Takke C, Campos-Ortega J A
Institut für Entwicklungsbiologie, Universität zu Köln, Germany.
Development. 1999 Jul;126(13):3005-14. doi: 10.1242/dev.126.13.3005.
During vertebrate embryonic development, the paraxial mesoderm becomes subdivided into metameric units known as somites. In the zebrafish embryo, genes encoding homologues of the proteins of the Drosophila Notch signalling pathway are expressed in the presomitic mesoderm and expression is maintained in a segmental pattern during somitogenesis. This expression pattern suggests a role for these genes during somite development. We misexpressed various zebrafish genes of this group by injecting mRNA into early embryos. RNA encoding a constitutively active form of notch1a (notch1a-intra) and a truncated variant of deltaD [deltaD(Pst)], as well as transcripts of deltaC and deltaD, the hairy-E(spl) homologues her1 and her4, and groucho2 were tested for their effects on somite formation, myogenesis and on the pattern of transcription of putative downstream genes. In embryos injected with any of these RNAs, with the exception of groucho2 RNA, the paraxial mesoderm differentiated normally into somitic tissue, but failed to segment correctly. Activation of notch results in ectopic activation of her1 and her4. This misregulation of the expression of her genes might be causally related to the observed mesodermal defects, as her1 and her4 mRNA injections led to effects similar to those seen with notch1a-intra. deltaC and deltaD seem to function after subdivision of the presomitic mesoderm, since the her gene transcription pattern in the presomitic mesoderm remains essentially normal after misexpression of delta genes. Whereas notch signalling alone apparently does not affect myogenesis, zebrafish groucho2 is involved in differentiation of mesodermal derivatives.
在脊椎动物胚胎发育过程中,轴旁中胚层会细分为称为体节的分节单位。在斑马鱼胚胎中,编码果蝇Notch信号通路蛋白同源物的基因在前体节中胚层中表达,并且在体节形成过程中以分节模式维持表达。这种表达模式表明这些基因在体节发育中发挥作用。我们通过向早期胚胎注射mRNA来错误表达该组中的各种斑马鱼基因。测试了编码Notch1a组成型活性形式(Notch1a-intra)和DeltaD截短变体[DeltaD(Pst)]的RNA,以及DeltaC和DeltaD、hairy-E(spl)同源物her1和her4以及groucho2的转录本对体节形成、肌发生以及假定下游基因转录模式的影响。在注射了这些RNA中的任何一种(groucho2 RNA除外)的胚胎中,轴旁中胚层正常分化为体节组织,但未能正确分节。Notch的激活导致her1和her4的异位激活。her基因表达的这种失调可能与观察到的中胚层缺陷有因果关系,因为注射her1和her4 mRNA导致的效应与注射Notch1a-intra所见效应相似。DeltaC和DeltaD似乎在体节中胚层细分后发挥作用,因为在错误表达Delta基因后,体节中胚层中的her基因转录模式基本保持正常。虽然单独的Notch信号显然不影响肌发生,但斑马鱼groucho2参与中胚层衍生物的分化。