MacKenzie McDowell

Jennifer J. Venditti, Ph.D.

William L. Coleman, Ph.D.
Abby Hare-Harris, Ph.D.

Bloomsburg University of Pennsylvania - Biological and Allied Health Sciences

Dissertation/Thesis

2020

English

electronic

60 leaves

Although alpha-L-fucosidase has been shown to be important during reproduction in many diverse organisms, the protein’s specific role remains unclear. To our knowledge, this would be the first report of alpha-L-fucosidase in zebrafish milt. Moreover, the utilization of zebrafish will allow for an evaluation of alpha-L-fucosidase’s potential role in embryonic development. Alpha-L-fucosidase activity in zebrafish sperm was quantified using the substrate 4-methylumbelliferyl-alpha-L-fucopyranoside (4-MU-fuc) to produce a fluorometric product known as 4-methyl-umbelliferone (4-MU). Fluorometric results suggested the presence of alpha-L-fucosidase in zebrafish milt. Partitioning assays were conducted to evaluate the presence of alpha-L-fucosidase in the sperm cells versus the liquid component of milt. Although not significant, a higher amount of alpha-L-fucosidase was detected in the pellet compared to the supernatant fraction. Treatment of zebrafish samples with deoxyfuconojirimycin (DFJ), a potent and specific inhibitor of alpha-L-fucosidase, inhibited enzyme activity. Egg extracts were prepared and evaluated for the presence of alpha-L-fucosidase and revealed minimal detection. In vitro fertilization experiments were conducted to evaluate embryonic success over 72 hours in the absence and presence of DFJ. A decrease in fertilization success was noted when comparing untreated versus DFJ treated experiments 0.75-2 hours post insemination (hpi). In both untreated and DFJ treated experiments, embryos successfully developed through 72 hpi. Understanding the role of alpha-L-fucosidase in reproduction could provide an understanding that can be translated to improving infertility in humans.

All rights reserved. Content may be used for educational and personal use only with full attribution to the author.

Zebra danio -- Reproduction; Enzymes; Catalysts; Proteins

Master of Science

Biological and Allied Health Sciences