Single-cell suction recordings from mouse cone photoreceptors.
Rod and cone photoreceptors in the retina are responsible for light detection. In darkness, cyclic nucleotide-gated (CNG) channels in the outer segment are open and allow cations to flow steadily inwards across the membrane, depolarizing the cell. Light exposure triggers the closure of the CNG channels, blocks the inward cation current flow, and thus results in cell hyperpolarization. Based on the polarity of photoreceptors, a suction recording method was developed in 1970s that, unlike the classic patch-clamp technique, does not require penetrating the plasma membrane. Drawing the outer segment into a tightly-fitting glass pipette filled with extracellular solution allows recording the current changes in individual cells upon test-flash exposure. However, this well-established "outer-segment-in (OS-in)" suction recording is not suitable for mouse cone recordings, because of the low percentage of cones in the mouse retina (3%) and the difficulties in identifying the cone outer segments. Recently, an inner-segment-in (IS-in) recording configuration was developed to draw the inner segment/nuclear region of the photoreceptor into the recording pipette. In this video, we will show how to record from individual mouse cone photoresponses using single-cell suction electrode.
Wang
Jin-Shan
JS
Department of Ophthalmology and Visual Sciences, Washington University in St. Louis, School of Medicine, USA.
Kefalov
Vladimir J
VJ
D000818:Animals; D004566:Electrodes; D051379:Mice; D017949:Retinal Cone Photoreceptor Cells / Q000166:cytology / Q000502:physiology*; D013396:Suction / Q000295:instrumentation / Q000379:methods