RESUMO
Cultures of adipose tissue explants are a valuable tool for studying the intracellular mechanisms involving hormones and nutrients. However, testing how fatty acids affect cells requires a carrier molecule; bovine serum albumin (BSA) has been used for this purpose. However, contaminants can alter the cellular response. Our objectives were to: 1) test BSA as a fatty acid carrier and 2) evaluate polyvinyl alcohol (PVA) as a replacement for BSA. Adipose tissue explants from nine pigs were cultured in medium 199 for 4, 12, 24, and 48 h, with the following treatments: control, PVA (100 mM PVA added) and PVA + pGH (100 mM PVA plus 0.1 mg/mL porcine growth hormone). After each culture period, explants were collected and assayed for lipogenesis. After 48 h in culture, explants were assayed for lipolysis. A preliminary study with different commercial sources and high concentrations showed that BSA affected lipogenic rates. On the other hand, there were no effects of PVA on lipid synthesis, while pGH (positive control) reduced glucose incorporation into lipids (P < 0.01) when compared to both control and PVA (P < 0.05). There was no difference between control and PVA for lipolysis rates. However, pGH increased lipolysis when compared to control (P < 0.01) and PVA (P < 0.05). We demonstrated that BSA can alter lipogenesis, which precludes its use as a carrier molecule. On the other hand, addition of PVA had no effect on lipolysis or lipogenesis. We suggest the use of PVA instead of BSA for adding bioactive fatty acids to cultures of adipose tissue.
Assuntos
Tecido Adiposo/metabolismo , Ácidos Graxos/metabolismo , Lipogênese/efeitos dos fármacos , Lipólise/efeitos dos fármacos , Álcool de Polivinil/farmacologia , Técnicas de Cultura de Tecidos/veterinária , Tecido Adiposo/efeitos dos fármacos , Animais , Bovinos , Masculino , Soroalbumina Bovina , Suínos , Fatores de Tempo , Técnicas de Cultura de Tecidos/métodosRESUMO
Cultures of adipose tissue explants are a valuable tool for studying the intracellular mechanisms involving hormones and nutrients. However, testing how fatty acids affect cells requires a carrier molecule; bovine serum albumin (BSA) has been used for this purpose. However, contaminants can alter the cellular response. Our objectives were to: 1) test BSA as a fatty acid carrier and 2) evaluate polyvinyl alcohol (PVA) as a replacement for BSA. Adipose tissue explants from nine pigs were cultured in medium 199 for 4, 12, 24, and 48 h, with the following treatments: control, PVA (100 mM PVA added) and PVA + pGH (100 mM PVA plus 0.1 mg/mL porcine growth hormone). After each culture period, explants were collected and assayed for lipogenesis. After 48 h in culture, explants were assayed for lipolysis. A preliminary study with different commercial sources and high concentrations showed that BSA affected lipogenic rates. On the other hand, there were no effects of PVA on lipid synthesis, while pGH (positive control) reduced glucose incorporation into lipids (P < 0.01) when compared to both control and PVA (P < 0.05). There was no difference between control and PVA for lipolysis rates. However, pGH increased lipolysis when compared to control (P < 0.01) and PVA (P < 0.05). We demonstrated that BSA can alter lipogenesis, which precludes its use as a carrier molecule. On the other hand, addition of PVA had no effect on lipolysis or lipogenesis. We suggest the use of PVA instead of BSA for adding bioactive fatty acids to cultures of adipose tissue
Assuntos
Animais , Masculino , Bovinos , Tecido Adiposo/metabolismo , Ácidos Graxos/metabolismo , Lipogênese/efeitos dos fármacos , Lipólise/efeitos dos fármacos , Álcool de Polivinil/farmacologia , Técnicas de Cultura de Tecidos/veterinária , Tecido Adiposo/efeitos dos fármacos , Soroalbumina Bovina , Suínos , Fatores de Tempo , Técnicas de Cultura de Tecidos/métodosRESUMO
Este estudo foi desenvolvido com o objetivo de avaliar as freqüências gênicas em diferentes geraçöes de bovinos da raça sintética Canchim (5/8 Charolês, 3/8 Zebu). Uma amostra de 154 animais, representando três classes de geraçöes de um rebanho da raça Canchim, foi analisada para sete marcadores moleculares. Uma amostra da raça Charolesa (N=36) foi incluída nas análises para permitir comparaçöes. Observou-se um aumento linear, altamente significativo (P < 0,01), na freqüência do alelo que codifica o aminoácido valina na posiçäo 127 do hormônio de crescimento, ao longo das geraçöes de Canchim. Este alelo foi observado na amostra da raça Charolesa e näo é encontrado em raças zebuínas nacionais. Quatro alelos foram observados para o microssatélite localizado na regiäo 5' näo transcrita do gene do fator de crescimento semelhante à insulina do tipo I (IGF-I), com tamanhos variando de 231 a 225 pb. A freqüência do alelo de 225 pb apresentou um aumento significativo (P < 0,05) näo linear ao longo das geraçöes. Este alelo näo foi observado na raça Charolesa e é predominante nas raças zebuínas. Para o microssatélite CSFM50, seis alelos foram observados na raça Canchim, tendo sido verificada uma reduçäo linear (P < 0,05) na freqüência do alelo de 168 bp. A hipótese de acasalamento preferencial com relaçäo ao polimorfismo do hormônio de crescimento foi reforçada pelos resultados da análise de estatística F de Wright. O valor estimado de Fis para este loco foi 0,59 (P < 0,01). O aumento simultâneo de um alelo do hormônio de crescimento característico de Bos taurus e de um alelo de IGF-I característico de Bos indicus sugere que a seleçäo fenotípica tenha favorecido regiöes diferentes de ambos os genomas que entraram na formaçäo dessa raça.
