bluuebunny/crossref_metadata_2025 huggingface dataset:
JAZ proteins promote growth and reproduction by restraining transcriptional programs that link primary and specialized metabolism
Authors: Guo Qiang , Major Ian , Yoshida Yuki , Howe Gregg
Published: June 2021

Plants constantly experience attack from surrounding organisms. Because they cannot escape from the environment, plants have evolved a sophisticated immune system to defend themselves against pests and pathogens. However, mounting a defense response is often accompanied by a decrease in growth. The antagonistic relationship between growth and defense has been interpreted as the plant's “dilemma” of how to distribute limited resources between these two fundamental processes. Jasmonic acid (JA) and its derivatives are a class of lipid‐derived hormones that mediate plant immunity against insect herbivores and necrotrophic pathogens, as well as growth, reproduction, and several other physiological processes. JA is perceived by its receptor CORONATINE INSENSITIVE 1 (COI1), which mediates the degradation of the JASMONATE ZIM DOMAIN (JAZ) transcriptional repressors by the 26S proteasome. JAZ degradation releases JA‐responsive transcription factors such as MYC2 from repression, thereby initiating JA responses. Despite the importance of JA in mediating growth‐defense tradeoffs, the mechanism by which JA effects carbon resource allocation between growth and defense is poorly understood. In this study we demonstrate strong constitutive activation of JA signaling pathway in an Arabidopsis mutant (jazD) that lacks ten members of JASMONATE ZIM DOMAIN (JAZ) family proteins. This mutant displayed elevated defense against insect herbivores, stunted growth of vegetative tissues, as well as compromised seed reproduction. Metabolic phenotypes of jazD discerned from global protein and transcript profiling were indicative of elevated carbon allocation to protein‐ and amino acid‐based defense compounds. The strong defense sink in jazD leaves was linked to increased respiration and hallmarks of carbon starvation, whereas area‐based photosynthetic efficiency was unaffected. These findings establish a role for JAZ proteins in promoting growth and reproductive success through restraint of transcriptional programs that link primary metabolism to specialized metabolism. Our data suggest that JAZs adjust growth‐defense balance to match anticipated changes in carbon availability, potentially avoiding detrimental effects of carbon limitation that arise during growth‐to‐defense transitions.

Support or Funding Information

This research was funded by the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, Office of Science, US Department of Energy through grant DE–FG02–91ER20021.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Sinorhizobium meliloti Bacteriophage HMSP‐1‐Susan: Genome Sequence, Molecular Characterization and Implications for Symbiosis
Authors: Fleagle Brennon , Imamovic Aldina , Martinez‐Vaz Betsy
Published: June 2021

Bacteria of the genus Sinorhizobium are plant‐growth promoting microbes that form symbiotic relationships with legumes. These bacteria are important in agriculture because of their ability to convert atmospheric nitrogen gas to ammonia. Ammonia is used to synthesize proteins that are essential for plant growth. Bacteriophages are bacterial viruses that decrease microbial populations in soils and other environments. Despite extensive research on the molecular mechanisms governing the process of nitrogen fixation in Sinorhizobium, little is known about the biotic factors that influence the persistence of these bacteria in soil and impact their ability to fix nitrogen. The presence of bacteriophages in soils has been shown to select for specific types of Sinorhizobium strains and to affect the populations nitrogen‐fixing bacteria. In spite of this observation, there are only a limited number of Sinorhizobium phages that have been characterized. The goal of this project was to carry out a molecular characterization of phage HMSP‐1, a bacteriophage isolated from an agricultural soil that infects strains of Sinorhizobium. Viral DNA was isolated and used to obtain the complete nucleotide sequence of HMSP‐1. Genome analyses revealed that HMSP‐1 is a double stranded DNA virus of 52 kb in size. The genome has a GC content of 52.5% and encodes 96 proteins. 83% percent of these proteins were classified as hypothetical proteins of unknown function. HMSP‐1 did not have significant DNA sequence homology to known bacteriophages; these data suggest HMSP‐1 is novel bacterial virus. We investigated the effects of bacteriophage HMSP‐1 on plant growth and the numbers of Sinorhizobium associated with the model legume, Medicago truncatula. No significant differences in growth or microbial numbers were found between the plants exposed to the phage and unexposed the controls. These data suggest that under the conditions tested, HMSP‐1 does not affect the bacterial populations associated with Medicago truncatula.

