Jin Nan Ge, Chuang Alice Z, Masson Philippe J, Ribelayga Christophe P
Ruiz Department of Ophthalmology and Visual Science, Medical School, The University of Texas Health Science Centre at Houston, 6431 Fannin Street, Suite MSB 7.024, Houston, TX, 77030, USA.
J Physiol. 2015 Apr 1;593(7):1597-631. doi: 10.1113/jphysiol.2014.284919. Epub 2015 Feb 19.
Rod single-photon responses are critical for vision in dim light. Electrical coupling via gap junction channels shapes the light response properties of vertebrate photoreceptors, but the regulation of rod coupling and its impact on the single-photon response have remained unclear. To directly address these questions, we developed a perforated patch-clamp recording technique and recorded from single rod inner segments in isolated intact neural mouse retinae, maintained by superfusion. Experiments were conducted at different times of the day or under constant environmental conditions, at different times across the circadian cycle. We show that rod electrical coupling is regulated by a circadian clock and dopamine, so that coupling is weak during the day and strong at night. Altogether, patch-clamp recordings of single-photon responses in mouse rods, tracer coupling, receptive field measurements and pharmacological manipulations of gap junction and dopamine receptor activity provide compelling evidence that rod coupling is modulated in a circadian manner. These data are consistent with computer modelling. At night, single-photon responses are smaller due to coupling, but the signal-to-noise ratio for a dim (multiphoton) light response is increased at night because of signal averaging between coupled rods.
视杆单光子反应对于暗光下的视觉至关重要。通过缝隙连接通道的电耦合塑造了脊椎动物光感受器的光反应特性,但视杆耦合的调节及其对单光子反应的影响仍不清楚。为了直接解决这些问题,我们开发了一种穿孔膜片钳记录技术,并在通过灌流维持的分离完整神经小鼠视网膜中的单个视杆内节进行记录。实验在一天中的不同时间或在恒定环境条件下、昼夜节律周期的不同时间进行。我们发现视杆电耦合受昼夜节律钟和多巴胺调节,因此耦合在白天较弱而在夜间较强。总之,小鼠视杆中单光子反应的膜片钳记录、示踪耦合、感受野测量以及缝隙连接和多巴胺受体活性的药理学操作提供了令人信服的证据,表明视杆耦合以昼夜节律方式受到调节。这些数据与计算机建模一致。在夜间,由于耦合,单光子反应较小,但由于耦合视杆之间的信号平均,暗光(多光子)光反应的信噪比在夜间增加。