Cell and Developmental Biology Commons^™

Phosphorylation Of Skeletal Muscle Acetyl-Coa Carboxylase By Ampk Enhances Palmitoyl-Coa Inhibition, Dustin S. Rubink

Theses and Dissertations

Acetyl-CoA carboxylase (ACC) catalyzes the formation of malnoyl-CoA, which in turn controls the rate of fatty acid metabolism. ACC beta or 2 has been shown to be localized on the mitochondria in close proximity to carnintine palmitoyl transferase 1 (CPT-1), the enzyme responsible for the influx of acyl-CoA into the matrix where beta oxidation takes place. CPT-1 is inhibited by malonyl-CoA produced by ACC. It has been well documented that AMP activated kinase (AMPK) when activated phosphorylates and inactivates ACC. ACC is controlled allosterically by citrate, which activates, and by palmitoyl-COA, which inhibits. In this study, we asked the question, …

Use Of Cre Recombinase And Sb Transposase To Engineer Transgenic Zebrafish, Jie Dong

All-Inclusive List of Electronic Theses and Dissertations

Current transgenesis has become easier for certain species due to the recent development of exogenous non site-specific recombination systems such as Sleeping Beauty (SB) transposon, and site-specific recombination systems such as Cre-loxP. Such recombination systems overcame two major problems caused by conventional transgenesis, position effect and copy number effect. SB is a reconstructed fish transposon that has shown to efficiently insert transgenes into heterologous mammalian chromosomes. The Cre-loxP system is frequently used to delete a given sequence from the genome or to target the integration of foreign DNA. The use of both systems in combination, however, has not yet been …

Waccnes Containing Bovine Herpe Svirus 1 Attenuated By Mutation In Latency-Related Gene, Clinton J. Jones

School of Veterinary and Biomedical Sciences: Faculty Publications

Vaccines for pathogenic Strains of bovine herpesvirus 1 (BHV-1) which are based on attenuated BHV-1 having a mutation in the latency-related gene are provided. Live, attenuated vaccines are also provided which express anti gens from other viral or bacterial pathogens and thus form the basis of a variety of vaccines.

The Bvg Virulence Control System Regulates Biofilm Formation In Bordetella Bronchiseptica, Yasuhiko Irie, Seema Mattoo, Ming H. Yuk

Biology Faculty Publications

Bordetella species utilize the BvgAS (Bordetella virulence gene) two-component signal transduction system to sense the environment and regulate gene expression among at least three phases: a virulent Bvg⁺ phase, a nonvirulent Bvg⁻ phase, and an intermediate Bvgⁱ phase. Genes expressed in the Bvg⁺ phase encode known virulence factors, including adhesins such as filamentous hemagglutinin (FHA) and fimbriae, as well as toxins such as the bifunctional adenylate cyclase/hemolysin (ACY). Previous studies showed that in the Bvgⁱ phase, FHA and fimbriae continue to be expressed, but ACY expression is significantly downregulated. In this report, we determine …

The Caenorhabditis Elegans F-Box Protein Sel-10 Promotes Female Development And May Target Fem-1 And Fem-3 For Degradation By The Proteasome, Sibylle Jager, Hillel T. Schwartz, H. Robert Horvitz, Barbara Conradt

Dartmouth Scholarship

The Caenorhabditis elegans F-box protein SEL-10 and its human homolog have been proposed to regulate LIN-12 Notch signaling by targeting for ubiquitin-mediated proteasomal degradation LIN-12 Notch proteins and SEL-12 PS1 presenilins, the latter of which have been implicated in Alzheimer's disease. We found that sel-10 is the same gene as egl-41, which previously had been defined by gain-of-function mutations that semidominantly cause masculinization of the hermaphrodite soma. Our results demonstrate that mutations causing loss-of-function of sel-10 also have masculinizing activity, indicating that sel-10 functions to promote female development. Genetically, sel-10 acts upstream of the genes fem-1, fem-2, and fem-3 and …

