Carbonic anhydrases in the reproductive system : with special emphasis on isoenzymes VI, IX, XII, and a novel nuclear nonclassical form
|Organizations:||University of Oulu, Faculty of Medicine, Department of Anatomy and Cell Biology
|Online Access:||PDF Full Text (PDF, 0.9 MB)|
|Persistent link:|| http://urn.fi/urn:isbn:9514266641
|Publish Date:|| 2002-05-17
|Thesis type:||Doctoral Dissertation
|Defence Note:||Academic Dissertation to be presented with the assent of the Faculty of Medicine, University of Oulu, for public discussion in the Auditorium of the Department of Anatomy and Cell Biology, on May 17th, 2002, at 12 noon.
Professor Jorma Paranko
Docent Jorma Toppari
Carbonic anhydrases (CAs) are a group of zinc-containing metalloenzymes that catalyze the interconversion of carbon dioxide and bicarbonate (CO2 + H2O ⇔ HCO3- + H+). They are present in almost all organs and are implicated in various biological functions, the most important of which is participation in the regulation of ion, water, and acid-base balance. Recently, some members of the CA gene family have been suggested to promote cell proliferation and to act as trophic growth factors.
The present study was undertaken to examine the distribution of CA isoenzymes in the reproductive system, to attain a more detailed view on their linkage to the reproductive processes and to neonatal development.
The expression of membrane-bound CA IX and CA XII was studied in the female and male reproductive tracts by immunohistochemistry and western blotting. CA XII was found to be expressed in the basolateral plasma membrane of luminal and glandular epithelia in human uterus. In human efferent ducts, it was located in the basolateral plasma membrane of luminal epithelium, where it coexpressed with Aquaporin-1. In epididymal duct, CA XII was only expressed in occasional epithelial cells. These cells coexpressed CA II, suggesting that they represent apical mitochondria-rich cells (AMRC). CA IX was also expressed in the basolateral plasma membrane of luminal epithelium in human efferent ducts, but its expression was not uniform among the tubules. These findings suggest that basolateral plasma membrane-associated CA IX and CA XII contribute, along with CA II and CA IV, to the regulation of acid-base balance and water transport in the reproductive tract.
Western blotting of rat Leydig tumor cells and testis for CA II revealed an unidentified 66-kDa polypeptide band. The polypeptide was successfully purified from several rat tissues using CA inhibitor affinity chromatography. The amino acid sequence of the polypeptide showed it to be identical to NonO/p54nrb, a non-POU domain-containing octamer-binding protein previously implicated in transcriptional regulation. The recombinant NonO/p54nrb was shown to display CA activity, and the antibody to it predominantly immunostained the nuclei in lymphocytes, where CA activity was also detected histochemically. Accordingly, the nuclear Leydig cell CA immunoreactivity represents NonO/p54nrb. It is classified as a novel, nonclassical CA, and it may participate in pH-related events in the nucleus.
Human and rat milk was found to contain CA VI by immunohistochemistry and western blotting. The enzyme purified from human milk by CA inhibitor affinity chromatography was confirmed by PNGase F digestion and amino acid sequence as CA VI. The CA VI concentrations in human colostral milk were approximately eight times higher than those in mature milk (34.7 mg/l vs. 4.5 mg/l). Secretion of CA VI into milk is suggested by its localization in the alveolar epithelium of the rat mammary gland. The structural and functional stability of CA VI in an acidic milieu, its suggested growth-supporting function in taste bud stem cells, and its high concentration in colostrum suggest that it is an essential factor for the growth and development of the newborn alimentary canal.
Acta Universitatis Ouluensis. D, Medica
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