Biology Commons^™

Gene Expression Studies For The Analysis Of Domoic Acid Production In The Marine Diatom Pseudo-Nitzschia Multiseries, Katie Boissonneault, Brooks M. Henningsen, Stephen S. Bates, Deborah L. Robertson, Sean Milton, Jerry Pelletier, Deborah A. Hogan, David E. Housman

Dartmouth Scholarship

Pseudo-nitzschia multiseries Hasle (Hasle) (Ps-n) is distinctive among the ecologically important marine diatoms because it produces the neurotoxin domoic acid. Although the biology of Ps-n has been investigated intensely, the characterization of the genes and biochemical pathways leading to domoic acid biosynthesis has been limited. To identify transcripts whose levels correlate with domoic acid production, we analyzed Ps-n under conditions of high and low domoic acid production by cDNA microarray technology and reverse-transcription quantitative PCR (RT-qPCR) methods. Our goals included identifying and validating robust reference genes for Ps-n RNA expression analysis under these conditions.

High-Throughput Rna Sequencing Of Pseudomonas-Infected Arabidopsis Reveals Hidden Transcriptome Complexity And Novel Splice Variants, Brian E. Howard, Qiwen Hu, Ahmet Can Babaoglu, Manan Chandra, Monica Borghi, Xiaoping Tan, Luyan He, Heike Winter-Sederoff, Walter Gassmann, Paola Veronese, Steffen Heber

Biology Faculty Publications

We report the results of a genome-wide analysis of transcription in Arabidopsis thaliana after treatment with Pseudomonas syringae pathovar tomato. Our time course RNA-Seq experiment uses over 500 million read pairs to provide a detailed characterization of the response to infection in both susceptible and resistant hosts. The set of observed differentially expressed genes is consistent with previous studies, confirming and extending existing findings about genes likely to play an important role in the defense response to Pseudomonas syringae. The high coverage of the Arabidopsis transcriptome resulted in the discovery of a surprisingly large number of alternative splicing (AS) events …

Subset Of Heat-Shock Transcription Factors Required For The Early Response Of Arabidopsis To Excess Light, Hou-Sung Jung, Peter A. Crisp, Gonzalo M. Estavillo, Benjamin Cole

Dartmouth Scholarship

Sunlight provides energy for photosynthesis and is essential for nearly all life on earth. However, too much or too little light or rapidly fluctuating light conditions cause stress to plants. Rapid changes in the amount of light are perceived as a change in the reduced/oxidized (redox) state of photosynthetic electron transport components in chloroplasts. However, how this generates a signal that is relayed to changes in nuclear gene expression is not well understood. We modified redox state in the reference plant, Arabidopsis thaliana, using either excess light or low light plus the herbicide DBMIB (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone), a well-known inhibitor of photosynthetic …

An Inverted Repeat In The Ospc Operator Is Required For Induction In Borrelia Burgdorferi, Dan Drecktrah, Laura S. Hall, Laura L. Hoon-Hanks, D. Scott Samuels

Biological Sciences Faculty Publications

Borrelia burgdorferi, the spirochete that causes Lyme disease, differentially regulates synthesis of the outer membrane lipoprotein OspC to infect its host. OspC is required to establish infection but then repressed in the mammal to avoid clearance by the adaptive immune response. Inverted repeats (IR) upstream of the promoter have been implicated as an operator to regulate ospC expression. We molecularly dissected the distal inverted repeat (dIR) of the ospC operator by site-directed mutagenesis at its endogenous location on the circular plasmid cp26. We found that disrupting the dIR but maintaining the proximal IR prevented induction of OspC synthesis by …

Germline Transgenic Methods For Tracking Cells And Testing Gene Function During Regeneration In The Axolotl, Shahryar Khattak, Maritta Schuez, Tobias Richter, Dunja Knapp, Saori L. Haigo, Tatiana Sandoval-Guzmán, Kristyna Hradlikova, Annett Duemmler, Ryan R. Kerney, Elly M. Tanaka

