Barth S W, Bathgate R A, Mess A, Parry L J, Ivell R, Grossmann R
Institute for Small Animal Research (FAL), Research Unit of Physiology, Celle, Germany.
J Neuroendocrinol. 1997 Oct;9(10):777-87. doi: 10.1046/j.1365-2826.1997.00643.x.
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.
本研究聚焦于鸡下丘脑内中催产素(MT)基因的结构与表达。采用锚定巢式逆转录聚合酶链反应(RT-PCR)策略,并结合环化快速扩增cDNA末端(RACE-PCR),获得了鸡MT cDNA的全长序列。该cDNA及其推导的氨基酸序列符合催产素样基因家族的结构。然而,与大多数哺乳动物不同,MT和精氨酸加压催产素(AVT)cDNA序列之间似乎不存在频繁的基因转换。通过对鸡基因组DNA进行Southern分析,检测到一个1.2 kb的单一特异性杂交信号,表明鸡基因组中只有一个基因拷贝。对下丘脑RNA进行Northern分析,在约0.6 kb处显示出一条单一的条带。使用相同的探针进行原位杂交组织化学分析,结果表明MT-mRNA主要定位于室旁核的小细胞、大细胞和室周亚群中,并且与同一区域中含有精氨酸加压催产素(AVT)-mRNA的神经元分布相比,外侧亚群中表达MT基因的神经元要少得多。在视上核的腹侧和外侧亚群中,仅发现少数散在的表达MT基因的神经元,而在连续切片中显示该区域许多神经元含有AVT转录本。在所研究的所有核团中,每个细胞的AVT和MT杂交信号强度大致相等。在终纹床核和伏隔核中未发现MT-mRNA的特异性标记。通过对AVT进行免疫细胞化学检测以及对表达MT-mRNA的神经元进行原位杂交,发现在室旁核中一些神经元同时含有AVT和MT基因产物,但在视上核中则没有。