Sexually Dimorphic Effects Of Prenatal Stress On Cognition, Hormonal Responses, And Central Neurotransmitters, Rachel E. Bowman, Neil J. Maclusky, Yessenia Sarmiento, Maya Frankfurt, Marisa Gordon, Victoria N. Luine

Psychology Faculty Publications

Exposure to stress during gestation results in physiological and behavioral alterations that persist into adulthood. This study examined the effects of prenatal stress on the postnatal expression of sexually differentiated cognitive, hormonal, and neurochemical profiles in male and female rats. Pregnant dams were subjected to restraint stress three times daily for 45 min during d 14-21 of pregnancy. The offspring of control and prenatally stressed dams were tested for anxiety-related and cognitive behaviors, stress and gonadal steroid hormone levels, as well as monoamines and metabolite levels in selected brain regions. Postnatal testosterone levels (measured at 1 and 5 d) did …

Thermal Adaptation Of Maternal And Embryonic Phenotypes In A Wide Spread Lizard (Sceloporus Undulatus), Christopher E. Oufiero

All-Inclusive List of Electronic Theses and Dissertations

Geographically widespread ectotherms encounter thermal gradients that generate variation in behavior, physiology and life-history. Because rates of physiological processes are very sensitive to temperature, cold environments are thought to favor genotypes that have relatively high capacities for growth despite environmental constraints. Two different adaptive responses can occur: (1) mothers in colder environments can allocate more energy to each offspring, or (2) offspring in colder environments make more efficient use of available resources. In this study, I examined both maternal adaptations and embryonic adaptations to environmental temperature. Evidence of maternal adaptations was assessed by examining the relationship between egg size and …

The Ultrastructure Of The Olfactory System In Two Species Of Short-Tailed Shrews, Blarina Brevicauda And Blarina Carolinensis, Lisa Johnson Byrum

Theses and Dissertations in Biomedical Sciences

Several studies of the fine structure of the olfactory system of rodents have been conducted, but very little research has been done on members of the Insectivora. The olfactory systems of the northern short-tailed shrew, Blarina brevicauda, and the southern short-tailed shrew, Blarina carolinensis, were examined by light and electron microscopy. These shrews were live trapped in the vicinity of Norfolk, Virginia throughout all months of the year. Olfactory tissues were processed following standard transmission and scanning electron microscopy protocols. The olfactory system structures investigated included the olfactory epitheliumlmucosa (OEM), main olfactory bulb (MOB), accessory olfactory bulb (AOB), …

Mechanism Governing The Cellular Susceptibility To Secretory Phospholipase A2, Lauren Blackburn Jensen

Theses and Dissertations

Secretory phospholipase A2 (sPLA2) is an important part of apoptosis and disposal of damaged and dying cells. However, healthy cells are not susceptible to attack by sPLA2. Recent studies have focused on membrane properties necessary to induce susceptibility in both artificial and biological membranes. Hydrolysis of phospholipids by sPLA2 requires at least two preliminary steps: first, adsorption of the enzyme to the cellular membrane, and second, movement of a phospholipid into the active site of the enzyme. We determined the effects of susceptibility on each of the two steps and determined the contributions changing the equilibrium constants have on susceptibility. …

Identification And Functional Analysis Of The Essential And Regulatory Light Chains Of The Only Type Ii Myosin Myo1p In Saccharomyces Cerevisiae, J. Luo, Elizabeth Ann Vallen, C. Dravis, S. E. Tcheperegine, B. Drees, E. Bi

Biology Faculty Works

Cytokinesis in Saccharomyces cerevisiae involves coordination between actomyosin ring contraction and septum formation and/or targeted membrane deposition. We show that Mlc1p, a light chain for Myo2p (type V myosin) and lqg1p (IQGAP), is the essential light chain for Myo1p, the only type 11 myosin in S. cerevisiae. However, disruption or reduction of Mlc1p-Myo1p interaction by deleting the Mlc1p binding site on Myo1p or by a point mutation in MLC1, mlc1-93, did not cause any obvious defect in cytokinesis. In contrast, a different point mutation, mlc1-11, displayed defects in cytokinesis and in interactions with Myo2p and lqg1p. These data suggest that …

