Biology Commons^™

Cellular And Organismal Ramifications Of De Novo Purine Synthesis Dysregulation, Randall Craig Mazzarino

Electronic Theses and Dissertations

Purines are a class of nitrogenous bases and are essential small molecules to life. Purines are used within the cell as genetic information carriers, energy currency, signaling molecules, and cofactors for multiple processes. They are formed through de novo and salvage pathways found in cells across the phylogenetic tree. The substrates of enzymes within de novo purine synthesis are known to influence other processes within the cell, such as energy homeostasis. In humans, de novo purine synthesis disorders are rare, with around 100 people identified. These patients exhibit a range of phenotypes, with varying degrees of mental retardation, seizure activity, …

Actin Regulation And Furrow Dynamics During Early Drosophila Embryogenesis, Yi Xie

Electronic Theses and Dissertations

Drosophila embryogenesis starts with a single nucleus undergo 13 rounds of nuclear divisions called syncytial cycles. Staring at cycle 10 when nuclei migrate to the surface of the embryo, massive and dynamic cortical actin structures and cleavage furrow formations occur. How actin regulators coordinate into an organized network directing three-dimension actin structures in the developing organisms is an unsolved question. Here, I present an in-depth characterization of actin cap dynamics: the actin caps go through expansion, stabilization, elongation and fragmentation phases in each cycle. Arp2/3 is the major contributor to actin cap formation. The functions of 7 different actin and …

Characterization Of Vps41 And Its Role In The Regulated Secretory Pathway, Christian Henry Burns

Electronic Theses and Dissertations

Insulin secretory granules (SGs) mediate the regulated secretion of insulin, which is essential for glucose homeostasis. The basic machinery responsible for this regulated exocytosis consists of specific membrane proteins present both at the plasma membrane and on insulin SGs. The protein composition of insulin SGs thus dictates their release properties, yet the mechanisms controlling insulin SG formation, which determines this molecular composition, remain poorly understood. VPS41, a component of the endolysosomal tethering HOPS complex, was recently identified as a cytosolic factor involved in the formation of neuroendocrine/neuronal granules. We now find that a stable pool of VPS41 exists outside of …

Gene Expression In The Choanoderm, Jesús Federico Peña

Electronic Theses and Dissertations

The body plan of sponges (phylum Porifera) is an outlier among modern animals and is thought to have special evolutionary significance. Sponges lack muscles, nerves and a gut. Instead, they are composed of few cell types and simple tissues that function to pump water through an internal canal network where bacterial prey are filtered by a specialized tissue called the choanoderm. The choanoderm is composed of cells with striking similarity to choanoflagellates, the unicellular relatives of animals. Thus, the traditional view is that the sponge choanoderm is a useful model of the first animal epithelial tissues. Using the freshwater sponge …

Development And Testing Of A Rapid Multiplex Assay For The Identification Of Biological Stains, Kevin M. Legg

Electronic Theses and Dissertations

While DNA profiling makes it possible to individualize biological stains, the identification of the stain itself can present forensic serologists with a significant challenge. Current antibody- and enzyme activity-based assays used by forensic practitioners for biological stain identification yield only presumptive results. Positive results with non-target body fluids or cross-reactivity with non-human sources has also been well documented. Some tests can consume unacceptable quantities of precious evidence while for some body fluids (vaginal fluid and menstrual blood) there are simply no available tests at all.

This research presented here aims to develop and rigorously test a fast, accurate, and sensitive …

Post-Transcriptional Shaping Of Neurons: The Role Of Mirnas And Fmrp-Interacting P-Body Components In Regulating Neuronal Structure, Breanna Symmes

Electronic Theses and Dissertations

Changes in synaptic structure in response to neuronal stimulation are believed to underlie the processes of learning and long-term memory. However, the mechanisms for these structural modifications are poorly understood. It is well-known that activity-dependent synaptic modifications rely upon new protein synthesis, and rapid new protein synthesis, at that. Therefore, it is widely believed that pools of messenger RNAs held in a state of translational repression are transcribed in a neuronal cell body prior to stimulation, and transported to the synapse, where they reside until stimulation occurs.

