Sim Ji-Young, Jung Eui-Man, Yoo Yeong-Min, Choi Kyung-Chul, Jeung Eui-Bae
Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea.
J Vet Sci. 2010 Mar;11(1):15-9. doi: 10.4142/jvs.2010.11.1.15.
Calbindin-D9k (CaBP-9k) is a cytosolic calcium-binding protein expressed in tissues in the intestine, uterus, placenta, kidney, pituitary gland and bone. Its exact function is unknown, but it is considered to regulate intracytoplasmic concentration and transport of free ions (Ca(2+)). CaBP-9k protein is involved in intestinal calcium absorption in the intestine and in the regulation of myometrial activity by intracellular calcium in the uterus. Renal CaBP-9k protein is expressed at the site of calcium re-absorption in the kidney and expressed in distal convoluted tubules, where it is thought to facilitate calcium re-absorption. Expression of the CaBP-9k gene has been explored in most mammalians except in a canine model. Presently, we elucidated the expression of CaBP-9k mRNA and protein in the duodenum, kidney and uterus in a canine model involving two adult (2.5-year-old) female beagles. To collect tissues, the dogs were euthanized and then the abdominal cavity was exposed by midline incision. The proximal duodenum, cortex of kidney and uterine horn were collected. Expression of CaBP-9k mRNA was confirmed by reverse transcriptionpolymerase chain reaction (RT-PCR) and real-time PCR. CaBP-9k protein expression and localization were ascertained by Western blot analysis and immunohistochemistry, respectively. CaBP-9k mRNA was detected in the duodenum, but not in the kidney and uterus. Its protein was expressed only in the enterocytes of the duodenum. Taken together, the results indicate that CaBP-9k mRNA and protein are highly expressed in the enterocytes of the duodenum of a canine model, consistent with findings in other mammalian species.
钙结合蛋白-D9k(CaBP-9k)是一种胞质钙结合蛋白,在肠道、子宫、胎盘、肾脏、垂体和骨骼等组织中表达。其确切功能尚不清楚,但被认为可调节游离离子(Ca(2+))的胞质内浓度和转运。CaBP-9k蛋白参与肠道中的钙吸收以及子宫中细胞内钙对子宫肌层活性的调节。肾脏中的CaBP-9k蛋白在肾脏钙重吸收部位表达,并在远曲小管中表达,据认为它有助于钙的重吸收。除了犬类模型外,在大多数哺乳动物中都对CaBP-9k基因的表达进行了研究。目前,我们阐明了CaBP-9k mRNA和蛋白在涉及两只成年(2.5岁)雌性比格犬的犬类模型的十二指肠、肾脏和子宫中的表达情况。为了收集组织,对犬实施安乐死,然后通过中线切口暴露腹腔。收集近端十二指肠、肾皮质和子宫角。通过逆转录聚合酶链反应(RT-PCR)和实时PCR确认CaBP-9k mRNA的表达。分别通过蛋白质印迹分析和免疫组织化学确定CaBP-9k蛋白的表达和定位。在十二指肠中检测到CaBP-9k mRNA,但在肾脏和子宫中未检测到。其蛋白仅在十二指肠的肠细胞中表达。综上所述,结果表明CaBP-9k mRNA和蛋白在犬类模型的十二指肠肠细胞中高表达,这与其他哺乳动物物种的研究结果一致。