Patch clamp recordings from mouse retinal neurons in a dark-adapted slice preparation.
Our visual experience is initiated when the visual pigment in our retinal photoreceptors absorbs photons of light energy and initiates a cascade of intracellular events that lead to closure of cyclic-nucleotide-gated channels in the cell membrane. The resulting change in membrane potential leads in turn to reductions in the amount of neurotransmitter release from both rod and cone synaptic terminals. To measure how the light-evoked change in photoreceptor membrane potential leads to downstream activity in the retina, scientists have made electrophysiological recordings from retinal slice preparations for decades. In the past these slices have been cut manually with a razor blade on retinal tissue that is attached to filter paper; in recent years another method of slicing has been developed whereby retinal tissue is embedded in low gelling temperature agar and sliced in cool solution with a vibrating microtome. This preparation produces retinal slices with less surface damage and very robust light-evoked responses. Here we document how this procedure can be done under infrared light to avoid bleaching the visual pigment.
Arman
A Cyrus
AC
Neurosciences Graduate Program, Zikha Neurogenetic Institute, Department of Physiology and Biophysics, Keck School of Medicine, University of Southern California, CA, USA.
Sampath
Alapakkam P
AP
D000818:Animals; D002462:Cell Membrane / Q000502:physiology; D003624:Darkness; D004566:Electrodes; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D008867:Microtomy / Q000379:methods*; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000295:instrumentation / Q000379:methods*; D020419:Photoreceptor Cells, Vertebrate / Q000502:physiology; D012160:Retina / Q000166:cytology / Q000502:physiology*