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Defining The Role Of Rbbp4 In Oocyte Maturation And Preimplantation Development Using Trim-Away, Holly L. Barletta 2021 University of Massachusetts Amherst

Defining The Role Of Rbbp4 In Oocyte Maturation And Preimplantation Development Using Trim-Away, Holly L. Barletta

Masters Theses

Retinoblastoma-binding protein 4 (RBBP4) is a subunit of chromatin remodeling factor 1 (CAF-1) and is essential for mammalian oocyte maturation, embryo survival, and embryo implantation. RBBP4 also localizes to the chromatin and is a ubiquitously expressed nuclear protein. Previous methods used to study this protein include short interfacing RNAs (siRNAs) and CRISPR/Cas9. These techniques have limitations such as determining an indirect depletion of proteins, may trigger compensatory mechanisms, and may not be useful in non-dividing primary cells. A new, acute, and rapid endogenous protein depletion technique called Trim-Away, can overcome these limitations. Trim-Away is also widely applicable since it ...


Molecular Mechanisms Underlying Cell Fate Choice Within Specific Retinal Lineages, Estie Schick 2021 The Graduate Center, City University of New York

Molecular Mechanisms Underlying Cell Fate Choice Within Specific Retinal Lineages, Estie Schick

Dissertations, Theses, and Capstone Projects

During development, retinal progenitor cells (RPCs) divide to form all of the cell types that make up the retina. Multipotent RPCs are competent to generate all retinal cell types, while restricted RPCs form specific lineages of cells. In particular, one genetically-defined RPC type preferentially gives rise to cone photoreceptors and horizontal cells. Many of the mechanisms that are responsible for directing cell fate choice within this lineage are unknown. This thesis largely focuses on examining the development of specific cell types and subtypes from restricted RPCs and on investigating the gene regulatory events that underlie cone photoreceptor and horizontal cell ...


Characterizing The Effects Of Pyrroloquinoline Quinone (Pqq) Supplementation On Skeletal Muscle Mitochondrial Function And Myogenesis During Oxidative Stress And Iugr., Allyson J. Wood 2021 The University of Western Ontario

Characterizing The Effects Of Pyrroloquinoline Quinone (Pqq) Supplementation On Skeletal Muscle Mitochondrial Function And Myogenesis During Oxidative Stress And Iugr., Allyson J. Wood

Electronic Thesis and Dissertation Repository

Intrauterine growth restriction (IUGR) affects 10-15% of births and is associated with placental insufficiency (PI), resulting in fetal oxidative stress (OS). This OS is a factor in the predisposition to postnatal noncommunicable disease (NCD) of which muscle mitochondrial dysfunctional is a key aspect. Pyrroloquinoline quinone (PQQ), an antioxidant-like compound, is capable of OS reduction and promotes mitochondrial function, though limited research has focused on its effects in in utero skeletal muscle. This study sought to investigate the impact of in vitro H2O2, a model of OS, and an in vivo model of OS associated IUGR, with PQQ ...


Creating A Protein Chimera To Study Regulation Of Muscle Diversity, Shannon Scarboro 2021 Kennesaw State University

Creating A Protein Chimera To Study Regulation Of Muscle Diversity, Shannon Scarboro

Symposium of Student Scholars

Creating a protein chimera to study regulation of muscle diversity.

Body muscles are made of many individual super-cells, called muscle fibers, that have distinct properties and determine every individual’s strength and endurance. Initially all muscle fibers have identical characteristics, but become differentiated into specific types in adults. The mechanism of such transition is not well understood, despite its obvious importance for shaping human physicality.

Remarkable conservation of the muscle tissue enables us to use fruit flies to study the mechanisms of muscle fiber diversity. We hypothesized that the transcription factor Mef2 acts as a molecular switch that activates structural ...


