|Budget Amount *help
¥3,200,000 (Direct Cost : ¥3,200,000)
Fiscal Year 1998 : ¥1,400,000 (Direct Cost : ¥1,400,000)
Fiscal Year 1997 : ¥1,800,000 (Direct Cost : ¥1,800,000)
The bfp (brain finger protein) is a member of the RING finger protein family which is highly expressed in the brain. We have previously shown that one copy of the human bfp gene, mapped at l7pll.2, was actually deleted in 6 out of 6 Smith-Magenis syndrome (SMS) patients. Now we have isolated the mouse bfp cDNA.Using in situ hybridization and irnmunohistochemistry, the distribution of mouse bfp mRNA and protein was especially identified in neural cells of the cerebral cortex, hippocampus, lateral amygdaloid nucleus and ventromedial hypothalamus (VMH). In primary culture of the whole brain in a neonatal mouse, the bfp protein was detected in both the neuron and glial cells and its subcellular localization was predominantly in the nucleus but some amounts were also found in the cytoplasm. The bfp mRNA was also expressed strongly on the marginal zone of brain vesicles, optic stalk, cartilage primordium, which are part of the critical tissues frequently involved in SMS patients, and such ti
ssues as nasal epithelium and primordium of follicles in a 13.5 d.p.c. embryo. Subsequently, its amount in the developing brain further increased during embryogenesis reaching the highest level in the adult brain. These findings suggest a possibility that bfp might be involved in the pathogenesis of SMS as a regulator protein related to the neural differentiation and function. This time we tried to generate the mice in which the bfp gene was deleted, in order to clarify biological functions of bfp and the relation with SMS.Jl ES cells were electroporated with the targeting vector and selected using G418.
Targeted ES clones got by homologous recombination were identified by Southern blot analysis using the probe a and the probe neo. Germ-line transmission of the bfp mutated allele was obtained for four independent targeted clones and heterozygous breeding was set up for Fl animals derived from each clone to produce homozygous mutants. The genotype of F2 offspring was performed by Southern blot and PCR analyses. Now analyses of phenotypes in bfp deficient mice are done. Less