Longitudinal in Vivo Imaging of Retinal Ganglion Cells and Retinal Thickness Changes Following Optic Nerve Injury in Mice

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2012 Jul 7

PMID 22768284

Citations 38

Authors

Balwantray C Chauhan

Kelly T Stevens

Julie M Levesque

Andrea C Nuschke

Glen P Sharpe

Neil OLeary

Michele L Archibald

Xu Wang

Affiliations

Soon will be listed here.

Abstract

Background: Retinal ganglion cells (RGCs) die in sight-threatening eye diseases. Imaging RGCs in humans is not currently possible and proof of principle in experimental models is fundamental for future development. Our objective was to quantify RGC density and retinal thickness following optic nerve transection in transgenic mice expressing cyan fluorescent protein (CFP) under control of the Thy1 promoter, expressed by RGCs and other neurons.

Methodology/principal Findings: A modified confocal scanning laser ophthalmoscopy (CSLO)/spectral-domain optical coherence tomography (SD-OCT) camera was used to image and quantify CFP+ cells in mice from the B6.Cg-Tg(Thy1-CFP)23Jrs/J line. SD-OCT circle (1 B-scan), raster (37 B-scans) and radial (24 B-scans) scans of the retina were also obtained. CSLO was performed at baseline (n = 11) and 3 (n = 11), 5 (n = 4), 7 (n = 10), 10 (n = 6), 14 (n = 7) and 21 (n = 5) days post-transection, while SD-OCT was performed at baseline and 7, 14 and 35 days (n = 9) post-transection. Longitudinal change in CFP+ cell density and retinal thickness were computed. Compared to baseline, the mean (SD) percentage CFP+ cells remaining at 3, 5, 7, 10, 14 and 21 days post-transection was 86 (9)%, 63 (11)%, 45 (11)%, 31 (9)%, 20 (9)% and 8 (4)%, respectively. Compared to baseline, the mean (SD) retinal thickness at 7 days post-transection was 97 (3)%, 98 (2)% and 97 (4)% for the circle, raster and radial scans, respectively. The corresponding figures at 14 and 35 days post-transection were 96 (3)%, 97 (2)% and 95 (3)%; and 93 (3)%, 94 (3)% and 92 (3)%.

Conclusions/significance: Longitudinal imaging showed an exponential decline in CFP+ cell density and a small (≤8%) reduction in SD-OCT measured retinal thickness post-transection. SD-OCT is a promising tool for detecting structural changes in experimental optic neuropathy. These results represent an important step towards translation for clinical use.

Citing Articles

Longitudinal Observation of Retinal Response to Optic Nerve Transection in Rats Using Visible Light Optical Coherence Tomography.

Pi S, Wang B, Gao M, Cepurna W, Lozano D, Morrison J Invest Ophthalmol Vis Sci. 2023; 64(4):17.

PMID: 37057973 PMC: 10117226. DOI: 10.1167/iovs.64.4.17.

Numerical method for axial motion artifact correction in retinal spectral-domain optical coherence tomography.

Ksenofontov S, Shilyagin P, Terpelov D, Gelikonov V, Gelikonov G Front Optoelectron. 2023; 13(4):393-401.

PMID: 36641561 PMC: 9743928. DOI: 10.1007/s12200-019-0951-0.

Longitudinal in vivo Ca imaging reveals dynamic activity changes of diseased retinal ganglion cells at the single-cell level.

Li L, Feng X, Fang F, Miller D, Zhang S, Zhuang P Proc Natl Acad Sci U S A. 2022; 119(48):e2206829119.

PMID: 36409915 PMC: 9889883. DOI: 10.1073/pnas.2206829119.

Light-Sheet Microscopy of the Optic Nerve Reveals Axonal Degeneration and Microglial Activation in NMDA-Induced Retinal Injury.

Ha Y, Ochoa L, Solomon O, Shi S, Villarreal P, Li S EC Ophthalmol. 2022; 12(11):23-31.

PMID: 36108311 PMC: 9450914.

Tools and Biomarkers for the Study of Retinal Ganglion Cell Degeneration.

Corral-Domenge C, de la Villa P, Mansilla A, Germain F Int J Mol Sci. 2022; 23(8).

PMID: 35457104 PMC: 9025234. DOI: 10.3390/ijms23084287.

