Comparative immunohistolocalization of carbonic anhydrase isozymes I, II and III in the equine and bovine digestive tract

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ABSTRACT. In the present study, we firstly demonstrated immunohistochemical expressions of secretory carbonic anhydrase (CA-VI) isozyme in bovine forestomach, large intestine and major salivary glands. CA-VI was detected in basal layer epithelial cells of esophageal and forestomach stratified epithelium, in mucous cells of upper glandular region of large intestine, in serous acinar cells of the parotid gland, in serous demilune cells and some ductal liner cells of mandibular, monostomatic sublingual and esophageal glands. These immunohistolocalizations suggested that bovine CA-VI plays various roles in pH regulation, maintenance of ion and fluid balance, and cell proliferation. KEY WORDS: bovine digestive organs, CA-VI, immunohistochemical localization.J. Vet. Med. Sci. 68(2): 131-135, 2006 Carbonic anhydrase isozyme-VI (CA-VI) is a unique secretory type CA isozyme. It was initially discovered and purified from ovine saliva [7], and then rat saliva [6,21], human saliva [13,20,22,25] and bovine saliva [5]. Fernley et al. [8] examined sheep CA-VI in 18 different tissues from sheep using biochemical analysis. They observed an immunoreactive signal for CA-VI only in parotid and mandibular glands, which suggested that CA-VI increased buffering capacity in the oral cavity. Some studies have identified the acinar cells of salivary glands as the origin of CA-VI in several species [5,8,21,22,25]. These studies have shown that CA-VI was present in acinar cells of the mammalian parotid and mandibular glands, from where CA-VI is secreted into the saliva. However, CA-VI expression in other segments of the alimentary canal has received little information and the precise physiological role of bovine CA-VI has never been revealed. The present study examines the immunohistochemical expression of CA-VI in bovine alimentary canal and in major salivary glands, and conjectures the function of CA-VI in the bovine digestive organs. MATERIALS AND METHODSTissue: Three adult Holstein cows were used in the present study. Small tissue blocks were prepared for microscopic observation. Samples were cut from the parotid, mandibular, and monostomatic sublingual glands, from cervical to thoracic segments of the esophagus, rumen, reticulum, omasum, abomasum, duodenum, jejunum, ileum, cecum, colon and rectum. The samples were immediately fixed in Bouin's solution, and then sliced into 3 µm-thick sections. CA-VI antibody:We purified CA-VI from bovine saliva as described previously [5]. A rabbit polyclonal antibody has been raised against purified bovine CA-VI and specific immunity to bovine CA-VI has been confirmed [5]. Briefly, the antibody against purified bovine CA-VI was produced in the rabbit. Rabbits were injected initially with 1 mg of purified CA-VI emulsified with an equal volume of Freund's complete adjuvant, followed by a weekly booster injection of a fixed amount of the enzyme for 5 weeks. The rabbits were bled via the auricular vein 10 days following the last injection. The specificity of the antiserum was confirmed by the doubl...

Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Immunohistochemistry of the Bovine Secretory Carbonic Anhydrase Isozyme (CA-VI) in Bovine Alimentary Canal and Major Salivary Glands

Kaseda

Ichihara

Nishita

et al. 2006

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“…We previously reported the immunohistochemical localization of CA-I, CA-II and CA-III in equine salivary glands [2]. We have also immunohistochemically localized CA isozymes to the equine digestive tract [15], but no previous studies have measured CA-I and CA-II levels in equine erythrocytes.…”

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confidence: 99%

Measurement of Erythrocyte Carbonic Anhydrase Isozymes (CA-I and CA-II) in Racehorses and Riding Horses

Nishita

Takahasi

Kasuya

et al. 2005

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ABSTRACT. Equine carbonic anhydrase isozymes (CA-I and CA-II) were purified from erythrocytes by several column chromatography. Polyclonal anti-CA-I and anti-CA-II sera were produced in rabbits. Sensitive competitive enzyme-linked immunosorbent assays (ELISA) were established to determine the developmental changes in CA-I and CA-II levels in equine erythrocytes. Concentrations of CA-I and CA-II in erythrocytes from 150 clinically normal thoroughbreds (123 racehorses and 27 riding horses) were determined by ELISA. Mean (± SD) concentrations of CA-I and CA-II in racehorses were 1.70 ± 0.48 and 0.94 ± 0.13 mg/g hemoglobin (Hb), respectively. Mean concentrations of CA-I and CA-II in riding horses were 2.34 ± 0.52 and 0.76 ± 0.08 mg/g Hb, respectively. When the CA levels in racehorses and riding horses were compared, the CA-I level in riding horses was higher than that in racehorses (p=0.01). The CA-II level in racehorses was higher than that in riding horses (p=0.02). These data suggest that the levels of CA isozymes in erythrocytes of racehorses were influenced by chronic physical stress. The CA-I concentration in erythrocytes of 2-month-old horses was approximately 0.25 mg/g Hb. The CA-I level noticeably increased during the first year of life and approached normal adult levels by 2 years. The CA-II level decreased slightly with age, indicating different regulation of CA-I and CA-II expression during development. KEY WORDS: carbonic anhydrase, equine, ELISA, erythrocyte.

“…The properties of epithelial cells in digestive tract (including expression of CA isoenzymes) vary according to animal species and region of the tract. CA isoenzymes are widely involved in the functions of the digestive tract, and investigators have reported different patterns of expressions of these enzymes in the digestive organs of cattle [6,14], horses [3], rats [4,7,9] and dogs [5], which represent significant variation in diet. However, no reports to date have investigated such expression in pigs.…”

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confidence: 99%

“…Carbonic anhydrase (CA) isoenzymes involved in the hydration of carbon dioxide are ubiquitous in the living world [8,[13][14][15][16][17]. In animals, there are at least 5 carbonic anhydrase isozymes: CA-I, CA-II, CA-III, CA-IV, and CA-VI, each with its own molecular structure.…”

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confidence: 99%

Immunohistochemical Localization of Carbonic Anhydrase Isoenzymes in Salivary Gland and Intestine in Adult and Suckling Pigs.

Amasaki

Nagasao

et al. 2001

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ABSTRACT. Localizations of carbonic anhydrase isoenzymes (CA I, CA II and CA III) were investigated immunohistochemically in the salivary glands and intestine of mature and suckling pigs. Carbonic anhydrase isoenzymes were not detected in the salivary glands of sucklings, but were present in the adult. Bicarbonate ion in saliva might be important for the digestion of solid foods in mature pigs, but unnecessary for the digestion of milk in sucklings. Expressions of CAI and CAII were detected strongly in the large intestine of the adult and sucklings, and faintly only at duodenum in the small intestine. CA I and CA II isoenzymes in the large intestine may be involved, at least in part, in ion absorption and water metabolism during digestion and absorption of milk in suckling pigs. In addition, CA I and CA II expression in the duodenal villus enterocyte may support the process of bicarbonate absorption in the duodenum. KEY WORDS: adult and suckling pig, carbonic anhydrase, intestine, salivary gland.