This study investigates the roles and interactions of translational repression mechanisms to better …

Rala And Rab8 Regulate Drosophila Embryogenesis, Zhongyuan Zuo

Electronic Theses and Dissertations

Pseudocleavage furrow dynamics initiate at cycle 10 to help with mitotic divisions that lack cytokinesis. Permanent furrows then form at the beginning of cellularization (cycle 14). Cytoskeletal networks such as F-actin and non-muscle Myosin II have been demonstrated to be required for furrow formation and provide anchor points for chromosomal segregation. However, what drives the onset of furrow formation and membrane addition remains largely unclear. In my thesis, I have explored RalA and Rab8 function during furrow formation at metaphase and cellularization stages. These furrows are critical for correct chromosomal segregation, while disruption of furrow formation results in severe developmental …

Role Of Camkii In Pancreatic Alpha Cells, Brooke A. Buckland

Electronic Theses and Dissertations

Calcium/calmodulin-dependent protein kinase II (CaMKII) has been identified as an important modulator in controlling electrical activity in neurons and the heart; however, a role for CaMKII in pancreatic alpha cell signaling has not been previously reported. Upon activation by calcium/calmodulin, CaMKII phosphorylates proteins involved in intracellular calcium homeostasis by inducing downstream effects such as an increase in AMPA-receptor single channel conductance, potentiation of L-type voltage dependent calcium channels, and enhanced surface expression of inhibitory GABA_A-receptors. In the pancreas, an increase in intracellular calcium drives secretion of glucagon from alpha cells within the Islets of Langerhans. α-cells contain many …

Vegetation Response After Invasive Tamarix Spp. Removal In The Riparian Zone And Semi-Arid Rangeland Ecosystems, Hisham Nagi El Waer

Electronic Theses and Dissertations

Removal of Tamarix spp. (a.k.a. tamarisk, saltcedar, Athel) invasion is often involved in restoration of Western, riparian habitat; however monitoring of vegetation after removal is often neglected and thus opportunity for adaptive management lost. To address this need, I have conducted three and half years of monitoring vegetation response after invasive Tamarix removal in twenty-five sites on the East and Western Colorado, starting fall 2009. I am also comparing six different methodologies: Point intercept, line transect, nearest neighbor, meter-square quadrats, nested Whittaker plots, and densitometer with the objective of developing monitoring protocols that can be used by …

Evolution Of Xenopus Tropicalis Mc2r: Expression And Activation, Perry Victoria Davis

Electronic Theses and Dissertations

The structure and functional relationship between the melanocortin-2 receptor (MC2R) and adrenocorticotropic hormone (ACTH) is the most complex of the melanocortin gene family. Prior studies had been done on amniote tetrapod MC2Rs (e.g., mammals); this study analyzed the expression and activation of MC2R by an anamniote tetrapod, Xenopus tropicalis (xtMC2R). An immunofluorescence approach, done on the expression of xtMC2R in Chinese Hamster Ovary cells (CHO cells), indicated that the trafficking of xtMC2R to the plasma membrane required co-expression with a tetrapod MRAP1(melanocortin-1 receptor accessory protein). A cAMP-reporter assay was used to show that xtMC2R can activated by human ACTH(1-24, but …

Mitochondrial Glutathione Transport: Implications For Bcl-2 And Neuronal Survival, Heather Marie Wilkins

Electronic Theses and Dissertations

Oxidative stress is a contributing factor to many neurodegenerative diseases. In particular, mitochondria are a key source of oxidative stress due to electron leakage at the level of the electron transport chain. To combat the endogenous production of reactive oxygen and reactive nitrogen species mitochondria are equipped with several redox-cycling systems, such as glutathione (GSH). Mitochondrial GSH has been shown to be a critical reservoir of this antioxidant, where selective depletion of mitochondrial GSH can induce apoptosis in several systems. Many studies have intricately linked Bcl-2 to cellular GSH status and it has been previously shown that Bcl-2 is a …

