Distribution of Carbonic Anhydrase Isozyme VI in the Developing Bovine Parotid Gland

Asari, Masao; Miura, Kohshi; Ichihara, Nobutsune; Nishita, Toshiho; Amasaki, Hajime

doi:10.1159/000016762

Cited by 20 publications

(38 citation statements)

References 22 publications

(34 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All preparations were incubated with 0.02% H 2 O 2 and 0.1% diaminobenzidine tetrahydrochloride in 0.05 M Tris-HCl (pH 7.6) for 5 min. Fixation with Bouin's fluids has been found to provide the maximum retention of affinity for the present antibody and the most consistent immunostaining by the previous studies [5].Western blotting: Ten % dodecyl sodium sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was applied in the analyzing of crude extracts from bovine tissues. Briefly, 10 µl samples from crude extracts of bovine colon, rumen and parotid gland, and low-molecular-weight protein marker (Pharmacia, Piscataway, NJ, U.S.A.) were directly boiled in 100 µl SDS-PAGE sample solvent with mercaptoethanol at 100°C for 5 min.…”

mentioning

confidence: 79%

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

Kaseda

Ichihara

Nishita

et al. 2006

The Journal of Veterinary Medical Science

Self Cite

View full text Add to dashboard Cite

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...

show abstract

mentioning

confidence: 79%

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

Kaseda

Ichihara

Nishita

et al. 2006

The Journal of Veterinary Medical Science

Self Cite

View full text Add to dashboard Cite

show abstract

“…The following findings indicate that this protein is CA-VI: 1) it bound to a sulfonamide inhibitor affinity column, from which it was eluted with carbonic anhydrase inhibitor NaN 3 ; 2) it showed CO 2 hydratase activity; 3) anti-bovine CA-VI serum (produced in our laboratories [1]), crossreacted with the purified protein (data not shown).…”

mentioning

confidence: 90%

“…The molecular mass of the cytoplasmic CA isozymes CA-I, CA-II and CA-III is approximately 30 kDa, whereas that of the CA-VI subunit is roughly 40 kDa (46 kDa, rat; 40-42 kDa, human; 42 kDa, cow) as measured by SDS-PAGE [1,3,10]. Fernley et al [5] reported that the molecular mass of sheep CA-VI is 540 kDa, according to gel filtration under non-denaturing conditions, and that its apparent subunit molecular mass is 45 kDa, as measured by SDS-PAGE.…”

mentioning

confidence: 96%

“…They concluded that human CA-VI and CA-II must share at least one antigenic site. Asari et al reported that anti-bovine CA-VI serum did not react with CA-I or CA-II [1].…”

mentioning

confidence: 98%

“…Salivary or secreted CA constituted a new class of CA (designated CA-VI), which is secreted into saliva by the serous acinar cells of the parotid and submandibular glands. CA-VI has been purified from saliva or parotid glands of humans [10], rats [3] and cattle [1,5]. It may participate in the regulation of salivary pH and buffer capacity and protect the upper alimentary canal against excess acidity [13].…”

mentioning

confidence: 99%

See 2 more Smart Citations

Purification of Carbonic Anhydrase Isozyme VI (CA-VT) from Swine Saliva.

Nishita

Sakomoto

Ikeda

et al. 2001

The Journal of Veterinary Medical Science

Self Cite

View full text Add to dashboard Cite

ABSTRACT. Salivary or secreted carbonic anhydrase (CA), which constitutes a new class of CA, designated CA-VI, was isolated. Swine CA-VI purified from swine saliva by inhibitor-affinity chromatography and ion exchange chromatography had a specific activity of 5,468 units/mg. The molecular weight was 250,000, as determined by gel filtration under non-denaturing conditions, and the subunit molecular weight was found to be 37,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that swine CA-VI consists of 7 subunits. The treatment of the enzyme with endo-N-acetylglucosaminidase F reduced its subunit molecular weight from 37,000 to 35,000 and 32,000. We raised a rabbit antibody against purified swine CA-VI. Double immunodiffusion showed that anti-swine CA-VI serum reacted with swine CA-VI and swine saliva, but not with hemolysate (containing CA-I and CA-II) or muscle extracts (containing CA-III). The concentration of CA-VI in swine saliva, measured using single radial immunodiffusion, was 0.027 ± 0.017 mg/mg total protein. KEY WORDS: carbonic anhydrase VI, saliva, swine.

show abstract

Cross-species comparison of mammalian saliva using an LC-MALDI based proteomic approach

Sousa-Pereira

Cova

Abrantes³

et al. 2015

Proteomics

View full text Add to dashboard Cite

Despite the importance of saliva in the regulation of oral cavity homeostasis, few studies have been conducted to quantitatively compare the saliva of different mammal species. Aiming to define a proteome signature of mammals' saliva, an in-depth SDS-PAGE-LC coupled to MS/MS (GeLC-MS/MS) approach was used to characterize the saliva from primates (human), carnivores (dog), glires (rat and rabbit), and ungulates (sheep, cattle, horse). Despite the high variability in the number of distinct proteins identified per species, most protein families were shared by the mammals studied with the exception of cattle and horse. Alpha-amylase is an example that seems to reflect the natural selection related to digestion efficacy and food recognition. Casein protein family was identified in all species but human, suggesting an alternative to statherin in the protection of hard tissues. Overall, data suggest that different proteins might assure a similar role in the regulation of oral cavity homeostasis, potentially explaining the specific mammals' salivary proteome signature. Moreover, some protein families were identified for the first time in the saliva of some species, the presence of proline-rich proteins in rabbit's saliva being a good example.

show abstract

Distribution of Carbonic Anhydrase Isozyme VI in the Developing Bovine Parotid Gland

Cited by 20 publications

References 22 publications

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

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

Purification of Carbonic Anhydrase Isozyme VI (CA-VT) from Swine Saliva.

Cross-species comparison of mammalian saliva using an LC-MALDI based proteomic approach

Contact Info

Product

Resources

About