References

Nickells R . Ganglion cell death in glaucoma: from mice to men. Vet Ophthalmol. 2007; 10 Suppl 1:88-94. DOI: 10.1111/j.1463-5224.2007.00564.x. View

van Velthoven M, Faber D, Verbraak F, van Leeuwen T, Smet M . Recent developments in optical coherence tomography for imaging the retina. Prog Retin Eye Res. 2006; 26(1):57-77. DOI: 10.1016/j.preteyeres.2006.10.002. View

Vidal-Sanz M, Bray G, Villegas-Perez M, Thanos S, Aguayo A . Axonal regeneration and synapse formation in the superior colliculus by retinal ganglion cells in the adult rat. J Neurosci. 1987; 7(9):2894-909. PMC: 6569122. View

Morrison J, Johnson E, Cepurna W . Rat models for glaucoma research. Prog Brain Res. 2008; 173:285-301. DOI: 10.1016/S0079-6123(08)01121-7. View

Chauhan B, Pan J, Archibald M, LeVatte T, Kelly M, Tremblay F . Effect of intraocular pressure on optic disc topography, electroretinography, and axonal loss in a chronic pressure-induced rat model of optic nerve damage. Invest Ophthalmol Vis Sci. 2002; 43(9):2969-76. View

Hoyt W, Frisen L, NEWMAN N . Fundoscopy of nerve fiber layer defects in glaucoma. Invest Ophthalmol. 1973; 12(11):814-29. View

Leung C, Lindsey J, Crowston J, Lijia C, Chiang S, Weinreb R . Longitudinal profile of retinal ganglion cell damage after optic nerve crush with blue-light confocal scanning laser ophthalmoscopy. Invest Ophthalmol Vis Sci. 2008; 49(11):4898-902. PMC: 5557086. DOI: 10.1167/iovs.07-1447. View

Schuman J, Hee M, Arya A, Puliafito C, Fujimoto J, Swanson E . Optical coherence tomography: a new tool for glaucoma diagnosis. Curr Opin Ophthalmol. 1995; 6(2):89-95. DOI: 10.1097/00055735-199504000-00014. View

Leung C, Lindsey J, Crowston J, Ju W, Liu Q, Bartsch D . In vivo imaging of murine retinal ganglion cells. J Neurosci Methods. 2007; 168(2):475-8. DOI: 10.1016/j.jneumeth.2007.10.018. View

10.

Leung C, Lindsey J, Chen L, Liu Q, Weinreb R . Longitudinal profile of retinal ganglion cell damage assessed with blue-light confocal scanning laser ophthalmoscopy after ischaemic reperfusion injury. Br J Ophthalmol. 2009; 93(7):964-8. PMC: 5499383. DOI: 10.1136/bjo.2008.150482. View

11.

Gabriele M, Ishikawa H, Schuman J, Ling Y, Bilonick R, Kim J . Optic nerve crush mice followed longitudinally with spectral domain optical coherence tomography. Invest Ophthalmol Vis Sci. 2011; 52(5):2250-4. PMC: 3080179. DOI: 10.1167/iovs.10-6311. View

12.

Jia L, Cepurna W, Johnson E, Morrison J . Patterns of intraocular pressure elevation after aqueous humor outflow obstruction in rats. Invest Ophthalmol Vis Sci. 2000; 41(6):1380-5. View

13.

Komaromy A, Brooks D, Kallberg M, Dawson W, Szel A, Lukats A . Long-term effect of retinal ganglion cell axotomy on the histomorphometry of other cells in the porcine retina. J Glaucoma. 2003; 12(4):307-15. DOI: 10.1097/00061198-200308000-00004. View

14.

Crish S, Calkins D . Neurodegeneration in glaucoma: progression and calcium-dependent intracellular mechanisms. Neuroscience. 2010; 176:1-11. PMC: 3040267. DOI: 10.1016/j.neuroscience.2010.12.036. View

15.

Morris R . Thy-1 in developing nervous tissue. Dev Neurosci. 1985; 7(3):133-60. DOI: 10.1159/000112283. View

16.

Quigley H . Glaucoma. Lancet. 2011; 377(9774):1367-77. DOI: 10.1016/S0140-6736(10)61423-7. View

17.

Wojtkowski M, Srinivasan V, Fujimoto J, Ko T, Schuman J, Kowalczyk A . Three-dimensional retinal imaging with high-speed ultrahigh-resolution optical coherence tomography. Ophthalmology. 2005; 112(10):1734-46. PMC: 1939719. DOI: 10.1016/j.ophtha.2005.05.023. View

18.

Srinivasan V, Ko T, Wojtkowski M, Carvalho M, Clermont A, Bursell S . Noninvasive volumetric imaging and morphometry of the rodent retina with high-speed, ultrahigh-resolution optical coherence tomography. Invest Ophthalmol Vis Sci. 2006; 47(12):5522-8. PMC: 1941766. DOI: 10.1167/iovs.06-0195. View

19.

Berkelaar M, Clarke D, Wang Y, Bray G, Aguayo A . Axotomy results in delayed death and apoptosis of retinal ganglion cells in adult rats. J Neurosci. 1994; 14(7):4368-74. PMC: 6577016. View

20.

Ruggeri M, Wehbe H, Jiao S, Gregori G, Jockovich M, Hackam A . In vivo three-dimensional high-resolution imaging of rodent retina with spectral-domain optical coherence tomography. Invest Ophthalmol Vis Sci. 2007; 48(4):1808-14. DOI: 10.1167/iovs.06-0815. View