Molecular Characterization and In Situ Localization of a Mouse Retinal Taurine Transporter

Authors

  • Shyamala Vinnakota,

    1. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas; and
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    • S. Vinnakota and X. Qian contributed equally to this study.

  • Xiaojun Qian,

    1. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas; and
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    • S. Vinnakota and X. Qian contributed equally to this study.

  • Hussein Egal,

    1. Department of Ophthalmology, Northwestern University Medical School, Chicago, Illinois, U.S.A.
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  • Vijay Sarthy,

    1. Department of Ophthalmology, Northwestern University Medical School, Chicago, Illinois, U.S.A.
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  • Hemanta K. Sarkar

    Corresponding author
    1. Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas; and
      Address correspondence and reprint requests to Dr. H. K. Sarkar at Department of Molecular Physiology and Biophysics, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, U.S.A.
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Address correspondence and reprint requests to Dr. H. K. Sarkar at Department of Molecular Physiology and Biophysics, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, U.S.A.

Abstract

Abstract: Various ocular tissues have a higher concentration of taurine than plasma. This taurine concentration gradient across the cell membrane is maintained by a high-affinity taurine transporter. To understand the physiological role of the taurine transporter in the retina, we cloned a taurine transporter encoding cDNA from a mouse retinal library, determined its biochemical and pharmacological properties, and identified the specific cellular sites expressing the taurine transporter mRNA. The deduced protein sequence of the mouse retinal taurine transporter (mTAUT) revealed >93% sequence identity to the canine kidney, rat brain, mouse brain, and human placental taurine transporters. Our data suggest that the mTAUT and the mouse brain taurine transporter may be variants of one another. The mTAUT synthetic RNA induced Na+- and Cl-dependent [3H]taurine transport activity in Xenopus laevis oocytes that saturated with an average Km of 13.2 µM for taurine. Unlike the previous studies, we determined the rate of taurine uptake as the external concentration of Cl was varied, a single saturation process with an average apparent equilibrium constant (KCl−) of 17.7 mM. In contrast, the rate of taurine uptake showed a sigmoidal dependence when the external concentration of Na+ was varied (apparent equilibrium constant, KNa+∼54.8 mM). Analyses of the Na+- and Cl-concentration dependence data suggest that at least two Na+ and one Cl are required to transport one taurine molecule via the taurine transporter. Varying the pH of the transport buffer also affected the rate of taurine uptake; the rate showed a minimum between pH 6.0 and 6.5 and a maximum between pH 7.5 and 8.0. The taurine transport was inhibited by various inhibitors tested with the following order of potency: hypotaurine > β-alanine > l-diaminopropionic acid > guanidinoethane sulfonate > β-guanidinopropionic acid > chloroquine > γ-aminobutyric acid > 3-amino-1-propanesulfonic acid (homotaurine). Furthermore, the mTAUT activity was not inhibited by the inactive phorbol ester 4α-phorbol 12,13-didecanoate but was inhibited significantly by the active phorbol ester phorbol 12-myristate 13-acetate, which was both concentration and time dependent. The cellular sites expressing the taurine transporter mRNA in the mouse eye, as determined by in situ hybridization technique, showed low levels of expression in many of the ocular tissues, specifically the retina and the retinal pigment epithelium. Unexpectedly, the highest expression levels of taurine transporter mRNA were found instead in the ciliary body of the mouse eye.

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