Transcript INM1 research poster_Brendon and Christina

The influence of bipolar drugs on the phospholipid biosynthetic
pathway in Saccharomyces cerevisiae
Brendon Ursomanno and Christina Moawad
This study investigates a specific yeast, Saccharomyces cerevisiae, to model the effect of Valproic acid and Lithium Chloride in humans. These drugs are
known to treat Bipolar disorder and affect genes that synthesize inositol in the phospholipid biosynthetic pathway. By testing different strands of cells
with the presence and absence of inositol, we were able to collect absorbance values (OD) using a spectrophotometer. With time, we will use the qPCR
treatment to quantify the gene expression of INM1 gene, which is responsible for the inositol production.
Introduction
Discussion
“Bipolar disorder affects approximately 5.7 million adult
Americans, or about 2.6% of the U.S. population age 18 and
older every year”(DBSA). This disorder can be treated with
drugs containing Valproic acid, Lithium Chloride and genes
that synthesize inositol. Specifically, these drugs affect genes
that are responsible for de novo synthesis of inositol in the
phospholipid biosynthetic pathway. The study will be
conducted using Saccharomyces cerevisiae as a model
organism. This yeast cell has genes in this pathway that are
evolutionarily conserved, making it comparable to the
human genome. We will be investigating INM1, a model
gene in this pathway because previous research has shown,
INM1 expression is affected by inositol, a carbon source and
growth stage, as well as lithium and Valproic acid.
Based on our optical density readings, we were able to
clearly see the effect that the two drugs, Lithium Chloride
and Valporic acid had on the specific yeast cell. We see that
Lithium Chloride has a greater effect than Valproic acid;
although both clearly do increase O.D. concentrations.
There is a direct relationship between increased time and
O.D. concentrations. But, eventually they will plateau
because all inositol in the media is used, or no more is
being produced de novo and the cells begin to die. For the
mutant cell, with SC- there is no inositol in the media nor
in the cell, making the O.D. concentrations approximately
zero for the entire experiment. Our study confirms the
effect that LiCl and Val have on the cells gene expression,
and further experiments can quantify these effects.
Methods
Conclusion
To conduct this experiment we took previously grown cells
and used a spectrophotometer to measure the Optical
Density. From this collected data, we were able to construct
various growth curves representing the Optical Density with
respect to time. Two main yeast strains that were used in this
analysis were, “#3” and “#27,” respectively modeling wild
type and ino2Δ. Wild type, strand #3, was used to represent
normal cells containing all genes in the presence of inositol.
Strand #27 deleted the ino2 protein, which deactivated the
synthesis of inositol, resulting in inositol absent media. SC+
denotes the addition of inositol in the media, while SCdenotes the absence of inositol in media. Both analyses were
performed with either of the two bipolar drugs.
Based on our results and data calculations, we will continue
our research in hopes of better understanding the gene
expression of INM1, by using First Strand Synthesis and
qPCR. These tests will quantify our results as it amplifies
our DNA copies. The lipid biosynthetic pathway is a
growing area of molecular biology research, and there is a
lot to gain when applying these experimental analyses to
various social applications.
References
#3 & #27 Growth Curve
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Data & Results
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Depression and bipolar support alliance. (n.d.). Retrieved from
http://www.dbsalliance.org/site/PageServer/Calendar/PageServer?pagename=
education_statistics_bipolar_disorder
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#27 SC+ val
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#27 SC+ LiCl
O.D.
We found that the greatest Optical Density was found in that of #3,
Lithium Chloride, and SC+. The lowest Optical density was found in
that of #27 Lithium Chloride and Valproic acid SC-. We also found out
that Optical Density will plateau around 30 hours, because cells begin
to die, regardless of inositol production.
(2013). Expression atlas. The European Bioinformatics Institute, Retrieved from
http://www.ebi.ac.uk/gxa/gene/YHR046C
#27 SC- val
#27 SC- LiCl
#3 SC+ val
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#3SC+ LiCl
Marlene Murray and Miriam L. Greenberg.(2000) Expression of yeast INM1
encoding inositol monophosphatase is regulated by inositol, carbon source and
growth stage and is decreased by lithium and valproate. Department of Biological
Sciences, Wayne State University
#3 SC- val
#3 SC- LiCl
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Acknowledgments
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Time (Hours)
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We would like to thank Dr. Shen and his entire lab,
including Paulina, Michelle, Goldie, and Gracie. Also, Dr.
Liu for being there for us with guidance and moral support
along the way.