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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
2011-6-28
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pubmed:abstractText |
Luminal 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.
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pubmed:grant |
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pubmed:commentsCorrections |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/1-(5-Isoquinolinesulfonyl)-2-Methylp...,
http://linkedlifedata.com/resource/pubmed/chemical/ADP Ribose Transferases,
http://linkedlifedata.com/resource/pubmed/chemical/Actins,
http://linkedlifedata.com/resource/pubmed/chemical/Amides,
http://linkedlifedata.com/resource/pubmed/chemical/Botulinum Toxins,
http://linkedlifedata.com/resource/pubmed/chemical/Enzyme Inhibitors,
http://linkedlifedata.com/resource/pubmed/chemical/Green Fluorescent Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Proton Pumps,
http://linkedlifedata.com/resource/pubmed/chemical/Pyridines,
http://linkedlifedata.com/resource/pubmed/chemical/Vacuolar Proton-Translocating...,
http://linkedlifedata.com/resource/pubmed/chemical/Y 27632,
http://linkedlifedata.com/resource/pubmed/chemical/exoenzyme C3, Clostridium botulinum,
http://linkedlifedata.com/resource/pubmed/chemical/fasudil,
http://linkedlifedata.com/resource/pubmed/chemical/rho-Associated Kinases,
http://linkedlifedata.com/resource/pubmed/chemical/rhoA GTP-Binding Protein
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
1522-1563
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:volume |
301
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
C31-43
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pubmed:meshHeading |
pubmed-meshheading:21411727-1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine,
pubmed-meshheading:21411727-ADP Ribose Transferases,
pubmed-meshheading:21411727-Actins,
pubmed-meshheading:21411727-Amides,
pubmed-meshheading:21411727-Animals,
pubmed-meshheading:21411727-Blotting, Western,
pubmed-meshheading:21411727-Botulinum Toxins,
pubmed-meshheading:21411727-Cytoskeleton,
pubmed-meshheading:21411727-Enzyme Inhibitors,
pubmed-meshheading:21411727-Epididymis,
pubmed-meshheading:21411727-Epithelial Cells,
pubmed-meshheading:21411727-Flow Cytometry,
pubmed-meshheading:21411727-Green Fluorescent Proteins,
pubmed-meshheading:21411727-Male,
pubmed-meshheading:21411727-Mice,
pubmed-meshheading:21411727-Mice, Transgenic,
pubmed-meshheading:21411727-Microvilli,
pubmed-meshheading:21411727-Proton Pumps,
pubmed-meshheading:21411727-Pyridines,
pubmed-meshheading:21411727-Rats,
pubmed-meshheading:21411727-Rats, Sprague-Dawley,
pubmed-meshheading:21411727-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:21411727-Signal Transduction,
pubmed-meshheading:21411727-Vacuolar Proton-Translocating ATPases,
pubmed-meshheading:21411727-rho-Associated Kinases,
pubmed-meshheading:21411727-rhoA GTP-Binding Protein
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pubmed:year |
2011
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pubmed:articleTitle |
Regulation of V-ATPase recycling via a RhoA- and ROCKII-dependent pathway in epididymal clear cells.
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pubmed:affiliation |
Center for Systems Biology, Program in Membrane Biology/Nephrology Division, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
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pubmed:publicationType |
Journal Article,
Research Support, N.I.H., Extramural
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