Culturing primary rat inner medullary collecting duct cells

Item Type:	Article
Title:	Culturing primary rat inner medullary collecting duct cells
Creators Name:	Faust, D. and Geelhaar, A. and Eisermann, B. and Eichhorst, J. and Wiesner, B. and Rosenthal, W. and Klussmann, E.
Abstract:	Arginine-vasopressin (AVP) facilitates water reabsorption by renal collecting duct principal cells and thereby fine-tunes body water homeostasis. AVP binds to vasopressin V2 receptors (V2R) on the surface of the cells and thereby induces synthesis of cAMP. This stimulates cellular signaling processes leading to changes in the phosphorylation of the water channel aquaporin-2 (AQP2). Protein kinase A phoshorylates AQP2 and thereby triggers the translocation of AQP2 from intracellular vesicles into the plasma membrane facilitating water reabsorption from primary urine. Aberrations of AVP release from the pituitary or AVP-activated signaling in principal cells can cause central or nephrogenic diabetes insipidus, respectively; an elevated blood plasma AVP level is associated with cardiovascular diseases such as chronic heart failure and the syndrome of inappropriate antidiuretic hormone secretion. Here, we present a protocol for cultivation of primary rat inner medullary collecting duct (IMCD) cells, which express V2R and AQP2 endogenously. The cells are suitable for elucidating molecular mechanisms underlying the control of AQP2 and thus to discover novel drug targets for the treatment of diseases associated with dysregulation of AVP-mediated water reabsorption. IMCD cells are obtained from rat renal inner medullae and are used for experiments six to eight days after seeding. IMCD cells can be cultured in regular cell culture dishes, flasks and micro-titer plates of different formats, the procedure only requires a few hours, and is appropriate for standard cell culture laboratories.
Keywords:	Bioengineering, Genetics, Molecular Biology, Biochemistry, Biomedical Engineering, Medicine, Pharmacology, Intercellular Signaling Peptides and Proteins, Exocytosis, Signal Transduction, Second Messenger Systems, Calcium Signaling, Cardiovascular Diseases, Hormones, Hormone Substitutes, Hormone Antagonists, Life Sciences (General), Water Reabsorption, Kidney, Principal Cells, Vasopressin, Cyclic AMP, Aquaporin, Animal Model, Cell Culture, Animals, Rats
Source:	Journal of Visualized Experiments
ISSN:	1940-087X
Publisher:	JoVE
Number:	76
Page Range:	e50366
Date:	21 June 2013
Official Publication:	https://doi.org/10.3791/50366
PubMed:	View item in PubMed

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