Muscle Physiology Laboratory, Department of Rehabilitation Sciences, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong.
Muscle Nerve. 2011 Sep;44(3):358-65. doi: 10.1002/mus.22060.
TRPC1 and TRPC3 proteins are widely expressed in skeletal muscles in forming calcium-permeable channels. Herein we characterize the expression pattern of TRPC transcripts during skeletal myogenesis in C2C12 myoblasts.
We used polymerase chain reaction and Western blotting to detect expression levels, immunohistochemistry for subcellular localization, and co-immunoprecipitation techniques to assess interaction.
TRPC1 localizes to the cytoplasm and is enriched in the perinuclear region in undifferentiated myoblasts. Expression of TRPC1 increases significantly during myogenesis and resides mainly in differentiated myocytes and myotubes. TRPC3 is absent in undifferentiated myoblasts, is dramatically upregulated in differentiated culture, and is preferentially expressed in myotubes. Physical interaction of TRPC1-TRPC3 was observed, suggesting the possible existence of heteromers.
Expression of TRPC1 and TRPC3 is tightly regulated during myogensis. Evidence of TRPC1-TRPC3 interaction was first demonstrated in a muscle cell line. The functional consequences of this interaction remain to be established.
TRPC1 和 TRPC3 蛋白广泛表达于骨骼肌中,形成钙通透性通道。本文作者在 C2C12 成肌细胞中研究了骨骼形成过程中 TRPC 转录本的表达模式。
作者采用聚合酶链反应和 Western blot 检测来评估表达水平,免疫组织化学进行亚细胞定位,以及免疫共沉淀技术来评估相互作用。
TRPC1 位于细胞质中,在未分化的成肌细胞中富含核周区域。在成肌过程中,TRPC1 的表达显著增加,主要存在于分化的肌细胞和肌管中。TRPC3 在未分化的成肌细胞中不存在,在分化培养中显著上调,并且优先在肌管中表达。观察到 TRPC1-TRPC3 的物理相互作用,表明可能存在异源二聚体。
TRPC1 和 TRPC3 的表达在成肌过程中受到严格调控。在肌肉细胞系中首次证明了 TRPC1-TRPC3 相互作用的证据。这种相互作用的功能后果仍有待确定。