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Calpains are non-lysosomal proteases involved in myofibrillar protein degradation. To facilitate studying the expression of the porcine calpain genes and their influence on protein accretion, we have cloned partial cDNAs for mu- and m-calpain from porcine skeletal muscle via PCR amplification. A 289 bp fragment for mu-calpain and a 629 bp fragment for m-calpain were cloned into the EcoRV site of pBluescript II KS+ vector. The nucleotide sequence for porcine mu-calpain and m-calpain were 92% and 90% identical to corresponding regions of rabbit mu- and m-calpain, respectively. The deduced amino acid sequences for both mu- and m-calpain share 94% identity with respective rabbit mu- and m-calpains. Isoform specificity was validated by Southern hybridization of mu- and m-calpain probes with cloned mu- and m-calpain fragments and Northern hybridization with pig muscle mRNA. These clones will be used to evaluate the role of calpain expression in muscle hypertrophy.

Aphidicolin (APC)-sensitive fragile sites were identified in chromosome preparations from peripheral lymphocyte cultures of 12 four-way crossbred pigs. A chi 2 analysis demonstrated that aphidicolin-induced breakage events and previously reported in vivo chromosome rearrangement events were not independent. Comparison of expected Poisson and negative binomial distributions to observed breakage patterns indicated that the negative binomial distribution provided a better fit to experimental data. The negative binomial distribution is consistent with a distribution of breakage rates, i.e., non-constant rates of breakage at chromosomal loci across the genome.

Vitellogenesis was induced in white sturgeon by administration of estrogen through silastic implants. Vitellogenin mRNA was identified by agarose gel electrophoresis and cell-free translation. A highly abundant 5.7-kb mRNA was induced in the liver of estrogen-treated sturgeon. Cell-free translation of poly(A)+ mRNA showed the induction of two high-molecular-weight proteins of 180 and 120 kDa. These two proteins, encoded by the 5.7-kb mRNA(s), were immunoprecipitated by antiserum to serum from vitellogenic sturgeon. Immunoprecipitations also showed the presence of four other serum proteins synthesized by the liver of estrogen-treated sturgeon. The induction of vitellogenesis by estrogen in sturgeon, which are a primitive teleost, was found to be similar to induction of vitellogenesis in amphibians, avians, and other teleosts. Estrogen treatment induced a highly abundant vitellogenin mRNA as well as several mRNAs for other serum proteins. However, the presence of two distinct vitellogenin monomers in the cell-free translation assay was significantly different from the results in other species.