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Bestrophin‐3‐associated Ca 2+ ‐activated Cl − channel in vascular smooth muscle and endothelial cells
Author(s) -
Matchkov Vladimir V.,
Eriksen Tina Fransgaard,
Larsen Per,
Pedersen Finn Skou,
Nilsson Holger,
Aalkjaer Christian
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.1018.8
Subject(s) - vascular smooth muscle , immunohistochemistry , microbiology and biotechnology , chemistry , downregulation and upregulation , messenger rna , medicine , biology , endocrinology , smooth muscle , biochemistry , gene
We have recently shown (1) that bestrophin‐3, a member of the Best family, is essential for the cGMP‐dependent Ca 2+ ‐activated Cl − current (I (Ca, cGMP)Cl ) (2) which has been shown in different types of smooth muscles (3). The I (Ca, cGMP)Cl has similar characteristics as the currents produced by heterologous expression of bestrophins and distributs similarly to Best‐3 expression. Furthermore, downregulation of Best‐3 with siRNA was associated with a significant reduction of the I (Ca, cGMP)Cl . To study the physiological significance of Best‐3 immunohistochemical (IHC) and siRNA approaches were used. Wistar rats were used in accordance with national guidelines for animal research. We characterized the Best‐3 specific antibody by expression of eGFP‐fusion‐protein with the epitope containing peptide. IHC showed a broad expression of Best‐3 (e.g. kidney, heart, brain and mesentery) primarily in the vasculature. Within the vascular wall Best‐3 was shown to be strongly expressed in both smooth muscle and endothelial cells. This was supported by PCR cloning. Specific siRNAs significantly suppressed Best‐3 expression and the I (Ca, cGMP)Cl . Our study demonstrates the functional significance of Best‐3 protein for the cGMP‐dependent Ca 2+ ‐activated Cl − channel in vascular tissue. 1. Matchkov et al., Circ Res 2008 2. Matchkov et al., J Gen Physiol 2004 3. Matchkov et al., Pflugers Arch 2005