Abstract
Analysis of how retinal ganglion cells change in retinal degeneration is critical for evaluating the potential of photoreceptor restorative therapies. Immunocytochemistry in combination with image analysis provides a way for quantifying not only the density of ganglion cells during disease, but also information about their morphology and an evaluation of excitatory and inhibitory synaptic inputs. Here, we describe how indirect immunofluorescence can be used in retinal whole mounts to obtain information about ganglion cells in retinal degeneration.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Chader GJ, Weiland J, Humayun MS (2009) Artificial vision: needs, functioning, and testing of a retinal electronic prosthesis. Prog Brain Res 175:317–332
O'Brien EE, Greferath U, Vessey KA et al (2012) Electronic restoration of vision in those with photoreceptor degenerations. Clin Exp Optom 95(5):473–483
Santos A, Humayun MS, de Juan E Jr et al (1997) Preservation of the inner retina in retinitis pigmentosa. A morphometric analysis. Arch Ophthalmol 115(4):511–515
Humayun MS, Prince M, de Juan E et al (1999) Morphometric analysis of the extramacular retina from postmortem eyes with retinitis pigmentosa. Invest Ophthalmol Vis Sci 40(1):143–148
Strettoi E, Pignatelli V (2000) Modifications of retinal neurons in a mouse model of retinitis pigmentosa. Proc Natl Acad Sci U S A 97(20):11020–11025
Strettoi E, Pignatelli V, Rossi C et al (2003) Remodeling of second-order neurons in the retina of rd/rd mutant mice. Vis Res 43(8):867–877
Strettoi E, Porciatti V, Falsini B et al (2002) Morphological and functional abnormalities in the inner retina of the rd/rd mouse. J Neurosci 22(13):5492–5504
Jeon CJ, Strettoi E, Masland RH (1998) The major cell populations of the mouse retina. J Neurosci 18(21):8936–8946
Downie LE, Hatzopoulos KM, Pianta MJ et al (2010) Angiotensin type-1 receptor inhibition is neuroprotective to amacrine cells in a rat model of retinopathy of prematurity. J Comp Neurol 518(1):41–63
Downie LE, Pianta MJ, Vingrys AJ et al (2007) Neuronal and glial cell changes are determined by retinal vascularization in retinopathy of prematurity. J Comp Neurol 504(4):404–417
Caruso DM, Owczarzak MT, Goebel DJ et al (1989) GABA-immunoreactivity in ganglion cells of the rat retina. Brain Res 476(1):129–134
Marc RE, Liu WL, Kalloniatis M et al (1990) Patterns of glutamate immunoreactivity in the goldfish retina. J Neurosci 10(12):4006–4034
Rodriguez AR, de Sevilla Muller LP, Brecha NC (2014) The RNA binding protein RBPMS is a selective marker of ganglion cells in the mammalian retina. J Comp Neurol 522(6):1411–1443
Xiang M, Zhou L, Macke JP et al (1995) The Brn-3 family of POU-domain factors: primary structure, binding specificity, and expression in subsets of retinal ganglion cells and somatosensory neurons. J Neurosci 15(7 Pt 1):4762–4785
Feng G, Mellor RH, Bernstein M et al (2000) Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 28(1):41–51
Damiani D, Novelli E, Mazzoni F et al (2012) Undersized dendritic arborizations in retinal ganglion cells of the rd1 mutant mouse, a paradigm of early-onset photoreceptor degeneration. J Comp Neurol 520(7):1406–1423
O'Brien EE, Greferath U, Fletcher EL (2014) The effect of photoreceptor degeneration on ganglion cell morphology. J Comp Neurol 522(5):1155–1170
Stasheff SF (2008) Emergence of sustained spontaneous hyperactivity and temporary preservation of OFF responses in ganglion cells of the retinal degeneration (rd1) mouse. J Neurophysiol 99(3):1408–1421
Stasheff SF, Shankar M, Andrews MP (2011) Developmental time course distinguishes changes in spontaneous and light-evoked retinal ganglion cell activity in rd1 and rd10 mice. J Neurophysiol 105(6):3002–3009
Schmitz F, Königstorfer A, Südhof TC (2000) RIBEYE, a component of synaptic ribbons: a protein's journey through evolution provides insight into synaptic ribbon function. Neuron 28:857–872
Saha S, Greferath U, Vessey KA et al (2016) Changes in ganglion cells during retinal degeneration. Neuroscience 329:1–11
Sun WZ, Li N, He SG (2002) Large-scale morophological survey of rat retinal ganglion cells. Vis Neurosci 19(4):483–493
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
1 Electronic Supplementary Material
“Eye dissection.mp4” provides a video of the procedure used to dissect the eye and posterior eye cup (MP4 256148 kb)
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Fletcher, E.L., Greferath, U., Saha, S., Anderson, E.E., Vessey, K.A. (2018). Ganglion Cell Assessment in Rodents with Retinal Degeneration. In: Tanimoto, N. (eds) Mouse Retinal Phenotyping. Methods in Molecular Biology, vol 1753. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7720-8_18
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7720-8_18
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7719-2
Online ISBN: 978-1-4939-7720-8
eBook Packages: Springer Protocols