The Effects Of Mapk Signaling On The Development Of Cerebellar Granule Cells, Kerry Morgan 2021 University of Connecticut

The Effects Of Mapk Signaling On The Development Of Cerebellar Granule Cells, Kerry Morgan

Honors Scholar Theses

The granule cells are the most abundant neuronal type in the human brain. Rapid proliferation of granule cell progenitors results in dramatic expansion and folding of the cerebellar cortex during postnatal development. Mis-regulation of this proliferation process causes medulloblastoma, the most prevalent childhood brain tumor. In the developing cerebellum, granule cells are derived from Atoh1-expressing cells, which arise from the upper rhombic lip (the interface between the roof plate and neuroepithelium). In addition to granule cells, the Atoh1 lineage also gives rise to different types of neurons including cerebellar nuclei neurons. In the current study, I have investigated the ...


Translational Fidelity And Its Role In Neuronal Homeostasis, Markus Terrey 2021 University of Maine

Translational Fidelity And Its Role In Neuronal Homeostasis, Markus Terrey

Electronic Theses and Dissertations

The process of translation, which refers to decoding genetic information from mRNA to protein, is vital for all cellular function. Translational fidelity starts at the level of aminoacylation of transfer RNAs (tRNA). This reaction is catalyzed by aminoacyl tRNA synthetases where each amino acid is transferred to its corresponding cognate tRNA. Because tRNAs harbor the anticodon sequence to decodes a particular mRNA codon, the specific aminoacylation of the tRNA with a cognate amino acid establishes the rules of decoding genetic code into proteins. Aminoacylated tRNAs are then delivered to ribosomes, where ribosomes in a highly organized manner need to accurately ...


Dynamics Of Hybrid Zones At A Continental Scale, Bradley T. Martin 2021 University of Arkansas, Fayetteville

Dynamics Of Hybrid Zones At A Continental Scale, Bradley T. Martin

Graduate Theses and Dissertations

Hybridization has traditionally been viewed as a happenstance that negatively impacts populations, but is now recognized as an important evolutionary mechanism that can substantially impact the evolutionary trajectories of gene pools, influence adaptive capacity, and contravene or reinforce divergence. Physiographic processes are important drivers of dispersal, alternately funneling populations into isolation, promoting divergence, or facilitating secondary contact of diverged populations, increasing the potential for hybridization. In North America, glacial-interglacial cycles and geomorphological changes have provided a dynamic backdrop over the last two million years that promoted such oscillations of population contraction and expansion. These biogeographic processes have resulted in regional ...


Determining The Effect Of A Cbc-Based Oxime On Cell Viability And Apoptosis In Sea Urchin Embryos, Nevin B. Hoenninger, Jack Preston 2021 Susquehanna University

Determining The Effect Of A Cbc-Based Oxime On Cell Viability And Apoptosis In Sea Urchin Embryos, Nevin B. Hoenninger, Jack Preston

Senior Scholars Day

Natural products are an important source of existing and potential new anti-cancer drugs. We have previously shown that cannabichromene (CBC) oxime and several CBC oxime esters have potent anti-mitotic activity when used in bioassays with sea urchin embryos. In this study we explored the effect of the CBC oxime on cell viability and apoptosis in sea urchin embryos. Newly fertilized or gastrula stage embryos were combined with 10µM or 50uM CBC oxime as well as a DMSO only control. At various timepoints, samples of the embryos were removed from the cultures and stained with three different fluorescent dyes. NucBlue (ThermoFisher ...


216— Loss Of Function Mutation For Tp53 Does Not Rescue The Chaf1bNt2 Small-Eye Phenotype In Danio Rerio, Alex Parks 2021 SUNY Geneseo

216— Loss Of Function Mutation For Tp53 Does Not Rescue The Chaf1bNt2 Small-Eye Phenotype In Danio Rerio, Alex Parks

GREAT Day

In Zebrafish, the chromosome assembly factor 1b (chaf1b) gene is in part responsible for the development of the eye. In homozygous chaf1bt24412 mutants retinal cell death is promoted through cell-death promoting activity of the gene, tumor suppressor protein p53 (tp53), resulting in a small-eye phenotype. Another allele chaf1bnt2, was found to also result in the small-eye phenotype when in a homozygous state. We found that knockdown of Tp53 protein via morpholino antisense oligonucleotide injection of 1-2 cell stage embryos failed to rescue retinal cell death of chaf1bnt2 homozygous mutants as detected by TUNEL labeling. Because morpholinos may fail to fully ...


