Differential expression of the GTL2 gene within the callipyge region of ovine chromosome 18.
The inheritance pattern of the skeletal muscle hypertrophy phenotype caused by the callipyge gene has been characterized as polar overdominance. We hypothesized that this trait may be caused by a gain or loss of gene expression because of the reversible nature of the phenotype in paternal vs. maternal inheritance. Suppression subtraction cDNA probes were made from skeletal muscle mRNA of normal (NN) and callipyge (C(Pat)N(Mat)) animals and hybridized to Southern blots containing bacterial artificial chromosomes (BACs) that comprise a physical contig of the callipyge region. The CN-NN probes hybridized to two ovine and seven bovine BACs. Sequence analysis of fragments within those BACs indicated short regions of similarity to mouse gene trap locus (gtl2). Northern blots analysis of RNA from hypertrophy-responsive muscles show a population of GTL2 mRNA centred around 2.4 kb that were abundantly expressed in 14-day prenatal NN and C(Pat)N(Mat) lambs but were down-regulated in day 14 and day 56 postnatal NN lambs. The expression of GTL2 remained elevated in 14- and 56-day-old C(Pat)N(Mat) lambs as well as in 56-day-old N(Pat)C(Mat) and CC lambs. Expression of GTL2 in the supraspinatus, which does not undergo hypertrophy, was very low for all genotypes and ages. Isolation of cDNA sequences show extensive alternative splicing and a lack of codon bias suggesting that GTL2 does not encode a protein. The mutation of the callipyge allele has altered postnatal expression of GTL2 in muscles that undergo hypertrophy and will help identify mechanisms involved in growth, genomic imprinting and polar overdominance.
Bidwell
C A
CA
Department of Animal Sciences, Purdue University, West Lafayette, IN 49707-1026, USA. cbidwell@purdue.edu
Shay
T L
TL
Georges
M
M
Beever
J E
JE
Berghmans
S
S
Cockett
N E
NE
D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002417:Cattle; D002874:Chromosome Mapping; D022202:Chromosomes, Artificial, Bacterial; D015342:DNA Probes; D018076:DNA, Complementary; D005260:Female; D005786:Gene Expression Regulation; D008297:Male; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000254:growth & development / Q000378:metabolism / Q000473:pathology*; D009154:Mutation; D022661:RNA, Untranslated / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D012756:Sheep / Q000235:genetics*; D012757:Sheep Diseases / Q000235:genetics* / Q000473:pathology
D015342:DNA Probes; D018076:DNA, Complementary; D022661:RNA, Untranslated