Multielectrode Array Recording of Mouse Retinas Transplanted with Stem Cell-Derived Retinal Sheets
Retinal multielectrode array (MEA) recording allows us to examine the action potentials of retinal ganglion cells and field potentials of photoreceptors and bipolar cells. In addition to studying the retinal circuitry, it has become one of the standard examination tools for the characterization of stem cell-derived retinal transplantation in degenerated retinas. Besides the detection of responses to simple light stimulation, it is also necessary to consider the spatial correlation of the graft and the electrodes, in order to unbiasedly reveal the locally reconstructed retinal circuitry after transplantation. Here, we introduce our newly developed protocol of MEA recording and analysis that may serve as a standard for evaluating transplanted retinas.
Key wordsMultielectrode array (MEA) Micro-electroretinogram (mERG) Spike sorting iPS/ES cell-derived 3D retina Retinal transplantation Metabotropic glutamate receptor type 6 (mGluR6) L-AP4 ON bipolar cells Retinal ganglion cells (RGCs) Bayesian inference
We would like to thank the Laboratory for Retinal Regeneration in RIKEN, Japan, especially for the retinal transplantation team led by Dr. Michiko Mandai. This methodology would not have been developed without the generous support from the team members.
- 1.Elstrott J, Anishchenko A, Greschner M, Sher A, Litke AM, Chichilnisky EJ, Feller MB (2008) Direction selectivity in the retina is established independent of visual experience and cholinergic retinal waves. Neuron 58(4):499–506. https://doi.org/10.1016/j.neuron.2008.03.013CrossRefPubMedPubMedCentralGoogle Scholar
- 2.Field GD, Gauthier JL, Sher A, Greschner M, Machado TA, Jepson LH, Shlens J, Gunning DE, Mathieson K, Dabrowski W, Paninski L, Litke AM, Chichilnisky EJ (2010) Functional connectivity in the retina at the resolution of photoreceptors. Nature 467(7316):673–677. https://doi.org/10.1038/nature09424CrossRefPubMedPubMedCentralGoogle Scholar
- 11.Iraha S, Tu HY, Yamasaki S, Kagawa T, Goto M, Takahashi R, Watanabe T, Sugita S, Yonemura S, Sunagawa GA, Matsuyama T, Fujii M, Kuwahara A, Kishino A, Koide N, Eiraku M, Tanihara H, Takahashi M, Mandai M (2018) Establishment of immunodeficient retinal degeneration model mice and functional maturation of human ESC-derived retinal sheets after transplantation. Stem Cell Reports 10(3):1059–1074. https://doi.org/10.1016/j.stemcr.2018.01.032CrossRefPubMedPubMedCentralGoogle Scholar
- 12.Tu HY, Watanabe T, Shirai H, Yamasaki S, Kinoshita M, Matsushita K, Hashiguchi T, Onoe H, Matsuyama T, Kuwahara A, Kishino A, Kimura T, Eiraku M, Suzuma K, Kitaoka T, Takahashi M, Mandai M (2018) Medium- to long-term survival and functional examination of human iPSC-derived retinas in rat and primate models of retinal degeneration. EBioMedicine. https://doi.org/10.1016/j.ebiom.2018.11.028CrossRefGoogle Scholar
- 13.Assawachananont J, Mandai M, Okamoto S, Yamada C, Eiraku M, Yonemura S, Sasai Y, Takahashi M (2014) Transplantation of embryonic and induced pluripotent stem cell-derived 3D retinal sheets into retinal degenerative mice. Stem Cell Reports 2(5):662–674. https://doi.org/10.1016/j.stemcr.2014.03.011CrossRefPubMedPubMedCentralGoogle Scholar
- 14.Mandai M, Fujii M, Hashiguchi T, Sunagawa GA, Ito SI, Sun J, Kaneko J, Sho J, Yamada C, Takahashi M (2017) iPSC-derived retina transplants improve vision in rd1 end-stage retinal-degeneration mice. Stem Cell Reports 8(1):69–83. https://doi.org/10.1016/j.stemcr.2016.12.008CrossRefPubMedPubMedCentralGoogle Scholar