In situ demonstration of kallikrein gene family menmbers in the mouse submandibular gland.

真正的/肾组织激肽释放酶，mK 1，是绝大多数生物活性物质之一，在小鼠下颌下腺（SMG）的颗粒曲小管（GCT）中检测到。然而，mK 1的确切分布一直是有问题的，因为激肽释放酶同工酶共享高氨基酸序列同源性，并且针对任何特定激肽释放酶的抗血清经常与该家族的其他成员交叉反应。在本项目期间，研究者制备了针对mK 1和地高辛标记的mK 1受体的特异性抗血清，并获得了以下结果：1）在两种性别中，仅对GCT细胞的一个亚群进行了免疫染色，证明了mK 1在GCT中分布的不寻常的性二态镶嵌模式。2）使用去势或垂体切除动物，mK 1细胞嵌合分布的性二型性受雄激素和甲状腺激素的协同调节。3）为了检测mK 1 mRNA的存在，（ACCATCACAGATCAGTGGG）和-down（CCTCCTGAATGAGCACACCC），采用北方印迹杂交技术，肾和肝组织中只出现一条与mK 1 mRNA相对应的条带。4）虽然作者多次尝试原位杂交，但至今未检测到满意的mK 1 mRNA信号。

A true/renal tissue kallikrein, mK1, is one of the vast majority of the biologically active substances, detected in the granular convoluted tubule (GCT) of mouse submandibular gland (SMG). However, the exact distribution of mK1 has been problematical, since the kallikrein isozymes share high amino acid sequence homology, and the antisera against any particular kallikrein frequently cross-react with other members of this family. The investigator prepared the specific antiserum against mK1 and digoxigenin-labeled mK1 oligonucleoides in the term of this project, and obtained the findings, as follows.1) Only a subset of GCT cells were immunostained in both sexes, demonstrating an unusual sexually dimorphic mosaic pattern of mK1 distribution in the GCTs.2) It is clarified, using castrated or hypophysectomized animals, that the sexual dimorphism of cellular mosaic dsitribution of mK1 is under synergistic regulation of androgens and thyroid hormones.3) To detect the presence of mK1 mRNA, the primer-up(ACCATCACAGATCAGTGGG) and -down(CCTCCTGAATGAGCACACCC) were designed, and Northern blot hybridization of mouse SMG, sublingual gland, kidney and liver revealed a sigle band corresponding to the mK1 mRNA.4) Though the author were trying in situ hybridization several times, any satisfactory signals for mK1 mRNA have not been detected till now.