Análisis del patrón de expresión de genes implicados en el desarrollo gonadal de Xenopus tropicalis
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2016-03-04
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Jaén: Universidad de Jaén
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[ES] En este trabajo se ha estudiado el desarrollo gonadal de la especie Xenopus tropicalis analizando el patrón de expresión de los genes kal1, ntn1 y cyp11a1, tres genes con expresión diferencial en experimentos de RNAseq. Además, se ha realizado un análisis comparativo de la expresión de cyp11a1 y hsd3b1 durante el desarrollo gonadal masculino, al presentar ambos genes un patrón de expresión similar en las células precursoras de la línea esteroidogénica. Para ello se procedió a la obtención, mediante fecundación in vivo, de huevos de X. tropicalis, al mantenimiento y recogida de larvas en diferentes estadios de desarrollo, la clonación de fragmentos y la síntesis de ribosondas específicas para dichos genes mediante técnicas de PCR y clonación in vivo, y al análisis de la expresión mediante la técnica de hibridación in situ sobre tejidos. Como resultado de estos estudios, se ha observado una expresión diferencial de los genes kal1 y ntn1, con expresión exclusivamente femenina. Por otro lado, cyp11a1 se expresa inicialmente en ambos sexos, para pasar a ser un gen específico de macho a partir del estadio NF 55.
[EN] Gonadal development of the species Xenopus tropicalis was studied by analyzing the expression pattern of kal1, ntn1 and cyp11a1 genes, three genes with differential gene expression in RNAseq experiments. Furthermore, a comparative analysis of the expression pattern of cyp11a1 and hsd3b1during male gonadal development was performed, as both genes has similar expression patterns in steroidogenic precursor cells. To this end we obtained Xenopus tropicalis eggs by in vivo fertilization; tadpole husbandry and dissection at specific developmental stages were done. Different regions from these genes were PCR-cloned and specific riboprobes were synthetized by in vitro transcription. Finally, expression analysis was done by whole-mount in situ hybridization. 2 As a result, female-specific expression was observed for kal1 and ntn1 genes. On the other side, cyp11a1 was initially expressed in both sexes, becoming male specific from stage NF 55.
[EN] Gonadal development of the species Xenopus tropicalis was studied by analyzing the expression pattern of kal1, ntn1 and cyp11a1 genes, three genes with differential gene expression in RNAseq experiments. Furthermore, a comparative analysis of the expression pattern of cyp11a1 and hsd3b1during male gonadal development was performed, as both genes has similar expression patterns in steroidogenic precursor cells. To this end we obtained Xenopus tropicalis eggs by in vivo fertilization; tadpole husbandry and dissection at specific developmental stages were done. Different regions from these genes were PCR-cloned and specific riboprobes were synthetized by in vitro transcription. Finally, expression analysis was done by whole-mount in situ hybridization. 2 As a result, female-specific expression was observed for kal1 and ntn1 genes. On the other side, cyp11a1 was initially expressed in both sexes, becoming male specific from stage NF 55.
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Biología molecular y clínica