Abstract
Obscurin and obscurin‐associated kinase are two products of the obscurin transcriptional unit that encodes a recently identified giant muscle‐specific protein obscurin. In this study, we characterized the developmental expression and cellular localization of obscurin and obscurin‐associated kinase in cardiac muscle cells. We cloned murine obscurin‐associated kinase and found that it is abundantly expressed in the heart as two isotypes encoded by 2.2 and 4.9 kb sequences. The 2.2 kb isotype of the kinase was more prominently expressed than the 4.9 kb isotype. Both obscurin and the kinase‐like domains were progressively upregulated since the early stages of cardiac development. Obscurin‐associated kinase was expressed at higher levels than obscurin at early stages of cardiomyogenesis. Increasing intensity of obscurin expression in the developing heart positively correlated with progressive cell differentiation and was higher in the ventricles compared to the atria. These data were supported by the results of experiments with primary cardiac cell cultures. Obscurin localization changed from a weakly immunopositive diffuse pattern in poorly differentiated cells to an intensely immunolabeled cross‐striated distribution at the level of mid‐A‐bands and Z‐disks during the assembly of the myofibrillar contractile apparatus. In dividing myocytes, unlike the interphase cells, obscurin translocated from disassembling myofibrils into a diffuse granulated pattern segregated separately from α‐actinin‐immunopositive aggregates. Obscurin‐associated kinase was localized mainly to cell nuclei with increasing incorporation into the Z‐disks during differentiation. Our results suggest that these two novel proteins are involved in the progression of cardiac myogenesis during the transition to advanced stages of heart development. J. Cell. Biochem. 103: 1621–1635, 2008. © 2007 Wiley‐Liss, Inc.