Expression of tropism-related genes in regenerating skeletal muscle of rats treated with cyclosporin-A.
This work was undertaken to provide further insights into the expression of tropism-related genes in regenerating skeletal muscle of adult rats treated with cyclosporin-A (CsA), a calcineurin inhibitor. Rats were treated with CsA for 5 days and, on the 6th day, were submitted to cryolesion of the soleus muscles. CsA treatment continued for 1, 10, and 21 days after cryolesion. Muscles were removed, frozen, and stored in liquid nitrogen. Body and muscle weights, histological sections stained with toluidine blue, and gene expression of the regeneration molecular markers, viz., desmin and neonatal myosin heavy chain, were assessed to confirm that cryolesion and CsA treatment were effective during the allowed regeneration time. Quantitative reverse transcription/polymerase chain reaction demonstrated that myostatin gene expression was not altered by either cryolesion or CsA treatment combined with cryolesion. Calpain-3 gene expression decreased at 1 day after cryolesion and also following CsA treatment combined with cryolesion. However, calpain-3 gene expression was strongly up-regulated (approximately five-fold) 10 days after cryolesion and returned to control levels at day 21. CsA treatment blocked calpain-3 gene expression rise induced by 10 days of cryolesion. Atrogin-1 gene expression was decreased at 1 day after cryolesion and following cryolesion combined with CsA treatment, returning to control levels at day 10. These results suggest that (1) calpain-3 has a differential role in the early and late stages of regeneration in a calcineurin-dependent manner, and (2) atrogin-1 is involved in the early stages of regeneration independently of calcineurin.
Miyabara
Elen H
EH
Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of Sao Paulo, 05508-900, Sao Paulo, Brazil.
Aoki
Marcelo S
MS
Soares
Antonio G
AG
Moriscot
Anselmo S
AS
D000818:Animals; D001835:Body Weight / Q000187:drug effects; D002154:Calpain / Q000235:genetics* / Q000378:metabolism; D003080:Cold Temperature / Q000009:adverse effects; D003452:Cryosurgery; D016572:Cyclosporine / Q000494:pharmacology*; D017931:DNA Primers / Q000737:chemistry; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects*; D007274:Injections, Intraperitoneal; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D008297:Male; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000293:injuries / Q000378:metabolism; D055435:Myostatin; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012038:Regeneration / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044843:SKP Cullin F-Box Protein Ligases / Q000235:genetics* / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; D004791:Enzyme Inhibitors; D007527:Isoenzymes; C525913:Mstn protein, rat; D009124:Muscle Proteins; D055435:Myostatin; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D016572:Cyclosporine; C486692:Fbxo32 protein, rat; D044843:SKP Cullin F-Box Protein Ligases; D002154:Calpain; C491372:Capn3 protein, rat