J Vet Sci.  2010 Mar;11(1):15-19. 10.4142/jvs.2010.11.1.15.

Transcriptional and translational expression of calbindin-D9k in the duodenum, kidney and uterus of a female canine model

• Sim JY
• Jung EM
• Yoo YM
• Choi KC
• Jeung EB

• KMID: 1106152
• DOI: http://doi.org/10.4142/jvs.2010.11.1.15

Abstract

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 (Ca2+). 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 transcription-polymerase 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.

Keyword

calbindin-D9k; canine model; duodenum; kidney; uterus

MeSH Terms

Animals
Blotting, Western/veterinary
Calcium-Binding Protein, Vitamin D-Dependent/*biosynthesis/genetics
Dogs/*physiology
Duodenum/*physiology
Female
Immunohistochemistry/veterinary
Kidney/*physiology
RNA, Messenger/biosynthesis/genetics
Reverse Transcriptase Polymerase Chain Reaction/veterinary
Transcription, Genetic
Uterus/*physiology

Figure

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  Fig. 1 Expression of CaBP-9k mRNA (A) and relative percentage of CaBP-9k/glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA (B) in duodenum, kidney and uterus.

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  Fig. 2 Expression of CaBP-9k protein in duodenum, kidney and uterus by Western blot analysis.

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  Fig. 3 Localization of CaBP-9k protein. The brush border (arrows) of duodenum showed positive immunoreactivities (A), while uterus (B) and kidney (C) showed negative immunoreactivities. Scale bar = 100 µm.

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