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Full-Text Articles in Research Methods in Life Sciences

Mitochondrial Aspects Of Neuronal Pathology In Triple-Transgenic Alzheimer’S Disease Mice, John Zachary Cavendish Jan 2021

Mitochondrial Aspects Of Neuronal Pathology In Triple-Transgenic Alzheimer’S Disease Mice, John Zachary Cavendish

Graduate Theses, Dissertations, and Problem Reports

Alzheimer’s disease (AD) is a fatal, progressive neurodegenerative disease afflicting millions of people in the United States alone and is the only one of the top leading causes of morbidity and mortality with no effective disease-modifying therapies. It is the most common form of dementia, affecting one in three people over the age of 85. While the hallmarks of the disease include accumulation of beta-amyloid-based extracellular plaques and hyperphosphorylated tau-based intracellular neurofibrillary tangles, treatment strategies centered on removing or mitigating these components of AD have all failed in humans. Mitochondrial dysfunction has been increasingly recognized as an early and ...


Practical Considerations In Regenerative Medicine Research: Iacucs, Ethics, And The Use Of Animals In Stem Cell Studies, Susan Vandewoude, Bernard E. Rollin Sep 2019

Practical Considerations In Regenerative Medicine Research: Iacucs, Ethics, And The Use Of Animals In Stem Cell Studies, Susan Vandewoude, Bernard E. Rollin

Bernard Rollin, PhD

The intent of US federal laws mandating IACUC review of animal-related activities was to satisfy contemporary socioethical concerns by introducing deliberations about ethics and animal welfare into the research process when animals are used. These laws and the system they chartered have worked well for the most part in providing opportunities for consideration of animal welfare as a vital part of animal research. As a result, investigators today are far less naïve about the ethical issues raised by research on animals and typically more sympathetic about the need for such consideration. As evidence of this growing awareness, the literature on ...


Beyond Pain—Controlling Suffering In Laboratory Animals, Bernard E. Rollin Sep 2019

Beyond Pain—Controlling Suffering In Laboratory Animals, Bernard E. Rollin

Bernard Rollin, PhD

No abstract provided.


The Animal Research Controversy: Protest, Process & Public Policy, Andrew N. Rowan, Franklin M. Loew, Joan C. Weer Sep 2019

The Animal Research Controversy: Protest, Process & Public Policy, Andrew N. Rowan, Franklin M. Loew, Joan C. Weer

Andrew N. Rowan, DPhil

The controversy today regarding the use of animals in research appears on the surface to be a strongly polarized struggle between the scientific community and the animal protection movement. However, there is a wide range of opinions and philosophies on both sides. Mistrust between the factions has blossomed while communication has withered. Through the 1960s, 1970s and early 1980s, the animal movement grew in numbers and financial resources, and developed much greater public recognition and political clout. The research community paid relatively little attention to the animal movement for much of this period but, alarmed by several public relations coups ...


“Everyday” Knowledge And A New Paradigm Of Animal Research, David Fraser, Jeffrey M. Spooner, Catherine A. Schuppli Sep 2019

“Everyday” Knowledge And A New Paradigm Of Animal Research, David Fraser, Jeffrey M. Spooner, Catherine A. Schuppli

David Fraser, PhD

Commentary on Marino and Allen (2017) The Psychology of Cows


Act On The Registration And Evaluation Of Chemicals (K-Reach) And Replacement, Reduction Or Refinement Best Practices, Soojin Ha, Troy Seidle, Kyung-Min Lim Aug 2019

Act On The Registration And Evaluation Of Chemicals (K-Reach) And Replacement, Reduction Or Refinement Best Practices, Soojin Ha, Troy Seidle, Kyung-Min Lim

Troy Seidle, PhD

Objectives - Korea’s Act on the Registration and Evaluation of Chemicals (K-REACH) was enacted for the protection of human health and the environment in 2015. Considering that about 2000 new substances are introduced annually across the globe, the extent of animal testing requirement could be overwhelming unless regulators and companies work proactively to institute and enforce global best practices to replace, reduce or refine animal use. In this review, the way to reduce the animal use for K-REACH is discussed. Methods - Background of the enforcement of the K-REACH and its details was reviewed along with the papers and regulatory documents ...