Zebrafish (Danio Rerio) Hoxb6: An Exploration Into The Divergence Of Genomic Dna Sequence And Gene Expression Across Teleost Fishes Post-Genome Duplication, Amber Lynn Rittgers, Pierre Le Pabic, Adam Davis 2021 University of Georgia

Zebrafish (Danio Rerio) Hoxb6: An Exploration Into The Divergence Of Genomic Dna Sequence And Gene Expression Across Teleost Fishes Post-Genome Duplication, Amber Lynn Rittgers, Pierre Le Pabic, Adam Davis

Georgia Journal of Science

Hoxb6 is an evolutionarily conserved developmental regulatory gene that functions, in part, to pattern several organs and organ systems within the embryonic trunk during vertebrate embryogenesis. The cis-regulatory circuitry mediating trunk expression in mouse (Mus musculus) may be conserved across gnathostome vertebrates, as several other species show similar trunk expression patterns, including chicken (Gallus gallus), dogfish shark (Scyliorhinus canicula), and several teleost fishes. A whole genome duplication event that occurred in the lineage leading to teleost fishes has generated at least two Hoxb6 genes, hoxb6a and b6b. Two teleost fishes of the superorder Acanthopterygii, Japanese medaka (Oryzias latipes) and ...


The Phoenix, Fernanda Perez-Alvarez 2021 College of the Holy Cross

The Phoenix, Fernanda Perez-Alvarez

Montserrat Annual Writing Prize

This article uses a mythical creature, the phoenix, to examine and illustrate the biological principles for generation of an adult body plan from a single cell. Using the study of developmental biology, it explores the cellular and molecular biology that underpins the massive complexity of creating an adult body plan. It also explores the similarities and differences between different embryos, and how nature and evolution have shaped the biology of those embryos to create different body plans.


Investigating The Role Of Alx4a In Mitfa Repression And Iridophore Fate Biasing, Melanie Cragan 2021 Rhode Island College

Investigating The Role Of Alx4a In Mitfa Repression And Iridophore Fate Biasing, Melanie Cragan

Honors Projects Overview

Investigating the role of the transcription factor alx4a on chromatophore fating in zebrafish.


Stability Of The Cytosine Methylome During Post-Testicular Sperm Maturation In Mouse, Carolina Galan, Ryan W. Serra, Fengyun Sun, Vera D. Rinaldi, Colin C. Conine, Oliver J. Rando 2021 University of Massachusetts Medical School

Stability Of The Cytosine Methylome During Post-Testicular Sperm Maturation In Mouse, Carolina Galan, Ryan W. Serra, Fengyun Sun, Vera D. Rinaldi, Colin C. Conine, Oliver J. Rando

Open Access Publications by UMMS Authors

Beyond the haploid genome, mammalian sperm carry a payload of epigenetic information with the potential to modulate offspring phenotypes. Recent studies show that the small RNA repertoire of sperm is remodeled during post-testicular maturation in the epididymis. Epididymal maturation has also been linked to changes in the sperm methylome, suggesting that the epididymis might play a broader role in shaping the sperm epigenome. Here, we characterize the genome-wide methylation landscape in seven germ cell populations from throughout the male reproductive tract. We find very few changes in the cytosine methylation landscape between testicular germ cell populations and cauda epididymal sperm ...


Germ Granule Dysfunction Is A Hallmark And Mirror Of Piwi Mutant Sterility, Maya Spichal, Bree Heestand, Katherine Kretovich Billmyre, Stephen Frenk, Craig C. Mello, Shawn Ahmed 2021 University of Massachusetts Medical School

Germ Granule Dysfunction Is A Hallmark And Mirror Of Piwi Mutant Sterility, Maya Spichal, Bree Heestand, Katherine Kretovich Billmyre, Stephen Frenk, Craig C. Mello, Shawn Ahmed

Open Access Publications by UMMS Authors

In several species, Piwi/piRNA genome silencing defects cause immediate sterility that correlates with transposon expression and transposon-induced genomic instability. In C. elegans, mutations in the Piwi-related gene (prg-1) and other piRNA deficient mutants cause a transgenerational decline in fertility over a period of several generations. Here we show that the sterility of late generation piRNA mutants correlates poorly with increases in DNA damage signaling. Instead, sterile individuals consistently exhibit altered perinuclear germ granules. We show that disruption of germ granules does not activate transposon expression but induces multiple phenotypes found in sterile prg-1 pathway mutants. Furthermore, loss of the ...


