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Using Crispr/Cas9 Gene Editing To Remove Twist2 To Determine Its Role During Somite Development In Zebrafish, Caleb Holdener 2020 Belmont University

Using Crispr/Cas9 Gene Editing To Remove Twist2 To Determine Its Role During Somite Development In Zebrafish, Caleb Holdener

Belmont Undergraduate Research Symposium (BURS)

Embryonic Mesoderm is a precursor to various types of tissues, such as the somatic mesoderm. Somites are a segmented tissue that gives rise to the sclerotome in vertebrates. This sclerotome further develops into the axial skeleton (Stickney, Barresi, and Devoto 2000). Somites are easily observable under low power microscopy during fish development. In fish, great strides have been made in studying the developmental fate of somites (Stoiber, Haslett, and Sänger 1999; Stickney, Barresi, and Devoto 2000). Zebrafish have been studied in many labs due to their similarities with higher vertebrates. These advantages include their inexpensive cost of care, external fertilization ...


Utilizing Crispr/Cas9 Gene Editing Determine The Role Of Cnr1 During Zebrafish Development, Ellis Chase 2020 Belmont University

Utilizing Crispr/Cas9 Gene Editing Determine The Role Of Cnr1 During Zebrafish Development, Ellis Chase

Belmont Undergraduate Research Symposium (BURS)

The CRISPR-Cas 9 system is a process used to create mutations, insertions, or deletions within a desired gene. It can be used to create mutant organisms necessary to study the function and impact of a particular gene1. Cannabinoids are active chemicals found in cannabis, the most common of which being cannabinol (CBD)2. Cannabinoid receptors are an integral part of the endocannabinoid system, recognizing cannabinoid signals to promote physiological processes like pain sensation, memory, mood, and appetite3. Studying these cannabinoid receptors can provide insight into how cannabis can affect bodily processes.

Zebrafish are small translucent freshwater fish residing ...


Utilizing Crispr/Cas9 Gene Editing Determine The Role Of Cnr1 During Zebrafish Development, Camme Sells 2020 Belmont University

Utilizing Crispr/Cas9 Gene Editing Determine The Role Of Cnr1 During Zebrafish Development, Camme Sells

Belmont Undergraduate Research Symposium (BURS)

In a world where cannabinoids are being used more frequently as for medicinal purposes, the importance for research on cannabinoid biology is steadily increasing. The goal of this study was to gain a greater understanding of the effect cannabinoids have on developing zebrafish embryos. Work in this area of developmental biology could give way to an understanding of the safety of a human fetus during pregnancy with a mother who uses cannabis medicinal therapy. The CRISPR/Cas 9 system was utilized in this study to genetically edit the zebrafish embryo genome to remove the function of the Cannabinoid Receptor 1 ...


Expression Of Tgf-Beta Ligands During Sclerotome Development, Estela Williams 2020 Belmont University

Expression Of Tgf-Beta Ligands During Sclerotome Development, Estela Williams

Belmont Undergraduate Research Symposium (BURS)

During early embryogenesis in vertebrates, somites are formed by the segmentation of the presomitic mesoderm (Gomez, Ozbudak, Wunderlich, Baumann. Lewis and Pourquie 2008). The somite possesses several domains within it and the ventral part of the somite is called the sclerotome. The sclerotome gives rise to the axial skeleton and tendons. Twist 1b and Twist 2 are genes that are expressed in the sclerotome during axial skeletal development. Our experiments were developed to help understand the roles of Twist1b and Twist2 in tendon development. By utilizing microinjection to alter gene expression, the Twist 1b and Twist 2 genes can be ...


Scleraxis Expression Within The Axial Tendon Population In Zebrafish, Jessica Bernaba 2020 Belmont University

Scleraxis Expression Within The Axial Tendon Population In Zebrafish, Jessica Bernaba

Belmont Undergraduate Research Symposium (BURS)

Zebrafish are a well-established model organism for vertebrate development. During embryonic development, a pattern of segmentation of mesoderm yields somites, which give rise to specific cell fates. Somites, bilateral blocks of mesoderm along the neural tube in developing vertebrate embryos, subdivide into sclerotome as the vertebrate matures. Depending on migratory position, sclerotome cells give rise to various components of the axial skeletal system, including bone, cartilage, and tendon. Each fate is determined by gene expressions within the sclerotome and occupy distinct domains of the body plan. By method of in situ hybridization, we studied the expression of the scleraxis (scxa ...