Drawing Lines In The Sand: Even Skipped Et Al. And Parasegment Boundaries., James B. Jaynes, Miki Fujioka

Department of Biochemistry and Molecular Biology Faculty Papers

The pair-rule segmentation gene even skipped (eve) is required to activate engrailed stripes and to organize odd-numbered parasegments (PSs). The protein product Eve has been shown to be an active repressor of transcription, and recent models for Eve function suggest that activation of engrailed is indirect, but these models have not been fully tested. Here we identify the forkhead domain transcription factor Sloppy-paired as the key intermediate in the initial activation of engrailed by Eve in odd-numbered parasegments. We also analyze the roles of the transcription factors Runt and Odd-skipped in this process. Detailed analysis of engrailed and pair-rule gene …

Development And Characterization Of Eukaryotic Biomimetic Liposomes, Bradley Jay Taylor

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

This study developed and characterized phospholipid vesicles, or liposomes, that mimic cell surfaces. Microemulsified liposomes contained biotinylated phosphatidylethanolamine, allowing them to be immobilized to avidin-coated glass. Laminin (LN), glycosphingolipids (GMl and GM3), and Escherichia coli's mechanosensitive channel of large conductance (EcoMscL) were embedded into liposome membranes. It was determined whether these embedded molecules exhibited their physiological roles of adhesion, cell recognition, and mechanosensation, respectively. Confocal laser scanning microscopy (CLSM) was employed to examine the interaction of fluorescently probed proteins, toxins, and bacteria with the immobilized microemulsified liposomes. Capture of individual and simultaneous multiple species of bacteria by GMl, GM3, …

Jnk Regulates Hipk3 Expression And Promotes Resistance To Fas-Mediated Apoptosis In Du 145 Prostate Carcinoma Cells, James Curtin, Thomas Cotter

Articles

Elevated endogenous JNK activity and resistance to Fas receptor-mediated apoptosis have recently been implicated in progression of prostate cancer and can promote resistance to apoptosis in response to chemotherapeutic drugs. In addition, JNK has been demonstrated to promote transformation of epithelial cells by increasing both proliferation and survival. Although numerous studies have reported a role for JNK in promoting Fas receptor-mediated apoptosis, there is a paucity in the literature studying the antiapoptotic function of JNK during Fas receptor-mediated apoptosis. Consequently, we have used the recently described specific JNK inhibitor SP600125 and RNA interference to inhibit endogenous JNK activity in the …

Drosophila As An Emerging Model To Study Metastasis, Madhuri Kango-Singh, Georg Halder

Biology Faculty Publications

Metastasis is the primary cause of human cancer-related deaths. Two recent studies describe a system for testing how multiple genetic events synergize to promote neoplastic growth and metastasis in Drosophila, paving the way for systematic approaches to understanding metastasis using the powerful tools of Drosophila genetics.

Transcriptional Profiling Of Caulobacter Crescentus During Growth On Complex And Minimal Media, Craig Stephens, Alison K. Hottes, Maliwan Meewan, Desiree Yang, Naomi Arana, Pedro Romero, Harley H. Mcadams.

Biology

Microarray analysis was used to examine gene expression in the freshwater oligotrophic bacterium Caulobacter crescentus during growth on three standard laboratory media, including peptone-yeast extract medium (PYE) and minimal salts medium with glucose or xylose as the carbon source. Nearly 400 genes (approximately 10% of the genome) varied significantly in expression between at least two of these media. The differentially expressed genes included many encoding transport systems, most notably diverse TonB-dependent outer membrane channels of unknown substrate specificity. Amino acid degradation pathways constituted the largest class of genes induced in PYE. In contrast, many of the genes upregulated in minimal …

Phylogenetic Analyses Of Texas Isolates Indicate An Evolving Subtype Of The Clade B Feline Immunodeficiency Viruses, Eric A. Weaver, Ellen W. Collisson, Margaret Slater, Guan Zhu

Nebraska Center for Virology: Faculty Publications

Rigorous phylogenetic analyses were used to compare the nucleotide sequences of feline immunodeficiency virus strains isolated from Texas and throughout the world. The envelope V3-V4 sequences and capsid gene of the Texas isolates formed a cluster between subtypes B and E. Statistical comparisons with other published sequences confirmed that the Texas group is a unique cluster, possibly a new subtype, arising from subtype B.

