College

Buchtel College of Arts and Sciences (BCAS)

Date of Last Revision

2020-05-07 11:28:33

Major

Chemistry - Biochemistry

Honors Course

3100:499

Number of Credits

2

Degree Name

Bachelor of Science

Date of Expected Graduation

Spring 2020

Abstract

I tested the hypothesis that recombinant leptin protein can be introduced to zebrafish in vivo through non-invasive soaking in a solution containing the protein. One way to study various molecules’ effects in vivo is through intraperitoneal or intracerebroventricular injections during the embryonic or larval stage, which is invasive, difficult to administer, and can have a high mortality rate. 48 hours post fertilization (hpf) zebrafish were soaked in a His-tagged recombinant leptin protein solution at 10 nM and 100 nM concentrations (produced by Genscript). After soaking, zebrafish larvae were washed extensively to remove all recombinant protein on their exterior before homogenization. The homogenate was evaluated for presence of His-tagged leptin protein using a western blot, which was quantified using ImageJ densitometry. Western blot results were indeterminate, and One-way ANOVA statistical analysis suggested there was no significant difference in mean protein uptake among larvae soaked in 0, 10 nM, or 100 nM leptin (p=0.5378). Relative target protein normalization with inconsistent β-actin loading control is likely to have affected results. Although results were inconclusive, this soaking method is still in its preliminary stages and should be explored in greater detail. Downstream applications of this technique include testing the effects of leptin on metabolic rate and whether leptin increases signal transduction in JAK-STAT pathways.

Research Sponsor

Richard Londraville

First Reader

Qin Liu

Second Reader

Catherine Konopka

Honors Faculty Advisor

Brian Bagatto

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.