Mental Stress From Animal Experiments: A Survey With Korean Researchers, Minji Kang, Ahram Han, Da-Eun Kim, Troy Seidle, Kyung-Min Lim, Seungjin Bae Aug 2019

Mental Stress From Animal Experiments: A Survey With Korean Researchers, Minji Kang, Ahram Han, Da-Eun Kim, Troy Seidle, Kyung-Min Lim, Seungjin Bae

Troy Seidle, PhD

Animal experiments have been widely conducted in the life sciences for more than a century, and have long been a subject of ethical and societal controversy due to the deliberate infliction of harm upon sentient animals. However, the harmful use of animals may also negatively impact the mental health of researchers themselves. We sought to evaluate the anxiety level of researchers engaged in animal use to analyse the mental stress from animal testing. The State Anxiety Scale of the State-Trait Anxiety Inventory (STAI) was used to evaluate how researchers feel when they conduct animal, as opposed to non-animal, based experiments ...


Non-Invasive Methods Of Identifying And Tracking Wild Squid, Ruth A. Byrne, James B. Wood, Roland C. Anderson, Ulrike Griebel, Jennifer A. Mather Aug 2019

Non-Invasive Methods Of Identifying And Tracking Wild Squid, Ruth A. Byrne, James B. Wood, Roland C. Anderson, Ulrike Griebel, Jennifer A. Mather

Jennifer Mather, PhD

The ability to identify individual free-living animals in the field is an important method for studying their behavior. Apart from invasive external or internal tags, which may cause injury or abnormal behavior, most cephalopods cannot be tagged, as their skin is too soft and delicate for tag retention. Additionally, cephalopods remove many types of tags. However, body markings have been successfully used as a non invasive method to identify individuals of many different species of animals, including whale sharks, grey whales, seals, and zebras. We developed methods to sex and individually identify Caribbean reef squid, Sepiotheuthis sepioidea. Males showed distinct ...


Animal Organs In Humans: Uncalculated Risks And Unanswered Questions, Gillian R. Langley, Joyce D'Silva Aug 2019

Animal Organs In Humans: Uncalculated Risks And Unanswered Questions, Gillian R. Langley, Joyce D'Silva

Gill Langley, PhD

This report, produced jointly by the British Union for the Abolition of Vivisection and Compassion in World Farming, fills a number of significant gaps in the current debate about xenotransplantation.

In this report we also summarise the ethical and welfare issues concerning experiments on animals for xenotransplant research and their possible use as source animals for organs. Both these aspects are responsible for much pain and distress caused to many animals. We prefer the term “source animals” to “donor animals”, because animals do not choose to donate their organs for xenotransplantation.


Rcrab: An R Analytics Tool To Visualize And Analyze The Movement Of Horseshoe Crabs In Long Island Sound, Ismael Youssef, Samah Senbel, Jo-Marie Kasinak, Jennifer Mattei Aug 2019

Rcrab: An R Analytics Tool To Visualize And Analyze The Movement Of Horseshoe Crabs In Long Island Sound, Ismael Youssef, Samah Senbel, Jo-Marie Kasinak, Jennifer Mattei

Samah Senbel

Mark-recapture programs are important for studying the ecology and population dynamics of wildlife. An R shiny analytics tool was developed to track the movement of horseshoe crabs in Long Island Sound based on tag and resight data. The crabs were tagged and recaptured by volunteers of Project Limulus, a community-based research program. The dataset contains tag and recapture location information for 14,065 horseshoe crabs over 18 years. The dataset was initially cleaned by removing records with missing, duplicate or incorrect data. A new data structure was developed to save the data and simplify processing: Three dimensions were used, one ...


The Impact Of Dietary Supplementation Of Arginine During Gestation In A Commercial Swine Herd: I. Gilt Reproductive Performance, Elizabeth A. Hines, Matthew R. Romoser, Zoë E. Kiefer, Aileen F. Keating, Lance H. Baumgard, Jarad Niemi, Nicholas K. Gabler, John F. Patience, Benjamin Haberl, Noel H. Williams, Brian J. Kerr, Kevin J. Touchette, Jason W. Ross Aug 2019

The Impact Of Dietary Supplementation Of Arginine During Gestation In A Commercial Swine Herd: I. Gilt Reproductive Performance, Elizabeth A. Hines, Matthew R. Romoser, Zoë E. Kiefer, Aileen F. Keating, Lance H. Baumgard, Jarad Niemi, Nicholas K. Gabler, John F. Patience, Benjamin Haberl, Noel H. Williams, Brian J. Kerr, Kevin J. Touchette, Jason W. Ross

