ABSTRACT
We studied the temporal and spatial distribution of the mRNA encoding for thymosin beta 4 (T beta 4), a small acidic actin-sequestering peptide, during the early postimplantation mouse development. Analysis of total embryo RNA demonstrated a strong activation of T beta 4 gene after gastrulation and coincident with neurulation. In situ hybridization showed that T beta 4 mRNA was strongly expressed in the central nervous system and peripheral ganglia, paralleling the gradient of neuronal differentiation. An intense signal was also observed in intraventricular macrophages and blood vessels. The role of T beta 4 in mammalian neuroembryogenesis is discussed.
Subject(s)
Neurons/physiology , Thalamus/embryology , Thymosin/genetics , Animals , Cell Differentiation , Embryonic and Fetal Development , Gene Expression Regulation, Developmental , In Situ Hybridization , Mice , Thalamus/physiologyABSTRACT
The Notch gene of Drosophila encodes a large transmembrane protein involved in cell fate determination during embryonic and larval development. This gene is evolutionarily conserved, and Notch homologs have been cloned from several vertebrate species. To examine the in vivo role of the Notch1 gene, a mouse homolog of Notch, a mutation was introduced by targeted disruption in embryonic stem cells, and these cells were used to generate mutant mice. Intercrosses of animals heterozygous for the Notch1 mutation yielded no live-born homozygous mutant offspring. Homozygous mutant embryos died before 11.5 days of gestation. Morphological and histological analysis of the homozygous mutant embryos indicated that pattern formation through the first nine days of gestation appeared largely normal. However, histological analysis of mutant embryos subsequent to this stage revealed widespread cell death. Death of mutant embryos did not appear to be attributable to defects in placentation or vascularization. Examination of the RNA expression pattern of the Notch2 gene, another Notch gene family member, indicated that it partially overlapped the Notch1 expression pattern. Genetic analysis of the Notch1 mutation also demonstrated that it was not allelic to a mouse mutation described previously, Danforth's short tail (Sd). These results demonstrate that the Notch1 gene plays a vital role during early postimplantation development in mice.
Subject(s)
Embryonic and Fetal Development/physiology , Membrane Proteins/genetics , Mice/genetics , Receptors, Cell Surface , Transcription Factors , Animals , Base Sequence , Biological Evolution , Conserved Sequence , Crosses, Genetic , DNA Primers , Drosophila/embryology , Drosophila/genetics , Embryo Implantation , Embryo, Nonmammalian/physiology , Female , Heterozygote , In Situ Hybridization , Larva , Male , Mice, Inbred C57BL , Mice, Mutant Strains , Molecular Sequence Data , Polymerase Chain Reaction , Receptor, Notch1 , Recombination, Genetic , Restriction Mapping , Stem Cells/cytology , Stem Cells/physiologyABSTRACT
The Notch gene of Drosophila encodes a large transmembrane protein involved in cell-cell interactions and cell fate decisions in the Drosophila embryo. We report here the isolation of cDNA clones encompassing the full-length coding sequence of Notch-1, a mouse homolog of Drosophila Notch. The predicted amino acid sequence of the Notch-1 protein retains all of the conserved amino acid motifs of Notch and the other vertebrate Notch homologs. The cDNA sequence predicts a 2531-amino-acid protein containing a signal peptide, 36 epidermal growth factor-like repeats, 3 Notch/lin-12 repeats, a transmembrane domain, and 6 cdc10/ankyrin repeats. The Notch-1 gene was localized to the proximal portion of mouse chromosome 2 by mapping with an interspecific backcross panel.
Subject(s)
Chromosome Mapping , Insect Hormones/genetics , Membrane Proteins/genetics , Receptors, Cell Surface , Transcription Factors , Amino Acid Sequence , Animals , Cloning, Molecular , DNA , Drosophila , Drosophila Proteins , Female , Male , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Receptor, Notch1 , Receptors, Notch , Sequence Homology, Amino AcidABSTRACT
The Notch gene of Drosophila encodes a large transmembrane protein involved in cell-cell interactions and cell fate decisions in the Drosophila embryo. To determine if a gene homologous to Drosophila Notch plays a role in early mouse development, we screened a mouse embryo cDNA library with probes from the Xenopus Notch homolog, Xotch. A partial cDNA clone encoding the mouse Notch homolog, which we have termed Motch, was used to analyze expression of the Motch gene. Motch transcripts were detected in a wide variety of adult tissues, which included derivatives of all three germ layers. Differentiation of P19 embryonal carcinoma cells into neuronal cell types resulted in increased expression of Motch RNA. In the postimplantation mouse embryo Motch transcripts were first detected in mesoderm at 7.5 days post coitum (dpc). By 8.5 dpc, transcript levels were highest in presomitic mesoderm, mesenchyme and endothelial cells, while much lower levels were detected in neuroepithelium. In contrast, at 9.5 dpc, neuroepithelium was a major site of Motch expression. Transcripts were also abundant in cell types derived from neural crest. These data suggest that the Motch gene plays multiple roles in patterning and differentiation of the early postimplantation mouse embryo.
