Mammalia

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    Protocols in Current Issue
    Isoelectric Focusing to Quantify Rhodopsin Phosphorylation in Mouse Retina
    Authors:  Sowmya Bekshe Lokappa, M. Carter Cornwall and Jeannie Chen, date: 07/20/2019, view: 1585, Q&A: 0
    [Abstract] Rhodopsin is a G-protein coupled receptor (GPCR) that mediates vision under dim light. Upon light exposure, rhodopsin is phosphorylated at multiple serine and threonine sites at its carboxyl-terminus by rhodopsin kinase (GRK1). This, in turn, reduces its ability to activate the visual G-protein transducin. Binding of light-activated, ...
    Partial Transection of Adult Rat Optic Nerve as a Model of Secondary Degeneration in the Central Nervous System
    Authors:  Carole A Bartlett and Melinda Fitzgerald, date: 12/20/2018, view: 1443, Q&A: 0
    [Abstract] Injury to the central nervous system is characterized by damage that spreads from the initial point of impact into the surrounding adjacent tissue, in a phenomenon referred to as secondary degeneration. The optic nerve can be used to effectively model injury and secondary degeneration to white matter tracts. Partial transection of the dorsal ...
    Embryonic Intravitreous Injection in Mouse
    Authors:  Liyuan Cui, Yupu Diao and Jiayi Zhang, date: 07/20/2018, view: 1905, Q&A: 0
    [Abstract] Axons of retinal ganglion cells (RGCs) relay visual information from the retina to lateral geniculate nucleus (LGN) and superior colliculus (SC), which are two major image-forming visual nuclei. Wiring of these retinal projections completes before vision begins. However, there are few studies on retinal axons at embryonic stage due to technical ...
    Murine Hair Follicle Derived Stem Cell Transplantation onto the Cornea Using a Fibrin Carrier
    [Abstract] The goal of this protocol is to establish a procedure for cultivating stem cells on a fibrin carrier to allow for eventual transplantation to the eye. The ability to transfer stem cells to a patient is critical for treatment for a variety of disorders and wound repair. We took hair follicle stem cells from the vibrissae of transgenic mice ...
    Mouse Müller Cell Isolation and Culture
    Authors:  Xiao Liu, Luosheng Tang and Yongqing Liu, date: 08/05/2017, view: 5776, Q&A: 0
    [Abstract] Müller cells are the major supportive and protective glial cells across the retina. Unlike in fish, they have lost the capacity to regenerate the retina in mammals. But, mammalian Müller cells still retain certain retinal stem cell properties with various degree of self-renewal and differentiation potentials, and thereby held a merit in cell-based ...
    Assessment of Murine Retinal Function by Electroretinography
    Authors:  Gillie Benchorin, Melissa A. Calton, Marielle O. Beaulieu and Douglas Vollrath, date: 04/05/2017, view: 8773, Q&A: 0
    [Abstract] The electroretinogram (ERG) is a sensitive and noninvasive method for testing retinal function. In this protocol, we describe a method for performing ERGs in mice. Contact lenses on the mouse cornea measure the electrical response to a light stimulus of photoreceptors and downstream retinal cells, and the collected data are analyzed to evaluate ...
    Heterochronic Pellet Assay to Test Cell-cell Communication in the Mouse Retina
    Authors:  Nobuhiko Tachibana, Dawn Zinyk, Randy Ringuette, Valerie Wallace and Carol Schuurmans, date: 02/05/2017, view: 6610, Q&A: 0
    [Abstract] All seven retinal cell types that make up the mature retina are generated from a common, multipotent pool of retinal progenitor cells (RPCs) (Wallace, 2011). One way that RPCs know when sufficient numbers of particular cell-types have been generated is through negative feedback signals, which are emitted by differentiated cells and must reach ...
    Mouse Corneal Stroma Fibroblast Primary Cell Culture
    Authors:  Yujin Zhang, Yen-Chiao Wang, Okada Yuka, Lingling Zhang and Chia-Yang Liu, date: 10/05/2016, view: 5069, Q&A: 0
    [Abstract] This protocol is developed for primary cell culture of cornea stromal keratocytes isolated from neonatal mouse eyeballs. It provides an optimal condition to isolate stromal keratocytes which maintain high viability for cell culture.
    Retinal Differentiation of Mouse Embryonic Stem Cells
    Author:  Anna La Torre, date: 07/05/2016, view: 8731, Q&A: 5
    [Abstract] Groundbreaking studies from Dr. Yoshiki Sasai’s laboratory have recently introduced novel methods to differentiate mouse and human Embryonic Stem Cells (mESCs and hESCs) into organ-like 3D structures aimed to recapitulate developmental organogenesis programs (Eiraku et al., 2011; Eiraku and Sasai, 2012; Nakano et al., 2012; ...
    Confocal Imaging of Myeloid Cells in the Corneal Stroma of Live Mice
    Authors:  Matilda F. Chan and Zena Werb, date: 07/05/2015, view: 7813, Q&A: 0
    [Abstract] The accessibility and transparency of the cornea makes it a good tissue model for monitoring immunological responses using in vivo real time imaging analysis (Lee et al., 2010; Tan et al., 2013). These corneal qualities have also allowed for high-resolution in vivo imaging of non-ocular tissue transplanted into ...



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