Measurement of carbonic anhydrase isozyme VI (CA-VI) in bovine sera, saliva, milk and tissues.
Concentrations of bovine carbonic anhydrase isozyme VI (CA-IV) in bovine serum, saliva, normal milk, colostrum, submandibular gland, liver, and mammary gland were determined. CA-VI was purified from bovine saliva and an antibody to CA-VI was generated. The concentrations of CA-VI in the saliva (7.8 +/- 7.9 microg/ml), serum (2.1+/- 5.7 ng/ml), milk (7.9 +/- 12.1 ng/ml), submandibular gland (284.7 microg/g protein), liver (921.0 +/- 180.7 ng/g protein) and mammary gland (399.6 +/- 191.2 ng/g protein) were determined by ELISA. No seasonal change in CA-VI levels was observed in normal milk. The concentration of CA-VI in colostrum (day 1 post partum) was 119 ng/ml and decreased rapidly by 1 month following birth. Mammary gland contained much smaller amounts than the submandibular gland. CA-VI mRNA was detected in the liver and mammary gland of cow by RT-PCR. The ELISA used in this study proved to be a precise and sensitive method for determining CA-VI concentrations in saliva, serum, milk and tissue specimens from cows. The ELISA may enable the study of changes in CA-VI associated with hereditary or metabolic disorders of the salivary gland, mammary gland and liver using small samples of saliva, serum or milk.
Nishita
T
T
Laboratory of Veterinary Physiology 1, School of Veterinary Medicine, Azabu University, Kanagawa, Japan. nishida@azabu-u.ac.jp
Tanaka
Y
Y
Wada
Y
Y
Murakami
M
M
Kasuya
T
T
Ichihara
N
N
Matsui
K
K
Asari
M
M
D000818:Animals; D002256:Carbonic Anhydrases / Q000032:analysis* / Q000097:blood / Q000235:genetics; D002417:Cattle / Q000378:metabolism*; D003126:Colostrum / Q000201:enzymology; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D005260:Female; D008099:Liver / Q000201:enzymology; D008321:Mammary Glands, Animal / Q000201:enzymology; D008892:Milk / Q000201:enzymology*; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012463:Saliva / Q000201:enzymology*; D013363:Submandibular Gland / Q000201:enzymology
D012313:RNA; D002256:Carbonic Anhydrases; C477051:carbonic anhydrase VI