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Am J Vet Res. 2002 Feb;63(2):229-35.

Isolation and measurement of carbonic anhydrase isoenzyme III in plasma, sera, and tissues of dogs.

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  • 1Laboratories of Veterinary Physiology, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan.

Abstract

OBJECTIVE:

To purify canine carbonic anhydrase isoenzyme III (CA-III) and determine plasma, serum, and tissue concentrations of CA-III in healthy dogs and dogs with experimentally induced muscle damage.

ANIMALS:

121 healthy Beagles.

PROCEDURE:

Muscle was obtained from 2 Beagles after euthanasia, and CA-III was purified and characterized by use of column chromatography and electrophoresis, respectively. A CA-III-specific ELISA was developed to determine concentrations of CA-III in plasma of 116 dogs and tissues of 1 dog. Serum creatine kinase (CK) activity and CA-III concentration were also determined before and after induction of muscle damage by IM injection of 2 ml of 10% lidocaine to 2 dogs.

RESULTS:

Canine CA-III had a molecular weight of 28 kd and an isoelectric point of 8.2. Mean (+/- SD) concentration of CA-III in plasma of healthy dogs was 16.91 +/- 9.55 ng/ml. The highest tissue concentration of CA-III was detected in skeletal muscle. Serum concentration of CA-III increased and peaked within the first 2 to 3 hours after induction of muscle damage. The increase in CA-III concentration was more rapid than that of CK activity, and concentration reached its maximum and returned to baseline sooner than did CK activity.

CONCLUSIONS AND CLINICAL RELEVANCE:

The CA-III ELISA we developed was a sensitive method for determining CA-III concentrations in plasma, serum samples, and tissue specimens of dogs. Use of this ELISA requires only a small volume of serum and may enable the study of changes in CA isoenzyme concentrations associated with muscle disorders in dogs.

PMID:
11843123
[PubMed - indexed for MEDLINE]
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