Aileen Keating

Supplemental Arg during gestation purportedly benefits fetal development. However, the benefits of a gestational Arg dietary strategy in commercial production are unclear. Therefore, objectives of this study examined Arg supplementation during different gestational stages and the effects on gilt reproductive performance. Pubertal gilts (n = 548) were allocated into four treatment groups: Control (n = 143; 0% supplemental Arg) or one of three supplemental Arg (1% as fed) treatments: from 15 to 45 d of gestation (n = 138; Early-Arg); from 15 d of gestation until farrowing (n = 139; Full-Arg); or from 85 d of gestation until farrowing (n = 128; Late-Arg). At farrowing ...


Ovarian Metabolism Of Xenobiotics, Poulomi Bhattacharya, Aileen F. Keating Aug 2019

Ovarian Metabolism Of Xenobiotics, Poulomi Bhattacharya, Aileen F. Keating

Aileen Keating

At birth, the mammalian ovary contains a finite number of primordial follicles, which once depleted, cannot be replaced. Xenobiotic exposures can destroy primordial follicles resulting in premature ovarian failure and, consequently, early entry into menopause. A number of chemical classes can induce premature ovarian failure, including environmental, chemotherapeutic and industrial exposures. While our knowledge on the mechanistic events that occur in the ovary with chemical exposures is increasing, our understanding of the ovary's capacity to metabolize such compounds is less established. This review will focus on three chemicals for which information on ovarian metabolism is known: trichloroethylene, 7,12-dimethylbenz ...


Rcrab: An R Analytics Tool To Visualize And Analyze The Movement Of Horseshoe Crabs In Long Island Sound, Ismael Youssef, Samah Senbel, Jo-Marie Kasinak, Jennifer Mattei Aug 2019

Rcrab: An R Analytics Tool To Visualize And Analyze The Movement Of Horseshoe Crabs In Long Island Sound, Ismael Youssef, Samah Senbel, Jo-Marie Kasinak, Jennifer Mattei

School of Computer Science & Engineering Faculty Publications

Mark-recapture programs are important for studying the ecology and population dynamics of wildlife. An R shiny analytics tool was developed to track the movement of horseshoe crabs in Long Island Sound based on tag and resight data. The crabs were tagged and recaptured by volunteers of Project Limulus, a community-based research program. The dataset contains tag and recapture location information for 14,065 horseshoe crabs over 18 years. The dataset was initially cleaned by removing records with missing, duplicate or incorrect data. A new data structure was developed to save the data and simplify processing: Three dimensions were used, one ...


Distribution And Responsiveness Of Rat Anti-Müllerian Hormone During Ovarian Development And Vcd-Induced Ovotoxicity, Connie J. Mark-Kappeler, Nivedita Sen, Aileen F. Keating, I. Glenn Sipes, Patricia B. Hoyer Jul 2019

Distribution And Responsiveness Of Rat Anti-Müllerian Hormone During Ovarian Development And Vcd-Induced Ovotoxicity, Connie J. Mark-Kappeler, Nivedita Sen, Aileen F. Keating, I. Glenn Sipes, Patricia B. Hoyer

Aileen Keating

Anti-Müllerian hormone (AMH) is produced by granulosa cells in primary to small antral follicles of the adult ovary and helps maintain primordial follicles in a dormant state. The industrial chemical, 4-vinylcyclohexene diepoxide (VCD) causes specific ovotoxicity in primordial and small primary follicles of mice and rats. Previous studies suggest that this ovotoxicity involves acceleration of primordial to primary follicle recruitment via interactions with the Kit/Kit ligand signaling pathway. Because of its accepted role in inhibiting primordial follicle recruitment, the present study was designed to investigate a possible interaction between AMH and VCD-induced ovotoxicity. Protein distribution of AMH was compared ...


