Naslov
Expression of organic anion transporter Oat3 in rat liver is androgen-dependent

Autori
Breljak, Davorka ; Ljubojević, Marija ; Balen, Daniela ; Kušan, Marija ; Anzai, Naohiko ; Burckhardt, Gerhard ; Sabolić, Ivan

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Book of abstracts of the HDBMB 2006 / Zrinka Kovarik - Zagreb : Hrvatsko društvo za biokemiju i molekularnu biologiju (HDBMB), 2006, 91-91

Skup
Congress of the Croatian Society of Biochemistry and Molecular Biology HDBMB 2006

Mjesto i datum
Vodice, Hrvatska, 03.10.2006. - 07.10.2006

Vrsta sudjelovanja
Poster

Vrsta recenzije
Domaća recenzija

Ključne riječi
sex differences; sex hormones; testosterone; hepatocyte

Sažetak
In the mammalian liver (L), various transporters mediate elimination of endogenous and exogenous organic anions, such as metabolic products, antibiotics and other drugs, and environmental toxins. Organic anion transporter 3 (Slc22a8), has been cloned from the rat L (rOat3) and its mRNA was defined as “ male predominant” due to strong gender differences in its expression ((males (M)>females (F)). However, the levels of expression and localization of rOat3 protein in the M and F rat L are unknown. In this study we examined in more details gender-dependent expression of rOat3 mRNA and protein in the rat L. Using the methods of RT-PCR, Western blotting (WB), and immunocytochemistry (IC) we studied, respectively, expression of mRNA in the tissue, protein abundance in total cell membranes (TCM) isolated from tissue homogenates, and localization of the protein in cryosections of the L from adult intact and gonadectomized M and F, and from testosterone (T)-, estradiol (E)- or progesterone (P)-treated castrated M. The RT-PCR data confirmed the male-predominant expression of rOat3 mRNA in the rat L. Castration markedly decreased the level of mRNA, whereas ovariectomy had no effect. Treatment of castrated M with T restored the expression of mRNA to the level of intact M, while E and P had no effect. In TCM, a polyclonal anti-rOat3 antibody labeled the protein band of ~48 kDa, that in various experimental conditions exhibited a density pattern comparable to that of mRNA. By IC, the antibody strongly stained some intracellular organelles in hepatocytes with the intensity pattern that corroborated the pattern of mRNA and WB data. The double-staining studies with the anti-rOat3 and organelle-specific marker antibodies indicated that rOat3 may be localized in lysosomes, but not in the plasma membranes, endocytic vesicles, mitochondria or peroxisomes. We conclude that in the rat L, Oat3 exhibits gender differences (M>F) at both mRNA and protein levels due to androgen stimulation, and might be localized in lysosomes.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

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