大鼠颌下腺黄体生成素及其受体的定位、核心片段的克隆及序列分析  

THE DISTRIBUTION,CLONING AND ANALYSIS OF PARTIAL SEQUENCE OF LH AND ITS RECEPTOR IN SUBMAXILLARY GLAND OF RATS

作　　者：陈蕾[1] 白宏伟[2] 孙绪德[3] 尹善德[1] 王蔼明[1] 张荣庆[4] 

机构地区：[1]北京海军总医院妇产科,北京100037 [2]北京海军总医院肝胆外科,北京100037 [3]第四军医大学唐都医院麻醉科,西安710038 [4]清华大学生命科学与技术系,北京100084

出　　处：《解剖学报》2007年第5期572-576,共5页Acta Anatomica Sinica

基　　金：中国博士后基金资助(2003)

摘　　要：目的探讨大鼠颌下腺组织中是否表达黄体生成素(LH)及其受体(LHR),为进一步研究LH对颌下腺的功能调节提供理论依据。方法采用免疫组织化学SABC和原位杂交方法进行定位研究;应用RT-PCR方法克隆获得LH及LHR基因的cDNA核心序列,并进行序列分析。结果大鼠颌下腺浆液性腺泡上皮细胞呈LH和LHR免疫反应阳性。阳性物质分布于胞浆,胞核呈阴性反应。上述细胞同样含有LH和LHR mRNA杂交信号,信号物质亦分布于胞浆内,胞核呈阴性反应。从大鼠颌下腺组织中扩增的LH及LHR基因的特异性条带经序列分析发现,扩增产物的LH基因序列与文献报道的大鼠腺垂体的完全一致;扩增产物的LHR基因序列与大鼠睾丸组织的完全一致。结论大鼠颌下腺浆液性腺泡上皮细胞既能产生LH,又能表达LHR,提示LH对颌下腺浆液性腺泡上皮细胞的作用可能是通过自分泌或旁分泌来实现的。Objective To study the localization of luteinizing hormone（LH）, its receptor （LHR） and their mRNA in submaxillary gland of rats, and to provide a theoretic evidence for further studying functional significance of the LH in submaxillary gland. Methods Immunohistochemical methods and in situ hybridization methods were used in the experiment. After isolation of the total RNA from the submaxillary gland, RT-PCR was conducted to obtain LH and LHR eDNA by using the degeneration primers. The products of PCR were analyzed by sequencing with Sanger＇ s method, respectively. Results The epithelial cells of serous acinus in submaxillary gland of rats showed LH and LHR immunoreactivity, which were located in cytoplasm with negative nuclei. LH and LHR mRNA hybridized signals were also detected in cytoplasm in the above cells. LH eDNA sequence was identical to that of the LH which has been reported in rat pituitary, and LHR eDNA sequence was identical to that of the LHR which has been reported in rat testis. Conclusion The epithelial cells of serous acinus in submaxillary gland of rats may not only produce LH but also be its target cells. LH may be involved in the functional regulation of submaxillary gland through autocrine or paracrine.

关 键 词：黄体生成素 黄体生成素受体 颌下腺 免疫组织化学 原位杂交 RT-PCR 大鼠 

分 类 号：R962[医药卫生—药理学]