Connexin-32 And Connexin-43 Immunoreactivity In Rodent Taste Buds, Amanda E. Bond

Electronic Theses and Dissertations

Studies indicate that ATP is one of the primary neurotransmitters in taste transduction. ATP release occurs from taste cells via specific hemichannels such as pannexin/connexin hemichannels (Huang et al., 2007; Romanov et al., 2007). We hypothesize that Type II (receptor) and possibly Type III (presynaptic) cells release ATP at sites containing pannexin/connexin hemichannels. In this study, we examine the presence of connexin–32–LIR (Like Immunoreactivity) and connexin–43–LIR in rodent taste buds through immunocytochemical analysis and DAB (Di–amino–benzidine) immunoelectron microscopy. We observed that connexin–32–LIR co–localizes with P2X2–LIR in nerve fibers and in a small subset of NCAM–LIR cells. Connexin–32–LIR does not co–localize …

Analyzing The Interaction Between Melanocortin 5 Receptor, Melanocortin Receptor Accessory Protein 1, And Melanocortin 2 Receptor In Rainbow Trout, Domicinda M. Hill

Electronic Theses and Dissertations

Using CHO cells we sought to explore and characterize the functional relationship of rainbow trout MC5 receptor (rtMC5R) with zebrafish MRAP1 (zfMRAP1) and rainbow trout MC2 receptor (rtMC2R), as well as how the effect of such relationships may play a significant role in Hypothalamic-Pituitary-Adrenal/Interrenal axis activation. This research demonstrated that rtMC5R can be successfully expressed and functionally activated in CHO cells and, in a manner similar to mammals, α-MSH is the preferred ligand for rtMC5R. The presence of MRAP1 does not seem to inhibit the expression or function of rtMC5R, rather it appears that it may increase expression and total …

Dopamine-Beta-Hydroxylase And Norepinephrine Transporter Immunoreactivity In Rat Taste Buds, Jenna Ashley Walton

Electronic Theses and Dissertations

Of the handful of neurotransmitters modulating taste response, norepinephrine (NE) may be the least understood. Although the physiological role of NE has been tested, it is still unclear whether the rat taste bud may serve as an endogenous source of neurotransmitter or if it must be taken up from outside the taste bud. Immunocytochemical analysis of DBH has shown that DBH–LIR is present in taste cells of the rat circumvallate papillae. Specifically, DBH–LIR is present in a subset of Type II taste cells. Nearly all (98%) DBH–LIR cells express PLCβ2–LIR, and about half (41%) of PLCβ2–LIR cells express DBH–LIR. DBH–LIR …

Nutraceutical Antioxidants And Their Therapeutic Potential In Neurodegeneration, Erika Kristine Ross

Electronic Theses and Dissertations

Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disease that affects motor neurons of the brain and spinal cord. Many studies indicate that mitochondrial oxidative stress (MOS) is a principal mechanism underlying the pathophysiology of this and other devastating neurodegenerative diseases. Here, we investigated a unique whey protein supplement (Immunocal®) to determine its neuroprotective efficacy in several in vitro models of MOS and in an in vivo mouse model of ALS. This non-denatured whey supplement contains cystine which is an oxidized form of cysteine, an essential precursor for synthesis of the endogenous antioxidant, glutathione (GSH). In primary cultured rat cerebellar …

Sequence Detection And Comparative Analysis Of The Hv1 And Hv2 Control Regions Of Human Mitochondrial Dna By Denaturing High-Performance Liquid Chromatography, Sarah E. Lewis

Electronic Theses and Dissertations

The objective of this research was the evaluation and forensic validation of Denaturing High-Performance Liquid Chromatography (DHPLC) as a sequencingindependent means of detecting the presence of sequence differences in pair-wise mixtures of non-concordant amplicons of human mitochondrial DNA (mtDNA). The reproducibility and efficacy of DHPLC results, including amplification reproducibility, injection reproducibility, and column-to-column reproducibility were measured, showing negligible assay-to-assay variability. In addition, cross-contamination on the DHPLC columns demonstrated very low level DNA carryover between a high-abundance sample and subsequent zero-volume injections.

