July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Quantification of amyloid-beta in eye tissues of APPswe/PS1ΔE9 mice
Author Affiliations & Notes
  • Wangfei Wang
    University of Illinois at Chicago, Chicago, Illinois, United States
  • Tara Nguyen
    University of Illinois at Chicago, Chicago, Illinois, United States
  • Rachana Mishra
    University of Illinois at Chicago, Chicago, Illinois, United States
  • Orly Lazarov
    University of Illinois at Chicago, Chicago, Illinois, United States
  • David R Pepperberg
    University of Illinois at Chicago, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Wangfei Wang, None; Tara Nguyen, None; Rachana Mishra, None; Orly Lazarov, None; David Pepperberg, None
  • Footnotes
    Support  BrightFocus Foundation, Research to Prevent Blindness, Search for Vision, NIH R01 AG051257
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 3090. doi:
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      Wangfei Wang, Tara Nguyen, Rachana Mishra, Orly Lazarov, David R Pepperberg; Quantification of amyloid-beta in eye tissues of APPswe/PS1ΔE9 mice. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3090.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Accumulations of amyloid-beta (Aβ) peptides, particularly as soluble oligomers and higher aggregates, are hallmarks of Alzheimer’s disease (AD). Aβ’s presence in eye tissues (e.g., ref. 1) raises the question of whether Aβ accumulation may promote the progression of age-related macular degeneration and/or other retina and retinal pigment epithelium (RPE) diseases. APPswe/PS1ΔE9 transgenic mice are a widely used model for AD-relevant Aβ studies of hippocampus (2). We determined the amounts of Aβ40 and Aβ42 (prominent Aβ forms; monomeric lengths 40- and 42-amino acids, respectively) in retina and in combined RPE/choroid of these mice. In the AD field, Aβ42 is widely viewed as the more toxic Aβ form.

Methods : Neural retina and RPE/choroid were separately recovered from enucleated eyes (1,3); hippocampus was dissected from the brain. Tissues were homogenized in PBS supplemented with 5 M guanidine HCl (GuHCl), pH 7.4, and 2% (v/v) EDTA-free protease inhibitor, incubated overnight, then centrifuged. After desalting, filtrates were analyzed for Aβ40 and Aβ42 (ELISA) and protein (Bradford) (3). Data were obtained from duplicate samples of pooled tissues from multiple eyes; n values below represent animal numbers.

Results : We observed a wide range in comparative amounts (mean ± SEM of pmol per g protein; pmol/g) of Aβ40 and Aβ42 (Table). Other than for 2-month old males, levels of both Aβ40 and Aβ42 in both retina and RPE/choroid of the transgenic mice were considerably smaller (less than ~55%) than those in hippocampus. Additional data obtained by whole-eye analysis (combined lens, vitreous, retina, and RPE/choroid) yielded (in pmol/g): for 6-month females (n=4), Aβ40 9.63 ± 0.83 and Aβ42 4.65 ± 0.43; for 1-year males (n=2), Aβ40 10.91 ± 0.20 and Aβ42 4.28 ± 0.16.

Conclusions : The present ELISA results, together with data reported by Gupta et al. (4), encourage larger-scope investigation of Aβ40/Aβ42 abundances in APPswe/PS1ΔE9 retina/RPE in relation to structural and functional properties.

References: (1) Parthasarathy et al. (2015) Exper. Eye Res. 138:134-144. (2) Shen et al. (2017) PLoS One 12:e0174763. (3) Wang et al. (2017) IOVS 58:3574a. (4) Gupta et al. (2016) Neurosci. Lett. 623:52-56.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

 

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