Buttgereit D, Paululat A, Renkawitz-Pohl R
FB Biologie, Marburg, Germany.
Int J Dev Biol. 1996 Feb;40(1):189-96.
In Drosophila beta tubulins are encoded by a small gene family whose members are differentially expressed in a highly cell and tissue specific manner. Here we focus on the expression of the beta 3 tubulin isotype during mesoderm differentiation and beta 1 tubulin expression in the apodemes during embryonic development. The beta 3 tubulin isotype is first detectable at the extended germband stage shortly before the separation of somatic and visceral derivatives. Comparing the distribution of the beta 3 mRNA and the beta 3 isotype shows that the transcription of the beta 3 tubulin gene is cell type specifically repressed during differentiation of individual mesodermal derivatives, from which the dorsal vessel remains transcriptionally active until shortly before hatching. In contrast the beta 3 tubulin protein is detectable in all mesodermal derivatives. The beta 3 tubulin is an excellent marker to study mesoderm differentiation on a regulatory and cellular level using both genetics and molecular biology. In the visceral mesoderm, the expression of the beta 3 tubulin gene is regulated by homeotic gene products, while other transactivators regulate expression in the dorsal vessel and the body wall musculature. In the somatic mesoderm, the beta 3 tubulin allows to visualize myotube formation and insertion into the epidermis. This contact to the epidermal attachment sites (apodemes) induces beta 1 tubulin expression, as can be seen in double staining experiments. We determined a 14bp cis-regulatory enhancer element guiding expression of the beta 1 tubulin gene in these attachment sites. Using the beta 1 and beta 3 tubulin isotypes as markers we started to isolate mutants which are disturbed in muscle formation.
在果蝇中,β微管蛋白由一个小基因家族编码,其成员以高度细胞和组织特异性的方式差异表达。在这里,我们重点关注中胚层分化过程中β3微管蛋白亚型的表达以及胚胎发育过程中表皮内突中β1微管蛋白的表达。β3微管蛋白亚型在体壁和内脏衍生物分离前不久的伸长胚带期首次可检测到。比较β3 mRNA和β3亚型的分布表明,在各个中胚层衍生物的分化过程中,β3微管蛋白基因的转录受到细胞类型特异性抑制,其中背血管在孵化前不久一直保持转录活性。相比之下,β3微管蛋白在所有中胚层衍生物中均可检测到。β3微管蛋白是利用遗传学和分子生物学在调控和细胞水平上研究中胚层分化的优秀标记物。在内脏中胚层中,β3微管蛋白基因的表达受同源异型基因产物调控,而其他反式激活因子则调控背血管和体壁肌肉组织中的表达。在体壁中胚层中,β3微管蛋白可使肌管形成和插入表皮可视化。如双重染色实验所示,与表皮附着位点(表皮内突)的这种接触诱导β1微管蛋白表达。我们确定了一个14bp的顺式调控增强子元件,它指导β1微管蛋白基因在这些附着位点的表达。我们以β1和β3微管蛋白亚型作为标记物开始分离在肌肉形成方面受到干扰的突变体。