Somatic Pirnas And Transposons Are Differentially Regulated During Skeletal Muscle Atrophy And Programmed Cell Death [Preprint], Junko Tsuji, Travis Thomson, Christine Brown, Subhanita Ghosh, William E. Theurkauf, Zhiping Weng, Lawrence M. Schwartz 2021 University of Massachusetts Medical School

Somatic Pirnas And Transposons Are Differentially Regulated During Skeletal Muscle Atrophy And Programmed Cell Death [Preprint], Junko Tsuji, Travis Thomson, Christine Brown, Subhanita Ghosh, William E. Theurkauf, Zhiping Weng, Lawrence M. Schwartz

University of Massachusetts Medical School Faculty Publications

PiWi-interacting RNAs (piRNAs) are small single-stranded RNAs that can repress transposon expression via epigenetic silencing and transcript degradation. They have been identified predominantly in the ovary and testis, where they serve essential roles in transposon silencing in order to protect the integrity of the genome in the germline. The potential expression of piRNAs in somatic cells has been controversial. In the present study we demonstrate the expression of piRNAs derived from both genic and transposon RNAs in the intersegmental muscles (ISMs) from the tobacco hawkmoth Manduca sexta. These piRNAs are abundantly expressed, are ~27 nt long, map antisense to transposons ...


Deficient Spermiogenesis In Mice Lacking Rlim, Feng Wang, Maria Gracia Gervasi, Ana Boskovic, Fengyun Sun, Vera D. Rinaldi, Jun Yu, Mary C. Wallingford, Darya A. Tourzani, Jesse Mager, Lihua Julie Zhu, Oliver J. Rando, Pablo E. Visconti, Lara Strittmatter, Ingolf Bach 2021 University of Massachusetts Medical School

Deficient Spermiogenesis In Mice Lacking Rlim, Feng Wang, Maria Gracia Gervasi, Ana Boskovic, Fengyun Sun, Vera D. Rinaldi, Jun Yu, Mary C. Wallingford, Darya A. Tourzani, Jesse Mager, Lihua Julie Zhu, Oliver J. Rando, Pablo E. Visconti, Lara Strittmatter, Ingolf Bach

Radiology Publications

The X-linked gene Rlim plays major roles in female mouse development and reproduction, where it is crucial for the maintenance of imprinted X chromosome inactivation in extraembryonic tissues of embryos. However, while females carrying a systemic Rlim knockout (KO) die around implantation, male Rlim KO mice appear healthy and are fertile. Here, we report an important role for Rlim in testis where it is highly expressed in post-meiotic round spermatids as well as in Sertoli cells. Systemic deletion of the Rlim gene results in lower numbers of mature sperm that contains excess cytoplasm, leading to decreased sperm motility and in ...


Annotation Of Chromatin States In 66 Complete Mouse Epigenomes During Development, Arjan van der Velde, Kaili Fan, Junko Tsuji, Jill E. Moore, Michael J. Purcaro, Henry E. Pratt, Zhiping Weng 2021 University of Massachusetts Medical School

Annotation Of Chromatin States In 66 Complete Mouse Epigenomes During Development, Arjan Van Der Velde, Kaili Fan, Junko Tsuji, Jill E. Moore, Michael J. Purcaro, Henry E. Pratt, Zhiping Weng

Open Access Publications by UMMS Authors

The morphologically and functionally distinct cell types of a multicellular organism are maintained by their unique epigenomes and gene expression programs. Phase III of the ENCODE Project profiled 66 mouse epigenomes across twelve tissues at daily intervals from embryonic day 11.5 to birth. Applying the ChromHMM algorithm to these epigenomes, we annotated eighteen chromatin states with characteristics of promoters, enhancers, transcribed regions, repressed regions, and quiescent regions. Our integrative analyses delineate the tissue specificity and developmental trajectory of the loci in these chromatin states. Approximately 0.3% of each epigenome is assigned to a bivalent chromatin state, which harbors ...


