Promotion of gastrulation by maternal growth factor in cultured rabbit blastocysts.
Rabbit blastocysts of day 6 post coitus were cultured in a chemically defined, protein-free medium for 24 h. Although the trophoblast continued to grow, the embryonic disc degenerated. Addition of basic fibroblast growth factor (FGF-2, of human recombinant or bovine origin, 10 ng/ml) to the culture medium resulted in significant developmental progress. The embryonic disc became pear-shaped showing a round anterior edge and a posterior node. The primitive streak and Hensen's node indicated that gastrulation had begun. Mesoderm formation was confirmed from histological sections and by localization of the expression of T-gene transcripts in whole-mount preparations. FGF-2 mRNA was detected in both day-6 endometrium and day 6-blastocysts using in vitro translation followed by immunoprecipitation with a monoclonal antibody to FGF-2. In the uterine secretions of day-6 pregnant and pseudopregnant animals, several proteins exhibiting FGF-2 antigenicity were detected on Western blots following two-dimensional gel electrophoresis. As day-6 blastocysts required exogenous FGF-2 in vitro and as FGF-2 of uterine origin is present in the uterine secretion, the maternal growth factor can promote gastrulation in vivo.
Hrabé de Angelis
M
M
Department of Biology, Philipps University, D-35032 Marburg, Germany.
Gründker
C
C
Herrmann
B G
BG
Kispert
A
A
Kirchner
C
C
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D003470:Culture Media; D004707:Endoderm / Q000187:drug effects; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005775:Gastrula / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D005819:Genetic Markers; D006801:Humans; D008648:Mesoderm / Q000187:drug effects; D011247:Pregnancy; D014176:Protein Biosynthesis; D011817:Rabbits; D011994:Recombinant Proteins / Q000494:pharmacology
D003470:Culture Media; D005819:Genetic Markers; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2