Support or Funding Information

The work was supported by grant 1237993 from the National Science Foundation and the Hamline University Ridgeway Fund for Undergraduate Collaborative Research.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
“Community Problem Based Learning to Prepare Students for Real‐World STEM Opportunities”
Authors: Tabor Aaron James , Woodruff Robert , Kesler Kenric M. , Bell Mindy , Ross Danielle K. , Neilson Brent , Gray Ron
Published: June 2021

iCREATE is an NSF‐funded project between the CAVIAT School District and Northern Arizona University's Center for Science Teaching and Learning. The project is designed to engage community partners in a dual‐enrollment (DE) high school bioscience course to positively impact the number of students interested in pursuing bioscience‐related careers and opportunities. The project integrates community collaborations, innovative course design, and modern science technologies to engage students in an authentic problem in their community. A high school level bioscience course has been designed and implemented through CAVIAT, a regional collaboration between school districts to provide high quality career and technical education (CTE) courses. Through the course, project partners engage students in examining community needs regarding the authentic problem of tracking the spread of influenza‐like‐illnesses (ILI) across the Northern Arizona region. Students work with project partners in an effort to design innovative, technology‐rich solutions to address the ILI problem. The university and industry partners bring current knowledge in bioscience and technological understanding of geospatial technologies to the classroom. In this study, we examine the ways in which the project impacts participating students' self‐efficacy, interest in STEM careers, and plan to take part in further internship opportunities. Initial pilot study findings indicate students enjoyed the course, the many challenges it offered, and the opportunity to interact with researchers and community members. Additionally, students' career interest and science motivation increased compared to the control group.

Support or Funding Information

National Science Foundation NSF‐1513198

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Establishing Practices Integrating Commuter Students – Year 1
Authors: Watters Michael K. , Capaldi Mindy , Bugajski Kristi , Schmitt Karl , Schoer Jon , Dahlke‐Goebbert Bonnie
Published: June 2021

The EPIC grant focuses on the integration of STM (Science, Technology, & Mathematics) commuter students into the Valparaiso University (VU) campus community through shared undergraduate research experiences, as well as skill‐building and social activities. Goals of the EPIC grant include improving recruiting and retention of STM students, increasing the number of STM graduates prepared for research careers, and increasing the number of research intensive majors within STM departments. Students are recruited with scholarships averaging $5,300 per year.

During year 0 the EPIC program developed the infrastructure and tools to recruit, select, enroll, and support the first cohort of students in fall 2017. We report the results of our recruitment initiatives on the poster. In year 1 we refined these practices to include earlier and more varied contact with high schools. Recruitment of the second cohort is underway.

Assessment procedures were initiated through the establishment of pre‐EPIC baselines measuring retention, academic, and demographic data for STM students at VU. Data were collected through institutional research and multiple student surveys. This data includes GPAs, STM student retention, graduation rates, and the impact of undergraduate research. Some data is broken down demographically according to gender, race, ethnicity, and commuter status. Preliminary analysis revealed several notable results, such as a 44% retention rate for STM students remaining in their original major and 25% of VU STM students from the 2012 freshman cohort not graduating at all. Additionally, VU commuters are less likely to be retained in a STM major and are less likely to graduate. This group of students is the primary focus of the EPIC program; we predict that our model will improve their retention numbers.

Support or Funding Information

This work was supported by award # 1564855 from the National Science Foundation.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Type III Polyketide Synthase Involved in Tropane and Granatane Alkaloid Biosynthesis
Authors: Kim Neill , D'Auria John C.
Published: June 2021

The lack of data concerning the enzymes and genes responsible for the production of tropane and granatane alkaloids has hindered endeavors to metabolically engineer important pharmacologically active specialized metabolites. It is important to understand how biochemical pathways perform when they originate from evolutionarily diverse families. It is hypothesized that a type III polyketide synthase enzyme is involved in the extension and subsequent cyclization of the second ring in the bicyclic tropane and grantane core structure. This represents a new class of type III polyketide synthases in which the heterocyclic starter molecule is charged and also lacks a Coenzyme A thioester. Putative polyketide synthase enzymes were identified in the Erythroxylum coca and Punica granatum cDNA transcriptome database and heterologously expressed in Pichia pastoris. LC‐MS/MS analysis has revealed the formation of a 4‐(1‐methyl‐2‐pyrrolidinyl)‐3‐oxobutanoate intermediate that is hypothesized to lead to the second ring formation. Understanding how evolutionary changes have occurred in type III polyketide synthases from different metabolic pathways that determine the development and expansion of chemical diversity to produce high valued compounds will be a powerful tool for bioengineering pharmaceuticals.

Support or Funding Information

National Science Foundation

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Nutrient sensing and mitochondrial Coenzyme Q biosynthesis: Are they connected by a phosphatase?
Authors: Awad Agape M. , Venkataramanan Srivats , Nag Anish , Galivanche Anoop R. , Bradley Michelle C. , Neves Lauren , Douglass Stephen , Johnson Tracy L. , Clarke Catherine F.
Published: June 2021