Non-Apoptotic Enteroblast-Specific Role Of The Initiator Caspase Dronc For Development And Homeostasis Of The Drosophila Intestine, Jillian L. Lindblad, Meghana Tare, Alla Amcheslavsky, Alicia Shields, Andreas Bergmann 2021 University of Massachusetts Medical School

Non-Apoptotic Enteroblast-Specific Role Of The Initiator Caspase Dronc For Development And Homeostasis Of The Drosophila Intestine, Jillian L. Lindblad, Meghana Tare, Alla Amcheslavsky, Alicia Shields, Andreas Bergmann

Open Access Publications by UMMS Authors

The initiator caspase Dronc is the only CARD-domain containing caspase in Drosophila and is essential for apoptosis. Here, we report that homozygous dronc mutant adult animals are short-lived due to the presence of a poorly developed, defective and leaky intestine. Interestingly, this mutant phenotype can be significantly rescued by enteroblast-specific expression of dronc(+) in dronc mutant animals, suggesting that proper Dronc function specifically in enteroblasts, one of four cell types in the intestine, is critical for normal development of the intestine. Furthermore, enteroblast-specific knockdown of dronc in adult intestines triggers hyperplasia and differentiation defects. These enteroblast-specific functions of Dronc do ...


Neuronal Post-Developmentally Acting Sax-7s/L1cam Can Function As Cleaved Fragments To Maintain Neuronal Architecture In C. Elegans [Preprint], Virginie E. Desse, Cassandra R. Blanchette, Paola N. Perrat, Claire Y. Benard 2021 Université du Québec à Montréal

Neuronal Post-Developmentally Acting Sax-7s/L1cam Can Function As Cleaved Fragments To Maintain Neuronal Architecture In C. Elegans [Preprint], Virginie E. Desse, Cassandra R. Blanchette, Paola N. Perrat, Claire Y. Benard

University of Massachusetts Medical School Faculty Publications

Whereas remarkable advances have uncovered mechanisms that drive nervous system assembly, the processes responsible for the lifelong maintenance of nervous system architecture remain poorly understood. Subsequent to its establishment during embryogenesis, neuronal architecture is maintained throughout life in the face of the animal’s growth, maturation processes, the addition of new neurons, body movements, and aging. The C. elegans protein SAX-7, homologous to the vertebrate L1 protein family, is required for maintaining the organization of neuronal ganglia and fascicles after their successful initial embryonic development. To dissect the function of sax-7 in neuronal maintenance, we generated a null allele and ...


Long First Exons And Epigenetic Marks Distinguish Conserved Pachytene Pirna Clusters From Other Mammalian Genes, Tianxiong Yu, Kaili Fan, Deniz M. Ozata, Gen Zhang, Yu Fu, William E. Theurkauf, Phillip D. Zamore, Zhiping Weng 2021 Tongji University

Long First Exons And Epigenetic Marks Distinguish Conserved Pachytene Pirna Clusters From Other Mammalian Genes, Tianxiong Yu, Kaili Fan, Deniz M. Ozata, Gen Zhang, Yu Fu, William E. Theurkauf, Phillip D. Zamore, Zhiping Weng

University of Massachusetts Medical School Faculty Publications

In the male germ cells of placental mammals, 26-30-nt-long PIWI-interacting RNAs (piRNAs) emerge when spermatocytes enter the pachytene phase of meiosis. In mice, pachytene piRNAs derive from ~100 discrete autosomal loci that produce canonical RNA polymerase II transcripts. These piRNA clusters bear 5' caps and 3' poly(A) tails, and often contain introns that are removed before nuclear export and processing into piRNAs. What marks pachytene piRNA clusters to produce piRNAs, and what confines their expression to the germline? We report that an unusually long first exon ( > /= 10 kb) or a long, unspliced transcript correlates with germline-specific transcription and piRNA ...


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