Cloning and functional characterization of two key enzymes of glycosphingolipid biosynthesis in the amphibian Xenopus laevis.
Gangliosides are a subfamily of complex glycosphingolipids (GSLs) with important roles in many biological processes. In this study, we report the cDNA cloning, functional characterization, and the spatial and temporal expression of Xlcgt and Xlgd3 synthase during Xenopus laevis development. Xlcgt was expressed both maternally and zigotically persisting at least until stage 35. Maternal Xlgd3 synthase mRNA could not be detected and showed a steady-state expression from gastrula to late tailbud stage. Xlcgt is mainly present in involuted paraxial mesoderm, neural folds, and their derivatives. Xlgd3 synthase transcripts were detected in the dorsal blastoporal lip, in the presumptive neuroectoderm, and later in the head region, branchial arches, otic and optic primordia. We determined the effect of glycosphingolipid depletion with 1-phenyl-2-palmitoyl-3-morpholino-1-propanol (PPMP) in mesodermal layer. PPMP-injected embryos showed altered expression domains in the mesodermal markers. Our results suggest that GSL are involved in convergent-extension movements during early development in Xenopus.
Luque
Melchor E
ME
CONICET, UNT, INSIBIO, Departamento de Biología del Desarrollo, San Miguel de Tucumán, Argentina.
Crespo
Pilar M
PM
Mónaco
María E
ME
Aybar
Manuel J
MJ
Daniotti
José L
JL
Sánchez
Sara S
SS
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D004798:Enzymes / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D005964:Glucosyltransferases / Q000145:classification / Q000235:genetics / Q000378:metabolism; D006028:Glycosphingolipids / Q000096:biosynthesis*; D017403:In Situ Hybridization; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009025:Morpholines / Q000494:pharmacology; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012799:Sialyltransferases / Q000145:classification / Q000235:genetics / Q000378:metabolism; D013107:Sphingolipids / Q000494:pharmacology; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism*
C095823:1-phenyl-2-palmitoylamino-3-morpholino-1-propanol; D018076:DNA, Complementary; D004798:Enzymes; D006028:Glycosphingolipids; D009025:Morpholines; D013107:Sphingolipids; D029867:Xenopus Proteins; D005964:Glucosyltransferases; C023246:ceramide glucosyltransferase; D012799:Sialyltransferases; C043662:alpha-N-acetylneuraminate alpha-2,8-sialyltransferase