University of Kentucky
RNA Degradation is Elevated with Age, but not Disuse-Associated Skeletal Muscle Atrophy
Institution
University of Kentucky
Faculty Advisor/ Mentor
Esther E. Dupont-Versteegden
Abstract
Aging and inactivity are both associated with decreased muscle size and protein content. The possible role of RNA degradation in the loss of protein has not yet been investigated. Therefore, we hypothesized that RNA degradation was elevated with muscle atrophy in aging and disuse. Brown Norway/Fisher344 male rats at 6 and 32 months were hindlimb suspended (HS) for 14 days to induce muscle atrophy or remained weight bearing (WB). Cytosolic extracts from gastrocnemius muscles were prepared for Western analysis of Dcp-2 protein (marker of pbodies) and RNA degradation assay. In vitro total RNA decay assay was performed using 30ug of total RNA (from tibialis anterior) incubated with 20ug of S15 extracts from gastrocnemius. RNA integrity was determined using the Agilent Technologies algorithm to calculate the RNA Integrity Number (RIN); decay rate and half-life were calculated for each sample. Results indicated an increase in Dcp-2 protein at 32 months of age in both HS and WB groups. In addition, an almost 2-fold increase in decay rate and 48% decrease in half-life of total RNA was observed in muscle from 32 month old rats. However, no significant difference in decay rate and half-life was observed with disuse at either 6 or 32 months. We conclude that muscle atrophy associated with aging, but not disuse, may be due to a decrease in total RNA because of increased RNA degradation. Supported by APS UGSRF and AG028925.
RNA Degradation is Elevated with Age, but not Disuse-Associated Skeletal Muscle Atrophy
Aging and inactivity are both associated with decreased muscle size and protein content. The possible role of RNA degradation in the loss of protein has not yet been investigated. Therefore, we hypothesized that RNA degradation was elevated with muscle atrophy in aging and disuse. Brown Norway/Fisher344 male rats at 6 and 32 months were hindlimb suspended (HS) for 14 days to induce muscle atrophy or remained weight bearing (WB). Cytosolic extracts from gastrocnemius muscles were prepared for Western analysis of Dcp-2 protein (marker of pbodies) and RNA degradation assay. In vitro total RNA decay assay was performed using 30ug of total RNA (from tibialis anterior) incubated with 20ug of S15 extracts from gastrocnemius. RNA integrity was determined using the Agilent Technologies algorithm to calculate the RNA Integrity Number (RIN); decay rate and half-life were calculated for each sample. Results indicated an increase in Dcp-2 protein at 32 months of age in both HS and WB groups. In addition, an almost 2-fold increase in decay rate and 48% decrease in half-life of total RNA was observed in muscle from 32 month old rats. However, no significant difference in decay rate and half-life was observed with disuse at either 6 or 32 months. We conclude that muscle atrophy associated with aging, but not disuse, may be due to a decrease in total RNA because of increased RNA degradation. Supported by APS UGSRF and AG028925.