Feline immunodeficiency virus (FIV) was initially isolated in 1987 from a cat in California with severe immunodeficiency and has been recognized as a common worldwide feline pathogen (11, 14, 19, 20, 33). FIV-infected cats exhibit a …

Phylogenetic Signal And The Utility Of 12s And 16s Mtdna In Frog Phylogeny, S. Hertwig, Rafael O. De Sá, A. Haas

Biology Faculty Publications

Genes selected for a phylogenetic study need to contain conserved information that reflects the phylogenetic history at the specific taxonomic level of interest. Mitochondrial ribosomal genes have been used for a wide range of phylogenetic questions in general and in anuran systematics in particular. We checked the plausibility of phylogenetic reconstructions in anurans that were built from commonly used 12S and 16S rRNA gene sequences. For up to 27 species arranged in taxon sets of graded inclusiveness, we inferred phylogenetic hypotheses based on different apriori decisions, i.e. choice of alignment method and alignment parameters, including/excluding variable sites, choice of reconstruction …

Androgen Receptor Expression In The Rat Prostate Is Down-Regulated By Dietary Phytoestrogens, Edwin D. Lephart, Trent D. Lund, Daniel J. Munson, Herman Aldercreutz, Robert J. Handa

Faculty Publications

Background- It is well established that the growth of the prostate gland is a hormone-dependent phenomenon involving both androgenic and estrogenic control. Proliferation of prostate cells is, at least in part, under control of estrogen receptor beta (ER-beta). Phytoestrogens bind ER-beta with high affinity and therefore may have antiproliferative effects in the prostate. Methods- The prostates of male Long-Evans rats fed a diet high in phytoestrogens (Phyto-600) or very low levels of phytoestrogens (Phyto-free) were analyzed to determine the impact of dietary phytoestrogens on prostate weight and androgen receptor (AR) expression in the prostate. Results- Dietary phytoestrogens significantly decreased post-pubertal …

Bcma Is Essential For The Survival Of Long-Lived Bone Marrow Plasma Cells, Brian P. O'Connor, Vanitha S. Raman, Loren D. Erickson, W. James Cook, Lehn K. Weaver, Cory Ahonen, Ling-Li Lin, George Mantchev, Richard J. Bram, Randolph J. Noelle

Dartmouth Scholarship

Long-lived humoral immunity is manifested by the ability of bone marrow plasma cells (PCs) to survive for extended periods of time. Recent studies have underscored the importance of BLyS and APRIL as factors that can support the survival of B lineage lymphocytes. We show that BLyS can sustain PC survival in vitro, and this survival can be further enhanced by inter- leukin 6. Selective up-regulation of Mcl-1 in PCs by BLyS suggests that this 􏰀-apoptotic gene product may play an important role in PC survival. Blockade of BLyS, via transmembrane activator and cyclophilin ligand interactor–immunoglobulin treatment, inhibited PC survival in …

Analysis Of Microtubule Sliding Patterns In Chlamydomonas Flagellar Axonemes Reveals Dynein Activity On Specific Doublet Microtubules, M. J. Wargo, Mark A. Mcpeek, Elizabeth F. Smith

