Connaughton Victoria P, Nelson Ralph, Bender Anna M
Department of Biology, American University, Washington, DC 20016, USA.
Vis Neurosci. 2008 Mar-Apr;25(2):139-53. doi: 10.1017/S0952523808080322.
To refine inhibitory circuitry models for ON and OFF pathways in zebrafish retina, GABAergic properties of zebrafish bipolar cells were studied with two techniques: whole cell patch responses to GABA puffs in retinal slice, and voltage probe responses in isolated cells. Retinal slices documented predominantly axon terminal responses; isolated cells revealed mainly soma-dendritic responses. In the slice, GABA elicited a conductance increase, GABA responses were more robust at axon terminals than dendrites, and Erev varied with [Cl(-)]in. Axon terminals of ON- and OFF-type cells were similarly sensitive to GABA (30-40 pA peak current); axotomized cells were unresponsive. Bicuculline-sensitive, picrotoxin-sensitive, and picrotoxin-insensitive components were identified. Muscimol was as effective as GABA; baclofen was ineffective. Isolated bipolar cells were either intact or axotomized. Even in cells without an axon, GABA or muscimol (but not baclofen) hyperpolarized dendritic and somatic regions, suggesting significant distal expression. Median fluorescence change for GABA was -0.22 log units (approximately -16 mV); median half-amplitude dose was 0.4 microM. Reduced [Cl(-)]out blocked GABA responses. GABA hyperpolarized isolated ON-bipolar cells; OFF-cells were either unresponsive or depolarized. Hyperpolarizing GABA responses in isolated cells were bicuculline and TPMPA insensitive, but blocked or partially blocked by picrotoxin or zinc. In summary, axon terminals contain bicuculline-sensitive GABAA receptors and both picrotoxin-sensitive and insensitive GABAC receptors. Dendritic processes express zinc- and picrotoxin-sensitive GABAC receptors.
为完善斑马鱼视网膜ON和OFF通路的抑制性电路模型,运用两种技术研究了斑马鱼双极细胞的GABA能特性:视网膜切片中对GABA微喷射的全细胞膜片钳反应,以及分离细胞中的电压探针反应。视网膜切片记录到的主要是轴突终末反应;分离细胞显示的主要是胞体 - 树突反应。在切片中,GABA引起电导增加,轴突终末的GABA反应比树突更强烈,且反转电位随细胞内Cl⁻浓度而变化。ON型和OFF型细胞的轴突终末对GABA同样敏感(峰值电流为30 - 40 pA);轴突切断的细胞无反应。鉴定出了荷包牡丹碱敏感、印防己毒素敏感和印防己毒素不敏感成分。蝇蕈醇与GABA效果相同;巴氯芬无效。分离的双极细胞分为完整的和轴突切断的。即使在没有轴突的细胞中,GABA或蝇蕈醇(而非巴氯芬)也会使树突和胞体区域超极化,表明有显著的远端表达。GABA的中位荧光变化为 -0.22对数单位(约 -16 mV);中位半数效应剂量为0.4 microM。降低细胞外Cl⁻浓度可阻断GABA反应。GABA使分离的ON双极细胞超极化;OFF细胞要么无反应,要么去极化。分离细胞中的超极化GABA反应对荷包牡丹碱和TPMPA不敏感,但被印防己毒素或锌阻断或部分阻断。总之,轴突终末含有荷包牡丹碱敏感的GABAA受体以及印防己毒素敏感和不敏感的GABAC受体。树突过程表达对锌和印防己毒素敏感的GABAC受体。