Coenzyme Q (also termed ubiquinone or Q) is an electron carrier in the mitochondrial respiratory chain that functions as an essential component in energy metabolism processes, while also serving as a vital lipid soluble antioxidant. Sufficient de novo Q biosynthesis is crucial for human health. Saccharomyces cerevisiae (baker's yeast) serves as an excellent model for studies on Q because of its powerful molecular genetics and the functional identity of yeast and human COQ genes. Coenzyme Q biosynthesis in S. cerevisiae occurs in the mitochondria and, currently, fourteen known mitochondrial proteins are responsible for facilitating this process—Coq1–Coq11, Yah1 (ferredoxin), Arh1 (ferredoxin reductase), and Hfd1 (aldehyde dehydrogenase). Many of the Coq proteins associate in a high molecular weight complex (“CoQ‐Synthome”) that localizes in the inner mitochondrial membrane. Previous work indicated that the stability of the Q biosynthetic complex (as well as the phosphorylation status of several Coq polypeptides, including Coq3, Coq5, Coq7, and potentially Coq6) relied on the presence of Coq8, the putative kinase of the system. Thus, the role of phosphorylation may provide novel information as to how the CoQ‐synthome is regulated. Indeed, intriguing links have been discovered that relate Coq polypeptides and phosphorylation status with nutrient availability and regulatory genes. Most recently, a link has been found between a nutrient sensing gene, SNF2, and the spicing efficiency of PTC7, the phosphatase responsible for controlling a key regulation site in Q biosynthesis. Our objectives are to further investigate these regulatory links. Such studies will aid our understanding and treatment of multiple diseases caused by CoQ‐deficiencies and provide novel insight in longevity and issues related to aging in humans.

Support or Funding Information

This work was supported by the National Science Foundation, Grant MCB‐1330803 and the National Institute of General Medical Sciences (GM‐085474), Whitcome Pre‐doctoral Fellowship in Molecular Biology, and the Ruth L. Kirschstein National Service Award GM‐007185.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
The PAH1‐encoded Phosphatidate Phosphatase Plays a Role in Lipogenesis in the Oleaginous Yeast Yarrowia lipolytica
Authors: Hardman Derell , Ukey Rahul , Fakas Stylianos
Published: June 2021

The PAH1 gene encodes for phosphatidate phosphatase (PAP), the enzyme that catalyzes the dephosphorylation of phosphatidic acid to yield diacylglycerol and Pi. PAP activity is essential for the biosynthesis of the storage lipid triacylglycerol (TAG) in several eukaryotes (e.g. Saccharomyces cerevisiae). In Yarrowia lipolytica, TAG accumulates during lipogenesis and is responsible for the increase in cellular lipid content under these conditions. We hypothesized that under lipogenic conditions, lack of the Y. lipolytica PAH1 homolog (i.e. pah1Δ) should result in a reduction of lipid content and diminished TAG levels. To examine this hypothesis, we cultivated the Y. lipolytica wild type and pah1Δ strains in a lipogenic (8% glucose) medium, and analyzed their growth and lipid profiles. The results showed that the pah1Δ mutant exhibited increased growth but a lower lipid content in comparison to the wild type. However, the pah1Δ mutant was able to synthesize TAG, although to a lesser extent than the wild type. These results indicate that the Y. lipolytica PAH1 homolog contributes to, but is not essential for, TAG biosynthesis during lipogenesis.

Support or Funding Information

This research was supported by Evans‐Allen Grant no.1005735 and Capacity Building Grants Program Grant no.1007503 from the USDA National Institute of Food and Agriculture, as well as the Strengthening Historically Black Graduate Institutions Program under Title III of the Higher Education Act of 1965.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Chemoproteomic discovery of ligand binding hotspots in the lipid kinome
Authors: Hsu Ku‐Lung
Published: June 2021

Diacylglycerol kinases (DGKs) are integral components of signal transduction cascades that regulate cell biology through phosphorylation of the ubiquitous secondary messenger diacylglycerol. Methods for direct evaluation of DGK activity in native biological systems are lacking and needed to develop chemical probes for studying isoform‐specific functions. Here, I will discuss efforts from my group to utilize ATP acyl phosphate activity‐based probes and quantitative mass spectrometry to map previously undefined ATP‐ and small molecule‐binding sites of representative members from all subtypes of the DGK family. We use our chemoproteomics strategy to discover an unusual binding mode for the DGK‐alpha (DGKA) inhibitor ritanserin, including interactions at a novel binding site remote from the ATP binding pocket. I will also describe our efforts towards discovery of fragment leads for future development of highly potent and selective DGKA inhibitors. Collectively, our studies illustrate the power of chemoproteomics to site‐specifically profile protein‐small molecule interactions and reveal key ligand binding sites for selective inactivation of the DGK family of lipid kinases.

