Life Sciences Commons^™

A Laser Pointer Driven Microheater For Precise Local Heating And Conditional Gene Regulation In Vivo. Microheater Driven Gene Regulation In Zebrafish, Mike Placinta, Meng-Chieh Shen, Marc Achermann, Rolf O. Karlstrom

Rolf O Karlstrom

Background - Tissue heating has been employed to study a variety of biological processes, including the study of genes that control embryonic development. Conditional regulation of gene expression is a particularly powerful approach for understanding gene function. One popular method for mis-expressing a gene of interest employs heat-inducible heat shock protein (hsp) promoters. Global heat shock of hsp-promoter-containing transgenic animals induces gene expression throughout all tissues, but does not allow for spatial control. Local heating allows for spatial control of hsp-promoter-driven transgenes, but methods for local heating are cumbersome and variably effective. Results - We describe a simple, highly controllable, …

The Acidic Domains Of The Toc159 Chloroplast Preprotein Receptor Family Are Instrinsically Disordered Protein Domains, Lynn G.L. Richardson, Masoud Jelokhani-Niaraki, Matthew D. Smith

Biology Faculty Publications

Background: The Toc159 family of proteins serve as receptors for chloroplast-destined preproteins. They directly bind to transit peptides, and exhibit preprotein substrate selectivity conferred by an unknown mechanism. The Toc159 receptors each include three domains: C-terminal membrane, central GTPase, and N-terminal acidic (A-) domains. Although the function(s) of the A-domain remains largely unknown, the amino acid sequences are most variable within these domains, suggesting they may contribute to the functional specificity of the receptors.

Results: The physicochemical properties of the A-domains are characteristic of intrinsically disordered proteins (IDPs). Using CD spectroscopy we show that the A-domains of two Arabidopsis Toc159 …

Mir319a Targeting Of Tcp4 Is Critical For Petal Growth And Development In Arabidopsis, Anwesha Nag, Stacey King, Thomas Jack

Dartmouth Scholarship

In a genetic screen in a drnl-2 background, we isolated a loss-of-function allele in miR319a (miR319a¹²⁹). Previously, miR319a has been postulated to play a role in leaf development based on the dramatic curled-leaf phenotype of plants that ectopically express miR319a (jaw-D). miR319a¹²⁹ mutants exhibit defects in petal and stamen development; petals are narrow and short, and stamens exhibit defects in anther development. The miR319a¹²⁹ loss-of-function allele contains a single-base change in the middle of the encoded miRNA, which reduces the ability of miR319a to recognize targets. Analysis of the expression patterns of the …

Sympatric Ecological Speciation Meets Pyrosequencing: Sampling The Transcriptome Of The Apple Maggot Rhagoletis Pomonella, Dietmar Schwarz, Hugh M. Robertson, Jeffrey L. Feder, Kranthi Varala, Matthew E. Hudson, Gregory J. Ragland, Daniel A. Hahn, Stewart H. Berlocher

Biology Faculty and Staff Publications

Background

The full power of modern genetics has been applied to the study of speciation in only a small handful of genetic model species - all of which speciated allopatrically. Here we report the first large expressed sequence tag (EST) study of a candidate for ecological sympatric speciation, the apple maggot Rhagoletis pomonella, using massively parallel pyrosequencing on the Roche 454-FLX platform. To maximize transcript diversity we created and sequenced separate libraries from larvae, pupae, adult heads, and headless adult bodies.

Results

We obtained 239,531 sequences which assembled into 24,373 contigs. A total of 6810 unique protein coding genes …

Effects Of Temperature On Gene Expression In Embryos Of The Coral Montastraea Faveolata, Christian Voolstra, Julia Schnetzer, Leonid Peshkin, Carly J. Randall, Alina M. Szmant, Mónica Medina

Biomedical Engineering and Sciences Faculty Publications

Coral reefs are expected to be severely impacted by rising seawater temperatures associated with climate change. This study used cDNA microarrays to investigate transcriptional effects of thermal stress in embryos of the coral Montastraea faveolata. Embryos were exposed to 27.5°C, 29.0°C, and 31.5°C directly after fertilization. Differences in gene expression were measured after 12 and 48 hours.Results: Analysis of differentially expressed genes indicated that increased temperatures may lead to oxidative stress, apoptosis, and a structural reconfiguration of the cytoskeletal network. Metabolic processes were downregulated, and the action of histones and zinc finger-containing proteins may have played a role in the …