Biology Faculty Publications

The salamander is the only tetrapod that regenerates complex body structures throughout life. Deciphering the underlying molecular processes of regeneration is fundamental for regenerative medicine and developmental biology, but the model organism had limited tools for molecular analysis. We describe a comprehensive set of germline transgenic strains in the laboratory-bred salamander Ambystoma mexicanum(axolotl) that open up the cellular and molecular genetic dissection of regeneration. We demonstrate tissue-dependent control of gene expression in nerve, Schwann cells, oligodendrocytes, muscle, epidermis, and cartilage. Furthermore, we demonstrate the use of tamoxifen-induced Cre/loxP-mediated recombination to indelibly mark different cell types. Finally, we inducibly …

Behavioral And Neurogenomic Transcriptome Changes In Wild-Derived Zebrafish With Fluoxetine Treatment, Ryan Y. Wong, Sarah E. Oxendine, John Godwin

Biology Faculty Publications

Background: Male and female vertebrates typically differ in a range of characteristics, from morphology to physiology to behavior, which are influenced by factors such as the social environment and the internal hormonal and genetic milieu. However, sex differences in gene expression profiles in the brains of vertebrates are only beginning to be understood. Fishes provide a unique complement to studies of sex differences in mammals and birds given that fish show extreme plasticity and lability of sexually dimorphic characters and behaviors during development and even adulthood. Hence, teleost models can give additional insight into sexual differentiation. The goal of …

Patterns Of Gene Expression From Human Costal Cartilage In Relation To The Chest Wall Deformity Pectus Carinatum, Janna E. Grubbs

Biological Sciences Theses & Dissertations

Cartilage deformities within the human chest wall, specifically pectus excavatum (PE) and pectus carinatum (PC) are common (1/400-1/1000) and yet, despite their clinical significance, are some of the least studied disorders pertaining to cartilage [1]. The costal cartilage connecting "false ribs" 8-10 to the sternum is often abnormally grown and can lead to formation of a severely sunken "funnel" chest (PE) or push outwards to form a "pigeon" chest (PC). Both conditions can have impact on the diaphragm, heart, lungs, and psychological function. An established ratio of PE and PC in males to females is 4:1, indicating a sex-linked male …

Comparison Of Th1 Cytokines And T Cell Markers Gene Expressions Between Virulent And An Attenuated Eiav Vaccine Strain, Talia R. Henkle

Lewis Honors College Capstone Collection

The equine infectious anemia virus (EIAV) is closely related to HIV and has been used as a model to identify protective mechanisms against lentivirus infection. In horses, EIA infection progresses for about a year before infected horses manage to control virus replication. This naturally-gained protection is absolutely dependent on active immune responses as evidenced by the fact that immunosuppressive drugs can induce the recurrence of disease. As the resolution of initial viremia correlates with the appearance of virus specific cytotoxic T lymphocytes (CTL), we believe that cellular immune responses play a key role in controlling EIAV in the horse. In …

Axolotl Paedomorphosis: A Comparison Of Juvenile, Metamorphic, And Paedomorphic Ambystoma Mexicanum Brain Gene Transcription, Carlena Johnson

Theses and Dissertations--Biology

Unlike many amphibians, the paedomorphic axolotl (Ambystoma mexicanum) rarely undergoes external morphological changes indicative of metamorphosis. However, internally, some axolotl tissues undergo cryptic metamorphic changes. A previous study examined interspecific patterns of larval brain gene expression and found that these species exhibited unique temporal expression patterns that were hypothesized to be morph specific. This thesis tested this hypothesis by examining differences in brain gene expression between juvenile (JUV), paedomorphic (PAED), and metamorphic (MET) axolotls. I identified 828 genes that were expressed differently between JUV, PAED, and MET. Expression estimates from JUV were compared to estimates from PAED and …

Activity Of Brassica Napus And Perilla Frutescens Microsomal \Omega-3 Desaturases Expressed In Yeast (Saccharomyces Cerevisiae), Mohammed Abdelreheem, David Hildebrand

Turkish Journal of Biology

Yeast transformations with Brassica napus and Perilla frutescens \omega-3 desaturases were carried out. Feeding experiments on exogenous 18:2 at 10 mg/L and 100 mg/L for both genes under this investigation showed the same activity levels over the 3 days of the incubation, producing almost the same amount of 18:3. To further investigate the characteristics of B. napus and P. frutescens \omega -3 desaturases, a time-course in vivo activity study was conducted. Both enzymes showed the same behavior over the time course, desaturating 18:2 into 18:3. Moreover, the use of 2 different temperatures for incubation (30 °C and 20 °C) did …