Support or Funding Information

This work was supported by the University of Virginia, the LaunchPad for Diabetes Program funded by the Manning Family Foundation at the University of Virginia, National Institutes of Health Grants (DA035864 and DA043571) and U.S. Department of Defense (W81XWH‐17‐1‐0487 to K.‐L.H.).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Modified HPLC method for detection of hydroxyoctadecadienoic acid with greater sensitivity
Authors: Chesmore Nathan J. , Zhang Wenjing , Richards Mark P. , Shanmuganayagam Dhanansayan
Published: June 2021

Hydroxyoctadecadienoic acid (HODE) is an oxidized fatty acid derivative of linoleic acid that is also a product of lipoxygenase‐mediated reactions. It has been implicated in the development of cardiovascular disease, differentiation of adipocytes, and several pro‐ and anti‐inflammatory mechanisms. HPLC, GC/MS, and LC/MS methods have been developed for the assay of HODE(s) from simple assay schemes to lipidomic approaches in fatty acid signaling, metabolism, and regulation research. Current HPLC methods used to quantify unmodified HODE(s) in fatty acid matrixes lack the required sensitivity necessary to detect levels of HODE found in tissue and serum. Sensitizing agents that bind HODE(s) or all sample fatty acids may be used to increase the signal of analytes by post‐column detectors. Here we describe the use of 2‐(2,3‐Naphthalimino)ethyl trifluoromethanesulfonate (NT) as a sensitizing fluorophore for the detection of 13(S)‐HODE in fatty acid matrixes using a c18 column for separation. NT fluorophore detection increases the sensitivity by more than 15‐fold compared to unmodified HODE conjugated diene absorbance at 234nm without modifications for fluorophore‐HODE conjugate stability. Pre‐column labeling of HODE with NT fluorophore in fatty acid matrixes also markedly increases the c18 column separation efficiency of the HODE‐fluorophore conjugate, reducing the amount of solvent and analysis time needed if shorter columns are used.

Support or Funding Information

The research was supported by the Biomedical & Genomic Research Group Discretionary Fund (University of Wisconsin‐Madison) and by USDA Formula Project Grant (WIS01972) National Institute of Food and Agriculture, USDA).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Personalized Diagnosis for Lafora Disease, a Fatal Epilepsy
Authors: Wayne Jeremiah L. , Brewer M. Kathryn , Gentry Matthew S.
Published: June 2021

Lafora disease (LD) is a fatal, genetic disorder characterized by progressive neurodegeneration, myoclonus (i.e. uncontrolled muscle spasms), and epilepsy. LD patients present with seizures in adolescence that become increasingly severe and frequent, suffer rapid cognitive decline, and typically die within ten years of onset. Abnormal carbohydrate deposits known as Lafora bodies are found in the brains of LD patients and have been shown to drive disease progression.

Approximately 50% of LD cases are caused by mutations in the Epilepsy progressive myoclonus 2A (EPM2A) gene that encodes a protein called laforin. Laforin is the only protein in humans that can release phosphate from carbohydrates. In LD patients, mutations in laforin lead to excess phosphate and abnormal branching in cellular carbohydrate stores, causing carbohydrate accumulation and toxicity. There are >50 different laforin mutations that have been described in LD patients, and some patients have milder forms of the disease. We have shown that not all laforin mutations are the same and that many mutations may have milder effects on protein function. Our biochemical analysis of disease mutations would allow us to predict disease outcome based on a patient's individual genetics.

Recently an unusual case of late‐onset LD was described in a patient who lived to the age of 59. This patient contained a novel laforin mutation, where a specific amino acid at position 321 was changed from a phenylalanine to a cysteine (F321C). The objective of this study was to determine the effect of the F321C mutation on laforin function. We generated and purified recombinant laforin F321C, tested its biochemical effects using activity, binding, and stability assays, and found that this mutation and a previously described LD patient mutation, F321S, only mildly alter the function of laforin, providing a biochemical explanation for the very mild clinical phenotype. Our studies also establish a biochemical avenue for rapid, personalized diagnoses of LD patients, enabling doctors to predict patient progression and design treatment schemes that are specific to patients.

Support or Funding Information

This study was supported by NIH Grants F31NS093892 (M.K.B), R01NS070899 and P01NS097197 (M.S.G.), and NSF Grant IIA‐1355438 (M.S.G.).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Cultural Analysis and Demographics of Caprine Consumption among Texas Residents
Authors: Dimgba Chibuikem Samuel , Copeland Beverly , Ojumu Oluwagbemiga
Published: June 2021

Background

According to report, nearly 1.5 million heads of goat are consumed annually within the US. The popularity of goat meat is increasing due to the increase in consumption from the immigrant population according to D'Rozario and Gaung, (2015), with this increase in demand for goat meat, the purpose of this research was to study the current and projected consumption patterns of Caprine products in Texas and identify target populations for the distribution of value‐added caprine products.

Methods

A survey was developed, validated and piloted to measure food acceptability, cultural influences, demographics, generational acceptance, and geographic influences. Communities were selected by stratified sampling. Individuals were selected to participate through community outreach activities. Data were collected by trained research personnel.

Data Analysis

The data were analyzed using the statistical package for the Social Sciences (SPSS)

Results

The sample size consisted of 79 study participants, with the majority (48%) being African Americans. When combined with participants who identified as African or Caribbean, blacks accounted for 58% of the total participants. This was followed by whites with 24% and Hispanics with 13% of the sample. Of the 79 participants who participated in the study, 44 (56%) reported having tasted goat meat before and 35 (44%) said they had never tasted it. Of the 44 who reported having tasted goat meat, only 15 (19%) reported actually eating it. The majority of the participants, 69, reported not seeing goat meat in their local grocery stores. Only 7 (9%) reported having seen goat meat in the grocery store.

