title: Pre- and post-synaptic effects of manipulating surface charge with divalent cations at the photoreceptor synapse.
abstract: Persistence of horizontal cell (HC) light responses in extracellular solutions containing low Ca2+ plus divalent cations to block Ca2+ currents (ICa) has been attributed to Ca2+-independent neurotransmission. Using a retinal slice preparation to record both ICa and light responses, we demonstrate that persistence of HC responses in low [Ca2+]o can instead be explained by a paradoxical increase of Ca2+ influx into photoreceptor terminals arising from surface charge-mediated shifts in ICa activation. Consistent with this explanation, application of Zn2+ or Ni2+ caused a hyperpolarizing block of HC light responses that was relieved by lowering [Ca2+]o. The same concentrations of Zn2+ and Ni2+ reduced the amplitude of ICa at the rod dark potential and this reduction was relieved by a hyperpolarizing shift in voltage dependence induced by lowering [Ca2+]o. Block of ICa by Mg2+, which has weak surface charge effects, was not relieved by low [Ca2+]o. Recovery of HC responses in low [Ca2+]o was assisted by enhancement of rod light responses. To bypass light stimulation, OFF bipolar cells were stimulated by steps to -40 mV applied to presynaptic rods during simultaneous paired recordings. Consistent with surface charge theory, the post-synaptic current was inhibited by Zn2+ and this inhibition was relieved by lowering [Ca2+]o. Nominally divalent-free media produced inversion of HC light responses even though rod light responses remained hyperpolarizing; HC response inversion can be explained by surface charge-mediated shifts in ICa. In summary, HC light responses modifications induced by low divalent cation solutions can be explained by effects on photoreceptor light responses and membrane surface charge without necessitating Ca2+-independent neurotransmission. Furthermore, these results suggest that surface charge effects accompanying physiological changing divalent cation levels in the synaptic cleft may provide a means for modulating synaptic output from photoreceptors.
authors.affiliation: Department of Ophthalmology, University of Nebraska Medical Center, 985540 Nebraska Medical Center, Omaha, NE 68198, USA.
authors.forename: L
authors.initials: L
authors.lastname: Cadetti
authors.forename: W B
authors.initials: WB
authors.lastname: Thoreson
authors.forename: M
authors.initials: M
authors.lastname: Piccolino
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020746:Calcium Channels, L-Type / Q000502:physiology; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D002413:Cations, Divalent / Q000494:pharmacology*; D004347:Drug Interactions; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D009433:Neural Inhibition / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D018408:Patch-Clamp Techniques / Q000379:methods; D010775:Photic Stimulation / Q000379:methods; D020419:Photoreceptor Cells, Vertebrate / Q000187:drug effects* / Q000502:physiology / Q000528:radiation effects; D017948:Retinal Rod Photoreceptor Cells / Q000502:physiology / Q000528:radiation effects; D013569:Synapses / Q000187:drug effects* / Q000528:radiation effects; D009435:Synaptic Transmission / Q000187:drug effects* / Q000502:physiology / Q000528:radiation effects; D014562:Caudata
chemical_list: D020746:Calcium Channels, L-Type; D002413:Cations, Divalent; D002118:Calcium