|Li, Ming - UNIVERSITY OF MINNESOTA|
Submitted to: Biology of Reproduction Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: April 23, 1997
Publication Date: N/A
Technical Abstract: Expression of FSHbeta was evaluated by an RNASE protection assay (RPA) that protected a fragment of 205 nucleotides at positions 759-963. When pituitaries of 24 crossbred boars (1/4 MS and 3/4 WC x 3/4 MS and 1/4 WC) were evaluated with this assay, the expected fragment was not detected in one boar; however, this boar had moderate plasma and pituitary FSH concentrations. To understand this unexpected observation, cDNA that had been reverse-transcribed from the anterior pituitary RNA of this boar was amplified by PCR with the same FSHbeta specific primers used for the creation of the riboprobe. The amplified RT-PCR product was subcloned, sequenced & determined to be lacking an 11 bp fragment (5'-CATTTGGAAAC-3') at nucleotide positions 807-817 of the 3'-untranslated region of FSHbeta mRNA (defined as deleted FSHbeta; delta) in comparison with the reported FSHbeta (defined as wild type; +) gene. These 24 crossbred boars were screened by RT-PCR and found that 8 were +/+, 15 were +/delta, and 1 was delta/delta. Straightbred pigs of MS and WC were examined at both the pituitary RNA and genomic DNA levels. Wild type FSHbeta mRNA was present in both breeds (4/6 MS vs. 6/6 WC) while the deleted FSHbeta mRNA was present only in MS pigs (P<0.01; 6/6 MS vs. 0/6 WC). We further demonstrated that these two FSHbeta mRNA species were transcribed from different FSHbeta alleles rather than from alternative splicing of a single gene (allele), and these two alleles segregated in Mendelian fashion. Also, we found that this deleted FSHbeta allele was present in two other Chinese pig breeds (Fengjing:3/5 and Minzhu:5/8). In summary, we identified a novel FSHbeta allele in Chinese MS pigs; this allele was uniquely present in other Chinese pig breeds but absent in WC pigs.