University of British Columbia, Department of Microbiology & Immunology

Are eyes more than just windows to the soul? Exploring ocular vascularization
and amyloid deposition in a mouse model of Alzheimer's disease.
Maria-Elizabeth Baeva, Chahaat S. B. Singh, Cheryl Pfeifer, Wilf Jefferies
maria.baeva@msl.ubc.ca

Introduction Hypothesis & Aims

• Alzheimer’s disease (AD) is the most common form of dementia and We hypothesize that the deposition of amyloid, increased
affects 10% of people over the age of 651. angiogenesis and increased retinal blood barrier permeability
• An often-overlooked early symptom of AD are visual changes such in the eye are pathological hallmarks of AD.
as decreased visual acuity, changes in contrast sensitivity, visual Aim 1: Quantify amyloid accumulation and distribution in the eyes of
agnosia, poor color discrimination and decreased motion Tg2576 mice.
perception.2-5 Aim 2: Investigate evidence of increased vascularization in the eyes
• Decrease in integrity of the blood brain barrier due to of Tg2576 mice.
neovascularization leads to an increase in amyloid accumulation in Aim 3: Investigate retinal-blood barrier permeability in the eyes of
the brain in Tg2576 mice and AD patients.6,7 Tg2576 mice.

Aim 1 Results: Human amyloid deposits are found in Tg2576 c. Tg2576 a, b. Western Blots of brain (a)
a. and eyes (b) of 10-month old WT
brain and eyes and are predominantly located near blood
* (n=2) and Tg2576 (n=2) mice.
vessels. Positive control (+) is Tg2576
Eyes brain homogenate (N=3,*p<0.05,
a. Brain b. unpaired t-test). c. Human amyloid
0.0004
0.15
* beta (huAβ) in WT (a, n=2) and
Human APP expression
Human APP expression

normalized to -actin
normalized to -actin

* Tg2576 (b, n=2) mice was

0.0003
0.10 visualized using
immunofluorescent microscopy.
0.0002
GL= ganglion cell layer, IPL= inner
0.05 plexiform layer, INL= inner nuclear
0.0001
layer, OPL= outer plexiform layer,
0.00 0.0000
ONL= outer nuclear layer, PR=
WT Tg2576 WT Tg2576 photoreceptors.

Aim 2 Results: Tg2576 eyes have increased microvessels in b. WT c. Tg2576

the choroid. a.
1500
CD31 sum fluorescence

a-c. Eyes of 9-month old WT (b, n=4) *

and Tg2576 (c, n=5) mice were
examined using immunofluorescent 1000
microscopy. Tg2576 choroid has more
(a.u.)

CD31+ microvessels as determined by

increased sum fluorescence of CD31 500
(*p<0.05, unpaired t test). White arrows
indicate microvessels along choroid.

0
WT Tg2576

a. b.
Aim 3 Results: Tg2576 retinal pigmental
4
epithelial barrier (RPE) demonstrates
signs of deterioration and permeability.
Occludin (ng/mL)

3
a. ELISA was used to quantify occludin from eyes of twenty- *
four 11-month old mice (WT=11, Tg2576=13). Tg2576 whole
eye homogenates have less occludin in the eyes compared to 2
wild-type (p<0.05, unpaired t test). Each point on graph
represents an image. b. Eyes of WT (n=2) and Tg2576 (n=2)
1
mice were examined using immunofluorescent microscopy.
Tg2576 RPE appear to have weaker DAPI mean
fluorescence (0.03754 vs 0.01714; p<0.0001, unpaired t test) 0
and ZO-1 mean fluorescence (1149 vs 1111; unpaired t test: WT Tg2576
p<0.0001).

Future Directions Bibliography

Alzheimer’s Association (2018). Alzheimer's & Dementia 14(3): p. 367-429; 2. Armstrong
• Quantify and qualify changes in vascular in vivo in Tg2576 eyes RA. (2019). Journal of Optometry 12(3): 103-111; 3. Kiyosawa M et al. (1989).
over time. Ophthalmology 96(7): 1077-1086; 4. Javaid FZ et al. (2016) Frontiers in Neurology 7: 55-
55; 5. Cerquera-Jaramillo MA et al. (2018). Neural Plast 2018: 2941783; 6. Biron KE et al.
• Investigate whether brain vasculature and amyloid deposition (2011). PLoS One 6(8): e23789; 7. Biron KE et al. (2013). Sci Rep 3: 1354.

correlates temporally with changes in retinal vasculature.

Acknowledgement
• Examine angiogenesis signaling pathway in the eyes of Tg2576
Michael Smith Laboratories student fund (MEB/WAJ); Centre for Blood Research (CBR)
mice. Graduate Award Program Scholarship (MEB & CSBS); CBR Summer Studentship award
(MEB); CIHR project grant MOP-133635 (WAJ); Canada Graduate Scholarship – Masters
award (MEB); All current and former members of the Jefferies Lab.