![]() ![]() There was also evidence for up regulation of three pathways involved in nucleotide biosynthesis. Proteins which were differentially abundant during CG (n = 39) were involved in cellular binding processes, oxidative phosphorylation and mitochondrial function. Proteins were isolated from samples and subjected to label-free mass spectrometry proteomic quantification. longissimus dorsi samples were collected at slaughter from all animals. Remaining animals (R2 and A2) were slaughtered following ad libitum access to feed for successive 55 days (Period 2). Following Period 1, all animals from R1 and A1 were slaughtered. A1 and A2 animals had ad libitum access to feed in Period 1. R1 and R2 groups were subjected to restricted feed allowance for 125 days (Period 1). Forty Holstein Friesian bulls were equally assigned to one of four groups. This studies objective was to identify key proteins involved in the expression of CG. *p < 0.05 (vs SA).Ĭompensatory growth (CG) is a naturally occurring physiological process whereby an animal has the ability to undergo enhanced growth following a period of restricted feeding. The statistical analysis of the results was carried out with SigmaPlot 11.0, from Systat Software, Inc., San Jose California USA, https ://The data were analysed using nonparametric single-factor dispersion analysis for repeated measurements (Kruskal-Wallis One Way Analysis of Variance on Ranks) followed with the pairwise comparison using the Tukey's test. (B) Bar graphs of titin phosphorylation level in striated muscles (Supplementary Tables S23, S24). The second (SA heart), fourth (HIB heart), fifth (IBA heart), seventh (SA LD), eighth (HIB LD) and ninth (IBA LD) tracks from the full-length gel are shown in (A). Full-length gel is shown in Supplementary Fig. The native level of protein phosphorylation was estimated in gels using Pro-Q Diamond fluorescent dye (ThermoFisher Scientific) (brightness, - 41 contrast, - 50). SA summer activity, HIB hibernation, IBA interbout arousal. # p < 0.05 (IBA vs HIB).Ĭhanges in titin phosphorylation levels in striated muscles of ground squirrels. ![]() ![]() The statistical analysis of the results was carried out with SigmaPlot 11.0, from Systat Software, Inc., San Jose California USA, https ://The data were analysed using nonparametric single-factor dispersion analysis for repeated measurements (Kruskal-Wallis One Way Analysis of Variance on Ranks) followed with the pairwise comparison by the Tukey's test. (B) Bar graphs of T1, T2 fragments and nebulin contents in striated muscles (Supplementary Tables S18-S22). High molecular weight titin isoforms (denoted as NT-titin) were found in striated muscles of mammals 63. NT, N2A, N2BA and N2B are isoforms of intact titin-1 (T1 MW, 3,000-3,700 kDa). T2 (MW, 2,000-2,200 kDa) are proteolytic fragments of titin. longissimus dorsi (LD, right) (brightness, - 9 contrast, + 30). (A) SDS-PAGE analysis of titin in the heart (left) and titin and nebulin in the m. Titin and nebulin contents in striated muscles of ground squirrels. These changes may contribute to muscle mass retention during hibernation. The results indicate that hyperactivation of calpain-1 in striated muscles of long-tailed ground squirrels during interbout arousal is accompanied by predominant synthesis of giant sarcomeric cytoskeleton proteins. longissimus dorsi muscle fibres, a trend towards a heart-muscle mass increase and a restoration of the normal titin content (reduced in the muscles during hibernation) were observed. The synthesis rates of calpain-1 substrates nebulin and titin in muscles did not differ during interbout arousal from those in active summer animals. longissimus dorsi (p < 0.01) during interbout arousal. The total protein synthesis rate determined by SUnSET declined 3.67-fold in the heart (p < 0.01) and 2.96-fold in m. longissimus dorsi (1.8-fold, p < 0.01) of ground squirrels during interbout arousal. Western blotting detected increased amounts of autolysed calpain-1 fragments in the heart (1.54-fold, p < 0.05) and m. Calpain-1 is known to be autolysed and activated in parallel. This work tested the assumption that protein synthesis hyperactivation during interbout arousal of the long-tailed ground squirrel Urocitellus undulatus should be accompanied by increased calpain-1 activity in striated muscles. Molecular mechanisms underlying muscle-mass retention during hibernation have been extensively discussed in recent years. ![]()
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