ABSTRACT
The Hedgehog signaling pathway is critical for a significant number of developmental patterning events. In this study, we focus on the defects in pharyngeal arch and cardiovascular patterning present in Sonic hedgehog (Shh) null mouse embryos. Our data indicate that, in the absence of Shh, there is general failure of the pharyngeal arch development leading to cardiac and craniofacial defects. The cardiac phenotype results from arch artery and outflow tract patterning defects, as well as abnormal development of migratory neural crest cells (NCCs). The constellation of cardiovascular defects resembles a severe form of the human birth defect syndrome tetralogy of Fallot with complete pulmonary artery atresia. Previous studies have demonstrated a role for Shh in NCC survival and proliferation at later stages of development. Our data suggest that SHH signaling does not act directly on NCCs as a survival factor, but rather acts to restrict the domains that NCCs can populate during early stages (e8.5-10.5) of cardiovascular and craniofacial development.
Subject(s)
Arteries/embryology , Body Patterning , Neural Crest/embryology , Trans-Activators/genetics , Animals , Branchial Region/embryology , Cell Death , Cell Proliferation , Endoderm/physiology , Female , Heart/embryology , Hedgehog Proteins , Intracellular Signaling Peptides and Proteins , Male , Membrane Proteins/biosynthesis , Mice , Mice, Knockout , Neural Crest/cytology , Neural Crest/metabolism , Patched Receptors , Receptors, Cell Surface , Signal TransductionABSTRACT
About 80% of all cases of Fanconi anemia (FA) can be accounted for by complementation groups A and C. To understand the relationship between these groups, we analyzed the expression pattern of the mouse FA group-A gene (Fanca) during embryogenesis and compared it with the known pattern of the group-C gene (Fancc). Northern analysis of RNA from mouse embryos at embryonic days 7, 11, 15, and 17 showed a predominant 4.5 kb band in all stages. By in situ hybridization, Fanca transcripts were found in the whisker follicles, teeth, brain, retina, kidney, liver, and limbs. There was also stage-specific variation in Fanca expression, particularly within the developing whiskers and the brain. Some tissues known to express Fancc (eg, gut) failed to show Fanca expression. These observations show that (1) Fanca is under both tissue- and stage-specific regulation in several tissues; (2) the expression pattern of Fanca is consistent with the phenotype of the human disease; and (3) Fanca expression is not necessarily coupled to that of Fancc. The presence of distinct tissue targets for FA genes suggests that some of the variability in the clinical phenotype can be attributed to the complementation group assignment.
Subject(s)
DNA-Binding Proteins , Embryonic and Fetal Development/genetics , Fanconi Anemia/genetics , Fetal Proteins/biosynthesis , Gene Expression Regulation, Developmental , Protein Biosynthesis , Ameloblasts/metabolism , Animals , Brain/embryology , Brain/metabolism , DNA, Complementary/genetics , Epithelium/metabolism , Extremities/embryology , Eye Proteins/biosynthesis , Eye Proteins/genetics , Fanconi Anemia/pathology , Fanconi Anemia Complementation Group A Protein , Fetal Proteins/genetics , Humans , In Situ Hybridization , Kidney/embryology , Kidney/metabolism , Liver/embryology , Liver/metabolism , Mesoderm/metabolism , Mice , Nerve Tissue Proteins/biosynthesis , Nerve Tissue Proteins/genetics , Odontoblasts/metabolism , Organ Specificity , Phenotype , Proteins/genetics , Vibrissae/embryology , Vibrissae/metabolismABSTRACT
A hemizygous deletion of LIS1, the gene encoding alphaLis1 protein, causes Miller-Dieker syndrome (MDS). MDS is a developmental disorder characterized by neuronal migration defects resulting in a disorganization of the cerebral and cerebellar cortices. alphaLis1 binds to two other proteins (beta and gamma) to form a heterotrimeric cytosolic enzyme which hydrolyzes platelet-activating factor (PAF). The existence of heterotrimers is implicated from copurification and crosslinking studies carried out in vitro. To determine whether such a heterotrimeric complex could be present in tissues, we have investigated whether the alphaLis1, beta, and gamma genes are coexpressed in the developing and adult brain. We have isolated murine cDNAs and show by in situ hybridization that in developing brain tissues alphaLis1, beta, and gamma genes are coexpressed. This suggests that alphaLis1, beta, and gamma gene products form heterotrimers in developing neuronal tissues. In the adult brain, alphaLis1 and beta mRNAs continue to be coexpressed at high levels while gamma gene expression is greatly diminished. This reduction in gamma transcript levels is likely to result in a decline of the cellular concentration of alphaLis1, beta, and gamma heterotrimers. The developmental expression pattern of alphaLis1, beta, and gamma genes is consistent with the neuronal migration defects seen in MDS; regions containing migrating neurons such as the developing cerebral and cerebellar cortices express these genes at a particularly high level. Furthermore, we uncovered a correlation between gamma gene expression, granule cell migration, and PAF hydrolytic activity in the cerebellum. In this tissue gamma gene expression and PAF hydrolysis peaked at Postnatal Days P5 and P15, a period during which neuronal migration in the cerebellum is most extensive. Mechanisms by which PAF could affect neuronal migration are discussed.
Subject(s)
Aging/metabolism , Brain/metabolism , Embryonic and Fetal Development , Gene Deletion , Gene Expression Regulation, Developmental , Microtubule-Associated Proteins , Neurons/physiology , Phospholipases A/biosynthesis , Protein Biosynthesis , Proteins/genetics , 1-Alkyl-2-acetylglycerophosphocholine Esterase , Amino Acid Sequence , Animals , Brain/embryology , Brain/growth & development , Cattle , Cell Movement , Humans , In Situ Hybridization , Mice , Mice, Knockout , Molecular Sequence Data , Organ Specificity , Phospholipases A/metabolism , Proteins/chemistry , RNA, Messenger/biosynthesis , Sequence Homology, Amino Acid , Transcription, GeneticABSTRACT
In this paper we describe the results of genetic analysis of the vestigial locus by studying its interactions with hairy and Notch loci in Drosophila melanogaster. Different vestigial alleles in homo- and heterozygous combination with different hairy alleles show synergism in increasing both cell death and formation of ectopic bristles and produce ectopic veins. Interactions between N and vg also show synergism in increasing cell death and formation of ectopic bristles. Only synergism in cell death is seen between h and N. The interactions indicate that vg product plays a role in the differentiation of epidermal and neural cells of the wing disc by interacting with N and h products either directly or indirectly. Mechanisms of molecular interactions among the three loci are discussed.
