mRNA levels of myogenic regulatory factors in rat slow and fast muscles regenerating from notexin-induced necrosis.

The transcript levels of the myogenic regulatory factors (myoD, myf5, myogenin and MRF4) were measured by RT PCR in rat soleus (slow) and EDL (fast) muscles which were regenerating from notexin-induced necrosis. Some muscle fibers in the EDL were more resistant to the toxin, therefore the necrosis and the dominance of myoblasts were delayed for two days in EDL compared to soleus. In spite of this shift in time-course of necrosis, both types of muscle presented roughly similar, although variable, changes in the expression pattern of MRF mRNA levels. For both muscles, the myoD mRNA was upregulated on the first day after administration of the toxin, whereas concomitantly myf-5 mRNA disappeared but showed a substantial increase in later stages of regeneration. In contrast, the mRNA levels of the late MRFs myogenin and MRF4 decreased on day one only in the soleus, then increased on day three in both types of muscle. Meanwhile in EDL the level of MRF4 mRNA remained relatively normal. Four weeks after administration of the toxin the mRNA levels for each of the MRFs returned to nearly control levels. This shows that in spite of the different time course of the necrosis and regeneration, also documented by the microscopical morphology and the skeletal actin mRNA levels of the muscles, the level of MRF transcripts changed according to a quite predictable pattern; the upregulation corresponded to myoblast activation and the downregulation to the reinnervation.