Isolation and characterization of a Xenopus gene (XMLP) encoding a MARCKS-like protein

We have identified a cDNA coding for a Xenopus MARCKS-like protein (XMLP) from a cDNA library prepared from activin-treated ectoderm. Using whole-mount in situ hybridization and RT-PCR, we found XMLP maternal transcripts during the cleavage stages. After MBT, the signals were restricted to the neural plate. Subsequently XMLP was expressed predominantly in the brain, somites and pronephros. Ectopic expression of XMLP resulted in eye and axis defects and in a change of the expression pattern of Krox 20, a neural marker for rhombomeres 3 and 5. Injected XMLP caused apoptosis. It was characterized by loss of intercellular adhesion contacts, transient plasma membrane ruffling at gastrula, and epithelial disruption attailbud stage. Overexpression of mutant XMLPs showed that this phenotype was correlated with its putative PSD domain and glycine at position 2. The embryos injected with a morpholino oligo complementaryto XMLPmRNA showed malformations of the anterior axis and eye defects. Extirpation experiments indicated that the phenotypes might be correlated with disturbed morphorgenetic movements rather than an inhibition of induction process. Overexpression of XCYP26 resulted in a shift of the expression pattern of XMLP. In the early tailbud stage (stage 20) the signal stripe in the XCYP26 injected half of the embryo got diffuse or even disappeared. This observation suggests that retinoic acid plays an important role in the regulation of XMLP. Our results suggest that XMLP might participate in pattern formation of the embryonic axis and the central nervous system.