PARP1 Expression in Animal Models of Stress

Authors' Affiliations

Liza Wills, Ph.D., Department of Biomedical Sciences, East Tennessee State University, Johnson City, TN.

Location

D.P. Culp Center Ballroom

Start Date

4-5-2024 9:00 AM

End Date

4-5-2024 11:30 AM

Poster Number

33

Name of Project's Faculty Sponsor

Michelle Chandley

Faculty Sponsor's Department

Biomedical Sciences

Classification of First Author

Undergraduate Student

Competition Type

Competitive

Type

Poster Presentation

Presentation Category

Health

Abstract or Artist's Statement

Major depressive disorder (MDD) is a mood disorder that affects millions of people every year around the world and is characterized by feelings of despair and loss of interest in daily activities that can ultimately greatly hinder one’s quality of life. Many current antidepressant medications lack efficacy in providing full remission for patients diagnosed with MDD. Previous studies have shown that there is an increased expression of oxidative stress markers and inflammation including poly (ADP-ribose) polymerase-1(PARP1) from human postmortem brain tissue in major depressive disorder when compared to normal control human brain tissue. It has also been shown that elevated levels of DNA oxidation have been found in rat models of stress. These findings suggest rat models may be useful for studying the effects of drug therapies on behavior and pathophysiology of stress. Our laboratory recently demonstrated that the PARP1-inhibitor 3-aminobezaminde (3-AB) impacted the behavior of animals exposed to stress. The aim of this study was to compare the protein expression levels of PARP1 and high-mobility group box 1 (HMGB1) from the prelimbic cortex of stressed rats with and without treatments to no-stress control animals. Immunoblotting was used to evaluate the expression of PARP1 and HMGB1 protein in four groups of rats: no stress control animals, stress-induced rats that received no treatment (saline), and two stress-induced groups that received treatment (Fluoxetine-FLX and 3-AB). Fluoxetine is a selective serotonin reuptake inhibitor that has been used for years in the treatment of MDD. Animals were exposed to chronic unpredictable stress and social defeat stress for 28 days. Animals were treated with FLX and 3AB prior to daily stress. Each brain was sectioned, punch-dissected, and stored in -80oC until use. Tissue was homogenized and subjected to a Lowry protein assay. The immunoblotting system from Protein Simple known as the Abby was used for protein band acquisition. Antibodies for target proteins, PARP1 and HMGB1 were used to probe protein expression levels. Glyceraldehyde 3-phosphate dehydrogenase (GADPH) and tubulin protein levels were averaged and used to normalize PARP1 and HMGB1 expression. No statistically significant differences in PARP1 and HMGB1 protein expression levels were found within the groups using a one-way ANOVA analysis. These data contribute to the ongoing investigations for novel drug targets in the treatment of stress-related disorders.

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Apr 5th, 9:00 AM Apr 5th, 11:30 AM

PARP1 Expression in Animal Models of Stress

D.P. Culp Center Ballroom

Major depressive disorder (MDD) is a mood disorder that affects millions of people every year around the world and is characterized by feelings of despair and loss of interest in daily activities that can ultimately greatly hinder one’s quality of life. Many current antidepressant medications lack efficacy in providing full remission for patients diagnosed with MDD. Previous studies have shown that there is an increased expression of oxidative stress markers and inflammation including poly (ADP-ribose) polymerase-1(PARP1) from human postmortem brain tissue in major depressive disorder when compared to normal control human brain tissue. It has also been shown that elevated levels of DNA oxidation have been found in rat models of stress. These findings suggest rat models may be useful for studying the effects of drug therapies on behavior and pathophysiology of stress. Our laboratory recently demonstrated that the PARP1-inhibitor 3-aminobezaminde (3-AB) impacted the behavior of animals exposed to stress. The aim of this study was to compare the protein expression levels of PARP1 and high-mobility group box 1 (HMGB1) from the prelimbic cortex of stressed rats with and without treatments to no-stress control animals. Immunoblotting was used to evaluate the expression of PARP1 and HMGB1 protein in four groups of rats: no stress control animals, stress-induced rats that received no treatment (saline), and two stress-induced groups that received treatment (Fluoxetine-FLX and 3-AB). Fluoxetine is a selective serotonin reuptake inhibitor that has been used for years in the treatment of MDD. Animals were exposed to chronic unpredictable stress and social defeat stress for 28 days. Animals were treated with FLX and 3AB prior to daily stress. Each brain was sectioned, punch-dissected, and stored in -80oC until use. Tissue was homogenized and subjected to a Lowry protein assay. The immunoblotting system from Protein Simple known as the Abby was used for protein band acquisition. Antibodies for target proteins, PARP1 and HMGB1 were used to probe protein expression levels. Glyceraldehyde 3-phosphate dehydrogenase (GADPH) and tubulin protein levels were averaged and used to normalize PARP1 and HMGB1 expression. No statistically significant differences in PARP1 and HMGB1 protein expression levels were found within the groups using a one-way ANOVA analysis. These data contribute to the ongoing investigations for novel drug targets in the treatment of stress-related disorders.