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Silicone oil-induced ocular hypertension and glaucomatous neurodegeneration in mouse

  1. Jie Zhang
  2. Liang Li
  3. Haoliang Huang
  4. Fang Fang
  5. Hannah C Webber
  6. Pei Zhuang
  7. Liang Liu
  8. Roopa Dalal
  9. Peter H Tang
  10. Vinit B Mahajan
  11. Yang Sun
  12. Shaohua Li
  13. Mingchang Zhang
  14. Jeffrey L Goldberg
  15. Yang Hu  Is a corresponding author
  1. Stanford University School of Medicine, United States
  2. Union Hospital, Huazhong University of Science & Technology, China
  3. The Second Xiangya Hospital, Central South University, China
  4. Veterans Affairs Palo Alto Health Care, United States
Tools and Resources
Cite this article as: eLife 2019;8:e45881 doi: 10.7554/eLife.45881
7 figures, 5 videos and 1 table

Figures

Figure 1 with 1 supplement
Silicone oil-induced ocular hypertension under-detected (SOHU) mouse model.

(A) Cartoon illustration of SO intracameral injection, pupillary block, closure of the anterior chamber angle, and reopening of the angle of anterior chamber after pupil dilation. (B) Representative anterior chamber OCT images of SOHU eyes in living animals showing the relative size of SO droplet (blue arrow) to pupil (black arrow) and the corresponding closure or opening of the anterior chamber angle before and after pupil dilation. Red curved arrow indicates the direction of aqueous humor flow. (C) Longitudinal IOP measurements at different time points before and after SO injection, and continuous measurements for 18 min after anesthesia with isoflurane at each time point. (D) The sizes of SO droplet and corresponding IOP measurements at different time points after SO injection; IOP measured 12–15 min after anesthesia. SO: SO injected eyes; CL: contralateral control eyes. Data are presented as means ± s.e.m, SO > 1.5 mm, n = 17; SO ≤ 1.5 mm, n = 6.

https://doi.org/10.7554/eLife.45881.002
Figure 1—figure supplement 1
Extended pupillary dilation lowers down IOP in the SOHU eyes.

Longitudinal IOP measurements at different time points after induction of anesthesia, and continuous measurements for 30 min. Data are presented as means ± s.e.m, n = 7.

https://doi.org/10.7554/eLife.45881.003
Dynamic changes in RGC morphology and visual function in living SOHU animals.

(A) Representative OCT images of mouse retina. Green circle indicates the OCT scan area surrounding ON head. GCC: ganglion cell complex, including RNFL, GCL and IPL layers; indicated by double end arrows. (B) Quantification of GCC thickness, represented as percentage of GCC thickness in the SO eyes, compared to the CL eyes. n = 10–20. (C) Visual acuity measured by OKR, represented as percentage of visual acuity in the SO eyes, compared to the CL eyes. n = 10–20. (D) Representative waveforms of PERG in the contralateral control (CL, black lines) and the SO injected (SO, red lines) eyes at different time points after SO injection. P1: the first positive peak after the pattern stimulus; N2: the second negative peak after the pattern stimulus. (E) Quantification of P1-N2 amplitude, represented as percentage of P1-N2 amplitude in the SO eyes, compared to the CL eyes. n = 13–15. Data are presented as means ± s.e.m, *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001, one-way ANOVA with Tukey’s multiple comparison test.

https://doi.org/10.7554/eLife.45881.008
Glaucomatous RGC soma and axon degeneration in SOHU eyes.

(A) Upper panel, confocal images of whole flat-mounted retinas showing surviving RBPMS-positive (red) RGCs at different time points after SO injection. Scale bar, 100 µm. Middle panel, confocal images of a portion of flat-mounted retinas showing surviving RBPMS-positive (red) RGCs at different time points after SO injection. Scale bar, 20 µm. Lower panel, light microscope images of semi-thin transverse sections of ON stained with PPD at different time points after SO injection. Scale bar, 10 µm. (B,C) Quantification of surviving RGCs in the peripheral retina (n = 11–13) and surviving axons in ON (n = 10–16) at different time points after SO injection, represented as percentage of SO eyes compared to CL eyes. Data are presented as means ± s.e.m. *p<0.05, **p<0.01, ***: p<0.001, ****: p<0.0001; one-way ANOVA with Tukey’s multiple comparison test.

https://doi.org/10.7554/eLife.45881.009
SO itself does not cause glaucomatous degeneration.

