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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）一种新的大鼠舌下腺表达蛋白：我们克隆了一种新的大鼠舌下腺表达基因，并将其命名为SLAMP（sublingual acinar membrane protein）。SLAMP与动物L型凝集素家族成员ERGIC-53-like蛋白的同源性为63%。SLAMP mRNA和蛋白主要表达于舌下腺粘液腺泡细胞。超微结构上，SLAMP定位于ER-高尔基体中间室（ERGIC），表明SLAMP在糖蛋白的早期分泌途径中起作用。2）一种时间更短、成本更低的大鼠多克隆抗体生产新方法：生产抗体通常需要较长的时间。我们建立了一种在不到一个月的时间内制备多克隆抗体的新方法，该方法基于制备重组寡肽作为抗原，然后免疫大鼠。用这种方法，我们生产了抗ERGIC-53的抗血清，并证实了它在ERGIC中的定位。3）一种新的小鼠下颌下腺雄激素抑制蛋白：我们鉴定了一个来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（SMARP）。在小鼠中，SMARP mRNA仅在SMG和眶外泪腺（LG）中表达。SMARP mRNA水平在SMG中雌性是雄性的36倍，而在LG中雄性是雌性的28倍，并且在给雌性或去势雄性睾酮后SMG中SMARP mRNA水平降低，而LG中SMARP mRNA水平升高。SMARP mRNA和蛋白主要定位于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中受雄激素负调控，在LG中受雄激素正调控。