Acute cold stress induces transient MuRF1 upregulation in the skeletal muscle of zebrafish

• Published in: Biochemical and biophysical research communications Vol. 608; pp. 59 - 65
• Main Authors: Tamai, Shinsuke, Fujita, Shin-ichiro, Komine, Ritsuko, Kanki, Yasuharu, Aoki, Kai, Watanabe, Koichi, Takekoshi, Kazuhiro, Sugasawa, Takehito
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 11.06.2022
• Subjects: Animals; Cold stress; Cold-Shock Response; Cryotherapy; Fatigue - metabolism; Fatigue - pathology; mRNA sequencing; Muscle, Skeletal - metabolism; Muscular Atrophy - metabolism; Skeletal muscle; SKP Cullin F-Box Protein Ligases - genetics; SKP Cullin F-Box Protein Ligases - metabolism; Temperature; Tripartite Motif Proteins - genetics; Tripartite Motif Proteins - metabolism; Ubiquitin-Protein Ligases - metabolism; Ubiquitins - metabolism; Up-Regulation; Zebrafish; Zebrafish - genetics; Zebrafish - metabolism; Zebrafish; Temperature; Cold stress; mRNA sequencing; Cryotherapy; Skeletal muscle
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2022.03.093

Cryotherapy is one of the most common treatments for trauma or fatigue in the field of sports medicine. However, the molecular biological effects of acute cold exposure on skeletal muscle remain unclear. Therefore, we used zebrafish, which have recently been utilized as an animal model for skeletal muscle, to comprehensively investigate and selectively clarify the time-course changes induced by cryotherapy. Zebrafish were exposed intermittently to cold stimulation three times for 15 min each. Thereafter, skeletal muscle samples were collected after 15 min and 1, 2, 4, and 6 h. mRNA sequencing revealed the involvement of trim63a, fbxo32, fbxo30a, and klhl38b in “protein ubiquitination” from the top 10 most upregulated genes. Subsequently, we examined the time-course changes of the four genes by quantitative PCR, and their expression peaked 2 h after cryotherapy and returned to baseline after 6 h. Moreover, the proteins encoded by trim63a and fbxo32 (muscle-specific RING finger protein 1 [MuRF1] and muscle atrophy F-box, respectively), which are known to be major genes encoding E3 ubiquitin ligases, were examined by western blotting, and MuRF1 expression displayed similar temporal changes as trim63a expression. These findings suggest that acute cold exposure transiently upregulates E3 ubiquitin ligases, especially MuRF1; thus, cryotherapy may contribute to the treatment of trauma or fatigue by promoting protein processing. •Acute cold stress affected the expression of multiple genes, mainly those related to E3 ubiquitin ligases.•The increase in gene expression was transiently induced, peaking at 2 h after cold exposure and returning to baseline by 6 h.•In particular, the protein expression of MuRF1 (encoded by trim63a) was significantly increased.•Cryotherapy may contribute to recovery from trauma or fatigue by promoting the processing of damaged or unfolded proteins.