Dual Protective Role For Glutathione S-Transferase Class Pi Against Vcd-Induced Ovotoxicity In The Rat Ovary, Aileen F. Keating, Nivedita Sen, I. Glenn Sipes, Patricia B. Hoyer Jul 2019

Dual Protective Role For Glutathione S-Transferase Class Pi Against Vcd-Induced Ovotoxicity In The Rat Ovary, Aileen F. Keating, Nivedita Sen, I. Glenn Sipes, Patricia B. Hoyer

Aileen Keating

The occupational chemical 4-vinylcyclohexene diepoxide (VCD) selectively destroys ovarian small pre-antral follicles in rats and mice via apoptosis. Detoxification of VCD can occur through glutathione conjugation, catalyzed by glutathione S-transferase (GST) enzymes. Further, GST class pi (GSTp) can negatively regulate JNK activity through protein:protein interactions in extraovarian tissues. Dissociation of this protein complex in the face of chemical exposure releases the inhibition of pro-apoptotic JNK. Increased JNK activity during VCD-induced ovotoxicity has been shown in isolated ovarian small pre-antral follicles following in vivo dosing of rats (80mg/ Kg/d; 15d, i.p). The present study investigated the pattern of ...


Inhibition Of Pik3 Signaling Pathway Members By The Ovotoxicant 4-Vinylcyclohexene Diepoxide In Rats, Aileen F. Keating, Shannon M. Fernandez, Connie J. Mark-Kappeler, Nivedita Sen, I. Glenn Sipes, Patricia B. Hoyer Jul 2019

Inhibition Of Pik3 Signaling Pathway Members By The Ovotoxicant 4-Vinylcyclohexene Diepoxide In Rats, Aileen F. Keating, Shannon M. Fernandez, Connie J. Mark-Kappeler, Nivedita Sen, I. Glenn Sipes, Patricia B. Hoyer

Aileen Keating

4-Vinylcyclohexene diepoxide (VCD), an occupational chemical that specifically destroys primordial and small primary follicles in the ovaries of rats and mice, is thought to target an oocyte-expressed tyrosine kinase receptor, Kit. This study compared the temporal effect of VCD on protein distribution of KIT and its downstream PIK3-activated proteins, AKT and FOXO3. Postnatal Day 4 Fischer 344 rat ovaries were cultured in control media ± VCD (30 μM) for 2–8 days (d2–d8). KIT, AKT, phosphorylated AKT, FOXO3, and pFOXO3 protein levels were assessed by Western blotting and/or immunofluorescence staining with confocal microscopy. Phosphorylated AKT was decreased (P < 0.05) in oocyte nuclei in primordial (39% decrease) and small primary (37% decrease) follicles within 2 days of VCD exposure. After d4, VCD reduced (P ...


Impact Of Obesity On 7,12-Dimethylbenz[A]Anthracene-Induced Altered Ovarian Connexin Gap Junction Proteins In Female Mice, Shanthi Ganesan, Jackson Nteeba, Aileen F. Keating Jul 2019

Impact Of Obesity On 7,12-Dimethylbenz[A]Anthracene-Induced Altered Ovarian Connexin Gap Junction Proteins In Female Mice, Shanthi Ganesan, Jackson Nteeba, Aileen F. Keating

Aileen Keating

The ovarian gap junction proteins alpha 4 (GJA4 or connexin 37; CX37), alpha 1 (GJA1 or connexin 43; CX43) and gamma 1 (GJC1 or connexin 45; CX45) are involved in cell communication and folliculogenesis. 7,12-dimethylbenz[a]anthracene (DMBA) alters Cx37 and Cx43 expression in cultured neonatal rat ovaries. Additionally, obesity has an additive effect on DMBA-induced ovarian cell death and follicle depletion, thus, we investigated in vivo impacts of obesity and DMBA on CX protein levels. Ovaries were collected from lean and obese mice aged 6, 12, 18, or 24 wks. A subset of 18 wk old mice (lean ...


High Fat Diet Induced Obesity Alters Ovarian Phosphatidylinositol-3 Kinase Signaling Gene Expression, J. Nteeba, J. W. Ross, J. W. Perfield Ii, A. F. Keating Jul 2019

High Fat Diet Induced Obesity Alters Ovarian Phosphatidylinositol-3 Kinase Signaling Gene Expression, J. Nteeba, J. W. Ross, J. W. Perfield Ii, A. F. Keating