Support or Funding Information

Evans Allen Capacity Funds, NIFA

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Nicotinic Receptor Dysfunction Underlies Abnormal Responses to Muscarinic Receptor Antagonist Treatment in DYT1 Dystonia
Authors: Downs Anthony M. , Fan Xueliang , Hess Ellen J.
Published: June 2021

DYT1 dystonia is an inherited dystonia caused by a mutation in the TOR1A gene, which encodes the torsinA protein. Trihexyphenidyl (THP) a nonselective muscarinic receptor antagonist is the small molecule drug of choice for the treatment of DYT1 dystonia, but it is poorly tolerated due to significant side effects. A better understanding of both the underlying pathology of DYT1 dystonia and the mechanism of action of THP is needed for the development of better treatments. Abnormalities in dopamine (DA) transmission have been implicated in DYT1 and other forms of dystonia. Previous studies have shown that Dyt1 knockin mice have reduced extracellular striatal DA in vivo and reduced striatal DA release ex vivo. Acetylcholine (ACh) is known to play a key role in regulating striatal DA release, and muscarinic receptor antagonists are used to treat dystonia which suggests the hypothesis that ACh signaling mediates the DA release deficiency in DYT1 dystonia. To test this hypothesis, we used ex vivo fast scan cyclic voltammetry (FSCV) and in vivo microdialysis to investigate ACh regulation of striatal DA release in Dyt1 knockin mice. We found that THP increased striatal DA release as assessed by ex vivo FSCV and in vivo microdialysis. However, THP was less effective at enhancing DA release in DYT1 mice relative to WT mice in the ex vivo FSCV experiment (mean increase WT: 65% vs DYT1: 35%). In striatum, blocking mACh autoreceptors on cholinergic interneurons increases ACh release. ACh then activates nicotinic acetylcholine receptors (nAChRs) located on DA terminals to enhance DA release. To determine if abnormal nAChR responses could account for the attenuated response to THP in Dyt1 knockin mice, we performed a dose response experiment with the α4 nAChR subunit selective antagonist dihydro‐β‐erythroidine. We found that Dyt1 mice are more sensitive to nAChR antagonism, which reduces DA release (IC50: WT= 29.46nM, DYT1= 12.26nM). These data suggest that nAChRs mediate the differential effects of THP in Dyt1 mice. These data also suggest that nAChRs may be suitable therapeutic targets for DYT1 dystonia. Future studies will evaluate nAChR positive allosteric modulators for DA‐enhancing effects.

Support or Funding Information

Supported by: DoD W81XWH‐15‐1‐0545 and NIH T32‐GM008602

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Identification of key regions mediating human melatonin type 1 receptor biased signaling revealed by natural variants
Authors: HEGRON Alan , PLOUFFE Bianca , BONNEFOND Amelie , GAO Wenwen , FROGUEL Philippe , LE GOUILL Christian , JOCKERS Ralf , BOUVIER Michel
Published: June 2021

Background

Melatonin is a circulating neurohormone mainly released from the pineal gland in a circadian manner regulating many physiological functions, such as glycemia homeostasis. A misregulation of the glycemia could lead to Type 2 Diabetes (T2D). Recently, a genetic variant (rs2119882) found on melatonin type 1 receptor (MT1) promoter has been associated to higher risks to develop T2D.

Hypothesis

Variants on the MT1 coding region could modify receptor signalling and consequently modulate incidence to develop T2D.

Methods

we performed a genome‐wide association studies with both normoglycemic individuals and T2D patients and 32 non synonymous polymorphisms of MT1 were identified. Our primary aim was to functionally characterize these variants to further verify if there is an association between specific impaired signaling pathways and higher risks to develop T2D. Affinity for melatonin (Kd) and cell surface expression of variants were measured using 125I‐melatonin binding assays and enzyme‐linked immunosorbent assay (ELISA) respectively to be sure that all receptors were expressed at the cell surface. Melatonin dose‐response curves were performed at equivalent cell surface expression to determine MT1s efficacy and potency to activate each G protein using a bioluminescence resonance energy transfer (BRET) assay monitoring proximity between G alpha and G gamma proteins as activation indicator. Finally, b‐arrestin 2 recruitment to cytoplasmic membrane upon MT1s stimulated with melatonin was measured by monitoring BRET between RlucII‐fused b‐arrestin and the cytoplasmic membrane targeted sensor (rGFP‐fused to CAAX box of KRas).

Results

Results obtained by the functional characterization indicate several types of signaling signatures. Indeed, some mutations show a biased signaling between G protein activation and b‐arrestin recruitment, while others show G protein isoform selectivity.

Conclusion

Association studies between patients having MT1 mutations and these signaling signatures will help us to understand in a better way the relationship between melatonin and insulin and represent an opportunity to identify new potential genetic markers of higher risks to develop T2D.

