title: Pharmacological inhibitions of glutamate transporters EAAT1 and EAAT2 compromise glutamate transport in photoreceptor to ON-bipolar cell synapses.
abstract: To maintain reliable signal transmission across a synapse, free synaptic neurotransmitters must be removed from the cleft in a timely manner. In the first visual synapse, this critical task is mainly undertaken by glutamate transporters (EAATs). Here we study the differential roles of the EAAT1, EAAT2 and EAAT5 subtypes in glutamate (GLU) uptake at the photoreceptor-to-depolarizing bipolar cell synapse in intact dark-adapted retina. Various doses of EAAT blockers and/or GLU were injected into the eye before the electroretinogram (ERG) was measured. Their effectiveness and potency in inhibiting the ERG b-wave were studied to determine their relative contributions to the GLU clearing activity at the synapse. The results showed that EAAT1 and EAAT2 plays different roles. Selectively blocking glial EAAT1 alone using UCPH101 inhibited the b-wave 2-24h following injection, suggesting a dominating role of EAAT1 in the overall GLU clearing capacity in the synaptic cleft. Selectively blocking EAAT2 on photoreceptor terminals had no significant effect on the b-wave, but increased the potency of exogenous GLU in inhibiting the b-wave. These suggest that EAAT2 play a secondary yet significant role in the GLU reuptake activity at the rod and the cone output synapses. Additionally, we have verified our electrophysiological findings with double-label immunohistochemistry, and extend the literature on the spatial distribution of EAAT2 splice variants in the mouse retina.
authors.affiliation: Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX, USA. Electronic address: ytse@bcm.edu.
authors.forename: Dennis Y
authors.initials: DY
authors.lastname: Tse
authors.affiliation: Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX, USA; Department of Ophthalmology, Gyeongsang National University, Jinju, Republic of Korea.
authors.forename: Inyoung
authors.initials: I
authors.lastname: Chung
authors.affiliation: Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX, USA.
authors.forename: Samuel M
authors.initials: SM
authors.lastname: Wu
mesh_terms: D000818:Animals; D001692:Biological Transport; D003623:Dark Adaptation / Q000187:drug effects; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004596:Electroretinography / Q000187:drug effects; D027341:Excitatory Amino Acid Transporter 1 / Q000037:antagonists & inhibitors / Q000502:physiology*; D027342:Excitatory Amino Acid Transporter 2 / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D050582:Excitatory Amino Acid Transporter 5 / Q000378:metabolism / Q000502:physiology*; D005971:Glutamates / Q000378:metabolism* / Q000494:pharmacology; D007150:Immunohistochemistry; D058449:Intravitreal Injections; D007608:Kainic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010786:Photoreceptor Cells / Q000187:drug effects / Q000378:metabolism; D012160:Retina / Q000187:drug effects / Q000502:physiology*; D051245:Retinal Bipolar Cells / Q000187:drug effects / Q000378:metabolism; D009435:Synaptic Transmission / Q000187:drug effects*
chemical_list: D027341:Excitatory Amino Acid Transporter 1; D027342:Excitatory Amino Acid Transporter 2; D050582:Excitatory Amino Acid Transporter 5; D005971:Glutamates; C493394:SLC1A3 protein, human; C493396:Slc1a2 protein, mouse; C493387:Slc1a7 protein, mouse; C020889:dihydrokainic acid; D007608:Kainic Acid
keywords: EAAT1; EAAT2; Electroretinogram; GLT1; Immunohistochemistry; Retina