TY - JOUR
T1 - Identification of renal transporters involved in sulfate excretion in marine teleost fish
AU - Kato, Akira
AU - Chang, Min Hwang
AU - Kurita, Yukihiro
AU - Nakada, Tsutomu
AU - Ogoshi, Maho
AU - Nakazato, Takeru
AU - Doi, Hiroyuki
AU - Hirose, Shigehisa
AU - Romero, Michael F.
PY - 2009/12
Y1 - 2009/12
N2 - Sulfate (SO42-) is the second most abundant anion in seawater (SW), and excretion of excess SO42- from ingested SW is essential for marine fish to survive. Marine teleosts excrete SO 42- via the urine produced in the kidney. The SO 42- transporter that secretes and concentrates SO 42- in the urine has not previously been identified. Here, we have identified and characterized candidates for the long-sought transporters. Using sequences from the fugu database, we have cloned cDNA fragments of all transporters belonging to the Slc13 and Slc26 families from mefugu (Takifugu obscurus). We compared Slc13 and Slc26 mRNA expression in the kidney between freshwater (FW) and SW mefugu. Among 14 clones examined, the expression of a Slc26a6 paralog (mfSlc26a6A) was the most upregulated (30-fold) in the kidney of SW mefugu. Electrophysiological analyses of Xenopus oocytes expressing mfSlc26a6A, mfSlc26a6B, and mouse Slc26a6 (mSlc26a6) demonstrated that all transporters mediate electrogenic Cl-/SO4 2-, Cl-/oxalate2-, and Cl-/nHCO 3- exchanges and electroneutral Cl-/ formate- exchange. Two-electrode voltage-clamp experiments demonstrated that the SO42--elicited currents of mfSlc26a6A is quite large (∼35 μA at +60 mV) and 50- to 200-fold higher than those of mfSlc26a6B and mSlc26a6. Conversely, the currents elicited by oxalate and HCO3- are almost identical among mfSlc26a6A, mfSlc26a6B, and mSlc26a6. Kinetic analysis revealed that mfSlc26a6A has the highest SO42- affinity as well as capacity. Immunohistochemical analyses demonstrated that mfSlc26a6A localizes to the apical (brush-border) region of the proximal tubules. Together, these findings suggest that mfSlc26a6A is the most likely candidate for the major apical SO42- transporter that mediates SO4 2- secretion in the kidney of marine teleosts.
AB - Sulfate (SO42-) is the second most abundant anion in seawater (SW), and excretion of excess SO42- from ingested SW is essential for marine fish to survive. Marine teleosts excrete SO 42- via the urine produced in the kidney. The SO 42- transporter that secretes and concentrates SO 42- in the urine has not previously been identified. Here, we have identified and characterized candidates for the long-sought transporters. Using sequences from the fugu database, we have cloned cDNA fragments of all transporters belonging to the Slc13 and Slc26 families from mefugu (Takifugu obscurus). We compared Slc13 and Slc26 mRNA expression in the kidney between freshwater (FW) and SW mefugu. Among 14 clones examined, the expression of a Slc26a6 paralog (mfSlc26a6A) was the most upregulated (30-fold) in the kidney of SW mefugu. Electrophysiological analyses of Xenopus oocytes expressing mfSlc26a6A, mfSlc26a6B, and mouse Slc26a6 (mSlc26a6) demonstrated that all transporters mediate electrogenic Cl-/SO4 2-, Cl-/oxalate2-, and Cl-/nHCO 3- exchanges and electroneutral Cl-/ formate- exchange. Two-electrode voltage-clamp experiments demonstrated that the SO42--elicited currents of mfSlc26a6A is quite large (∼35 μA at +60 mV) and 50- to 200-fold higher than those of mfSlc26a6B and mSlc26a6. Conversely, the currents elicited by oxalate and HCO3- are almost identical among mfSlc26a6A, mfSlc26a6B, and mSlc26a6. Kinetic analysis revealed that mfSlc26a6A has the highest SO42- affinity as well as capacity. Immunohistochemical analyses demonstrated that mfSlc26a6A localizes to the apical (brush-border) region of the proximal tubules. Together, these findings suggest that mfSlc26a6A is the most likely candidate for the major apical SO42- transporter that mediates SO4 2- secretion in the kidney of marine teleosts.
KW - Mefugu
KW - Oxalate
KW - Proximal tubule
KW - Slc13
KW - Slc26
KW - Sulfate homeostasis
UR - http://www.scopus.com/inward/record.url?scp=72449167894&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=72449167894&partnerID=8YFLogxK
U2 - 10.1152/ajpregu.00228.2009
DO - 10.1152/ajpregu.00228.2009
M3 - Article
C2 - 19812358
AN - SCOPUS:72449167894
VL - 297
SP - R1647-R1659
JO - American Journal of Physiology - Regulatory Integrative and Comparative Physiology
JF - American Journal of Physiology - Regulatory Integrative and Comparative Physiology
SN - 0363-6119
IS - 6
ER -