Analyses Of Chromosome Copy Number And Expression Level Of Four Genes In The Ciliate Chilodonella Uncinata Reveal A Complex Pattern That Suggests Epigenetic Regulation, Laure Bellec, Laura A. Katz

Biological Sciences: Faculty Publications

Chilodonella uncinata, like all ciliates, contains two distinct nuclei in every cell: a germline micronucleus and a somatic macronucleus. The macronucleus develops from the zygotic nucleus through a series of chromosomal rearrangements. Macronuclear development in C. uncinata yields a nucleus with highly amplified gene-sized chromosomes. The macronucleus is transcriptionally active during vegetative growth while there is no expression in the micronucleus except during a brief period following conjugation. Gene family evolution in ciliates occurs through complex processes including gene duplication and an alternative processing of scrambled genes. Here we use quantitative PCR to compare relative expression levels of eight genes …

Hypoxia-Sensitive Gene Expression In The Gastrocnemius Muscle Following Chronic Hind Limb Ischemia, Andrew Tilton

Biological Sciences

Chronic ischemia, caused by the formation atherosclerotic plaque occlusions in major conduit arteries, is the leading cause of morbidity and mortality in western societies. Vascular remodeling can help compensate for the adverse effects of atherosclerotic plaque formation. Vascular remodeling relies heavily on vascular endothelial growth factor (VEGF), a critical protein that contributes to all forms of vascular formation and remodeling including angiogenesis, arteriogenesisand vasculogenesis. VEGF itself is up-regulated by the transcription factor, hypoxia inducible factor 1 alpha (HIF-1α), which becomes activated in low oxygen environments.

Through the use of animal chronic hindlimb ischemia models, these genes can be evaluated as …

Selfish Little Circles: Transmission Bias And Evolution Of Large Deletion-Bearing Mitochondrial Dna In Caenorhabditis Briggsae Nematodes, Katie A. Clark, Dana K. Howe, Kristin Gafner, Danika Kusuma, Sita Ping, Suzanne Estes

Biology Faculty Publications and Presentations

Selfish DNA poses a significant challenge to genome stability and organismal fitness in diverse eukaryotic lineages. Although selfish mitochondrial DNA (mtDNA) has known associations with cytoplasmic male sterility in numerous gynodioecious plant species and is manifested as petite mutants in experimental yeast lab populations, examples of selfish mtDNA in animals are less common. We analyzed the inheritance and evolution of mitochondrial DNA bearing large heteroplasmic deletions including nad5 gene sequences (nad5Δ mtDNA), in the nematode Caenorhabditis briggsae. The deletion is widespread in C. briggsae natural populations and is associated with deleterious organismal effects. We studied the inheritance patterns of nad5Δ …

Consequences Of Stochastic Mrna Synthesis In A Gene Regulatory Pathway, Khyati Shah

Dissertations

Gene expression is a stochastic process, with elements of randomness present in both transcription and translation. This stochasticity results in cell-to-cell variation in the amounts of gene products, mRNAs and proteins, and is observed in organisms ranging from bacteria and yeast to higher eukaryotes. Randomness in the activation and inactivation of a gene is the preliminary cause of this variation. At the level of proteins, these variations are buffered compared to levels of mRNA, due to the longer lifespan of proteins. Nevertheless, there is substantial variation observed at the level of proteins, resulting in phenotypic diversity among genetically identical cells. …

Gld-1 Represses Its Puf Mrna Targets Prior To/At Initiation Of Translation In The C.Elegans Germline, Gautham Sarathy

Legacy Theses & Dissertations (2009 - 2024)

The C.elegans germline offers an ideal system to study posttranscriptional regulation of gene expression as it is a major mechanism through which the control over gene expression is achieved. GLD-1 (defective in GermLine Development) is a maxi-KH motif containing RNA binding protein that controls various aspects of germline development from decision over germcell proliferation vs. meiotic entry to the production of mature gametes suggesting that GLD-1 likely controls many mRNA targets.

