HIGH RESOLUTION ANALYSIS OF DIFFERENTIAL GENE EXPRESSION DURING SKELETAL MUSCLE ATROPHY AND PROGRAMMED CELL DEATH

Skeletal muscles can undergo atrophy and/or programmed cell death (PCD) during development or in response to a wide range of insults, including immobility, cachexia, and spinal cord injury. However, the protracted nature of atrophy and the presence of multiple cell types within the tissue complicates molecular analyses. One model that does not suffer from these limitations is the intersegmental muscle (ISM) of the tobacco hawkmoth Manduca sexta. Three days before the adult eclosion (emergence) at the end of metamorphosis, the ISMs initiate a non-pathological program of atrophy that results in a 40% loss of mass. The ISMs then generate the eclosion behaviour and initiate a non-apoptotic PCD during the next 30 hours. We have performed a comprehensive transcriptomics analysis of all mRNAs and microRNAs throughout ISM development in order to better understand the molecular mechanisms that mediate atrophy and death. Atrophy involves enhanced protein catabolism and reduced expression of the genes involved in respiration, adhesion and the contractile apparatus. In contrast, PCD involved the induction of numerous proteases, DNA methylases, membrane transporters, ribosomes, and anaerobic metabolism. These changes in gene expression are largely repressed when insects are injected with the insect steroid hormone 20-hydroxyecdysone, which delays death. The expression of the death-associated proteins may be greatly enhanced by reductions in specific microRNAs that function to repress translation. This study not only provides fundamental new insights into basic developmental processes, it may also represent a powerful resource for identifying potential diagnostic markers and molecular targets for therapeutic intervention.

Keywords: Manduca sexta; autophagy; metamorphosis; proteasome; ubiquitin.