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


METHODS
121 healthy Beagles.


METHODS
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
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.
Nishita
Toshiho
T
Laboratories of Veterinary Physiology, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan.
Matsuura
Katsuko
K
Ichihara
Nobutune
N
Asari
Masao
M
D000818:Animals; D024403:Carbonic Anhydrase III / Q000032:analysis / Q000302:isolation & purification*; D002850:Chromatography, Gel / Q000662:veterinary; D004285:Dogs / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D008297:Male; D009132:Muscles / Q000201:enzymology; D011817:Rabbits; D012016:Reference Values
D024403:Carbonic Anhydrase III