Research on role in uterus and ovary of receptors LGR homologous to gonadotropin receptors

促性腺激素（LH和FSH）受体和TSH受体是七种跨膜受体的成员，这些受体具有大的N-末端细胞外结构域，含有富含亮氨酸的重复序列（LGR：富含亮氨酸的重复序列，含有G蛋白偶联受体）。在本研究中使用了同型半胱氨酸受体LGR 4、LGR 5、LGR 6、LGR 7和LGR 8。LGR 4、LGR 5和LGR 6是孤儿受体。将M1表位添加到受体蛋白的N-末端区域，因为不能确认通过受体的配体结合的细胞膜表达，允许通过DNA转染在293 T细胞的细胞表面上通过M1抗体确认。当受体被引入几种氨基酸突变，导致LH受体中cAMP产生的组成性激活时，尽管通过M1表位检测到受体细胞表面表达，但cAMP产生的基础水平并没有增加，这表明cAMP不太可能是第二信使的候选者。表达 ...更多信息 将cAMP反应性启动子驱动基因表达的荧光素酶蛋白表达载体与LGR 7表达载体共转染293 T细胞，建立了以受体系统代替免疫反应性松弛素检测活性松弛素的检测体系。用RT-PCR方法检测卵巢、颗粒细胞、子宫内膜、子宫肌瘤、输卵管和绒毛组织中松弛素、INSL 3、LGR 7和LGR 8的表达。松弛素和INSL 3在所有细胞中表达。另一方面，受体LGR 7仅在子宫内膜中表达，而LGR 8在各组织中均未检测到。用TESE收集的睾丸组织进行RT-PCR。松弛素和INSL 3的检测，无论精子的存在。LGR 7表达相似，但当标本中未发现精子时，LGR 4不能被检测到。提出了在存在LRG 8的情况下精子的存在是可预测的可能性。少

The gonadotropin (LH and FSH) receptor and TSH receptor are members of seven transmembrane receptors that have large N-terminal extracellular domain containing leucine-rich repeats (LGR : leucine-rich repeats containing G-protein coupled receptor). Homologous receptors, LGR4, LGR5, LGR6, LGR7, and LGR8 were used in the present study. LGR4, LGR5, and LGR6 are the orphan receptors. The M1 epitope was added to the N-terminal region of the receptor protein because the cell membrane expression by the ligand binding of the receptor was not able to be confirmed, allowing to be confirmed by the M1 antibody on cell surface of 293T cells by DNA transfection. When the receptors were introduced several kinds of the amino acid mutation that causes constitutive activation of cAMP production in the LH receptor, basal level of cAMP production was not increased in spite of receptor cell surface expression detected by M1 epitope, suggesting cAMP unlikely the candidate of second messenger. The expression … More of LGR4 and LGR5 was able to be confirmed by RT-PCR by using the endometrium.The luciferase protein expression vector in which cAMP responsive promoter drives gene expression was co-transfected in 293T cell with expression vector of LGR7 for the establishment of the assay system to measure bioactive relaxin by receptor sysytem instead of measuring immunoreactive relaxin. Luciferase resposiveness was too low to measure clinical samples.The expression of relaxin, INSL3, LGR7, and LGR8 was examined with RT-PCR by using the ovary, the granulosa cell, the endometrium, the myoma, the oviduct, and the villus. Relaxin and INSL3 were expressed in all cells. On the other hand, LGR8 was not ditected in each organization though receptors LGR7 were expressed only in the endometrium.The expression these gene was similarly examined by using the testis tissue. RT-PCR was done by using the testis tissue that had been gathered by TESE. Relaxin and INSL3 were detected regardless of the presence of the sperm. LGR4 could not be detected when sperm was not found in the specimen though LGR7 expression was similar. The possibility that the presence of the sperm is predictable in the presence of the appearance of LRG8 was suggested. Less