Support or Funding Information

This work was supported by a grant from CIHR to MB, from FRM, ANR and Université Paris 5 Descartes to RJ and from NIH, NSF and DARPA to OL. PhD student fellowship from Université de Montréal to AH, postdoctoral fellowships from CIHR and from Diabetes Canada to BP and from FRM to AK also funded this work.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Can Inhibition of Soluble Epoxide Hydrolase Ameliorate Increased Susceptibility to DSS Induced Colitis in Soybean Oil Fed Mice?
Authors: Deol Sonia Poonamjot , Yang Jun , Yu Jacqueline , Trivedi Amol , Morisseau Christophe , Hammock Bruce D. , Sladek Frances M.
Published: June 2021

The incidence of inflammatory bowel disease (IBD) in the U.S. has increased significantly since 1980, preceded by a remarkable increase in the prevalence of soybean oil (SO) in the American diet. Although the role of SO in the etiology of IBD is ambiguous, one of its major components, linoleic acid (LA), is associated with an increased risk of ulcerative colitis (UC). LA is metabolized into oxylipins (also referred to as leukotoxins) via a two‐step reaction involving cytochrome P450 enzymes and the enzyme soluble epoxide hydrolase (sEH). We have recently shown that a diet high in SO (and thus LA) increases the accumulation of oxylipins generated by the cytochrome P450 (CYP)/soluble epoxide hydrolase (sEH) pathway in the liver (PMID: 28970503). Our lab has previously shown that LA decreases the protein levels of the transcription factor hepatocyte nuclear factor 4 alpha (HNF4α) in wild type mouse livers (PMID: 19440305) and that a proper balance of HNF4α isoforms is required for the development and proper functioning of the gut (PMID: 27166517). To investigate the effect of dietary soybean oil on intestinal health, mice were fed either a low‐fat, high fiber vivarium chow diet (Viv) or a high soybean oil plus fiber diet (SO+f) with 19 kcal% LA and the same amount of fiber as Viv. Both male and female C57BL/6N (WT) mice and HNF4α exon‐swap mice (genetically engineered to express just the P1‐form of HNF4α so that in tissues, such as intestine, where both P1 and P2 promoters are active, only HNF4α1/2 is expressed instead of both HNF4α1/2 and HNF4α7/8 (PMID: 16498401)) were put on the diets at weaning. Colitis was induced by Dextran Sodium Sulfate (DSS) treatment eight to nine weeks after weaning. Increased weight loss was observed in both male and female HNF4α (A1HMZ) transgenic mice compared to WT mice, regardless of diet. Differences in other parameters, such as spleen weight, colonic and small intestinal length, that could be attributed to differences in diet and/or HNF4α isoform were also observed. Experiments with an inhibitor of soluble epoxide hydrolase are underway to determine whether soybean oil affects the balance between the HNF4α isoforms via LA diols and whether blocking the production of these oxylipins will make the mice more resistant to DSS induced colitis.

Support or Funding Information

Supported by CCFA CDA 454808 (P.D.), USDA NIFA Hatch project CA‐R‐NEU‐5680 (F.M.S.), NIEHS grant R01‐ES002710 (B.D.H.), Superfund Research Program P42 ES04699 (B.D.H.).

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Targeting the Right Ventricle as a Treatment Strategy for Pulmonary Arterial Hypertension
Authors: Gorr Matthew W. , Liang Wenjing , Muthusamy Abinaya , Sriram Krishna , Insel Paul A.
Published: June 2021

Pulmonary arterial hypertension (PAH) is characterized by remodeling of the pulmonary artery within the lung that leads to increased pulmonary vascular resistance (PVR). This increase in PVR can result in progressive remodeling of the right ventricle (RV) and eventual right heart failure. Current therapy aims to decrease PVR via vasodilation of the PA, but this strategy does not work for some patients, and is not a cure for the disease. This study aims to uncover novel therapeutic targets by assessing the RV for expression of G protein‐coupled receptors (GPCRs) during PAH. GPCRs are the target of more than 30% of approved drugs and regulate multiple cellular processes by activation on the cell membrane. We utilized a rat model of PAH in which Sprague Dawley rats were placed in hypoxia (10% O2) for two weeks, or normoxia as a control (n=3–4/treatment). Placement in hypoxia resulted in RV remodeling evident by an increase in the RV weight/left ventricle (LV) + septum weight (hypoxia‐treatment: 0.458 ± 0.036 vs. control rats: 0.210 ± 0.020; mean ± standard deviation, Student's t‐test p<0.0001). We then utilized RNA sequencing (RNA‐seq) of the RV and the left ventricle (LV) tissue coupled with differential expression analysis to identify GPCRs expressed with significantly (false discovery rate <0.1) altered expression in the RV of hypoxia‐treated rats compared to controls. In the RV, 37 GPCRs were altered in expression in hypoxia‐treated rats vs. control rats compared to 7 GPCRs altered in the LV. To examine the cell‐specific expression of these targets, cardiac myocytes and fibroblasts were isolated from a mouse model of PAH (10% O2 for 3 weeks with weekly injection of SU5416). qPCR of the target GPCRs revealed that most of the differentially expressed GPCRs were altered in the cardiac myocytes but not in the fibroblasts. These results indicate that a subset of genes is differentially expressed in the RV compared to the LV both in normoxia and hypoxia, particularly in the cardiac myocytes. This raises the possibility that treatments for PAH could be directed at GPCRs selectively expressed in RV remodeling, thus providing a novel means of treating an unmet need for patients with RV hypertrophy and failure.

