Development and analysis of a Fanconi anemia group A mouse model.
Published: 2004
Language: eng
Number of pages: 161
Description: Fanconi Anemia (FA) is an autosomal recessive disorder characterized by cellular hypersensitivity to DNA cross-linking agents. Despite the cloning of six disease-associated genes for FA and the identification of  BRCA2 as the gene mutated in complementation groups B and D1, the precise role of the FA pathway remains largely unknown. The mouse homolog of the human FANCA cDNA was cloned and characterized to facilitate the study of FA complementation group A using the mouse as a model system. The mouse cDNA (Fanca) encodes a 161-kDa protein that shares 65% amino acid sequence identity with human FANCA. Expression of the mouse cDNA in human FA-A cells restores the cellular drug sensitivity to normal levels, affirming that the function of FANCA is conserved in the mouse. To study the in vivo  role of Fanca, gene-targeting techniques were used to generate Fancatm1Hsc mice in which  Fanca exons 1 to 6 were replaced by a beta-galactosidase reporter gene. Fancatm1.1Hsc mice were then generated by Cre-mediated removal of the neomycin selection cassette of Fanca tm1Hsc mice. Fancatm1.1Hsc homozygotes displayed FA-like phenotypes including hypogonadism, growth retardation, microphthalmia, and bone marrow hypersensitivity to mitomycin C. Manifestation of specific phenotypes, including microphthalmia and hypogonadism, was affected by the genetic background. Since germ cell development in  Fancatm1.1Hsc homozygotes was clearly abnormal, it was investigated in detail. Diminished populations of primordial germ cells in the gonadal ridges were apparent by E11.5 in Fancatm1.1Hsc  homozygotes, leading to a reduced germ cell reserve and premature reproductive senescence. Very high levels of Fanca expression was observed in pachytene spermatocytes, and spermatocytes from  Fancatm1Hsc homozygous males exhibited an elevated frequency of mispaired meiotic chromosomes and increased apoptosis, implicating a previously unrecognized role for Fanca in meiotic recombination. However, the localization of proteins that associate with the meiotic chromosomes during meiotic recombination, including Rad51, Brca1, Fancd2 and Mlh1, appeared normal on Fancatm1Hsc homozygous meiotic chromosomes. Taken together, these results emphasize that the FA pathway plays a role in the maintenance of reproductive germ cells and in meiotic recombination. These findings document the utility of Fancatm1.1Hsc mice as an in vivo model for the study of FA.
Notes: Adviser:  Manuel Buchwald.

Thesis (Ph.D.)--University of Toronto, 2004.

Electronic version licensed for access by U. of T. users.

Source: Dissertation Abstracts International, Volume: 65-05, Section: B, page: 2221.