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

小鼠颌下腺中激肽释放酶基因家族成员的原位演示。

基本信息

批准号：
11671820
负责人：
KURABUCHI Shingo
金额：
$ 2.3万
依托单位：
The Nippon Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671820/
关键词：
kallikrein gene family true renal kallikrein mK1 RT-PCR in situ hybridyzation immunocytochemistry submandibular gland mouse

项目摘要

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.

真实/肾脏组织Kallikrein MK1是在小鼠下颌腺（SMG）的颗粒状杂质小管（GCT）中检测到的绝大多数生物活性物质之一。但是，由于Kallikrein同工酶具有高氨基酸序列同源性，并且针对任何特定的Kallikrein经常与该家族的其他成员进行反应，因此MK1的确切分布是有问题的。研究者在该项目的一项中准备了针对MK1和非氧基素标记的MK1寡核苷的特异性抗血清，并获得了以下发现，如下所示。1）仅在两性中都对GCT细胞进行了免疫染色，证明了异常性的性型性型摩擦模式。 2）使用cast割或下生理切除的动物来澄清MK1的细胞镶嵌性二依屈的性二态性在雄激素和甲状腺激素的协同调节下。设计了Primer -Up（AccAcagatCagtggg）和-down（Cctcctgaatgagcacacccc），并设计了鼠标SMG，舌下腺体，肾脏和肝脏的北印迹杂交，揭示了与Mk1 mRNA相对应的SIGLE。时间，直到现在尚未检测到MK1 mRNA的任何令人满意的信号。