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  • Myo1c is an unconventional myosin required for zebrafish glomerular development.

Myo1c is an unconventional myosin required for zebrafish glomerular development.

Kidney international (2013-05-30)
Ehtesham Arif, Babita Kumari, Mark C Wagner, Weibin Zhou, Lawrence B Holzman, Deepak Nihalani
ABSTRACT

The targeting and organization of podocyte slit diaphragm proteins nephrin and neph1 is critical for development and maintenance of a functional glomerular filtration barrier. Myo1c is a non-muscle myosin motor protein that interacts directly with nephrin and neph1, and mediates their intracellular transport to the podocyte intercellular junction. Here we investigated the necessity of Myo1c in podocyte development using zebrafish as a model system. Immunofluorescence microscopy and in situ RNA hybridization analysis of zebrafish embryos showed that Myo1c is widely expressed in various tissues including the zebrafish glomerulus. Knockdown of the Myo1c gene in zebrafish using antisense morpholino derivatives resulted in an abnormal developmental phenotype that included pericardial edema and dilated renal tubules. Ultrastructural analysis of the glomerulus in Myo1c-depleted zebrafish showed abnormal podocyte morphology and absence of the slit diaphragm. Consistent with these observations, the glomerular filter permeability appeared altered in zebrafish in which Myo1c expression was attenuated. The specificity of Myo1c knockdown was confirmed by a rescue experiment in which co-injection of Myo1c morpholino derivatives with orthologous Myo1c mRNA prepared from mouse cDNA lessened phenotypic abnormalities including edema in Myo1c morphants. Thus, our results demonstrate that Myo1c is necessary for podocyte morphogenesis.

MATERIALS
Product Number
Brand
Product Description

Roche
DIG RNA Labeling Kit (SP6/T7), sufficient for 2 x 10 labeling reactions, kit of 1 (12 components), suitable for hybridization, suitable for Southern blotting
Roche
DIG-High Prime DNA Labeling and Detection Starter Kit II, sufficient for 12 labeling reactions (10 ng to 3 μg per assay), sufficient for 24 blots (blots of 100 cm2)