Defining The Role Of Syndecan-4 In Mechanotransduction Using Surfacemodification Approaches, Robert M. Bellin, James D. Kubicek, Matthew J. Frigault, Andrew J. Kamien, Robert L. Steward, Hillary M. Barnes, Michael B. Digiacomo, Luke J. Duncan, Christina K. Edgerly, Elizabeth M. Morse, Chan Young Park, Jeffrey J. Fredberg, Chao Min Cheng, Philip R. Leduc

Biology Department Faculty Scholarship

The ability of cells to respond to external mechanical stimulation is a complex and robust process involving a diversity of molecular interactions. Although mechanotransduction has been heavily studied, many questions remain regarding the link between physical stimulation and biochemical response. Of significant interest has been the contribution of the transmembrane proteins involved, and integrins in particular, because of their connectivity to both the extracellular matrix and the cytoskeleton. Here, we demonstrate the existence of a mechanically based initiation molecule, syndecan-4. We first demonstrate the ability of syndecan-4 molecules to support cell attachment and spreading without the direct extracellular binding of …

Inferring Dispersal Of Aquatic Invertebrates From Genetic Variation: A Comparative Study Of An Amphipod (Talitridae Hyalella Azteca) And Mayfly (Baetidae Callibaetis Americanus) In Great Basin Springs, Heather Lynn Stutz

Theses and Dissertations

Whether active or passive, dispersal accompanied by gene flow shapes the population genetics and evolutionary divergence of species. Indirect methods which use genetic markers have the ability to assess effective dispersal—that which resulted in gene flow. My objective was to see if an aquatic insect and an obligate aquatic invertebrate show similar phylogeographic patterns and genetic uniqueness. Hyalella azteca and Callibaetis americanus were collected from 4-5 springs in each of six basins in the Great Basin of western North America. No dispersal or genetic studies of C. americanus have been conducted to date. However, several studies focusing on mtDNA diversity …

The Gaas Metagenomic Tool And Its Estimations Of Viral And Microbial Average Genome Size In Four Major Biomes, Florent E. Angly, Dana Willner, Alejandra Prieto-Davo, Robert A. Edwards, Robert Schmieder, Rebecca Vega-Thurber, Dionsios A. Antonopoulos, Katie Barott, Matthew T. Cottrell, Christelle Desnues, Elizabeth A. Dinsdale, Mike Furlan, Matthew Haynes, Matthew R. Henn, Yongfei Hu, David L. Kirchman, Tracey Mcdole, John D. Mcpherson, Folker Meyer, R. Michael Miller, Egbert Mundt, Robert K. Naviaux, Beltran Rodriguez-Mueller, Rick Stevens, Linda Wegley, Lixin Zhang, Baoli Zhu, Forest Rohwer

Department of Biological Sciences

Metagenomic studies characterize both the composition and diversity of uncultured viral and microbial communities. BLAST-based comparisons have typically been used for such analyses; however, sampling biases, high percentages of unknown sequences, and the use of arbitrary thresholds to find significant similarities can decrease the accuracy and validity of estimates. Here, we present Genome relative Abundance and Average Size (GAAS), a complete software package that provides improved estimates of community composition and average genome length for metagenomes in both textual and graphical formats. GAAS implements a novel methodology to control for sampling bias via length normalization, to adjust for multiple BLAST …

Environmental Dna Sequencing Primers For Eutardigrades And Bdelloid Rotifers, Byron J. Adams, Jeremy Whiting, Elizabeth K. Costello, Kristen R. Freeman, Andrew P. Martin, Michael S. Robeson, Steve K. Schmidt

Faculty Publications

Background: The time it takes to isolate individuals from environmental samples and then extract DNA from each individual is one of the problems with generating molecular data from meiofauna such as eutardigrades and bdelloid rotifers. The lack of consistent morphological information and the extreme abundance of these classes makes morphological identification of rare, or even common cryptic taxa a large and unwieldy task. This limits the ability to perform large-scale surveys of the diversity of these organisms. Here we demonstrate a culture-independent molecular survey approach that enables the generation of large amounts of eutardigrade and bdelloid rotifer sequence data directly …

The Ecological Importance Of Extrinsic And Intrinsic Drivers Of Animal Movement, Josh Earl Rasmussen

