Impact of Ethylene Glycol on Glycine Receptor Functionality

Contamination of groundwater with components of fracking fluid has garnered heightened levels of concern in recent years due to increased utilization, poor regulation, and limited longitudinal studies on health impacts. This project examines the impact of groundwater contamination with components of fracking fluid on glycine receptor function. To do this, glycine receptors, responsible for central nervous system function, will be expressed in Xenopus Laevis oocytes. The expression of glycine receptors will be accomplished through microinjecting RNA that encodes glycine receptors directly into the cytoplasm of the Xenopus oocyte. Once injected and expressed, the current across the membranes of the Xenopus oocytes will be recorded using Two Voltage Clamp Electrophysiology (TVEC). Ethylene glycol, a common sealing agent in fracking fluid, will be used as an exemplary component of fracking fluid due to its solubility in water and relative non-toxicity. Concentrations of 1000 μM, 333μM, and 111μM will be perfused over the Xenopus oocytes and recorded using TVEC. Ultimately, these recordings will demonstrate how ethylene glycol modifies glycine receptor and central nervous system activity. Our hypothesis is that concentrations of ethylene glycol greater than 333μM will inhibit normal glycine receptor functionality. This information provides us with the foundational knowledge necessary to explore how components of fracking fluid affect neurodevelopment in future experimentation. 

Hello.

My name is Casey Barry, and my

project is titled the Impact of Ethylene

Glycol on Glycine Receptor Functionality.

The second stage here is the abstract for

anyone who wishes to look at it.

Given the historic discovery of the Marcellus and

Barnett Shale deposits, recent geostrategic developments in the

Russia Ukraine conflict, and technological improvements, fracking has

become the cornerstone of domestic natural gas production

in the United States.

However, multifaceted political and economic views of

fracking have made exploring the potential impact

of fracking utilization on human health difficult.

Principally, the impact of groundwater contamination from components

of fracking fluid on receptor function has been

largely unexplored up until this point.

Figure one is providing an overview of

the fracking process, including the transportation, the

drilling, the storage, and the refinement.

My project examines the impact of

groundwater contamination with components of fracking

fluid on receptor function.

To do this, we are expressing

glycine receptors responsible for central nervous

system function in xenopus laevis oocytes.

Figure three shows the xenopus oocyte

in the upper right corner.

This is done by injecting RNA that encodes for

glycine receptors directly into the cytoplasm of the oocyte.

Once injected, the current across the membranes of

the oocyte can be recorded from using an

experimental technique called two voltage clamp electrophysiology.

Figure two shows the TVECor two voltage clamp electrophysiology

apparatus that we use in our lab, and that figure

is located in the left side of the page.

As the membrane current is recorded, different concentrations

of ethylene glycol stand in for components of

fracking fluid are profuse over the oocyte.

Ultimately, these recordings provide us with a

picture of how ethylene glycol modifies glycin

receptor and central nervous system activity.

This information provides us with the

foundational knowledge necessary to explore how

components of fracking fluid affect neurodevelopment.

In future experimentation, several tasks were performed

to gather data, including preparing salt solutions,

testing the perfusion system, pulling voltage and

current electrodes, measuring RNA stocks using a

nano drop, assessing RNA purity using gel

electrophoresis injecting xenopus oocytes with RNA and

recording from these oocytes using TVEC.

Figure four, in the upper left hand corner shows

the data from perfusion testing that we did.

Figure five, in the bottom left corner

shows the data obtained from pulling voltage

and current electrodes in their respective resistances.

Figure six in the upper right hand

corner shows data obtained from assessing the

quantity of RNA using the nano drop.

And figure seven in the bottom right hand

corner shows the results obtained from testing the

quality of RNA with gel electrophoresis.

Currently, we are in the process of

recording from the xenopus laevis oocytes to see

how ethylene glycol modifies glycine receptor activity.

In the coming weeks, we hope to collect data

from several xenopus oocytes and plot a curve of

glycine receptor activity in the presence of ethylene glycol.

Ultimately, this research is important to me

because I aspire to become a neurologist

and treat patients with neurodevelopmental conditions.

I began my undergraduate studies as a

geology major and have a particular interest

in understanding how environmental conditions contribute to

neurological health and development.

I hope to use this project as a

starting point for future investigations into the effect

of climate change on neurological health and development.

Finally, I would like to thank my mentor, dr.

Greta Ann Herin.

Committee members Dr.

Saleet Jeffrey and Dr.

Wendy Williams.

Lab partners Fae Jensen, Chanel

Green and Abigail Polanski.

I would also like to thank the Office

of Student Scholarship, creative Activities and Research for

their generous funding of my Undergraduate Research Scholars

Program and Mason Impact Minigrant scholarships.

Finally, I would like to thank the Interdisciplinary

Program of a Neuroscience, the Krasnow Institute, and

Miss Jeannie Scott for their support. Thank you.