RESUMO
In long germ embryos, all body segments are specified simultaneously during the blastoderm stage. In contrast, in short germ embryos, only the anterior segments are specified during the blastoderm stage, leaving the rest of the body plan to be specified later. The striking embryological differences between short and long germ segmentation imply fundamental differences in patterning at the molecular level. To gain insights into the segmentation mechanisms of short germ insects, we have investigated the role of the homologue of the Drosophila gap gene hunchback (hb) in a short germ insect Locusta migratoria manilensi by paternal RNA interference (RNAi). Phenotypes resulting from hb knockdown were categorized into three classes based on severity. In the most extreme case, embryos developed the most anterior structures only, including the labrum, antennae and eyes. The following conclusions were drawn: (i) L. migratoria manilensis hb (Lmm'hb) controls germ band morphogenesis and segmentation in the anterior region; (ii) Lmm'hb may function as a gap gene in a wide domain including the entire gnathum and thorax; and (iii) Lmm'hb is required for proper growth of the posterior germ band. These findings suggest a more extensive role for L. migratoria manilensis hunchback in anterior patterning than those described in Drosophila.
Assuntos
Padronização Corporal/fisiologia , Proteínas de Insetos/fisiologia , Locusta migratoria/fisiologia , Interferência de RNA/fisiologia , Fatores de Transcrição/fisiologia , Animais , Padronização Corporal/genética , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Embrião não Mamífero/embriologia , Embrião não Mamífero/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Locusta migratoria/embriologia , Locusta migratoria/genética , Masculino , Dados de Sequência Molecular , Fenótipo , Análise de Sequência de DNA , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Dedos de ZincoRESUMO
The transfection of mutated human insulin gene was studied using the complex of different proportion of plasmid and Liposome. Hepatic cell was used as the target cell in vitro,isolation of Hepatic cell including insulin gene was carried out by G418, the expression level of insulin in medium was measured by RIA method. The portal vein was cannulated with therapeutic gene in vivo, the blood glucose of the model was regularly examined and the insulin level was detected on the seventh day after transfection. The results showed that the target gene was transferred into the hepatic cell, expression of mature insulin was detected both in vivo and in vitro. It reached the peak 10.45microIU/ml on the 24th hour after transfection with the proportion 1:6 of plasmid and Liposome in hepatic cell. Diabetic symptom of the model was improved after transgene, the blood glucose could decrease 55% at the most.