Dartmouth Scholarship

Generating the complex waveforms characteristic of beating eukaryotic cilia and flagella requires spatial regulation of dynein-driven microtubule sliding. To generate bending, one prediction is that dynein arms alternate between active and inactive forms on specific subsets of doublet microtubules. Using an in vitro microtubule sliding assay combined with a structural approach, we determined that ATP induces sliding between specific subsets of doublet microtubules, apparently capturing one phase of the beat cycle. These studies were also conducted using high Ca2+ conditions. InChlamydomonas, high Ca2+ induces changes in waveform which are predicted to result from regulating dynein

activity on specific microtubules. Our …

Semi-Biosynthesis Of Dna Nanostructures, Aoune Barhoumi

Theses, Dissertations and Capstones

Nanotechnology refers to all technologies aiming to build objects, make measurements, and carry out processes on the nanometer length scale. In particular molecular nanotechnology exemplifies the so-called "bottom up" approach, which is briefly defined as the ability to build useful nanostructures with molecular precision, such as molecular machinery. Such capability for controlling matter at the molecular scale has always been the dream of scientists.

All living things are nanofoundries. Billions of years ago, nature perfectly provided all living things with the most accurate biological nanotechnology systems. Cellular internal dynamics, communicative resonance in protein conformational states, viruses as microreplicators, nanoscale life …

Changes In Gdnf Protein Content Of Skeletal Muscle Following Involuntary Exercise Are Long Lived, Kyle R. Kinnell, Monica J. Czarnopys, John Spitsbergen

Biological Sciences Faculty and Graduate Student Research

Glial-cell line derived neurotrophic factor (GDNF) is a neurotrophic factor produced by skeletal muscle. GDNF has been shown to be potent survival factor for motor neurons. We have previously shown that GDNF content in skeletal muscle is altered following exercise and GDNF levels 24 hours after the last bout of exercise are significantly different than those measured immediately after exercise. We sought to determine how long after exercise do GDNF protein levels remain altered. We removed extensor digitorum longus (EDL) and Soleus (SOL), 24 hr, 72 hr and 1 week following the last bout of involuntary exercise. Muscles were processed …

Loss-Of-Function Analysis Of Epha Receptors In Retinotectal Mapping, David Feldheim, Masaru Nakamoto, Miriam Osterfield, Nicholas Gale, Thomas Dechiara, Rajat Rohatgi, George Yancopoulos, John Flanagan

Biology Faculty Publications

EphA tyrosine kinases are thought to act as topographically specific receptors in the well-characterized projection map from the retina to the tectum. Here, we describe a loss-of-function analysis of EphA receptors in retinotectal mapping. Expressing patches of a cytoplasmically truncated EphA3 receptor in chick retina caused temporal axons to have reduced responsiveness to posterior tectal repellent activity in vitro  and to shift more posteriorly within the map in vivo . A gene disruption of mouse EphA5, replacing the intracellular domain with beta-galactosidase, reduced in vitro  responsiveness of temporal axons to posterior target membranes. It also caused map abnormalities in vivo …

The Evolving Role Of Lipid Rafts And Caveolae In G Protein-Coupled Receptor Signaling: Implications For Molecular Pharmacology, Rennolds S. Ostrom, Paul A. Insel

Pharmacy Faculty Articles and Research

The many components of G-protein-coupled receptor (GPCR) signal transduction provide cells with numerous combinations with which to customize their responses to hormones, neurotransmitters, and pharmacologic agonists. GPCRs function as guanine nucleotide exchange factors for heterotrimeric (α, β, γ) G proteins, thereby promoting exchange of GTP for GDP and, in turn, the activation of ‘downstream’ signaling components. Recent data indicate that individual cells express mRNA for perhaps over 100 different GPCRs (out of a total of nearly a thousand GPCR genes), several different combinations of G-protein subunits, multiple regulators of G-protein signaling proteins (which function as GTPase activating proteins), and various …

Nitric Oxide Inhibition Of Adenylyl Cyclase Type 6 Activity Is Dependent Upon Lipid Rafts And Caveolin Signaling Complexes, Rennolds S. Ostrom, Richard A. Bundey, Paul A. Insel

