The Effect of Including the C2 Insert of Nonmuscle Myosin II-C on Neuritogenesis |
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Authors: | Shekhar Saha Sumit K Dey Arunima Biswas Provas Das Mahua R Das Siddhartha S Jana |
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Institution: | From the ‡Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata-32, India and ;the §Infectious Diseases and Immunology Division, Indian Institute of Chemical Biology, Kolkata-32, India |
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Abstract: | The functional role of the C2 insert of nonmuscle myosin II-C (NM II-C) is poorly understood. Here, we report for the first time that the expression of the C2 insert-containing isoform, NM II-C1C2, is inducible in Neuro-2a cells during differentiation both at mRNA and protein levels. Immunoblot and RT-PCR analysis reveal that expression of NM II-C1C2 peaks between days 3 and 6 of differentiation. Localization of NM II-C1C2 in Neuro-2a cells suggests that the C2 insert-containing isoform is localized in the cytosol and along the neurites, specifically at the adherence point to substratum. Inhibition of endogenous NM II-C1C2 using siRNA decreases the neurite length by 43% compared with control cells treated with nonspecific siRNA. Time lapse image analysis reveals that neurites of C2-siRNA-treated cells have a net negative change in neurite length per minute, leading to a reduction of overall neurite length. During neuritogenesis, NM II-C1C2 can interact and colocalize with β1-integrin in neurites. Altogether, these studies indicate that NM II-C1C2 may be involved in stabilizing neurites by maintaining their structure at adhesion sites. |
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Keywords: | Alternative Splicing Cell Biology Molecular Cell Biology Myosin Neurodifferentiation |
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