The localization of VAMP5 in skeletal and cardiac muscle

Abstract

Vesicle-associated membrane protein 5 (VAMP5) is a member of the SNARE protein family, which is generally thought to regulate the docking and fusion of vesicles with their target membranes. This study investigated the expression and localization of the VAMP5 protein. Immunoblotting analyses detected the VAMP5 protein in skeletal muscle, heart, spleen, lung, liver, and kidney tissue, but not in brain or small intestine tissue. Through the immunofluorescence microscopy of skeletal muscle, we found that the expression level of VAMP5 varies among fibers. Most of the fibers with high expression levels of VAMP5 were categorized as type IIa fibers on the basis of their myosin heavy chain subtypes. In addition, the expression patterns of VAMP5 and glucose transporter 4 (GLUT4) were similar. In cardiac muscle, we determined that VAMP5 was localized to the vicinity of intercalated discs. These results suggest that VAMP5 plays local roles in membrane trafficking in skeletal and cardiac muscle.

Loss of VAMP5 in mice results in duplication of the ureter and insufficient expansion of the lung

“…As an immunohistochemical control, lung sections were treated with anti-VAMP5 antibody in the presence of 0.01 mg/ml recombinant VAMP5 protein (Takahashi et al, 2013). Sections 10 mm thick were cut using a Leica CM3050S cryostat (Leica Microsystems K.K., Tokyo, Japan), and placed on poly-L-lysine-coated glass slides (Matsunami Glass, Osaka, Japan).…”

“…The following reagents were used for immunostaining: rabbit anti-VAMP5 antibody (1:1,000; Synaptic Systems), mouse anti-pan keratin antibody (1:400; Cell Signaling), goat anti-CC-10 (1:50; Santa Cruz, Dallas, TX), Cy5-conjugated donkey antibody against rabbit IgG (1:500; Jackson ImmunoResearch), Alexa Fluor-488-conjugated phalloidin (1:50; Invitrogen), and DAPI (4 0 ,6-diamidine-2-phenylidole-dihydrochloride; 1:200; 10 mg/ml saturated solution). As an immunohistochemical control, lung sections were treated with anti-VAMP5 antibody in the presence of 0.01 mg/ml recombinant VAMP5 protein (Takahashi et al, 2013).…”

“…Little is known about the roles of VAMP5 and its associations with other molecules. Knockdown of VAMP5 using C2C12 myoblasts had no effect on cell-cell fusion and muscle differentiation (Takahashi et al, 2013). Fusogenic pairings of VAMP5 with other SNARE proteins have not been identified (Hasan et al, 2010).…”

“…VAMP5 was thought to be a muscle-specific isoform, as its mRNA was abundant in cardiac and skeletal muscle (Zeng et al, 1998). VAMP5 was also found in nonmuscle tissue, including the kidneys, lungs, liver, and spleen (Takahashi et al, 2013). Little is known about the roles of VAMP5 and its associations with other molecules.…”

“…VAMP5 was also found in nonmuscle tissue, including the kidneys, lungs, liver, and spleen (Takahashi et al, 2013). Knockdown of VAMP5 using C2C12 myoblasts had no effect on cell-cell fusion and muscle differentiation (Takahashi et al, 2013). Knockdown of VAMP5 using C2C12 myoblasts had no effect on cell-cell fusion and muscle differentiation (Takahashi et al, 2013).…”