a contracted dna repeat in lhx3 intron 5 is associated with aberrant splicing and pituitary dwarfism in german shepherd dogs简约的dna重复lhx3内含子5与德国牧羊犬异常剪接和垂体性侏儒症.pdfVIP

A Contracted DNA Repeat in LHX3 Intron 5 Is Associated with Aberrant Splicing and Pituitary Dwarfism in German Shepherd Dogs 1 1 1 1 Annemarie M. W. Y. Voorbij , Frank G. van Steenbeek , Manon Vos-Loohuis , Ellen E. C. P. Martens , 1 2 1 1 Jeanette M. Hanson-Nilsson , Bernard A. van Oost , Hans S. Kooistra , Peter A. Leegwater * 1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands, 2 Department of Biochemistry, American University of the Caribbean, Cupecoy, St. Maarten, Netherlands Antilles Abstract Dwarfism in German shepherd dogs is due to combined pituitary hormone deficiency of unknown genetic cause. We localized the recessively inherited defect by a genome wide approach to a region on chromosome 9 with a lod score of 9.8. The region contains LHX3, which codes for a transcription factor essential for pituitary development. Dwarfs have a deletion of one of six 7 bp repeats in intron 5 of LHX3, reducing the intron size to 68 bp. One dwarf was compound heterozygous for the deletion and an insertion of an asparagine residue in the DNA-binding homeodomain of LHX3, suggesting involvement of the gene in the disorder. An exon trapping assay indicated that the shortened intron is not spliced efficiently, probably because it is too small. We applied bisulfite conversion of cytosine to uracil in RNA followed by RT-PCR to analyze the splicing products. The aberrantly spliced RNA molecules resulted from either skipping of exon 5 or retention of intron 5. The same splicing defects were observed in cDNA derive