The accuracy with which DHPLC technology can be used to screen both evidence and control samples in the context …

Camp And Calcium Interactions In Endocrine Cells, Colette Crane

Electronic Theses and Dissertations

Inhibitors of cAMP phosphodiestereses (PDEs) promoted an increase in intracellular [cAMP]_c and an influx of Ca²⁺, in both MMQ pituitary cells and α-Pancreatic cells. The specific targets for PDEs and the domains of PDEs are still unknown. Pituitary MMQ cells were used as a model to test modulation of calcium influx through the L-type calcium channel with PDE inhibitors. cAMP analogs were used to determine cAMP molecular targets. 8-cpt-cAMP initiated an Epac mediated pathway and had strong effects on calcium influx in the cell; suggesting, a relatively strong cAMP target. Experiments of α-Pancreatic cells were done in …

Hepatitis C Virus-Induced Hepatocellular Carcinoma: Cancer Stem Cell And Gene Therapy, Heba Samir Esmaeil Allam

Electronic Theses and Dissertations

Hepatitis C virus (HCV) is an important human pathogen that causes chronic hepatitis cirrhosis, steatosis and hepatocellular carcinoma (HCC) worldwide. Although current FDA-approved drugs are partially effective in the treatment of HCV, methods for cure and vaccine against the virus are still awaited. My thesis work presented here is focused on finding mechanism (s) of HCV-induced hepatocarcinogenesis and devising novel means to combat the HCV infection.

Herein, we have shown that long-term HCV expression results in the acquisition of cancer stem-like cells (CSCs) traits in liver derived cell lines. These traits include enhanced expression of putative stem cell markers DCAMKL-1, …

Altered Calcium Handling In Cerebellar Purkinje Neurons With The Malignant Hyperthermia Mutation, Ryr1-Y522s/+, George C. Talbott

Electronic Theses and Dissertations

To investigate the etiology of malignant hyperthermia and central core disease, mouse models have recently been generated and characterized (Chelu et al., 2006). These RyRY522S/+ knock-in mutant mice provide an excellent tool to investigate calcium dysregulation, its pathological consequences, and potential therapeutic approaches. Skeletal muscle harboring this mutation exhibits calcium leak from internal stores and an increased sensitivity to activation by caffeine, voltage, and temperature (Durham et al., 2008). Although alterations in RyR1 channel function and resultant changes in cellular function have been characterized in skeletal muscle, the effects of MH mutations in RyR1 on central nervous system function have …

Neuronal Calcium Regulation And Cellular Stress In A Malignant Hyperthermia Disease Model, Jason Santiago

Electronic Theses and Dissertations

Calcium ions (Ca2+) are essential signaling molecules and must be carefully regulated to preserve specificity in function. One indispensible Ca2+ signaling protein is the ryanodine receptor (RyR) calcium release channel. RyR1 is essential for muscle contraction, and RyR1mutations can lead to severe muscle disorders such as malignant hyperthermia (MH). Recently, a mouse model of human MH (Y522S-RyR1) has been developed. In skeletal muscle, Y522S-RyR1 exhibits an increased sensitivity to activation resulting in Ca2+ leak, mitochondrial disorganization, and cellular stress. Although RyR1 expression is limited in the brain, the cerebellum may be particularly vulnerable to this disease because RyR1 is highly …

Effects Of Participation In Inquiry Science Workshops And Follow-Up Activities On Middle School Science Teachers' Content Knowledge, Teacher-Held Misconceptions, And Classroom Practices, Linda F. Cepeda

Electronic Theses and Dissertations

An important aspect of developing science literacy for all students is developing science-literate teachers. With the implementation of the No Child Left Behind Act, many middle school teachers found themselves in a position where they were no longer qualified to teach middle school science. This study was designed to help science teachers increase their science content knowledge, identify and resolve misconceptions/errors they may have, and assist them in their teaching by providing strategies for inquiry-based teaching, science laboratory exercises, and science equipment.