Support or Funding Information

Supported by NIH Grant: T32HL007444‐34 and DoD Grant: W81XWH‐14‐1‐0372

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Endothelial Cell Caveolin‐1 Depletion Modulates TGF and Notch Signaling Associated with Pulmonary Arterial Hypertension
Authors: Oliveira Suellen DArc Santos , Chen Jiwang , Castellon Maricela , Chernaya Olga , Colamonici Oscar , Bonini Marcelo G. , Machado Roberto , Minshall Richard
Published: June 2021

Reduced expression of caveolin‐1 (Cav1) in endothelial cells (ECs) is associated with pulmonary vascular remodeling. Pulmonary vascular pathology in global Cav1−/− mice is prevented when eNOS is also deleted. Thus, we hypothesized that eNOS‐derived oxidants in absence of Cav‐1 result in dysfunctional ECs and pathological vascular remodeling. Hypoxia exposure following VEGFRII inhibition in Sprague‐Dawley rats produced a significant reduction in pulmonary Cav‐1 expression. Moreover, C57BL6 WT (Tie2.Cre−;Cav1lox/lox) mice exposed for 1 month to hypoxia exhibited an increase in the right ventricular systolic pressure (RVSP) from 25.33 to 31.68 ± 1.75 mmHg which was even higher in EC‐Cav1−/− (Tie2.Cre+;Cav1lox/lox) mice (27.95 to *38.48 ± 1.76 mmHg) when compared to normoxia. The increase in pulmonary arterial pressure induced by hypoxia was associated mild cardiac hypertrophy, increased plasma TGF‐β levels, and increased microvascular area/thickness indicative of vascular remodeling. Interestingly, under normoxic conditions, we observed reduced VEGF level in the lungs of EC‐Cav1−/− mice (8998 ± 1062 and 4927 ± 505.4 pg/mL, n=3–7 mice) suggesting Cav‐1 expression may be required to maintain normal VEGF signaling. Since VEGFRII inhibition may promote EC injury, we hypothesized that in the absence of endothelial Cav‐1, a defect in VEGF‐dependent vascular repair after injury may be the mechanism underlying pathological vascular remodeling. Then, mouse lung ECs (MLECs) isolated from WT, Cav1−/−, and Cav1−/−;eNOS−/− mice were used to assess gene and protein expression, and to quantify indices of angiogenesis. We have observed a defect in tube formation in Cav1−/− MLECs which was abo‐lished with genetic ablation of eNOS−/−. Moreover, in support of these findings, we observed increased Notch‐2 and ‐3 gene expression, and reduced Notch‐1 cleavage in Cav1−/− but not Cav1−/−;eNOS−/− MLECs. Finally, our data suggest the absence of endothelial Cav‐1 may promote pathological vascular remodeling by modulating TGF‐β and Notch signaling.

Support or Funding Information

CNPq Fellowship‐CsF/Brazil; HL125356; DOD W911NF‐15‐R‐002.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Implementation of a place‐based, interdisciplinary, and research‐oriented teaching model to increase retention in STEM
Authors: Frederich Markus , Roese Ursula , Bymers Leah , Zeeman Stephan
Published: June 2021

STEM education is becoming increasingly important for succeeding in a society that depends on science and technology. However, STEM persistence rates in undergraduate programs are low, nationally in the range of 40%. A pedagogical shift towards more experiential, project‐based education has shown to be successful in increasing STEM persistence rates, enhancing understanding of science, and increasing student's motivation to learn. Building on a strong track record in undergraduate research we are implementing a place‐ and theme‐based interdisciplinary project that spans all 4 years of undergraduate education in all STEM majors. We use our location at the mouth of the Saco River and the surrounding ecosystem as the overarching theme: TURBO: The Undergraduate Saco River Biodiversity Observatory. This project is modeled after an LTER‐research site. Multiple existing and newly developed lab activities and field trips in Biology, Marine Biology, Environmental Science, Chemistry and Physics, and Mathematics are focused on the Saco River Ecosystem and new activities are implemented to investigate in an LTER‐style the fresh and seawater systems surrounding the campus. Students collect data on water parameters, invasive species, invertebrates, DNA barcoding, fish, algae, plankton, marsh diversity, plants, invasive species, and more. All data are available to students and faculty in a central database, developed by students. Statistics courses will use student‐generated data instead of standard text book examples. Our annual assessment tools revealed actually a slight drop in confidence in STEM comparing the time points before starting college and after the first year. This drop potentially represents an initially idealized view of STEM, that becomes more realistic during the first college year. This highlights the importance of engaging and authentic STEM activities early in the college career to fully capture them. This place‐ and project‐based interdisciplinary approach can be used as a blueprint for other programs and schools.