Aileen Keating

Insulin regulates ovarian phosphatidylinositol-3-kinase (PI3K) signaling, important for primordial follicle viability and growth activation. This study investigated diet-induced obesity impacts on: 1) insulin receptor (Insr) and insulin receptor substrate 1 (Irs1); 2) PI3K components (Kit ligand (Kitlg), kit (c-Kit), protein kinase B alpha (Akt1) and forkhead transcription factor subfamily 3 (Foxo3a)); 3) xenobiotic biotransformation (microsomal epoxide hydrolase (Ephx1), Cytochrome P450 isoform 2E1 (Cyp2e1), Glutathione S-transferase (Gst) isoforms mu (Gstm) and pi (Gstp)) and 4) microRNA’s 184, 205, 103 and 21 gene expression. INSR, GSTM and GSTP protein levels were also measured. Obese mouse ovaries had decreased Irs1, Foxo3a, Cyp2e1 ...


Heat Stress Alters Ovarian Insulin-Mediated Phosphatidylinositol-3 Kinase And Steroidogenic Signaling In Gilt Ovaries, Jackson Nteeba, M. Victoria Sanz-Fernandez, Robert P. Rhoads, Lance H. Baumgard, Jason W. Ross, Aileen F. Keating Jul 2019

Heat Stress Alters Ovarian Insulin-Mediated Phosphatidylinositol-3 Kinase And Steroidogenic Signaling In Gilt Ovaries, Jackson Nteeba, M. Victoria Sanz-Fernandez, Robert P. Rhoads, Lance H. Baumgard, Jason W. Ross, Aileen F. Keating

Aileen Keating

Heat Stress (HS) compromises a variety of reproductive functions in several mammalian species. Inexplicably, HS animals are frequently hyperinsulinemic despite marked hyperthermia-induced hypophagia. Our objectives were to determine the effects of HS on insulin signaling and components essential to steroid biosynthesis in the pig ovary. Female pigs (35±4 kg) were exposed to constant thermal neutral (TN; 20°C; 35-50% humidity; n = 6) or HS conditions (35°C; 20-35% humidity; n = 6) for either 7 (n = 10) or 35 d (n = 12). After 7d, HS increased (P < 0.05) ovarian mRNA abundance of the insulin receptor (INSR), insulin receptor substrate 1 (IRS1), protein kinase B subunit 1 (AKT1), low density lipoprotein receptor (LDLR), luteinizing hormone receptor (LHCGR), and aromatase (CYP19a). After 35d, HS increased INSR, IRS1, AKT1, LDLR, LHCGR, CYP19a, and steroidogenic acute regulatory protein (STAR) ovarian mRNA abundance. In addition, after 35d, HS increased ovarian phosphorylated IRS1 (pIRS1), phosphorylated AKT (pAKT), STAR and CYP19a protein abundance. Immunostaining analysis revealed similar localization of INSR and pAKT1 in the cytoplasmic membrane and oocyte cytoplasm, respectively, of all stage follicles, and in theca and granulosa cells. Collectively, these results demonstrate that HS alters ovarian insulin mediated-PI3K signaling pathway members which likely impacts follicle activation and viability. In summary, environmentally-induced HS is an endocrine disrupting exposure that modifies ovarian physiology and potentially compromises production of ovarian hormones essential for fertility and pregnancy maintenance.


A Comparison Of Inflammatory And Oxidative Stress Markers In Adipose Tissue From Weight-Matched Obese Male And Female Mice, Karen J. Nickelson, Kelly L. Stromsdorfer, R. Taylor Pickering, Tzu-Wen Liu, Laura C. Ortinau, Aileen F. Keating, James W. Perfield Ii Jul 2019

A Comparison Of Inflammatory And Oxidative Stress Markers In Adipose Tissue From Weight-Matched Obese Male And Female Mice, Karen J. Nickelson, Kelly L. Stromsdorfer, R. Taylor Pickering, Tzu-Wen Liu, Laura C. Ortinau, Aileen F. Keating, James W. Perfield Ii

Aileen Keating

Expansion of intra-abdominal adipose tissue and the accompanying inflammatory response has been put forward as a unifying link between obesity and the development of chronic diseases. However, an apparent sexual dimorphism exists between obesity and chronic disease risk due to differences in the distribution and abundance of adipose tissue. A range of experimental protocols have been employed to demonstrate the role of estrogen in regulating health benefits; however, most studies are confounded by significant differences in body weight and adiposity. Therefore, the purpose of this study was to compare weight-matched obese male and female mice to determine if the sex-dependent ...


