Senior Honors Projects, 2010-current

Date of Award

Spring 2016

Document Type

Dissertation/Thesis

Degree Name

Bachelor of Science (BS)

Department

Department of Integrated Science and Technology

Advisor(s)

Robert McKown, Ph. D.

Abstract

Keratoconjunctivitis sicca (KCS), the deficiency of tears also known as dry eye, is a prevalent disease that affects both humans and canines. The current treatment for dye eye, cyclosporine (Restatis®), only provides temporary relief, is often associated with discomfort and is inconsistently effective. Lacritin is a naturally occurring tear glycoprotein secreted from the human and canine lacrimal glands. It has been shown that lacritin stimulates basal tearing in rabbits when applied topically. This study characterized the amount and form of lacritin found in the tears of dogs with healthy and dry eyes—information which may be applied to the development of a lacritin-inspired therapeutic for humans and canines. In collaboration with the Virginia-Maryland College of Veterinarian Medicine, tear samples were collected from dogs being treated at the veterinary clinic and transported to JMU for analysis. At JMU, the lacritin and other proteins found in the canine tears were analyzed by indirect Enzyme-Linked Immunosorbent Assay (ELISA), SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE), and western blot. A total of 64 tear samples were analyzed with 32 samples from healthy dogs and 32 samples from dogs clinically diagnosed with dry eye. ELISA revealed that canines with KCS had a significant decrease in tear lacritin. Western blot analysis detected prominent bands in healthy tears at approximately 18 kDa (corresponding to monomeric canine lacritin) that were absent or faintly observed in tears from dry eye dogs. This study provides clinical data reinforces the hypothesis that lacritin replacement may be an effective therapeutic for dry eye.

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