Exploring The Effects Of Hypoxia On Sulfate Reducing Anaerobes, Annaliese K. Jones

Senior Honors Projects

Exploring the Effects of Hypoxia on Sulfate Reducing Anaerobes

Annaliese K. Jones

Sponsor: Bethany Jenkins, Cell and Molecular Biology

As a student about to graduate with a degree in Biological Sciences, I find myself faced with the need to find my own independent research interests and scientific voice. As a result of my interests in the fields of both microbiology and ecology, I am drawn to questions surrounding the role and behavior of microorganisms in the environment. With climate change being an issue capturing the attention of a large portion of the scientific community, I have chosen to focus my …

Tissue-Specific Thyroid Hormone Regulation Of Gene Transcripts Encoding Iodothyronine Deiodinases And Thyroid Hormone Receptors In Striped Parrotfish (Scarus Iseri), Kaitlin M. Johnson, Sean C. Lema

Biological Sciences

In fish as in other vertebrates, the diverse functions of thyroid hormones are mediated at the peripheral tissue level through iodothyronine deiodinase (dio) enzymes and thyroid hormone receptor (tr) proteins. In this study, we examined thyroid hormone regulation of mRNAs encoding the three deiodinases dio1, dio2 and dio3 – as well as three thyroid hormone receptors trαA, trαB and trβ – in initial phase striped parrotfish (Scarus iseri). Parrotfish were treated with dissolved phase T₃ (20 nM) or methimazole (3 mM) for 3 days. Treatment with exogenous T₃ elevated circulating T₃, while the …

Growth Phase-Dependent Gene Regulation In Vivo In Sulfolobus Solfataricus, Melissa Deyoung, Michelle Thayer, John Van Der Oost, Kenneth M. Stedman

Biology Faculty Publications and Presentations

Ribosomal genes are strongly regulated dependent on growth phase in all organisms, but this regulation is poorly understood in Archaea. Moreover, very little is known about growth phase-dependent gene regulation in Archaea. SSV1- based lacS reporter gene constructs containing the Sulfolobus 16S/23S rRNA gene core promoter, the TF55a core promoter, or the native lacS promoter were tested in Sulfolobus solfataricus cells lacking the lacS gene. The 42-bp 16S/23S rRNA gene and 39-bp TF55a core promoters are sufficient for gene expression in S. solfataricus. However, only gene expression driven by the 16S/23S rRNA gene core promoter is dependent on …

Analysis Of Common And Specific Mechanisms Of Liver Function Affected By Nitrotoluene Compounds, Youping Deng, Sharon A. Meyer, Xin Guan, Barbara Lynn Escalon, Junmei Ai, Mitchell S. Wilbanks, Ruth Welti, Natàlia Garcia-Reyero, Edward J. Perkins

Faculty Publications

Background: Nitrotoluenes are widely used chemical manufacturing and munitions applications. This group of chemicals has been shown to cause a range of effects from anemia and hypercholesterolemia to testicular atrophy. We have examined the molecular and functional effects of five different, but structurally related, nitrotoluenes on using an integrative systems biology approach to gain insight into common and disparate mechanisms underlying effects caused by these chemicals.

Methodology/Principal Findings: Sprague-Dawley female rats were exposed via gavage to one of five concentrations of one of five nitrotoluenes [2,4,6-trinitrotoluene (TNT), 2-amino-4,6-dinitrotoluene (2ADNT) 4-amino-2,6-dinitrotoulene (4ADNT), 2,4-dinitrotoluene (2,4DNT) and 2,6-dinitrotoluene (2,6DNT)] with necropsy …

Molecular Cloning, Characterization, And Expression Analysis Of A Gene Encoding A Ran Binding Protein (Ranbp) In Cucumis Melo L., Mehmet Cengi̇z Baloğlu, Florence Negre-Zakharov, Hüseyi̇n Avni̇ Öktem, Ayşe Meral Yücel