Support or Funding Information

Funded by NSF grant# DUE‐IUSE 1431955.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
3D Contrast Techniques for Visualizing Anatomy and Their Application for Human Education, Vertebrate Biomechanics and Paleobiology
Authors: Holliday Casey Monahan , McGetchie Faye , Johnson Laura , Hill Cheryl , Sellers Kaleb , Middleton Kevin , Sullivan Samuel , Schiffbauer James
Published: June 2021

New contrast techniques for visualizing 3D anatomy of soft tissues using tomography are revealing outstanding new opportunities for understanding the internal anatomy of humans and other animals. Iodine‐contrast (DiceCT) combined with 3D modeling and computational approaches now enables new directions in anatomy education, body composition, muscle fiber architecture, biomechanics and evolutionary biology. Here we present several new applications of DiceCT‐derived soft‐tissue data used in education and research approaches. First we illustrate new methods using human material that reveal new data on complicated, high‐yield soft‐tissue centric regions including the infratemporal fossa and ear, throat, pelvic floor and wrist. Second we share new methods in applying 3D muscle fiber architecture estimates to biomechanical investigations of vertebrate function. Finally we show how soft‐tissue data help test taphonomic and paleobiological hypotheses of body composition in soft‐bodied fossils derived from conservat‐lagerstatten. These vignettes offer incredible utility for hypothesis testing, biomechanical modeling and anatomy education.

Support or Funding Information

NSF IOS 1457319, NSF EAR 163753, Missouri Research Council, NSF CAREER 1652351, NSF IF 1636643

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Spatial and temporal processes involved in the naturally occurring sarcomere disassembly during electric organ regeneration in the teleost Sternopygus macrurus
Authors: Unguez Graciela Alexandra
Published: June 2021

Exciting new findings are increasing our understanding of the protein composition of sarcomeres – the highly ordered contractile units of striated muscle – and the common pathways of sarcomere assembly in both heart and skeletal muscle. Although disruption s in these pathways are implicated in muscle diseases, events involved in the disassembly of sarcomeres remain largely unknown. The electric fish S. macrurus has the unique ability to regenerate all tail tissues after amputation including skeletal muscle and the muscle ‐ derived cells of the electric organ, i.e., electrocytes. During regeneration, some muscle fibers fuse and disassemble their sarcomeres to convert into electrocytes. To study changes in the spatial and temporal expression of protein components of distinct sarcomeric regions during the muscle ‐ to ‐ electrocyte conversion, we have begun to use immunofluorescence and ultrastructural approaches. Specifically, we are using markers of the Z ‐ disk (EA53 antibody), inter ‐ Z ‐ disk region (E398P antibody), A ‐ b ands (MF20 antibody), and I ‐ bands (phalloidin stain) to describe this naturally occurring process of sarcomere disassembly during electrocyte differentiation. Our preliminary data suggest a pattern of sarcomere breakdown characterized by dissociation of the I ‐ band followed by loss of A ‐ band then Z ‐ disks structures while the inter ‐ Z ‐ disk components are conserved. This pattern of sarcomere disassembly differs from a dedifferentiation process predicted by a current model of sarcomere formation during muscle differentiation (Ehler and Gautel, Adv. Exp. Med. Biol., 2008).

Support or Funding Information

The following grants funded this work: NIH SCORE Grant 1SC1GM092297‐01A1 (GAU), NSF Grant CNS‐1248109 (GAU), and Howard Hughes Medical Institute Science Education Grant 5200693.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.
Effects of the Organophosphate Pesticide Chlorpyrifos on Muscle Architecture in 72 hour Zebrafish Embryos
Authors: Willrich Robert C. , Watanabe Taylor H. , Fradinger Erica A.
Published: June 2021

Chlorpyrifos (CPF) and other organophosphate pesticides (OP) are commonly used in the agricultural industries of the United States and developing countries. High concentrations of OP's in rivers and lakes have led to concerns for its negative effects on the wildlife and humans. These substances have been documented to act as neurotoxins that lead to the buildup of acetylcholinesterase in cholinergic neurons. This study was performed to determine the effects of CPF on the development of muscles for zebrafish embryos 72 hours post‐fertilization (hpf) in a calcium‐depleted medium. Using confocal microscopy of wholemount MF20 immuno‐labeled embryos, several morphological changes were observed. There was an increase in the length of muscle myomeres and width of individual muscle fibers in CPF exposed embryos. Additionally, some embryos showed strong muscle fiber disorganization. These observations show that CPF has a clear effect to muscle development and architecture possibly due to increased cholinergic stimulation during development.

Support or Funding Information

Department of Defense, ARO 66367‐RT‐REP; Keck Foundation Undergraduate Fellowship to R.C.W.; James Irvine Chair in Biological Sciences to E.A.F.

This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.