RESUMEN
BACKGROUND: Our interests lie in determining the genes and genetic pathways that are important for establishing and maintaining maternal-fetal interactions during pregnancy. Mutation analysis targeted to a 34 Mb domain flanked by Trp53 and Wnt3 demonstrates that this region of mouse chromosome 11 contains a large number of essential genes. Two mutant alleles (l11Jus1 and l11Jus4), which fall into the same complementation group, survive through implantation but fail prior to gastrulation. RESULTS: Through a positional cloning strategy, we discovered that these homozygous mutant alleles contain non-conservative missense mutations in the Notchless homolog 1 (Drosophila) (Nle1) gene. NLE1 is a member of the large WD40-repeat protein family, and is thought to signal via the canonical NOTCH pathway in vertebrates. However, the phenotype of the Nle1 mutant mice is much more severe than single Notch receptor mutations or even in animals in which NOTCH signaling is blocked. To test the hypothesis that NLE1 functions in multiple signaling pathways during pre-implantation development, we examined expression of multiple Notch downstream target genes, as well as select members of the Wnt pathway in wild-type and mutant embryos. We did not detect altered expression of any primary members of the Notch pathway or in Notch downstream target genes. However, our data reveal that Cdkn1a, a NOTCH target, was upregulated in Nle1 mutants, while several members of the Wnt pathway are downregulated. In addition, we found that Nle1 mutant embryos undergo caspase-mediated apoptosis as hatched blastocysts, but not as morulae or blastocysts. CONCLUSIONS: Taken together, these results uncover potential novel functions for NLE1 in the WNT and CDKN1A pathways during embryonic development in mammals.
Asunto(s)
Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Desarrollo Embrionario/genética , Ratones/genética , Proteínas de Microfilamentos/genética , Preñez/genética , Vía de Señalización Wnt , Animales , Etilnitrosourea/toxicidad , Femenino , Regulación del Desarrollo de la Expresión Génica , Mutagénesis , Mutación , EmbarazoRESUMEN
Polymorphisms such as single-nucleotide polymorphisms (SNPs) and insertions/deletions (Indels) can be associated with phenotypic traits and be used as markers for disease diagnosis. Identification of these genetic variations within laboratory mice is crucial to improve our understanding of the genetic background of the mice used for research. As part of a positional cloning project, we sequenced six genes (Mettl16, Evi2a, Psmd11, Cct6d, Rffl, and Ap2b1) within a 6.8-Mb domain of mmu chr 11 in the C57BL/6J and 129S6/SvEvTac inbred strains. Although 129S6/SvEvTac is widely used in the mouse community, there is very little current (or projected future) sequence information available for this strain. We identified 6 Indels and 21 novel SNPs and confirmed genotype information for 114 additional SNPs in these 6 genes. Mettl16 and Ap2b1 contained the largest numbers of variants between the C57BL/6J and 129S6/SvEvTac strains. In addition, we found five new SNPs between 129S6/SvEvTac and 129S1/SvImJ within the Ap2b1 locus. Although we did not detect differences between C57BL/6J and 129S6/SvEvTac within Evi2a, this locus contains a relatively high SNP density compared with the surrounding sequence. Our study highlights the genetic differences among three inbred mouse strains (C57BL/6J, 129S6/SvEvTac, and 129S1/SvImJ) and provides valuable sequence information that can be used to track alleles in genomics-based studies.
Asunto(s)
Cromosomas de los Mamíferos , Polimorfismo Genético , Animales , Secuencia de Bases , Cruzamiento , Ratones , Ratones EndogámicosRESUMEN
Five strains representing a novel family within the Gammaproteobacteria were isolated from the estuarine grasses Spartina alterniflora and Juncus roemerianus. All strains were facultatively anaerobic, Gram-negative, short, motile, polar monotrichous rods that were mesophilic, oxidase-negative, catalase-positive, had DNA G+C contents of 41.5-44.4 mol% and required seawater salts or NaCl. Growth was observed at pH 3.5-8.0. Polar lipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, aminophospholipid, phospholipids and unidentified aminolipids were found in the representative strain S-G2-2(T). The major menaquinone and ubiquinone were MK-8 (100 %) and Q-8 (93 %), respectively. Predominant fatty acids present were C(12 : 0) aldehyde and/or unknown fatty acid 10.9525 (MIDI designation) and/or iso-C(16 : 1) I/C(14 : 0) 3-OH, C(16 : 1)ω7c/C(16 : 1)ω6c, C(16 : 0), C(17 : 0) cyclo and C(18 : 1)ω7c and/or C(18 : 1)ω6c. The nearly full-length 16S rRNA gene sequences of the strains were very similar (99-100 % similarity), and the strains were identified as members of the same species by DNA-DNA relatedness measurements. 16S rRNA gene sequence analysis revealed that the strains formed a monophyletic lineage within the order Alteromonadales. All five strains fixed N(2). Analysis of partial nifH gene sequences also revealed a monophyletic lineage within the Gammaproteobacteria, and the sequences were dissimilar to those of any previously described diazotroph. Differences between the novel strains and other members of the Alteromonadales include the inability to produce cytochrome oxidase. The novel strains were metabolically versatile. On the basis of the information described above, the new genus and species Celerinatantimonas diazotrophica gen. nov., sp. nov. are proposed to accommodate the five strains within a new family, Celerinatantimonadaceae fam. nov. The type strain of Celerinatantimonas diazotrophica is S-G2-2(T) (â=âATCC BAA-1368(T) â=âDSM 18577(T)).