Turkish Journal of Biology

Ran binding proteins (RanBPs) are highly conserved members of the GTP-binding protein family that are involved in nuclear protein export between the nucleus and the cytoplasm. In this study, a CmRanBP gene from a melon was isolated (Cucumis melo L.) using the RACE (rapid amplification of cDNA ends) method. The 778 basepair long melon, with a RanBP cDNA encoding consisting of 197 amino acids (22.2 kDa protein), was characterized (GenBank accession no: EU853459). The predicted amino acid sequence of CmRanBP was found to be 70% identical to VvRanBP, PtRanBP, and RcRanBP from Vitis vinifera, Populus trichocarpa, and Ricinus communis, respectively. …

Genetics And Genomics Of Complex Diseases;Mouse Atherosclerosis Modifier Genes And Human Gene Expression After Bariatric Surgery, Stela Z. Berisha

ETD Archive

The overall goal of our research was to study and identify genes involved in the development of complex diseases in mice and humans through integrative genetic/genomic approaches. Two complex diseases were the focus of research, atherosclerosis and type 2 diabetes. Mouse models of atherosclerosis, in which the genetic background and the environment can be controlled for, were used to identify and validate atherosclerosis modifier genes. We studied an atherosclerosis quantitative trait locus (QTL) on the distal end of chromosome 5, called Ath24, and generated interval-specific congenic mice on the atherosclerosis resistant AKR and susceptible DBA/2 apoE-deficient strains. To date we …

A Comparative Study Of Larval Gene Expression Between A Paedomorphic And Metamorphic Species Of Ambystomatid Salamander, Meredith A. Boley

University of Kentucky Master's Theses

Ambystoma tigrinum undergoes an obligatory metamorphosis while A. mexicanum fails to metamorphose and exhibits paedomorphosis. While it is clear that salamander paedomorphosis is associated with genetic changes that delay developmental timing, it is not clear when and how these changes manifest during development. It is possible that paedomorphic and metamorphic larvae show equivalent patterns of developmental until late in the larval period, when brain regions become competent to stimulate the release of metamorphic hormones. To test this hypothesis, I compared gene expression patterns between the brains of A. mexicanum and A. t. tigrinum larvae. In support of the developmental equivalence …

Human Cardiac-Specific Cdna Array For Idiopathic Dilated Cardiomyopathy: Sex-Related Differences, Georges E. Haddad, Lori J. Saunders, Seth D. Crosby, Maria Carles, Federica Del Monte, Kindra King, Michael R. Bristow, Francis G. Spinale, Thomas E. Macgillivray, Marc J. Semigran, G. William Dec, Steven A. Williams, Roger J. Hajjar, Judith K. Gwathmey

Biological Sciences: Faculty Publications

Idiopathic dilated cardiomyopathy (IDCM) constitutes a large portion of patients with heart failure of unknown etiology. Up to 50% of all transplant recipients carry this clinical diagnosis. Female-specific gene expression in IDCM has not been explored. We report sex-related differences in the gene expression profile of ventricular myocardium from patients undergoing cardiac transplantation. We produced and sequenced subtractive cDNA libraries, using human left ventricular myocardium obtained from male transplant recipients with IDCM and nonfailing human heart donors. With the resulting sequence data, we generated a custom human heart failure microarray for IDCM containing 1,145 cardiac-specific oligonucleotide probes. This array was …

Inference Of Genetic Regulatory Networks With Recurrent Neural Network Models Using Particle Swarm Optimization, Rui Xu, Donald C. Wunsch, Ronald L. Frank

Electrical and Computer Engineering Faculty Research & Creative Works

Genetic regulatory network inference is critically important for revealing fundamental cellular processes, investigating gene functions, and understanding their relations. The availability of time series gene expression data makes it possible to investigate the gene activities of whole genomes, rather than those of only a pair of genes or among several genes. However, current computational methods do not sufficiently consider the temporal behavior of this type of data and lack the capability to capture the complex nonlinear system dynamics. We propose a recurrent neural network (RNN) and particle swarm optimization (PSO) approach to infer genetic regulatory networks from time series gene …

Identification Of Factors Affecting Bovine Somatic Cell Nuclear Transfer Efficiency And Characterization Of Transcriptional Profiles Of Nuclear Transfer Embryos And Cotyledons, Kenneth Ivan Aston

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Since the production of the first sheep by somatic cell nuclear transfer a great deal of effort has been made to improve efficiency and to understand nuclear reprogramming mechanisms. Unfortunately efficiency remains low, and nuclear reprogramming mechanisms remain uncharacterized. The objectives of this research were to identify factors associated with somatic cell nuclear transfer efficiency and to analyze the transcriptome of blastocyst-stage clone and control embryos and cotyledonary tissue in an effort to elucidate mechanisms responsible for the low developmental efficiency and high post-implantation losses.

