequences. A single specific hybridization signal of 1.2 kb was detected by Southern analysis of chicken genomic DNA, indicating only a single gene copy in the chicken genome, Northern analysis of hypothalamic RNA revealed a single band at approximately 0.6 kb, Using the same probe for in situ hybridization histochemistry, MT-mRNA was demonstrated to be predominantly localized in the parvocellular, magnocellular and periventricular subgroups of the paraventricular nucleus and, when compared to the distribution of neurons containing arginine-vasotocin (AVT)-mRNA in the same region, with far fewer neurons expressing the MT gene in the lateral subgroups. Only few and scattered neurons expressing the MT gene were found in the ventral and external subgroups of the supraoptic nucleus in which many neurons contain AVT transcripts, as demonstrated in consecutive sections. In all nuclei investigated, the intensity of AVT and MT hybridization signals per cell was approximately equal, No specific labelling for MT-mRNA was found in the bed nucleus of the stria terminalis, nor the nucleus accumbens, Using immunocytochemical detection of AVT and in situ hybridization for neurons expressing MT-mRNA, some neurons were found to contain both AVT and MT gene products in the paraventricular nucleus but not in the supraoptic nucleus.