Organic anion transporter 6 (Slc22a20) specificity and sertoli cell-specific expression provide new insight on potential endogenous roles

Gloriane W. Schnabolk, Bhawna Gupta, Aditi Mulgaonkar, Mrugaya Kulkarni, Douglas H. Sweet

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Organic anion transporter 6 (Oat6; Slc22a20), a member of the OAT family, was demonstrated previously to mediate the transport of organic anions (Am J Physiol Renal Physiol 291:F314-F321, 2006). In the present study, we sought to further delineate the function of murine Oat6 (mOat6) by analyzing the effect of select organic anions on mOat6-mediated transport by using a Chinese hamster ovary (CHO) cell line stably expressing mOat6 (CHO-mOat6). When examined, kinetic analysis demonstrated that the mechanism of inhibition of mOat6 and mOat3 was competitive. Homovanillic acid, 5-hydroxyindole acetic acid, 2,4-dihydroxyphenylacetate, hippurate, and dehydroepiandrosterone sulfate (DHEAS) each significantly reduced mOat6 activity with inhibitory constant (Ki) values of 3.0 ± 0.5, 48.9 ± 10.3, 61.4 ± 7.1, 59.9 ± 4.9, and 38.8 ± 3.1 μM, respectively. Comparison to Ki values determined for mOat3 (67.8 ± 7.2, 134.5 ± 27.0, 346.7 ± 97.9, 79.3 ± 4.0, and 3.8 ± 1.1 μM, respectively) revealed that there are significant differences in compound affinity between each transporter. Fluoroquinolone antimicrobials and reduced folates were without effect on mOat6-mediated uptake. Investigation of testicular cell type-specific expression of mOat6 by laser capture microdissection and quantitative polymerase chain reaction revealed significant mRNA expression in Sertoli cells, but not in Leydig cells or spermatids. Overall, these data should aid further refinements to the interpretation and modeling of the in vivo disposition of OAT substrates. Specifically, expression in Sertoli cells suggests Oat6 may be an important determinant of blood-testis barrier function, with Oat6-mediated transport of estrone sulfate and DHEAS possibly representing a critical step in the maintenance of testicular steroidogenesis.

Original languageEnglish (US)
Pages (from-to)927-935
Number of pages9
JournalJournal of Pharmacology and Experimental Therapeutics
Volume334
Issue number3
DOIs
StatePublished - Sep 2010
Externally publishedYes

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Organic Anion Transporters
Dehydroepiandrosterone Sulfate
Sertoli Cells
Cricetulus
Anions
Ovary
Blood-Testis Barrier
Laser Capture Microdissection
Homovanillic Acid
Spermatids
Leydig Cells
Fluoroquinolones
Folic Acid
Acetic Acid
Maintenance
Kidney
Cell Line
Polymerase Chain Reaction
Messenger RNA
5-hydroxyindole

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

Cite this

Organic anion transporter 6 (Slc22a20) specificity and sertoli cell-specific expression provide new insight on potential endogenous roles. / Schnabolk, Gloriane W.; Gupta, Bhawna; Mulgaonkar, Aditi; Kulkarni, Mrugaya; Sweet, Douglas H.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 334, No. 3, 09.2010, p. 927-935.

Research output: Contribution to journalArticle

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abstract = "Organic anion transporter 6 (Oat6; Slc22a20), a member of the OAT family, was demonstrated previously to mediate the transport of organic anions (Am J Physiol Renal Physiol 291:F314-F321, 2006). In the present study, we sought to further delineate the function of murine Oat6 (mOat6) by analyzing the effect of select organic anions on mOat6-mediated transport by using a Chinese hamster ovary (CHO) cell line stably expressing mOat6 (CHO-mOat6). When examined, kinetic analysis demonstrated that the mechanism of inhibition of mOat6 and mOat3 was competitive. Homovanillic acid, 5-hydroxyindole acetic acid, 2,4-dihydroxyphenylacetate, hippurate, and dehydroepiandrosterone sulfate (DHEAS) each significantly reduced mOat6 activity with inhibitory constant (Ki) values of 3.0 ± 0.5, 48.9 ± 10.3, 61.4 ± 7.1, 59.9 ± 4.9, and 38.8 ± 3.1 μM, respectively. Comparison to Ki values determined for mOat3 (67.8 ± 7.2, 134.5 ± 27.0, 346.7 ± 97.9, 79.3 ± 4.0, and 3.8 ± 1.1 μM, respectively) revealed that there are significant differences in compound affinity between each transporter. Fluoroquinolone antimicrobials and reduced folates were without effect on mOat6-mediated uptake. Investigation of testicular cell type-specific expression of mOat6 by laser capture microdissection and quantitative polymerase chain reaction revealed significant mRNA expression in Sertoli cells, but not in Leydig cells or spermatids. Overall, these data should aid further refinements to the interpretation and modeling of the in vivo disposition of OAT substrates. Specifically, expression in Sertoli cells suggests Oat6 may be an important determinant of blood-testis barrier function, with Oat6-mediated transport of estrone sulfate and DHEAS possibly representing a critical step in the maintenance of testicular steroidogenesis.",
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T1 - Organic anion transporter 6 (Slc22a20) specificity and sertoli cell-specific expression provide new insight on potential endogenous roles

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AU - Gupta, Bhawna

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AU - Kulkarni, Mrugaya

AU - Sweet, Douglas H.

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