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首页> 外文期刊>Life sciences >Distribution, cloning and sequencing of GnRH, its receptor, and effects of gastric acid secretion of GnRH analogue in gastric parietal cells of rats.
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Distribution, cloning and sequencing of GnRH, its receptor, and effects of gastric acid secretion of GnRH analogue in gastric parietal cells of rats.

机译：GnRH及其受体的分布，克隆和测序，以及GnRH类似物在大鼠胃壁细胞中胃酸分泌的作用。

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摘要

Our objective was to study the distribution of gonadotropin-releasing hormone (GnRH) and its receptor, cloning and sequencing of GnRH and its receptor gene in cultured gastric parietal cells of rats. The distribution of GnRH and its receptor mRNA were investigated through immunocytochemical ABC methods and in situ hybridization methods in cultured gastric parietal cells of rats. After isolation of the total RNA from the parietal cells, RT-PCR was conducted to obtain GnRH and its receptor cDNA. Then, the products of PCR was purified, digested by the restriction enzyme of Hind III and EcoR I, and DNA fragments of interests were cloned into pUC19 vector. The products of PCR were analyzed by sequencing with Sanger's method after identified by PCR and digestion of restriction enzyme. Gastric parietal cells showed GnRH and its receptor immunoreactivity; positive material was located in cytoplasm other than in nuclei. GnRH and its receptor mRNA hybridized signals were also detected in cytoplasm with negative nuclei. The specific amplified band of GnRH and its receptor sequences were detected through Agarose gel electrophoresis, and GnRH gene sequence is identical to that of GnRH which has been reported in rat hypothalamus and GnRH receptor sequence is identical to that of the pituitary of rat. GnRH analogue (Alarelin) could inhibit the gastric acid secretion both by direct actions on parietal cells and by inhibiting vagous function. Our data suggest that GnRH could be produced by gastric parietal cells of rats and may modulate physiological function of gastric parietal cells of rats through autocrinal and paracrinal way.
机译：我们的目的是研究促性腺激素释放激素（GnRH）及其受体的分布，在培养的大鼠胃壁细胞中GnRH及其受体基因的克隆和测序。通过免疫细胞化学ABC法和原位杂交方法研究了大鼠胃壁细胞中GnRH及其受体mRNA的分布。从壁细胞分离总RNA后，进行RT-PCR以获得GnRH及其受体cDNA。然后，纯化PCR产物，用Hind III和EcoRI限制性酶消化，并将目的DNA片段克隆到pUC19载体中。通过PCR鉴定和限制性内切酶消化后，通过Sanger法测序对PCR产物进行分析。胃壁细胞显示GnRH及其受体免疫反应性。阳性物质位于细胞质中，而不是细胞核中。在核阴性的细胞质中也检测到了GnRH及其受体mRNA杂交信号。通过琼脂糖凝胶电泳检测到GnRH及其受体序列的特异性扩增带，GnRH基因序列与大鼠下丘脑中报道的GnRH序列相同，GnRH受体序列与大鼠垂体相同。 GnRH类似物（Alarelin）可以通过直接作用于壁细胞和抑制阴道功能来抑制胃酸分泌。我们的数据表明，GnRH可能由大鼠的胃壁细胞产生，并可能通过自体和旁壁方式调节大鼠胃壁细胞的生理功能。

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