(A) IOP measurements at different time points after intravitreal SO injection. n = 15. (B) Visual acuity measured by OKR, represented as percentage of visual acuity in the SO eyes, compared to the CL eyes. n = 13–15. (C) Quantification of P1-N2 amplitude of PERG, represented as percentage of P1-N2 amplitude in the SO eyes, compared to the CL eyes. n = 12–15. (D) Quantification of GCC thickness measured by OCT, represented as percentage of GCC thickness in the SO eyes, compared to the CL eyes. n = 11–13. (E) Upper panel, confocal images of portions of flat-mounted retinas showing surviving RBPMS-positive (red) RGCs at 8wpi after intravitreal SO injection and contralateral naive eye. Scale bar, 20 µm. Lower panel, light microscope images of semi-thin transverse sections of ON stained with PPD at 8wpi after intravitreal SO injection and contralateral naive eye. Scale bar, 10 µm. (F) Quantification of surviving RGCs (n = 10) and surviving axons in ON (n = 10) at 8wpi after intravitreal SO injection, represented as percentage of SO eyes compared to the CL eyes. Data are presented as means ± s.e.m, Student t-test. (G) Upper panel, confocal images of portion of flat-mounted retinas showing surviving RBPMS positive (red) RGCs at 8wpi after intracameral SO injection (small size of SO droplet,≤1.5 mm) and contralateral naive eye. Scale bar, 20 µm. Lower panel, light microscope images of semi-thin transverse sections of ON stained with PPD at 8wpi after intracameral SO injection and contralateral naive eye. Scale bar, 10 µm. (H) Quantification of surviving RGCs (n = 12) and surviving axons in ON (n = 13) at 8wpi, represented as percentage of SO eyes compared to the CL eyes. Data are presented as means ± s.e.m, Student t-test.

https://doi.org/10.7554/eLife.45881.010
SOHU is reversible by SO removal.

(A) Representative images of SOHU eyes before and after SO removal, and anterior chamber OCT images in living animals showing the relative size of SO droplet to pupil and the corresponding closure or opening of the anterior chamber angle before and after SO removal. (B) IOP measurements before and after SO removal at different time points. n = 16.

https://doi.org/10.7554/eLife.45881.011
Author response image 1
Author response image 2

Videos

Video 1
Intracameral SO injection.

Demonstration of the anterior chamber SO injection with a glass pippet and the SO droplet formation on top of iris to block pupil.

https://doi.org/10.7554/eLife.45881.004
Video 2
Dye migration from vitreous chamber to anterior chamber in naïve eyes.

DiI injected into the posterior chamber of the naïve eye and migrated into the anterior chamber.

https://doi.org/10.7554/eLife.45881.005
Video 3
Dye migration blocked in SOHU eyes.

DiI injected into the posterior chamber of the SOHU eye and there was no DiI detected in the anterior chamber.

https://doi.org/10.7554/eLife.45881.006
Video 4
SO droplet flows away from pupil after dilation.

After pupil dilation, the SO droplet was pushed away from the pupil and iris by aqueous humor flooded into the anterior chamber.

https://doi.org/10.7554/eLife.45881.007
Video 5
SO removal from SOHU eyes.

To remove SO from the anterior chamber, one needle is used to flush normal saline into the anterior chamber from one side of the cornea and another glass pippet was used to suck away the SO from the anterior chamber.

https://doi.org/10.7554/eLife.45881.012

Tables

Key resources table
Reagent type (species)
or resource
DesignationSource or
reference
IdentifiersAdditional
information
Strain, strain background (Mus musculus)C57BL/6JJackson Laboratories000664
Antibodyanti-RBPMS (guinea pig polyclonal)Custom-made by ProSci1:4000
AntibodyCy3 Goat anti-Guinea Pig IgGJackson ImmunoResearch106-165-0031:200
Chemical compound, drugSilicone oilAlcon Laboratories1,000 mPa.s, Silikon
Software, algorithmGraphpad prism6GraphPad Software
Software, algorithmVolocity softwareQuorum Technologies

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for all the figures.

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