Theses and Dissertations

The movement of individuals is foundational to many ecological processes. For example, the movement of an organism from one place to another alters population density at both sites and has potential for affecting the genetic dynamics within the new population. Individual movement events may be in synchrony with overall trends in populations, e.g. spawning migrations, or may be atypical (asynchronous). This latter movement type can affect population and metapopulation dynamics, depending on its prevalence within a population. Nevertheless, given the complexity of interactions, the causative factors of movement are understood vaguely, much less for aquatic organisms. Drivers of movement are …

Varriation Of Growth Rates In Yellow-Bellied Marmots, Carmen M. Salsbury, K. B. Armitage

Carmen M. Salsbury

Growth rates of yellow-bellied marmot (Marmota flaviventris) populations over a 32-year period (1965 -1996) varied Significantly with sex, age, location, and year. Overall, males had higher growth rates than females and young and yearlings generally had higher growth rates compared to adults at all locations. The locations varied with respect to elevation and the relationship between elevation and growth rate was complex and likely weather dependent. Low rainfall in late summer was often associated with low growth rates at high elevations where the active season is constrained and delayed by late spring snowmelt compared to low elevations. Growth rates and …

Varriation Of Growth Rates In Yellow-Bellied Marmots, Carmen M. Salsbury, K. B. Armitage

Carmen M. Salsbury

Growth rates of yellow-bellied marmot (Marmota flaviventris) populations over a 32-year period (1965 -1996) varied Significantly with sex, age, location, and year. Overall, males had higher growth rates than females and young and yearlings generally had higher growth rates compared to adults at all locations. The locations varied with respect to elevation and the relationship between elevation and growth rate was complex and likely weather dependent. Low rainfall in late summer was often associated with low growth rates at high elevations where the active season is constrained and delayed by late spring snowmelt compared to low elevations. Growth rates and …

Zooplankton Abundance, Community Composition And Grazing In The James River Estuary (Virginia, Usa), Laura E. Barry

Theses and Dissertations

This study examined the extent to which zooplankton in the James River Estuary (Virginia, USA) exploit the available algal-food resources. Zooplankton abundance, community composition and grazing rates were assessed at three locations which included a site with high algal production (near the VCU Rice Center) and two sites where algal production was lower. Grazing rates were measured by determining the rate of phytoplankton decline (as chlorophyll a) during 48-hour incubation experiments in the presence and absence of zooplankton. Significant differences in zooplankton abundance were observed among the three sites, with the greatest average zooplankton density (434 ± 69 ind/L) occurring …

Examining The Effects Of Penning On Juvenile Eastern Box Turtles (Terrapene Carolina Carolina), Nicolas Frederick

Theses and Dissertations

Box turtle (Terrapene carolina) populations have been declining over the last several decades, and one major cause is increasing urbanization. As a result of habitat fragmentation, wildlife managers are frequently turning to new and alternative management strategies. Traditional box turtle management has included relocation, which has been met with limited success. This study aims to combine these strategies with another less-studied one: forcing turtles to overwinter on site by penning them in an outdoor enclosure. Two sets of juvenile box turtles were released at the Virginia Commonwealth University Rice Center: one penned on site in a pen for one year, …

Conservation Genetics And Systematics Of Several Turtles Species In The Southeastern United States, Joshua Robert Ennen

Dissertations

Chelonians (i.e., turtles) are an imperiled group of reptiles with about 66% of the recognized species listed as threatened by the IUCN. Most chelonian species have a unique set of life history traits (i.e., longevity, delayed sexual maturity, and low juvenile survivorship), which makes their populations exceedingly sensitive to increases in adult and juvenile moralities. With numerous anthropogenic effects (e.g., habitat alteration, exploitation, and over harvesting) negatively influencing mortality rates, chelonians have experienced global precipitous declines and extinctions.