Exploring The Role Of Tgfbr2 During Zebrafish Sclerotome Development, Nooralhuda Feron 2020 Belmont University

Exploring The Role Of Tgfbr2 During Zebrafish Sclerotome Development, Nooralhuda Feron

Belmont Undergraduate Research Symposium (BURS)

Zebrafish, Danio rerio, are used as a source model organism for human development of the tendons, bones, and ligaments. The sclerotome is an embryonic structure known as the somite. The sclerotome gives rise to axial bones and tendons. During development, different genes are involved and affect how these skeletal tissues develop. Specifically, within this project, the effects of Twist1b and Twist2 were studied to see their effects on tendon formation, as they are expressed within the developing sclerotome tissue. Transforming growth factor beta receptor type-2 (Tgfbr2) has been shown to be involved in tendon formation (Subramanian, Kanzaki, Galloway and Schilling ...


Enhancement Of Nkx3.2 Expression After Twist1b And Twist2 Knockdown In Zebrafish, Peyton Yearick 2020 Belmont University

Enhancement Of Nkx3.2 Expression After Twist1b And Twist2 Knockdown In Zebrafish, Peyton Yearick

Belmont Undergraduate Research Symposium (BURS)

Somitogenesis is a developmental event patterned by cellular signaling during gastrulation, ending around 30-hours post fertilization. Somites are segmented regions of mesoderm along the dorsal side of the body which form paraxial mesoderm in Danio rerio, zebrafish. Somite development results in the formation of the subcompartment called sclerotome, which differentiates to form the axial skeleton and associated tendons (Stickney et al., 2000). Further differentiation allows for sclerotome progenitor cell to develop into bone, muscle, tendon, and cartilage through sclerotome migration along the mesoderm. A portion of sclerotome cells migrate anterior towards the neural tube, where they will form the vertebral ...


Twist1b Knockout Generation In Zebrafish Using Crispr-Cas9, Grace Hurley 2020 Belmont University

Twist1b Knockout Generation In Zebrafish Using Crispr-Cas9, Grace Hurley

Belmont Undergraduate Research Symposium (BURS)

The Twist gene is found in numerous organisms, including humans, mice and zebrafish (Yeo et al., 2009). It is a vital gene in development and is needed for the organism to survive (Yeo et al., 2009). Specifically, the Twist gene family aids in specialization and differentiation of mesodermal cells, telling somites, segmented blocks of mesoderm, what to become (Germanguz et al., 2007). Over time, the Twist gene has duplicated into paralogs, each having differences in their functions (Germanguz et al., 2007). Twist1b is a protein coding gene that is a paralog to the human Twist1 gene (ZFIN, 2020). In zebrafish ...


Loss Of C-Jun N-Terminal Kinase 1 And 2 Function In Liver Epithelial Cells Triggers Biliary Hyperproliferation Resembling Cholangiocarcinoma, Francisco Javier Cubero, Mohamed Ramadan Mohamed, Marius M. Woitok, Gang Zhao, Maximilian Hatting, Yulia A. Nevzorova, Chaobo Chen, Johannes Haybaeck, Alain de Bruin, Matias A. Avila, Mark V. Boekschoten, Roger J. Davis, Christian Trautwein 2020 Complutense University of Madrid

Loss Of C-Jun N-Terminal Kinase 1 And 2 Function In Liver Epithelial Cells Triggers Biliary Hyperproliferation Resembling Cholangiocarcinoma, Francisco Javier Cubero, Mohamed Ramadan Mohamed, Marius M. Woitok, Gang Zhao, Maximilian Hatting, Yulia A. Nevzorova, Chaobo Chen, Johannes Haybaeck, Alain De Bruin, Matias A. Avila, Mark V. Boekschoten, Roger J. Davis, Christian Trautwein

University of Massachusetts Medical School Faculty Publications

Targeted inhibition of the c-Jun N-terminal kinases (JNKs) has shown therapeutic potential in intrahepatic cholangiocarcinoma (CCA)-related tumorigenesis. However, the cell-type-specific role and mechanisms triggered by JNK in liver parenchymal cells during CCA remain largely unknown. Here, we aimed to investigate the relevance of JNK1 and JNK2 function in hepatocytes in two different models of experimental carcinogenesis, the dethylnitrosamine (DEN) model and in nuclear factor kappa B essential modulator (NEMO)(hepatocyte-specific knockout (Deltahepa)) mice, focusing on liver damage, cell death, compensatory proliferation, fibrogenesis, and tumor development. Moreover, regulation of essential genes was assessed by reverse transcription polymerase chain reaction, immunoblottings ...