Pharmacy Faculty Articles and Research

Several cell types, including cardiac myocytes and vascular endothelial cells, produce nitric oxide (NO) via both constitutive and inducible isoforms of NO synthase. NO attenuates cardiac contractility and contributes to contractile dysfunction in heart failure, although the precise molecular mechanisms for these effects are poorly defined. Adenylyl cyclase (AC) isoforms type 5 and 6, which are preferentially expressed in cardiac myocytes, may be inhibited via a direct nitrosylation by NO. Because endothelial NO synthase (eNOS and NOS3), β-adrenergic ( AR) receptors, and AC6 all can localize in lipid raft/caveolin-rich microdomains, we sought to understand the role of lipid rafts in …

Phosphorylation Of Vesicular Stomatitis Virus Phosphoprotein P Is Indispensable For Virus Growth†, Subash C. Das, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

The phosphoprotein (P) of vesicular stomatitis virus (VSV) is an essential subunit of the viral RNAdependent RNA polymerase (RdRp) complex. It is phosphorylated at two different domains. Using defective interfering (DI) RNA or minigenomic RNA templates, we previously demonstrated that phosphorylation within the amino-terminal domain I is essential for transcription, whereas phosphorylation within the carboxyterminal domain II is necessary for replication. For the present study, we examined the role of the phosphorylation of residues in these domains in the life cycle of VSV. Various mutant P coding sequences were inserted into a full-length cDNA clone of VSV, and the virus …

The Effects Of Cucurbitacin E And Genistein In Mcf-7 Breast Cancer Cells, Angela Carlson

The Journal of Undergraduate Research

No abstract provided.

The Effects Of A One-Tesla Magnet On Human Fibroblast Cell Growth, Ashley Chaplin

Theses and Dissertations

The controversy regarding the ill effects of electromagnetic fields began in 1979 when Nancy Wertheimer and Ed Leeper claimed to have found a connection between childhood leukemia and power lines. In 1997, a group of researchers led by Martha Linet, M. D., attempted to provide evidence that there was no link between cancer and electromagnetic fields. The study showed that the risk of acute lymphoblastic leukemia did not escalate with increasing electromagnetic field levels in the children's homes. When compared to the Earth's static magnetic field of 0.5 Gauss, these fields were extremely small. Within the past decade, magnets have …

Visualizing The Needle In The Haystack: In Situ Hybridization With Fluorescent Dendrimers, Jacquelyn Gerhart, M. Baytion, J. Perlman, C. Neely, B. Hearon, T. Nilsen, R. Getts, J. Kadushin, Mindy George-Weinstein

PCOM Scholarly Works

In situ hybridization with 3DNA dendrimers is a novel tool for detecting low levels of mRNA in tissue sections and whole embryos. Fluorescently labeled dendrimers were used to identify cells that express mRNA for the skeletal muscle transcription factor MyoD in the early chick embryo. A small population of MyoD mRNA positive cells was found in the epiblast prior to the initiation of gastrulation, two days earlier than previously detected using enzymatic or radiolabeled probes for mRNA. When isolated from the epiblast and placed in culture, the MyoD mRNA positive cells were able to differentiate into skeletal muscle cells. These …

Morphological Characteristics And Species Separation Of Hawaiian Postlarval Amphidromous Fishes, M. Eric Benbow, Albert J. Burky, Carl M. Way

Biology Faculty Publications

There are five Hawaiian amphidromous fishes (Gobiidae: Lentipes concolor, Awaous guamensis, Sicyopterus stimpsoni, Stenogobius hawaiiensis; Eleotridae: Eleotris sandwicensis). Amphidromous adults deposit eggs on the stream bottom. After hatching, larvae drift to the ocean for growth followed by postlarval migration back into the streams. Postlarvae were collected to construct a dichotomous identification key based on 12 morphological measures and ray counts from four fins. Overall, the presense of fused pelvic fins, standard length (SL), and fin ray numbers were the most useful in species separation. Gobies were separated from the eleotrid by the former having fused …