This dissertation focuses on species within two chelonian genera, Gopherus and Graptemys. Although these two genera are vastly different ecologically, they are …

Structural Basis Of Caspase-3 Substrate Specificity Revealed By Crystallography, Enzyme Kinetics, And Computational Modeling, Bin Fang

Biology Dissertations

Caspase-3 is a cysteine protease that hydrolyzes diverse intracellular proteins during programmed cell death (known as apoptosis). It has been a popular target for drug design against abnormal cell death for more than a decade. No approved caspase based drug, however, is available so far. Therefore, structural insights about the substrate recognition of caspase-3 are needed for the future development of caspase-3 based inhibitors and drugs. In this study, crystal structures of recombinant caspase-3 in complex with seven substrate analog inhibitors, including acetyl (Ac)-DEVD-aldehyde (Cho), Ac-DMQD-Cho, Ac-IEPD-Cho, Ac-YVAD-Cho, Ac-WEHD-Cho, Ac-VDVAD-Cho, and tert-butoxycarbonyl (Boc)-D-fluoromethylketone (Fmk), have been analyzed in combination with …

Distribution And Diversity Of Archaeal And Bacterial Ammonia Oxidizers In Salt Marsh Sediments, Nicole S. Moin, Katelyn A. Nelson, Alexander Bush, Anne E. Bernhard

Biology Faculty Publications

Diversity and abundance of ammonia-oxidizing Betaproteobacteria (β-AOB) and archaea (AOA) were investigated in a New England salt marsh at sites dominated by short or tall Spartina alterniflora (SAS and SAT sites, respectively) or Spartina patens (SP site). AOA amoA gene richness was higher than β-AOB amoA richness at SAT and SP, but AOA and β-AOB richness were similar at SAS. β-AOB amoA clone libraries were composed exclusively of Nitrosospira-like amoA genes. AOA amoA genes at SAT and SP were equally distributed between the water column/sediment and soil/sediment clades, while AOA amoA sequences at SAS were primarily affiliated with the …

The Ecology Of The Peaks Of Otter Salamander (Plethodon Hubrichti) In Sympatry With The Eastern Red-Backed Salamander (Plethodon Cinereus), Norman Reichenbach, Andrew Kniowski

Faculty Publications and Presentations

The Peaks of Otter Salamander, Plethodon hubrichti, is found along a 19 km length of the Blue Ridge Mountains, Virginia, USA, often in sympatry with the Eastern Red-backed Salamander, P. cinereus. In a sympatric area of Bedford County, Virginia, we conducted a mark-recapture study on a 10 × 10 m site. Surface densities of salamanders increased as the number of days without precipitation prior to a collection event increased. This suggests vertical movements in response to surface moisture. When salamanders returned to the surface after rain, individuals appeared to “shuffle” between rocks and likely, leaf litter. That is, we were …

First Record Of The Extinct Sawfish Propristis Schweinfurthi Dames, 1883 (Batoidea: Pristiformes: Pristidae) From The Middle Eocene Of Spain, Francesc Farrés, Harry L. Fierstine

Biological Sciences

A partial rostrum of Propristis schweinfurthi Dames, 1883 was collected in the Bartonian strata of northeastern Spain. The specimen represents the second record of the species from Europe, and the second occurrence of a sawfish (Mesopristis osonensis Farrés, 2003) from the Vic-Manlleu Marls Formation. In spite of some evidence to the contrary, Propristis probably preferred ecological conditions similar to extant sawfishes, i.e., inhabiting near shore tropical to subtropical seas with occasional excursions into freshwater.

Remarks On The Stability Of Some Size-Structured Population Models V: The Case When The Death Rate Depends On Adults Only And The Growth Rate Depends On Size Only, M. El-Doma

Applications and Applied Mathematics: An International Journal (AAM)

We continue our study of size-structured population dynamics models when the population is divided into adults and juveniles, started in El-Doma (To appear). We concentrate our efforts in the special case when the death rate depends on adults only, the growth rate depends on size only and the maximum size for an individual in the population is infinite. Three demographic parameters are identified and are shown to determine conditions for the (in)stability of a nontrivial steady state. We also give examples that illustrate the stability results. The results in this paper generalize previous results, for example, see Calsina, et al. …

Phylogeography And Postglacial Dispersal Of Smallmouth Bass (Micropterus Dolomieu) Into The Great Lakes, W. Calvin Borden, Robert A. Krebs

Biological, Geological, and Environmental Faculty Publications

Refugia and dispersal routes of smallmouth bass (Micropterus dolomieu) into the Great Lakes were identified using 427 mitochondrial sequences from across their native range. Overall, smallmouth bass accessed the Great Lakes via long-distance dispersal from multiple refugia, and the distribution of genetic variation reflected the consequences of vicariant, dispersal, and paleogeological events. Proximity of lakes to glacial outlets had a greater predictive power on the resulting distribution of mitochondrial diversity relative to interbasin migration. Populations in the Eastern and Interior Highlands contained the most divergent and oldest haplotypes, reflecting their role as glacial refugia and subsequent high incidence of endemicity. …