Impact Of Obesity On Ovotoxicity Induced By 7,12-Dimethylbenz[A]Anthracene In Mice, Jackson Nteeba, Shanthi Ganesan, Aileen F. Keating Jul 2019

Impact Of Obesity On Ovotoxicity Induced By 7,12-Dimethylbenz[A]Anthracene In Mice, Jackson Nteeba, Shanthi Ganesan, Aileen F. Keating

Aileen Keating

Insulin, elevated during obesity, regulates xenobiotic biotransformation enzymes, potentially through phosphatidylinositol 3-kinase (PI3K) signaling, in extraovarian tissues. PI3K regulates oocyte viability, follicular activation, and ovarian chemical biotransformation. 7,12-Dimethylbenz[a]anthracene (DMBA), a carcinogen and ovotoxicant, destroys all stages of follicles, leading to premature ovarian failure. Obesity has been reported to promote DMBA-induced tumors, but it remains unknown whether obesity affects ovarian xenobiotic metabolism. Therefore, we investigated ovarian expression of xenobiotic metabolism genes—microsomal epoxide hydrolase (Ephx1), glutathione S-transferase (GST) class Pi (Gstp1) and class mu 1 (Gstm1), and PI3K-signaling members (protein kinase B [AKT] alpha [Akt1], beta [Akt2], and ...


Short Amylin Receptor Antagonist Peptides Improve Memory Deficits In Alzheimer’S Disease Mouse Model, Rania Soudy, Ryoichi Kimura, Aarti Patel, Wen Fu, Kamaljit Kaur, David Westaway, Jing Yang, Jack Jhamandas Jul 2019

Short Amylin Receptor Antagonist Peptides Improve Memory Deficits In Alzheimer’S Disease Mouse Model, Rania Soudy, Ryoichi Kimura, Aarti Patel, Wen Fu, Kamaljit Kaur, David Westaway, Jing Yang, Jack Jhamandas

Pharmacy Faculty Articles and Research

Recent evidence supports involvement of amylin and the amylin receptor in the pathogenesis of Alzheimer’s disease (AD). We have previously shown that amylin receptor antagonist, AC253, improves spatial memory in AD mouse models. Herein, we generated and screened a peptide library and identified two short sequence amylin peptides (12–14 aa) that are proteolytically stable, brain penetrant when administered intraperitoneally, neuroprotective against Aβ toxicity and restore diminished levels of hippocampal long term potentiation in AD mice. Systemic administration of the peptides for five weeks in aged 5XFAD mice improved spatial memory, reduced amyloid plaque burden, and neuroinflammation. The common ...


Phosphoramide Mustard Exposure Induces Dna Adduct Formation And The Dna Damage Repair Response In Rat Ovarian Granulosa Cells, Shanthi Ganesan, Aileen F. Keating Jul 2019

Phosphoramide Mustard Exposure Induces Dna Adduct Formation And The Dna Damage Repair Response In Rat Ovarian Granulosa Cells, Shanthi Ganesan, Aileen F. Keating

Aileen Keating

Phosphoramide mustard (PM), the ovotoxic metabolite of the anti-cancer agent cyclophosphamide (CPA), destroys rapidly dividing cells by forming NOR-G-OH, NOR-G and G-NOR-G adducts with DNA, potentially leading to DNA damage. A previous study demonstrated that PM induces ovarian DNA damage in rat ovaries. To investigate whether PM induces DNA adduct formation, DNA damage and induction of the DNA repair response, rat spontaneously immortalized granulosa cells (SIGCs) were treated with vehicle control (1% DMSO) or PM (3 or 6 μM) for 24 or 48 h. Cell viability was reduced (P < 0.05) after 48 h of exposure to 3 or 6 μM PM. The NOR-G-OH DNA adduct was detected after 24 h of 6 μM PM exposure, while the more cytotoxic G-NOR-G DNA adduct was formed after 48 h by exposure to both PM concentrations. Phosphorylated H2AX (γH2AX), a marker of DNA double stranded break occurrence, was also increased by PM exposure, coincident with DNA adduct formation. Additionally, induction of genes (Atm, Parp1, Prkdc, Xrcc6, and Brca1) and proteins (ATM, γH2AX, PARP-1, PRKDC, XRCC6, and BRCA1) involved in DNA repair were observed in both a time- and dose-dependent manner. These data support that PM induces DNA adduct formation in ovarian granulosa cells, induces DNA damage and elicits the ovarian DNA repair response.


