Expression of myosin VI within the early endocytic pathway in adult and developing proximal tubules

American Journal of Physiology. Renal Physiology
Daniel BiemesderferTama Hasson

Abstract

Myosin VI is a reverse-direction molecular motor implicated in membrane transport events. Because myosin VI is most highly expressed in the kidney, we investigated its renal localization by using high-resolution immunocytochemical and biochemical methods. Indirect immunofluorescence microscopy revealed myosin VI at the base of the brush border in proximal tubule cells. Horseradish peroxidase uptake studies, which labeled endosomes, and double staining for clathrin adapter protein-2 showed that myosin VI was closely associated with the intermicrovillar (IMV) coated-pit region of the brush border. Localization of myosin VI to the IMV region was confirmed at the electron microscopic level by colloidal gold labeling of ultrathin cryosections. In addition, antigen retrieval demonstrated a small but significant pool of myosin VI on the microvilli. To confirm the association of myosin VI with the IMV compartment, these membranes were separated from other membrane compartments by using 15-25% OptiPrep density gradients. Immunoblotting of the gradient fractions confirmed that myosin VI was enriched with markers for the IMV microdomain of the brush border, suggesting that myosin VI associates with proteins in this compartment. Finally, w...Continue Reading

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Citations

Jul 15, 2003·Molecular Biology of the Cell·Laura AschenbrennerTama Hasson
Aug 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Samia N NaccacheArie Horowitz
Sep 25, 2009·American Journal of Physiology. Cell Physiology·Judith BlaineMoshe Levi
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