Protocols in Current Issue
    Isolation, Culture, and Identification of Primary Müller Cells from Human Retina
    Authors:  Yingying Chen, Ting Zhang, Shaoxue Zeng, Michelle Yam, Mark C. Gillies and Ling Zhu, date: 10/05/2021, view: 1263, Q&A: 0

    Müller cells, the major glial cells of the retina, play vital roles in maintaining redox homeostasis and retinal metabolism. An immortalized human Müller cell line (MIO-M1) is widely used as an in vitro model to study Müller cells’ function, but they may not be exactly the same as primarily cultured human Müller cells. The use of human

    In vitro Measurement of Membrane Attack Complex in RPE Cells
    Authors:  Kelly Mulfaul and Sarah L. Doyle, date: 02/20/2021, view: 2477, Q&A: 0

    Initiation of the complement system results in the formation of a multiprotein pore termed the membrane attack complex (MAC, C5b-C9). MAC pores accumulate on a cell surface and can result in cell lysis. The retinal pigment epithelium (RPE) is a single monolayer of pigmented epithelial cells located at the posterior poll of the eye that forms the

    Precise Targeting of Single Microelectrodes to Orientation Pinwheel Centers
    Authors:  Xue Mei Song, Ming Li, Tao Xu, Dewen Hu and Anna Wang Roe, date: 06/05/2020, view: 1973, Q&A: 0
    [Abstract] In the mammalian visual system, early stages of visual form perception begin with orientation selective neurons in primary visual cortex (V1). In many species (including humans, monkeys, tree shrews, cats, and ferrets), these neurons are organized in pinwheel-like orientation columns. To study the functional organization within orientation ...
    Quantitative Determination of Ca2+-binding to Ca2+-sensor Proteins by Isothermal Titration Calorimetry
    Authors:  Seher Abbas and Karl-Wilhelm Koch, date: 04/05/2020, view: 3826, Q&A: 0
    [Abstract] Diverse and complex molecular recognitions are central elements of signal transduction cascades. The strength and nature of these interaction modes can be determined by different experimental approaches. Among those, Isothermal titration calorimetry (ITC) offers certain advantages by providing binding constants and thermodynamic parameters from ...
    Protocols for Processing and Interpreting cryoEM Data Using Bsoft: A Case Study of the Retinal Adhesion Protein, Retinoschisin
    Author:  J. Bernard Heymann, date: 01/20/2020, view: 3151, Q&A: 0
    [Abstract] The goal of cryoEM is to determine the structures of biomolecules from electron micrographs. In many cases the processing is straightforward and can be handled with routine protocols. In other cases, the properties and behavior of the specimen require adaptions to properly interpret the data. Here I describe the protocols for examining the higher ...
    In vitro Differentiation of Human iPSC-derived Retinal Pigment Epithelium Cells (iPSC-RPE)
    Authors:  Agnieszka D’Antonio-Chronowska, Matteo D’Antonio and Kelly A. Frazer, date: 12/20/2019, view: 5186, Q&A: 0
    [Abstract] Induced Pluripotent Stem Cells (iPSCs) serve as an excellent model system for studying the molecular underpinnings of tissue development. Human iPSC-derived retinal pigment epithelium (iPSC-RPE) cells have fetal-like molecular profiles. Hence, biobanks like iPSCORE, which contain iPSCs generated from hundreds of individuals, are an invaluable ...
    Small Molecule-Based Retinal Differentiation of Human Embryonic Stem Cells and Induced Pluripotent Stem Cells
    Authors:  Jie Zhu and Deepak A. Lamba, date: 06/20/2018, view: 5981, Q&A: 0
    [Abstract] Retinal degeneration leads to loss of light-sensing photoreceptors eventually resulting in vision impairment and impose a heavy burden on both patients and the society. Currently available treatment options are very limited and mainly palliative. Ever since the discovery of human pluripotent stem cell technologies, cell replacement therapy has ...

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