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21411727

Source:http://linkedlifedata.com/resource/pubmed/id/21411727

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pubmed-article:21411727pubmed:abstractTextLuminal acidification in the epididymis is critical for sperm maturation and storage. Clear cells express the vacuolar H(+)-ATPase (V-ATPase) in their apical membrane and are major contributors to proton secretion. We showed that this process is regulated via recycling of V-ATPase-containing vesicles. We now report that RhoA and its effector ROCKII are enriched in rat epididymal clear cells. In addition, cortical F-actin was detected beneath the apical membrane and along the lateral membrane of "resting" clear cells using a pan-actin antibody or phalloidin-TRITC. In vivo luminal perfusion of the cauda epididymal tubule with the ROCK inhibitors Y27632 (10-30 ?M) and HA1077 (30 ?M) or with the cell-permeable Rho inhibitor Clostridium botulinum C3 transferase (3.75 ?g/ml) induced the apical membrane accumulation of V-ATPase and extension of V-ATPase-labeled microvilli in clear cells. However, these newly formed microvilli were devoid of ROCKII. In addition, Y27632 (30 ?M) or HA1077 (30 ?M) decreased the ratio of F-actin to G-actin detected by Western blot analysis in epididymal epithelial cells, and Y27632 also decreased the ratio of F-actin to G-actin in clear cells isolated by fluorescence activated cell sorting from B1-enhanced green fluorescence protein (EGFP) transgenic mice. These results provide evidence that depolymerization of the cortical actin cytoskeleton via inhibition of RhoA or its effector ROCKII favors the recruitment of V-ATPase from the cytosolic compartment into the apical membrane in clear cells. In addition, our data suggest that the RhoA-ROCKII pathway is not locally involved in the elongation of apical microvilli. We propose that inhibition of RhoA-ROCKII might be part of the intracellular signaling cascade that is triggered upon agonist-induced apical membrane V-ATPase accumulation.lld:pubmed
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pubmed-article:21411727pubmed:articleTitleRegulation of V-ATPase recycling via a RhoA- and ROCKII-dependent pathway in epididymal clear cells.lld:pubmed
pubmed-article:21411727pubmed:affiliationCenter for Systems Biology, Program in Membrane Biology/Nephrology Division, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.lld:pubmed
pubmed-article:21411727pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:21411727pubmed:publicationTypeResearch Support, N.I.H., Extramurallld:pubmed
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