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. 2017 Apr 10;7:766. doi: 10.1038/s41598-017-00774-9

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Metabolic labeling and electrophysiology of retina cups. Brightfield image of D300 IMR90.4 derived RC shown at (a) low and (b) high magnification. Immunolabeling of ultrathin plastic sections of RCs with antibodies against (c,d) L-aspartate, (e,f) GABA, (e,g) Glutamate and (e,h) Glycine on D300 RCs. Panel d is a higher magnification of the inset box from (c). Panel e is a triple label of GABA, glutamate and glycine. A DIC image of a RC in panel (i) illustrates a recording pipette used for capacitance measurements. (j) Upper panel: Lock-in membrane capacitance measurement of a cone photoreceptor from a PSC derived eyecup showed a capacitance “jump” upon a brief depolarization (indicated by the black bar) from −60 mV to −10 mV. The yellow highlights are regions before and after the stimulation from which the capacitance values are averaged. Middle and lower panels show stable membrane resistance and series resistance during the course of the recording, suggesting that the capacitance jump in the upper panel is not an artifact of conductance change, rather it reflects synaptic vesicle release. (k) In a whole cell recording of a cone photoreceptor, the voltage was held at −60 mV, then stepped to −100 mV, and increased to 0 mV at a ramp speed of 100 mV/250 ms (0.4 mV/ms). The arrow points to the peak calcium current at −20 mV, which is typical of L-type calcium currents in mammalian photoreceptors. Scale bars b = 40 μm.