Computational Chemistry - Ulk 101, Michaela Montpas 2020 Olivet Nazarene University

Computational Chemistry - Ulk 101, Michaela Montpas

Scholar Week 2016 - present

Autophagy is a process that generates the necessary building components for cells by cytoplasmic breakdown of unnecessary materials (Martin, Celano, Solitro, Gunaydin, Scott, et. al., 2018). This is a survival technique for cells in times of stress, especially during periods of nutrient starvation. Cancer cells, unfortunately, benefit from this process due to their ability to flourish in nutrient-starved environments, becoming resistant to therapy. The primary protein in mammals responsible for this process is a serine/threonine kinase called ULK 1 (unc-51 like autophagy initiating kinase 1). As such, inhibitors of ULK 1 can be used in cancer therapies in order ...


Identification And Characterization Of The Lactating Mouse Mammary Gland Citrullinome, Guangyuan Li, Coleman H. Young, Bryce Snow, Amanda O. Christensen, M. Kristen Demoruelle, Venkatesh V. Nemmara, Paul R. Thompson, Heather M. Rothfuss, Brian D. Cherrington 2020 University of Wyoming

Identification And Characterization Of The Lactating Mouse Mammary Gland Citrullinome, Guangyuan Li, Coleman H. Young, Bryce Snow, Amanda O. Christensen, M. Kristen Demoruelle, Venkatesh V. Nemmara, Paul R. Thompson, Heather M. Rothfuss, Brian D. Cherrington

Open Access Articles

Citrullination is a post-translational modification (PTM) in which positively charged peptidyl-arginine is converted into neutral peptidyl-citrulline by peptidylarginine deiminase (PAD or PADI) enzymes. The full protein citrullinome in many tissues is unknown. Herein, we used mass spectrometry and identified 107 citrullinated proteins in the lactation day 9 (L9) mouse mammary gland including histone H2A, alpha-tubulin, and beta-casein. Given the importance of prolactin to lactation, we next tested if it stimulates PAD-catalyzed citrullination using mouse mammary epithelial CID-9 cells. Stimulation of CID-9 cells with 5 microg/mL prolactin for 10 min induced a 2-fold increase in histone H2A citrullination and a ...


Histone Deacetylases 1 And 2 Silence Cryptic Transcription To Promote Mitochondrial Function During Cardiogenesis, Zachary J. Milstone, Sherin Saheera, Lauren Bourke, Tomer Shpilka, Cole M. Haynes, Chinmay M. Trivedi 2020 University of Massachusetts Medical School

Histone Deacetylases 1 And 2 Silence Cryptic Transcription To Promote Mitochondrial Function During Cardiogenesis, Zachary J. Milstone, Sherin Saheera, Lauren Bourke, Tomer Shpilka, Cole M. Haynes, Chinmay M. Trivedi

Open Access Articles

Cryptic transcription occurs widely across the eukaryotic genome; however, its regulation during vertebrate development is not understood. Here, we show that two class I histone deacetylases, Hdac1 and Hdac2, silence cryptic transcription to promote mitochondrial function in developing murine hearts. Mice lacking Hdac1 and Hdac2 in heart exhibit defective developmental switch from anaerobic to mitochondrial oxidative phosphorylation (OXPHOS), severe defects in mitochondrial mass, mitochondrial function, and complete embryonic lethality. Hdac1/Hdac2 promotes the transition to OXPHOS by enforcing transcriptional fidelity of metabolic gene programs. Mechanistically, Hdac1/Hdac2 deacetylates histone residues including H3K23, H3K14, and H4K16 to suppress cryptic transcriptional initiation ...