The experiments reported here identify factors including oocyte source and timing of activation following nuclear …

Analysis Of Distal-Less Gene Expression In Ciona Intestinalis, Michelle Cangiano

Senior Honors Projects

Several homeobox-containing genes related to Distal-less (Dlx/Dll) have been isolated from a wide variety of organisms and have been shown to function as developmental regulators. In Drosophila only one Distal-less gene has been identified so far, and in vertebrates many components of the Distal-less family have been characterized. This suggests that, during the evolution of the Chordate phylum, the Dlx genes arose from an ancestral Distal-less gene via gene duplication. Three Dll homeoboxes have been isolated from the protochordate Ciona intestinalis and their clustered arrangement has been described. Since Ciona is regarded as one of the most primitive extant chordates, …

Contributions Of Heterosis And Epistasis To Hybrid Fitness, Mitchell B. Cruzan

Biology Faculty Publications and Presentations

Early-generation hybrid fitness is difficult to interpret because heterosis can obscure the effects of hybrid breakdown. We used controlled reciprocal crosses and common garden experiments to distinguish between effects of heterosis and nuclear and cytonuclear epistasis among morphotypes and advanced-generation hybrid derivative populations in the Piriqueta caroliniana (Turneraceae) plant complex. Seed germination, growth, and sexual reproduction of first-generation hybrids, inbred parental lines, and outbred parental lines were compared under field conditions. Average vegetative performance was greater for hybrids than for inbred lines, and firstseason growth was similar for hybrids and outbred parental lines. Hybrid survival surpassed that of inbred lines …

Monitoring Immediate-Early Gene Expression Through Firefly Luciferase Imaging Of Hrs/J Hairless Mice, Michael E. Geusz, Anne M. Collaco

Biological Sciences Faculty Publications

Background: Gene promoters fused to the firefly luciferase gene (luc) are useful for examining gene regulation in live transgenic mice and they provide unique views of functioning organs. The dynamics of gene expression in cells and tissues expressing luciferase can be observed by imaging this enzyme's bioluminescent oxidation of luciferin. Neural pathways involved in specific behaviors have been identified by localizing expression of immediate-early genes such as c-fos. A transgenic mouse line with luc controlled by the human c-fos promoter (fos::luc) has enabled gene expression imaging in brain slice cultures. To optimize imaging of immediate-early gene expression throughout intact mice, …

Enhancer Trapping Reveals Widespread Circadian Clock Transcriptional Control In Arabidopsis, Todd P. Michael, C Robertson Mcclung

Dartmouth Scholarship

The circadian clock synchronizes the internal biology of an organism with the environment and has been shown to be widespread among organisms. Microarray experiments have shown that the circadian clock regulates mRNA abundance of about 10% of the transcriptome in plants, invertebrates, and mammals. In contrast, the circadian clock regulates the transcription of the virtually all cyanobacterial genes. To determine the extent to which the circadian clock controls transcription in Arabidopsis, we used in vivo enhancer trapping. We found that 36% of our enhancer trap lines display circadian-regulated transcription, which is much higher than estimates of circadian regulation based on …

Quorum Sensing In Gram-Negative Bacteria, Gülgün Boşgelmez Tinaz

Turkish Journal of Biology

It has become increasingly and widely recognised that bacteria do not live as isolated entities but instead exist as communities that exploit elaborate systems of intercellular communication to facilitate their adaptation to changing environmental conditions. A well-characterised example of such intercellular communication is quorum sensing. Quorum sensing depends on the production of diffusible signal molecules termed autoinducers or pheromones, which enable a bacterium to monitor its own cell population density. A variety of physiological processes in a range of bacterial species is regulated by quorum sensing. Examples include bioluminescence, antibiotic biosynthesis, swarming, biofilm differentiation, conjugation and the production of virulence …