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Lui et al., 2013 Wang & Wiggs, 2014 Dengler-Crish et al., 2014 Smith et al., 2018 Wildtype CYP1B1-/- For Structure For function Immunohistochemistry with Epifluorescent Microscopy To assess retinal ganglion cell (RGC) soma,

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Wang & Wiggs, 2014 Lui et al., 2013

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Dengler-Crish et al., 2014 Smith et al., 2018

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Wildtype CYP1B1-/-

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Immunohistochemistry with Epifluorescent Microscopy

  • To assess retinal ganglion cell (RGC) soma, synapse, axonal

transport and axons structural integrity

  • Use of specific antibodies chemically conjugated to fluorescent

dyes that bind directly to cellular antigens. Allows visualization

  • f proteins/biomolecules in post-mortem fixed tissue

Electron Microscopy

  • To assess the ultrastructural composition of RGC axons.

For function For Structure

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Results: Cyp1b1-/- show poorer visual acuity

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  • CYP1B1-/- mice show reduced P1 component

amplitude indicative of reduced RGC responsive to visual stimulus.

  • P1 effect is similar to what is seen clinically and in

in other animal models with glaucoma.

  • No significant difference was seen in N2

amplitude nor with regard to the peak onset latency of both the P1 and N2 components.

  • Maintained N2 amplitude unusual
  • P1 and N2 amplitudes typically decrease with
  • nset of RGC degeneration in glaucoma---

however---can be influenced by pre-degenerative mechanisms.

Results: Cyp1b1-/- show reduced RGC Activity

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  • Optic nerve cross sections reveal discontinuous

focal myelin separations and increased cytoskeletal density /compaction in Cyp1b1-/- axons.

  • Cyp1b1-/- nodes of Ranvier appear absent of

major morphometric changes in the node (Nav1.6, red) and paranode (Caspr, green).

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  • SC lack signs of axonal transport, synapse or axon loss that typically

hallmarks glaucomatous pathology

  • Intraocular injection of cholera toxin-B conjugated -alexafluor488 (CTB488,

green) VGlut2: red, RGC presynaptic terminals Estrogen related receptor-B: magenta, RGC axon + presynaptic axon terminals

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  • Node of Ranvier alterations in CYP1b1-/-

post ocular microbead occlusion.

  • Increase in node length + reduction in

paranode length. Sodium channels remain normally distributed.

  • Node of Ranvier changes NOT normally

seen as a result of IOP elevation post- microbead occlusion.

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Acknowledgments

Smith Lab PI: Matthew Smith, PhD Brian Foresi, B.S. (U.Akron) Other Collaborators Rachida Bouhenni, PhD (Akron Children Hospital)