RESUMO
Fishes adapt to salinity changes primarily through osmotic pressure regulation, a process often associated with several genes, including 14-3-3a, NKCCla, APO-14, and Na+-K+-ATPaseß. The present study investigated the differential expression of genes 14-3-3a, NKCCla, APO-14, and Na+-K+-ATPaseß in the gill tissue of Mugil cephalus acclimated to low salinity. Susceptibility relationships between the four gene expressions levels and salinity were detected and analyzed using polymerase chain reaction-restriction fragment length polymorphism. Homology analysis results indicated significant differences in the correlation between gene expression and salinity. Under low-salt conditions, expression levels for genes Na+-K+-ATPaseß and NKCC1a were significantly elevated (P < 0.05), whereas those of genes 14-3-3a and APO-14 were significantly reduced (P < 0.05). Thus, when compared to 14-3-3a and APO-14, Na+-K+-ATPaseß, and NKCC1a may be better suited to promoting the development of osmotic-regulation mechanisms and increased resistance to environmental stress under low-salt conditions. Furthermore, Na+-K+-ATPaseß and NKCC1a were identified as suitable potential molecular biomarkers for regulating and controlling genes in low-salinity aquatic environments.
Assuntos
Proteínas de Peixes/genética , Peixes/genética , Expressão Gênica , Brânquias/metabolismo , Proteínas 14-3-3/genética , Adaptação Fisiológica , Animais , Proteínas de Peixes/metabolismo , Peixes/crescimento & desenvolvimento , Regulação da Expressão Gênica , SalinidadeRESUMO
This study evaluated the effect of muscle satellite cells (MSCs) overexpressing myogenin (MyoG) on denervated muscle atrophy. Rat MSCs were isolated and transfected with the MyoG-EGFP plasmid vector GV143. MyoG-transfected MSCs (MTMs) were transplanted into rat gastrocnemius muscles at 1 week after surgical denervation. Controls included injections of untransfected MSCs or the vehicle only. Muscles were harvested and analyzed at 2, 4, and 24 weeks post-transplantation. Immunofluorescence confirmed MyoG overexpression in MTMs. The muscle wet weight ratio was significantly reduced at 2 weeks after MTM injection (67.17±6.79) compared with muscles injected with MSCs (58.83±5.31) or the vehicle (53.00±7.67; t=2.37, P=0.04 and t=3.39, P=0.007, respectively). The muscle fiber cross-sectional area was also larger at 2 weeks after MTM injection (2.63×10³±0.39×10³) compared with MSC injection (1.99×10³±0.58×10³) or the vehicle only (1.57×10³±0.47×10³; t=2.24, P=0.049 and t=4.22, P=0.002, respectively). At 4 and 24 weeks post-injection, the muscle mass and fiber cross-sectional area were similar across all three experimental groups. Immunohistochemistry showed that the MTM group had larger MyoG-positive fibers. The MTM group (3.18±1.13) also had higher expression of MyoG mRNA than other groups (1.41±0.65 and 1.03±0.19) at 2 weeks after injection (t=2.72, P=0.04). Transplanted MTMs delayed short-term atrophy of denervated muscles. This approach can be optimized as a novel stand-alone therapy or as a bridge to surgical re-innervation of damaged muscles.
Assuntos
Transplante de Células , Denervação Muscular/reabilitação , Músculo Esquelético/inervação , Atrofia Muscular/reabilitação , Miogenina/metabolismo , Células Satélites de Músculo Esquelético/transplante , Animais , Imunofluorescência , Expressão Gênica , Masculino , Atrofia Muscular/etiologia , Miogenina/genética , Tamanho do Órgão/genética , Plasmídeos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Células Satélites de Músculo Esquelético/citologia , Células Satélites de Músculo Esquelético/metabolismo , Neuropatia Ciática/reabilitação , TransfecçãoRESUMO
This study evaluated the effect of muscle satellite cells (MSCs) overexpressing myogenin (MyoG) on denervated muscle atrophy. Rat MSCs were isolated and transfected with the MyoG-EGFP plasmid vector GV143. MyoG-transfected MSCs (MTMs) were transplanted into rat gastrocnemius muscles at 1 week after surgical denervation. Controls included injections of untransfected MSCs or the vehicle only. Muscles were harvested and analyzed at 2, 4, and 24 weeks post-transplantation. Immunofluorescence confirmed MyoG overexpression in MTMs. The muscle wet weight ratio was significantly reduced at 2 weeks after MTM injection (67.17±6.79) compared with muscles injected with MSCs (58.83±5.31) or the vehicle (53.00±7.67; t=2.37, P=0.04 and t=3.39, P=0.007, respectively). The muscle fiber cross-sectional area was also larger at 2 weeks after MTM injection (2.63×103±0.39×103) compared with MSC injection (1.99×103±0.58×103) or the vehicle only (1.57×103±0.47×103; t=2.24, P=0.049 and t=4.22, P=0.002, respectively). At 4 and 24 weeks post-injection, the muscle mass and fiber cross-sectional area were similar across all three experimental groups. Immunohistochemistry showed that the MTM group had larger MyoG-positive fibers. The MTM group (3.18±1.13) also had higher expression of MyoG mRNA than other groups (1.41±0.65 and 1.03±0.19) at 2 weeks after injection (t=2.72, P=0.04). Transplanted MTMs delayed short-term atrophy of denervated muscles. This approach can be optimized as a novel stand-alone therapy or as a bridge to surgical re-innervation of damaged muscles.