Protective Role For Ovarian Glutathione S-Transferase Isoform Pi During 7,12-Dimethylbenz[A]Anthracene-Induced Ovotoxicity, Poulomi Bhattacharya, Aileen F. Keating Jul 2019

Protective Role For Ovarian Glutathione S-Transferase Isoform Pi During 7,12-Dimethylbenz[A]Anthracene-Induced Ovotoxicity, Poulomi Bhattacharya, Aileen F. Keating

Aileen Keating

7,12-dimethylbenz[a]anthracene (DMBA) destroys ovarian follicles at all developmental stages. This study investigated a role for the glutathione S-transferase (Gst) isoforms alpha (a), mu (m) and pi (p) and the transcription factors, Ahr and Nrf2, during DMBA-induced ovotoxicity, and their regulation by phosphatidylinositol-3 kinase (PI3K) signaling. Negative regulation of JNK by GSTP during DMBA exposure was also studied. Post-natal day (PND) 4 Fischer 344 rat ovaries were exposed to vehicle control (1% DMSO) ± DMBA (1 μM) or vehicle control (1% DMSO) ± LY294002 (PI3K inhibitor; 20 μM) for 1, 2, 4, or 6 days. Total RNA or protein was ...


Mechanisms Of Ovarian Atresia Induced By Xenobiotic Exposures, Jason W. Ross, Aileen F. Keating Jul 2019

Mechanisms Of Ovarian Atresia Induced By Xenobiotic Exposures, Jason W. Ross, Aileen F. Keating

Aileen Keating

The focus of this chapter is the mechanisms of apoptosis that occur during “normal” (physiologically-induced) or “abnormal” (chemical- or pathology-induced) attrition of ovarian oocytes. One of the primary functions of the ovary is development and maturation of oocytes, which occur within a follicular structure. During fetal development of the ovary, primordial germ cells (oogonia) are formed and become oocytes when they stop dividing, and are arrested at the diplotene stage (prophase) of the first meiotic division. At this stage the oocyte is surrounded by a single layer of flattened somatic cells (pre-granulosa cells) and a basement membrane to form primordial ...


Ovarian Expressed Microsomal Epoxide Hydrolase: Role In Detoxification Of 4-Vinylcyclohexene Diepoxide And Regulation By Phosphatidylinositol-3 Kinase Signaling, Poulomi Bhattacharya, Nivedita Sen, Patricia B. Hoyer, Aileen F. Keating Jul 2019

Ovarian Expressed Microsomal Epoxide Hydrolase: Role In Detoxification Of 4-Vinylcyclohexene Diepoxide And Regulation By Phosphatidylinositol-3 Kinase Signaling, Poulomi Bhattacharya, Nivedita Sen, Patricia B. Hoyer, Aileen F. Keating

Aileen Keating

4-vinylcyclohexene diepoxide (VCD) is a metabolite of 4-vinylcyclohexene (VCH) which has the potential to be formed in the ovary through CYP2E1 activity. VCD specifically destroys primordial and small primary follicles in the rodent ovary. Mouse ovaries exposed to VCD demonstrate increased mRNA and protein expression of microsomal epoxide hydrolase (mEH), and an inactive tetrol metabolite (4-(1,2-dihydroxy)ethyl-1,2-dihydroxycyclohexane) can be formed in mouse ovarian follicles, potentially through detoxification action of mEH. In contrast, mEH can bioactivate another ovotoxic chemical, 7,12-dimethylbenz[a]anthracene (DMBA) to a more toxic compound, DMBA-3,4-diol-1,2-epoxide. Thus, the present study evaluated a ...


