www.fgks.org   »   [go: up one dir, main page]

Purification of swine carbonic anhydrase isoenzyme III and measurement of its levels in tissues and plasma

J Anim Physiol Anim Nutr (Berl). 2014 Feb;98(1):119-27. doi: 10.1111/jpn.12054. Epub 2013 Feb 8.

Abstract

The changes in the levels of carbonic anhydrase isozyme III (CA-III) in swine plasma and urine have not been previously determined or reported. CA-III is relatively specific to skeletal muscles, and should therefore be a useful diagnostic marker for muscle diseases. We isolated CA-III from swine muscle tissues and determined CA-III levels in the plasma and urine from both healthy and diseased pigs. The levels of CA-III in the tissues of female swine (age, 3 months) and plasma of young swine (age, 1-5 months) and adult female pigs (age, 2-3 years) were determined using the ELISA system for swine CA-III. The mean (± SD) levels of CA-III in the skeletal muscles were 3.8 ± 3.2 mg/g (wet tissue), and in the plasma, 230 ± 193 ng/ml at 1 month, 189 ± 208 ng/ml at 2 months, 141 ± 148 ng/ml at 3 months, 78 ± 142 ng/ml at 4 months and 53 ± 99 ng/ml at 5 months. The mean level of CA-III in the plasma samples from 2- to 3-year-old pigs was 18 ± 60 ng/ml. CA-III in the plasma samples was found to decrease from 1 month until 3 years of age (p < 0.01). We performed far-western blotting to clarify the cause of the observed decrease in CA-III in plasma. Our results demonstrated that CA-III is bound to the transferrin and albumin. In addition, we determined that the levels of CA-III in plasma and urine samples were higher in diseased swine compared with the healthy pigs.

Keywords: ELISA; carbonic anhydrase III; far-western blotting; muscle; serum; swine.

MeSH terms

  • Aging
  • Animals
  • Blotting, Western / veterinary
  • Carbonic Anhydrase III / genetics
  • Carbonic Anhydrase III / isolation & purification
  • Carbonic Anhydrase III / metabolism*
  • Female
  • Gene Expression Regulation, Enzymologic / physiology*
  • Muscle, Skeletal / enzymology*
  • Swine / blood*
  • Swine / metabolism
  • Swine Diseases / blood*
  • Swine Diseases / metabolism

Substances

  • Carbonic Anhydrase III