title: Lactate-H⁺ transport is a significant component of the in vivo corneal endothelial pump.
abstract: OBJECTIVE
  To confirm the expression of monocarboxylate transporters (MCT) 1, 2, and 4 in rabbit CE and to test the hypothesis that cellular buffering contributed by HCO₃⁻, NBCe1, and carbonic anhydrase (CA) activity facilitates lactate-H⁺ efflux thereby controlling corneal hydration in vivo.
  
  
  METHODS
  MCT1-4 expression of rabbit endothelium was examined by Western blotting and immunofluorescence staining. Lactate-induced acidification (LIA) was measured in perfused CE in the presence and absence of HCO₃⁻ and acetazolamide (ACTZ) using tissue treated with siRNA specific to MCT1, 2, and 4. Corneal thickness and lactate concentration were measured in New Zealand White rabbits treated with the topical CA inhibitor Azopt, and from eyes that were injected intracamerally with ouabain, disodium 4,4'-diisothiocyanatostilbene-2,2'-disulfonate (DIDS), and shRNA specific to the 1Na⁺:2HCO₃⁻ cotransporter NBCe1.
  
  
  RESULTS
  MCT1 and MCT4 are localized to the lateral membrane, while MCT2 is apical. Cell pH measurements showed LIA in response to 40 mM lactate in bicarbonate free (BF) Ringer's that was inhibited by niflumic acid and by MCT siRNA knockdown, and significantly reduced in the presence of HCO₃⁻. Lactate-dependent proton flux in vitro was not significantly greater in the presence of HCO₃⁻ or reduced by ACTZ. However, when active transport, NBCe1, or CA activity was disrupted in vivo, corneal edema ensued and was associated with significant corneal lactate accumulation.
  
  
  CONCLUSIONS
  MCT1, 2, and 4 are expressed in rabbit CE on both the apical and basolateral surfaces and function to transport lactate-H⁺. Lactate-H⁺ flux is facilitated by active transport, HCO₃⁻ transport and CA activity, disruption of which causes corneal edema in vivo and indicates that facilitation of lactate efflux is a component of the endothelial pump.
authors.affiliation: Indiana University School of Optometry, Bloomington, Indiana 47405, USA.
authors.forename: Tracy T
authors.initials: TT
authors.lastname: Nguyen
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Bonanno
mesh_terms: D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D015153:Blotting, Western; D004728:Endothelium, Corneal / Q000166:cytology / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D017136:Ion Transport; D019344:Lactic Acid / Q000378:metabolism*; D027501:Monocarboxylic Acid Transporters / Q000096:biosynthesis*; D009124:Muscle Proteins / Q000096:biosynthesis*; D011817:Rabbits; D027981:Symporters / Q000096:biosynthesis*; D000099233:Monocarboxylate Transport Protein 1
chemical_list: D019344:Lactic Acid; D027501:Monocarboxylic Acid Transporters; D009124:Muscle Proteins; D027981:Symporters; D000099233:Monocarboxylate Transport Protein 1; C406962:SLC16A4 protein, human