Inorganic Mercury vs Brain Health Project

Human toxicology science fair projects
look at what compounds are hazardous for humans...

Brain Toxicology

The human brain is extremely sensitive. It can be negatively impacted by a number of factors including toxins.

Objectives/Goals

To determine if inorganic mercury is able to penetrate the cell membrane of mammalian neurons.

Methods / Materials

Materials: The Mutant GFP and mercury (HgCl2)The fluorometer
fluorescence microscope
sonicator
centrifuges
UV spectrophotometer
pH meter
rat brain neurons.

We will use a fluorescence microscope to visualize the light emitted from the rat neurons. The fluorescence microscope will be used at UCSB. Mercury (HgCl2), Buffer, Salt, Culture media, Virus.

Methods: A. Treatment of Purified GFP with Mercury

Measure Fluorescence Emission (Using a fluorometer)
. Ignite the xenon lamp with 40,000 volts
. Load GFP sample into cuvette
. Load cuvette into fluorometer
. Determine the excitation wavelength maximum by detecting fluorescence emission at 512nm
   and then scanning the excitation wavelengths from between 350nm to 500nm (Fig. 7)
. Set excitation wavelength to 466 nm for all experiments.
. Add mercury to GFP in different amounts
. Record the fluorescence emission at 513 nM (Fig. 8)
. Treatment of Neurons with Mercury

Expression of GFP in Mammalian Neurons
. GFP gene is cloned into a virus for infection of neurons
. Neurons are taken from the brains of rat fetuses and kept alive in cell culture
. Then neurons are infected with the GFP-containing virus
. The GFP protein is expressed in the neurons
. Mercury is added to the neurons and GFP fluorescence was then examined under a fluorescence microscope (Figs. 10 & 11)
. Pictures were taken and analyzed

Results

We found that the inorganic mercury was not able to penetrate in the 5 minute time limit we gave it but only after 9 hours did the mercury get into the neurons.

Conclusions/Discussion

the mercury was not able to penetrate the cell membrane 3rd party contributor