Teachers enrolled in biology courses offered by the Rocky Mountain Middle School Math and Science Partnership participated in …

Structure-Function Analysis Of Endoproteolytic Cleavage: Site-Directed Mutagenesis Studies Of The Α-Msh Cleavage Site In Silurana Tropicalis Pomc, Quinn Kun Chen

Electronic Theses and Dissertations

The maturation process of many prohormone molecules typically requires endoproteolytic cleavage C-terminal of dibasic residues, such as the K¹⁴¹R¹⁴²↓ site N-terminal of the α-melanocyte stimulating hormone (α-MSH) sequence in Silurana tropicalis proopiomelanocortin (POMC). In order to determine the absolute requirement of basic amino acid residues in the cleavage process, site-directed mutagenesis was employed to substitute alanine for the wild-type residues in the frog POMC open reading frame. Specifically, the following underlined residues were individually targeted for alanine substitution: R ¹³⁷ Q ¹³⁸ E ¹³⁹ N ¹⁴⁰ K ¹⁴¹ R ¹⁴²↓. The mouse pancreatic αTC1.9 cell …

Role Of Pro-Apoptotic Bcl2-Homology-3 Domain (Bh3)-Only Proteins In The Mutant Sod1 Mouse Model Of Als, Anna George Andrianakos

Electronic Theses and Dissertations

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease involving rapid degeneration of motor neurons in the spinal cord and retraction of their axonal projections to the neuromuscular junctions. Several known mutations linked to some familial cases of ALS have been linked to mutations in Cu/Zn superoxide dismutase (SOD1), resulting in mitochondrial oxidative stress and intrinsic apoptosis. Transgenic mice expressing a G93A mutant of SOD1 provide an in vivo model to investigate motor neuron death during disease progression. The principal regulators of intrinsic apoptosis are the Bcl-2 family proteins. While some members of this family are pro-survival, the Bcl-2 homology-3 …

Quantifying Soil And Groundwater Chemistry In Areas Invaded By Tamarix Spp. Along The Middle Rio Grande, New Mexico, Michelle Kelly Ohrtman

Electronic Theses and Dissertations

Tamarix spp. (a.k.a. saltcedar, tamarisk) invasion is considered a major ecological threat at both national and global levels, with supposed impacts on soil and water chemistry. One of the most often cited mechanisms of ecosystem change by Tamarix is through its ability to deposit salty exudates and salt-rich leaf litter. The degree to which Tamarix relates to elevated soil and groundwater salinity, however, has not been adequately quantified, especially in the context of environmental factors that may also influence salinity. If Tamarix does elevate localized salinity by means of uptake, concentration and exudation by plant tissues, then we might expect …

Purinergic Receptors In The Circumvallate Papillae Of Rats, Alana Marselle Montoya

Electronic Theses and Dissertations

P2X2 receptors and P2Y4 receptors are ATP-activated cell surface receptors that gate movement of K+, Na+, and Ca2+. We used immunocytochemistry for P2X2 and P2Y4 receptors as well as taste cell type markers to learn if P2X2 immunoreactivity (IR) is present in nerve processes in close apposition to Type II and/or Type III taste cells. We also tested to see if P2Y4 IR is present in Type II and/or Type III taste cells. Our results indicate that P2X2 is present only in intragemmal nerve processes. P2X2-LIR nerve processes form close contacts with Type II and Type III taste cells. P2Y4 …