Complex cytogenetic analysis of early lethality mouse embryos

Qi Tian, Amy E. Hanlon Newell, Yingming Wang, Susan B. Olson, Lev M. Fedorov

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

An increasing interest in the molecular mechanisms governing cell division has resulted in the discovery of several groups of genes that participate in the regulation of mitosis and meiosis in eukaryotes. Inactivation of these genes in mice often leads to early embryonic lethality. To show direct causality between mutations of these genes, chromosomal instability and embryonic death, a technique enabling detailed cytogenetic analysis of embryonic cells is required. Here, we develop and test a comprehensive approach that allows complex analysis of individual early postimplantation embryos and combines polymerase chain reaction genotyping with the preparation and detailed karyotypic inspection of cells at the metaphase and anaphase stages. The method enables good chromosomal spreading and scattering of nuclei to perform routine cytogenetics (i.e., standard stain and G-banding). It also permits the application of specialized techniques such as fluorescence in situ hybridization to detect particular chromosomes and to verify the integrity of individual chromosomes. Utility of the new method is demonstrated by an analysis of embryonic day E7.5-E9.5 tissue from mice deficient in the spindle checkpoint gene Bub1b.

Original languageEnglish (US)
Pages (from-to)567-574
Number of pages8
JournalChromosome Research
Volume19
Issue number4
DOIs
StatePublished - May 2011

Keywords

  • Chromosomes
  • Mitotic checkpoints
  • Mouse Embryos

ASJC Scopus subject areas

  • Genetics

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