Expression of caspase family and muscle- and apoptosis-specific genes during skeletal myogenesis in mouse embryo
2009
Ikeda, T.(Hokkaido Univ., Sapporo (Japan)) | Kanazawa, T. | Otsuka, S. | Ichii, O. | Hashimoto, Y. | Kon, Y.
The caspases (Casps) are a family of cysteine proteases that are known to regulate apoptotic signaling. Apoptosis by activation of Casp is strongly associated with embryonal development and regeneration in many organs, therefore indicating that disorders caused by homozygous mutation in Casp genes can result in embryonic lethality. In the present study, the authors investigated the causative relationship between skeletal myogenesis and the activation of Casps by analyzing their dynamics during mouse embryogenesis. Individual myogenetic tissues were obtained from C57BL/6 mouse embryos aged 12.5-17.5 days post-conception (dpc), and the expression of Casps was analyzed by histochemical and molecular biological methods. Immunoreactions for Casp-3, -9 and -12 were detected first in myoblasts, increasing according to embryonal development, as a result of which myoblasts differentiated into myotube cells. On the other hand, the immunoreaction for ssDNA, which is well-known as an apoptosis marker, was little detected during the skeletal myogenesis. Quantification analysis for Casp mRNA expression by RT-PCR as well as by in situ hybridization showed a peak at 15.5 dpc but a decrease at 17.5 dpc. Similar dynamics were detected for Myod1 mRNA, one of the muscle regulatory factors, but not for Fasl, Bax and Rock1, apoptosis-associated factors during skeletal myogenesis. These results suggest that the activation of Casps in skeletal myogenesis is deeply associated with myoblast differentiation, but not directly related to apoptosis.
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