Regenerative Medicine Therapy: Adipose Derived Extracellular Vesicles In Viral Myocarditis, David Gorelov, Damian N. Di Florio, Gary R. Salomon, Angita Jain, Nick E. Saikaili, Danielle J. Beetler, Swikriti Shrestha, Ming Tian, Joy Wolfram PhD, DeLisa Fairweather PhD, Katelyn Bruno PhD, Judith D. Ochrietor PhD 2020 University of North Florida

Regenerative Medicine Therapy: Adipose Derived Extracellular Vesicles In Viral Myocarditis, David Gorelov, Damian N. Di Florio, Gary R. Salomon, Angita Jain, Nick E. Saikaili, Danielle J. Beetler, Swikriti Shrestha, Ming Tian, Joy Wolfram Phd, Delisa Fairweather Phd, Katelyn Bruno Phd, Judith D. Ochrietor Phd

Showcase of Osprey Advancements in Research and Scholarship (SOARS)

Objective: Myocarditis, inflammation of the heart muscle, is an autoimmune heart disease that can be caused by viruses, bacteria and toxins. Myocarditis can lead to dilated cardiomyopathy (DCM) and heart failure. Currently there are no disease-specific therapies for treating myocarditis or preventing progression to DCM. Adipose Extracellular Vesicles (AEVs) are lipid bilayer nanoparticles that are released into the outside environment of adipocytes and provide promising regenerative potential for inflammatory diseases like myocarditis.

Methods: Lipoaspirate was obtained from women and men and AEVs isolated from the lipoaspirate using tangential flow filtration. We injected wild type male BALB/c mice with 250uL ...


Structure, Function And Metabolism Of Hepatic And Adipose Tissue Lipid Droplets: Implications In Alcoholic Liver Disease, Sathish Kumar Natarajan, Karuna Rasineni, Murali Ganesan, Dan Feng, Benita L. McVicker, Mark A. McNiven, Natalia A. Osna, Justin L. Mott, Carol A. Casey, Kusum K. Kharbanda 2020 University of Nebraska - Lincoln

Structure, Function And Metabolism Of Hepatic And Adipose Tissue Lipid Droplets: Implications In Alcoholic Liver Disease, Sathish Kumar Natarajan, Karuna Rasineni, Murali Ganesan, Dan Feng, Benita L. Mcvicker, Mark A. Mcniven, Natalia A. Osna, Justin L. Mott, Carol A. Casey, Kusum K. Kharbanda

Papers in Veterinary and Biomedical Science

For more than 30 years, lipid droplets (LDs) were considered as an inert bag of lipid for storage of energy-rich fat molecules. Following a paradigm shift almost a decade ago, LDs are presently considered an active subcellular organelle especially designed for assembling, storing and subsequently supplying lipids for generating energy and membrane synthesis (and in the case of hepatocytes for VLDL secretion). LDs also play a central role in many other cellular functions such as viral assembly and protein degradation. Here, we have explored the structural and functional changes that occur in hepatic and adipose tissue LDs following chronic ethanol ...


Innate Immunity In The C. Elegans Intestine Is Programmed By A Neuronal Regulator Of Awc Olfactory Neuron Development, Kyle J. Foster, Hilary K. Cheesman, Pengpeng Liu, Nicholas D. Peterson, Sarah M. Anderson, Read Pukkila-Worley 2020 University of Massachusetts Medical School

Innate Immunity In The C. Elegans Intestine Is Programmed By A Neuronal Regulator Of Awc Olfactory Neuron Development, Kyle J. Foster, Hilary K. Cheesman, Pengpeng Liu, Nicholas D. Peterson, Sarah M. Anderson, Read Pukkila-Worley

Open Access Articles

Olfactory neurons allow animals to discriminate nutritious food sources from potential pathogens. From a forward genetic screen, we uncovered a surprising requirement for the olfactory neuron gene olrn-1 in the regulation of intestinal epithelial immunity in Caenorhabditis elegans. During nematode development, olrn-1 is required to program the expression of odorant receptors in the AWC olfactory neuron pair. Here, we show that olrn-1 also functions in AWC neurons in the cell non-autonomous suppression of the canonical p38 MAPK PMK-1 immune pathway in the intestine. Low activity of OLRN-1, which activates the p38 MAPK signaling cassette in AWC neurons during larval development ...


