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Agricultural runoff and industrial discharge are known to contaminate ground and surface waters and are not typically removed during water purification. These compounds may have significant effects on biological systems through the disruption of normal endocrine function. This study aims to determine the sex specific effects of phenothiazine (PTZ), a potential endocrine disruptor, on markers for endocrine disruption and stress in Fundulus heteroclitus. The following steroid hormone receptors were assessed: glucocorticoid receptor (GR), androgen receptor (AR), progesterone receptor (PR), estrogen related receptor alpha (ERα), estrogen receptor beta A (ERβA), and estrogen receptor beta B (ERβB). Phenothiazine was detected at levels between 1–2ppm via gas chromatography‐mass spectrophotometry (GC‐MS) in river water sampling conducted in southern Connecticut, but the biological impact of the compound is unknown. Phenothiazine has been used previously as an antipsychotic medication, as well as, industrially in plastics to add softness and flexibility. To assess the impact of phenothiazine, female and male fish were exposed to environmentally relevant concentrations of phenothiazine for one week and sacrificed. Brain, liver, and gonad tissues were removed and pooled from each treatment group. Quantitative PCR (qPCR) was conducted on tissues and analysis showed significant changes in mRNA expression and both a sex‐specific and tissue specific response. There were no changes in the testis and the female liver. In male brain there was a decrease in GR at 1ppm (p<0.05), and an increase in ERβB at 2ppm (p<0.01); in comparison to female brain which had an increase in PR (p< 0.01), AR (p<0.05), ERα (p< 0.0001) and ERβB (p<0.05). In male liver there was an upregulation in GR (p<0.05), and AR (p<0.05) mRNA expression and a down regulation of ERβB (p<0.0001). Lastly, within the ovary there is an increase in AR (p<0.05) and ERβA (p< 0.01) mRNA expression and a decrease in GR (p<0.05) mRNA expression. These findings indicate that phenothiazine may alter endocrine function through the steroid hormone receptors. Collectively, these tissue specific results demonstrate how phenothiazine disrupts normal fish physiology. Additionally, serum cortisol levels were assayed via enzyme linked immunosorbent assay (ELISA) and no significant changes in cortisol levels following phenothiazine treatment were observed.Support or Funding InformationConnecticut Community Foundation, Quinnipiac River FundThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Agricultural runoff and industrial discharge are known to contaminate ground and surface waters and are not typically removed during water purification. These compounds may have significant effects on biological systems through the disruption of normal endocrine function. This study aims to determine the sex specific effects of phenothiazine (PTZ), a potential endocrine disruptor, on markers for endocrine disruption and stress in Fundulus heteroclitus. The following steroid hormone receptors were assessed: glucocorticoid receptor (GR), androgen receptor (AR), progesterone receptor (PR), estrogen related receptor alpha (ERα), estrogen receptor beta A (ERβA), and estrogen receptor beta B (ERβB). Phenothiazine was detected at levels between 1–2ppm via gas chromatography‐mass spectrophotometry (GC‐MS) in river water sampling conducted in southern Connecticut, but the biological impact of the compound is unknown. Phenothiazine has been used previously as an antipsychotic medication, as well as, industrially in plastics to add softness and flexibility. To assess the impact of phenothiazine, female and male fish were exposed to environmentally relevant concentrations of phenothiazine for one week and sacrificed. Brain, liver, and gonad tissues were removed and pooled from each treatment group. Quantitative PCR (qPCR) was conducted on tissues and analysis showed significant changes in mRNA expression and both a sex‐specific and tissue specific response. There were no changes in the testis and the female liver. In male brain there was a decrease in GR at 1ppm (p<0.05), and an increase in ERβB at 2ppm (p<0.01); in comparison to female brain which had an increase in PR (p< 0.01), AR (p<0.05), ERα (p< 0.0001) and ERβB (p<0.05). In male liver there was an upregulation in GR (p<0.05), and AR (p<0.05) mRNA expression and a down regulation of ERβB (p<0.0001). Lastly, within the ovary there is an increase in AR (p<0.05) and ERβA (p< 0.01) mRNA expression and a decrease in GR (p<0.05) mRNA expression. These findings indicate that phenothiazine may alter endocrine function through the steroid hormone receptors. Collectively, these tissue specific results demonstrate how phenothiazine disrupts normal fish physiology. Additionally, serum cortisol levels were assayed via enzyme linked immunosorbent assay (ELISA) and no significant changes in cortisol levels following phenothiazine treatment were observed. Support or Funding Information Connecticut Community Foundation, Quinnipiac River Fund This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The National Academies and the American Association for the Advancement of Science (AAAS) have acknowledged that undergraduate education could be improved by more opportunities in authentic research (1, 2). In addition, there appears to be strong correlations between research‐based laboratories fostering an increased interest in STEM disciplines and improved grades, over traditional “cookbook” laboratories. This is especially true for underrepresented minorities and given our changing demographics at Quinnipiac, designing courses that better engage and support the student body is an essential component to our mission (3). Authentic research experiences are being implemented at many colleges and universities, as a means of giving students hands on experience conducting real science. Since there are limited research opportunities for our students to conduct faculty‐mentored research, developing a course where students can have an authentic research experience can address these shortages and obstacles (4). Additionally, in a senior exit survey completed by students within the College of Arts and Science, they expressed an interest in having more authentic lab experiences, more independent research opportunities and they wished they participated in these opportunities earlier in their career. To address some aspects of these shortages an authentic research course was developed to give students the opportunity to design and carry out experiments using one of four model organisms. Drosophila melanogaster (Fruit Fly), Caenorhabditis elgans (Roundworm), Dugesia tigrina (Planaria), or Danio rerio (Zebrafish), were all selected as model organisms because of their adaptability to undergraduate research and breadth of research topics. Student groups designed a novel experiment and generated a hypothesis, based on the current literature. The students were responsible for the animal husbandry of the models; C. elegans, D. tigrina and D. melanogaster throughout the course and performed experiments for over 50% of the semester. Once the experimental design and materials list were submitted students conducted experiments in weeks 4–6 and then analyzed their data. Following this data analysis, student groups either: repeated the same experiment, refined their experiment or generated a new question and then performed experimentation in weeks 8–11. Repeating experiments is a necessary component of scientific research, one that is typically lost in a traditional cookie‐cutter laboratory design. Students spent the remaining weeks of the semester working in their groups, analyzing data and preparing their oral presentation. The course has ran twice (Fall 2015 and Fall 2017) since it was developed and student survey response reports general satisfaction with the course.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

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