GENE EXPRESSION DURING THE DUCT CELL DIFFERENTIATION IN SALIVARY GLANDS

唾液腺导管细胞分化过程中的基因表达

基本信息

批准号：
17590154
负责人：
ISEKI Shoichi
金额：
$ 2.3万
依托单位：
KANAZAWA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

1)A novel protein expressed in rat sublingual gland : We cloned a novel rat gene expressed primarily in the sublingual gland and named the predicted 503 amino-acid protein SLAMP (sublingual acinar membrane protein). SLAMP had 63% homology with human ERGIC-53-like protein, a member of the family of animal L-type lectins. SLAMP mRNA and protein were expressed predominantly in the mucous acinar cells of sublingual gland. Ultrastructurally, SLAMP was localized to the ER-Golgi intermediate compartment (ERGIC), suggesting that SLAMP functions in the early secretory pathway of glycoproteins. 2)A new method of producing rat polyclonal antibodies with shorter time and lower cost: Producing antibodies usually takes a long time. We developed a new method of producing polyclonal antibodies in less than a month, based on preparation of the recombinant oligopeptides as antigen followed by immunization of rats. Using this method, we produced antisera against ERGIC-53 and confirmed its localization in the ERGIC. 3)A novel submandibular androgen-repressed protein of the mouse: We characterized a cDNA clone derived from the female mouse submandibular gland (SMG). The protein product of this clone had a 165-amino acid protein with a putative signal sequence and was named submandibular androgen-repressed protein (SMARP). In the mouse SMARP mRNA was expressed only in the SMG and exorbital lacrimal gland (LG). The level of SMARP mRNA was 36 times higher in female than male SMG, whereas it was 28 times higher in male than female LG. Furthermore, it was reduced in SMG but increased in LG after administration of testosterone to females or castrated males. SMARP mRNA and protein were predominantly localized in acinar cells in both SMG and lacrimal gland. These results suggested that SMARP is a secretory protein whose expression is regulated by androgens negatively in the SMG and positively in the LG.

1）大鼠舌下腺表达的新蛋白：我们克隆了一种主要在舌下腺表达的新型大鼠基因，并将预测的503个氨基酸蛋白命名为SLAMP（舌下腺泡膜蛋白）。 SLAMP 与人类 ERGIC-53 样蛋白（动物 L 型凝集素家族的成员）有 63% 同源性。 SLAMP mRNA和蛋白主要在舌下腺粘液腺泡细胞中表达。从超微结构上看，SLAMP 定位于 ER-高尔基中间室 (ERGIC)，表明 SLAMP 在糖蛋白的早期分泌途径中发挥作用。 2）一种生产大鼠多克隆抗体的新方法，时间更短，成本更低：生产抗体通常需要很长时间。我们开发了一种在不到一个月的时间内生产多克隆抗体的新方法，其基础是制备重组寡肽作为抗原，然后对大鼠进行免疫。使用这种方法，我们生产了针对 ERGIC-53 的抗血清，并确认了其在 ERGIC 中的定位。 3)一种新型小鼠下颌下雄激素抑制蛋白：我们鉴定了源自雌性小鼠下颌下腺(SMG)的cDNA克隆。该克隆的蛋白质产物具有 165 个氨基酸的蛋白质，具有推定的信号序列，被命名为下颌下雄激素抑制蛋白质（SMARP）。在小鼠中，SMARP mRNA 仅在 SMG 和眶外泪腺 (LG) 中表达。女性SMG中的SMARP mRNA水平比男性SMG高36倍，而男性LG中的SMARP mRNA水平比女性LG高28倍。此外，在给女性或阉割的男性注射睾酮后，SMG 减少，但 LG 增加。 SMARP mRNA 和蛋白主要位于 SMG 和泪腺的腺泡细胞中。这些结果表明 SMARP 是一种分泌蛋白，其表达在 SMG 中受雄激素负调节，在 LG 中受雄激素正调节。