Npgreat: Hybrid Assembly Of Human Subtelomeres With The Use Of Nanopore And Linked-Read Datasets, Eleni Adam, Desh Ranjan, Harold Riethman 2020 Old Dominion University

Npgreat: Hybrid Assembly Of Human Subtelomeres With The Use Of Nanopore And Linked-Read Datasets, Eleni Adam, Desh Ranjan, Harold Riethman

College of Sciences Posters

The telomeres are vitally important regions that are located at the tips of the chromosomes. Their dysfunction, caused by length shortening can lead to senescent cells, which in turn cause age-related diseases, including cancer. The subtelomeres, located next to the telomeres, possess the critical role of regulating the adjacent telomere lengths. Even after many years of research, human subtelomeres have proven to be very hard to assemble due to their morphology. In order to overcome these problems, the hybrid assembly method we develop utilizes two of the latest available types of data, which complement each other: Linked-Reads and ultralong Nanopore ...


Tip60 Regulation Of Δnp63Α Is Associated With Cisplatin Resistance, Akshay Hira, Andrew Stacy, Jin Zhang, Michael P. Craig, Madhavi Kadakia 2020 Wright State University - Main Campus

Tip60 Regulation Of Δnp63Α Is Associated With Cisplatin Resistance, Akshay Hira, Andrew Stacy, Jin Zhang, Michael P. Craig, Madhavi Kadakia

Symposium of Student Research, Scholarship, and Creative Activities Materials

About 5.4 million basal and squamous cell skin cancers are diagnosed every year in the US. ΔNp63a, a member of the p53 transcription factor family, is overexpressed in non-melanoma skin cancer and regulates cell survival, migration and invasion. TIP60 is histone acetyltransferase (HAT) which mediates cellular processes such as transcription and the DNA damage response (DDR). Previous studies in our lab have shown that overexpression of TIP60 induces ΔNp63a protein stabilization in a catalytic-dependent manner. Since ΔNp63a is known to transcriptionally regulate several DDR genes and promote cisplatin resistance, its stabilization by TIP60 may contribute to the failure of ...


Can Protozoa Prove The Beginning Of The World?, Karina L. Burton 2020 Southeastern University - Lakeland

Can Protozoa Prove The Beginning Of The World?, Karina L. Burton

Classical Conversations

Protozoa are magnificent creatures. They exhibit all of the functions intrinsic to living organisms: irritability, metabolism, growth and reproduction. Within these functions, there are numerous examples of mutations that occur in order for organisms to adapt to their given environments. Irritability is demonstrated in protozoa by their use of pseudopodia, flagella, or cilia for motility; it has been shown that such locomotors exhibit diversity while maintaining similar protein and chemical structures that appear to be a result of evolutionary processes. Metabolism in protozoa is similar to that of larger animals, but their diet is unique. They primarily feast upon bacteria ...


Dgts Production As A Phosphate Starvation Response In The Human Fungal Pathogen Candida Albicans, Caleb Wehling 2020 University of Nebraska - Lincoln

Dgts Production As A Phosphate Starvation Response In The Human Fungal Pathogen Candida Albicans, Caleb Wehling

Dissertations and Theses in Biological Sciences

Betaine lipids are a class of membrane lipids with betaine head groups. Three betaine lipids are known - diacylglyceryltrimethylhomoserine (DGTS), diacylglycerylhydroxymethylalanine (DGTA), and diacylglycerylcarboxymethylcholine (DGCC). Betaine lipids are most common in algae, although DGTS, the most common betaine lipid, is also found in many bacteria and fungi. Organisms which produce betaine lipids (especially DGTS) often don’t produce phosphatidylcholine (PtdCho), and DGTS structure resembles PtdCho structure without any phosphorous, leading to the hypothesis that betaine lipids may substitute for phospholipids in some organisms. This has been confirmed by discoveries that some organisms are capable of switching their membrane composition from PtdCho ...


Mechanisms Of Insulin Resistance Related To White, Beige, And Brown Adipocytes, Michael P. Czech 2020 University of Massachusetts Medical School

Mechanisms Of Insulin Resistance Related To White, Beige, And Brown Adipocytes, Michael P. Czech

Open Access Articles

BACKGROUND: The diminished glucose lowering effect of insulin in obesity, called "insulin resistance," is associated with glucose intolerance, type 2 diabetes, and other serious maladies. Many publications on this topic have suggested numerous hypotheses on the molecular and cellular disruptions that contribute to the syndrome. However, significant uncertainty remains on the mechanisms of its initiation and long-term maintenance.

SCOPE OF REVIEW: To simplify insulin resistance analysis, this review focuses on the unifying concept that adipose tissue is a central regulator of systemic glucose homeostasis by controlling liver and skeletal muscle metabolism. Key aspects of adipose function related to insulin resistance ...


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