Glutathione S-Transferase Class Mu Regulation Of Apoptosis Signal-Regulating Kinase 1 Protein During Vcd-Induced Ovotoxicity In Neonatal Rat Ovaries, Poulomi Bhattacharya, Jill A. Madden, Nivedita Sen, Patricia B. Hoyer, Aileen F. Keating Jul 2019

Glutathione S-Transferase Class Mu Regulation Of Apoptosis Signal-Regulating Kinase 1 Protein During Vcd-Induced Ovotoxicity In Neonatal Rat Ovaries, Poulomi Bhattacharya, Jill A. Madden, Nivedita Sen, Patricia B. Hoyer, Aileen F. Keating

Aileen Keating

4-vinylcyclohexene diepoxide (VCD) destroys ovarian primordial and small primary follicles via apoptosis. In mice, VCD exposure induces ovarian mRNA expression of glutathione S-transferase (GST) family members, including isoform mu (Gstm). Extra-ovarian GSTM negatively regulates pro-apoptotic apoptosis signal-related kinase 1 (ASK1) through protein complex formation, which dissociates during stress, thereby initiating ASK1-induced apoptosis. The present study investigated the ovarian response of Gstm mRNA and protein to VCD. Induction of Ask1 mRNA at VCD-induced follicle loss onset was determined. Ovarian GSTM:ASK1 protein complex formation was investigated and VCD exposure effects thereon evaluated. Phosphatidylinositol-3 kinase (PI3K) regulation of GSTM protein was also ...


Disruption Of Female Reproductive Function By Endotoxins, K. L. Bidne, M. J. Dickson, J. W. Ross, L. H. Baumgard, A. F. Keating Jul 2019

Disruption Of Female Reproductive Function By Endotoxins, K. L. Bidne, M. J. Dickson, J. W. Ross, L. H. Baumgard, A. F. Keating

Aileen Keating

Endotoxemia can be caused by obesity, environmental chemical exposure, abiotic stressors, and bacterial infection. Circumstances that deleteriously impact intestinal barrier integrity can induce endotoxemia and controlled experiments have identified negative impacts of lipopolysaccharide (LPS; an endotoxin mimetic) on folliculogenesis, puberty onset, estrus behavior, ovulation, meiotic competence, luteal function and ovarian steroidogenesis. In addition, neonatal LPS exposures have transgenerational female reproductive impacts, raising concern about early life contacts to this endogenous reproductive toxicant. Aims of this review are to identify physiological stressors causing endotoxemia, to highlight potential mechanism(s) by which LPS compromises female reproduction, and identify knowledge gaps regarding how ...


Involvement Of A Volatile Metabolite During Phosphoramide Mustard-Induced Ovotoxicity, Jill A. Madden, Patricia B. Hoyer, Patrick J. Devine, Aileen F. Keating Jul 2019

Involvement Of A Volatile Metabolite During Phosphoramide Mustard-Induced Ovotoxicity, Jill A. Madden, Patricia B. Hoyer, Patrick J. Devine, Aileen F. Keating

Aileen Keating

The finite ovarian follicle reserve can be negatively impacted by chemical exposures including the anti-neoplastic agent, cyclophosphamide (CPA). CPA requires bioactivation to phosphoramide mustard (PM) to elicit its therapeutic effects however; in addition to being the tumor-targeting metabolite, PM is also ovotoxic. In addition, PM can break down to a cytotoxic, volatile metabolite, chloroethylaziridine (CEZ). The aim of this study was initially to characterize PMinduced ovotoxicity in growing follicles. Using PND4 Fisher 344 rats, ovaries were cultured for 4 days before being exposed once to PM (10 or 30 μM). Following eight additional days in culture, relative to control (1 ...


Impact Of 7,12-Dimethylbenz[A]Anthracene Exposure On Connexin Gap Junction Proteins In Cultured Rat Ovaries, Shanthi Ganesan, Aileen F. Keating Jul 2019

Impact Of 7,12-Dimethylbenz[A]Anthracene Exposure On Connexin Gap Junction Proteins In Cultured Rat Ovaries, Shanthi Ganesan, Aileen F. Keating

Aileen Keating

7,12-dimethylbenz[a]anthracene (DMBA) destroys ovarian follicles in a concentration-dependent manner. The impact of DMBA on connexin (CX) proteins that mediate communication between follicular cell types along with pro-apoptotic factors p53 and Bax were investigated. Postnatal day (PND) 4 Fisher 344 rat ovaries were cultured for 4 days in vehicle medium (1% DMSO) followed by a single exposure to vehicle control (1% DMSO) or DMBA (12.5 nM or 75 nM) and cultured for 4 or 8 days. RT-PCR was performed to quantify Cx37, Cx43, p53 and Bax mRNA level. Western blotting and immunofluorescence staining were performed to determine ...