Remodeling Of The Tight Junction During Recovery From Exposure To Hydrogen Peroxide In Kidney Epithelial Cells, Jeannette E. Gonzalez, R. J. Digeronimo, D'Ann E. Arthur, Jonathan M. King

Biology Faculty Research

Renal ischemia-reperfusion injury results in oxidative stress-induced alterations in barrier function. Activation of the mitogen-activated protein (MAP) kinase pathway during recovery from oxidative stress may be an effector of oxidant-induced tight junction reorganization. We hypothesized that tight junction composition and barrier function would be perturbed during recovery from oxidative stress. We developed a model of short-term H₂O₂ exposure followed by recovery using Madin Darby canine kidney (MDCK II) cells. H₂O₂ perturbs barrier function without a significant cytotoxic effect except in significant doses. ERK-1/2 and p38, both enzymes of the MAP kinase pathway, were activated …

A New North American Genus Of Hetaeraiinae (Coleoptera: Histeridae), With Descriptions Of Six New Species From The U.S.A. And Mexico, Michael S. Caterino, Alexey K. Tishechkin

Publications

Renclasea n. gen., a genus of the Hetaeriinae (Histeridae), with six species, R. skelleyi n. sp. (United States: Florida and Georgia), R. falli n. sp. (United States: California), R. helavai n. sp. (United States: Arizona), R. mexicana n. sp. (Mexico: Hidalgo), R. occidentalis n. sp. (United States: Arizona and New Mexico) and R. cazieri n. sp. (United States: Arizona), are described, illustrated and diagnosed. The status of two female specimens of Renclasea from western Texas, apparently closely related to R. occidentalis, remains uncertain. The only host record available for the genus is an association of R. mexicana with undetermined species …

Remote Sensing Of Harmful Algal Blooms In The Mississippi Sound And Mobile Bay: Modelling And Algorithm Formation, Dan Martin Holiday

Dissertations

The incidence and severity of harmful algal blooms have increased in recent decades, as have the economic effects of their occurrence./The diatom Pseudo-nitzschia spp. caused fisheries closures in Mobile Bay during 2005 due to elevated levels of domoic acid. In the previous 4 years Karenia brevis counts of >5,000 cells L"1 have occurred in Mobile Bay and the Mississippi Sound. Population levels of this magnitude had previously been recorded only in 1996. Increases in human populations, urban sprawl, development of shoreline properties, sewage effluent and resultant changes in NP ratios of discharge waters, and decline in forest and marsh lands, …

Two-Layered Model Of Blood Flow Through Composite Stenosed Artery, Padma Joshi, Ashutosh Pathak, B. K. Joshi

Applications and Applied Mathematics: An International Journal (AAM)

In this paper a steady, axisymmetric flow, with a constricted tube has been studied. The artery has been represented by a two-layered model consisting of a core layer and a peripheral layer. It has been shown that the resistance to flow and wall shear stress increases as the peripheral layer viscosity increases. The results are compared graphically with those of previous investigators. It has been observed that the existence of peripheral layer is useful in representation of diseased arterial system.

Remarks On The Stability Of Some Size-Structured Population Models Vi: The Case When The Death Rate Depends On Juveniles Only And The Growth Rate Depends On Size Only And The Case When Both Rates Depend On Size Only, M. El-Doma

Applications and Applied Mathematics: An International Journal (AAM)

We continue our study of size-structured population dynamics models when the population is divided into adults and juveniles, started in El-Doma (to appear 1) and continued in El-Doma (to appear 2). We concentrate our efforts in two special cases, the first is when the death rate depends on juveniles only and the growth rate depends on size only, and, the second is when both the death rate and the growth rate depend on size only. In both special cases we assume that the maximum size for an individual in the population is infinite. We identify three demographic parameters and show …

Remarks On The Stability Of Some Size-Structured Population Models Iv: The General Case Of Juveniles And Adults, M. El-Doma

Applications and Applied Mathematics: An International Journal (AAM)

The stability of some size-structured population dynamics models is investigated when the population is divided into adults and juveniles. We determine the steady states and study their stability. We also give examples that illustrate the stability results. The results in this paper generalize previous results, for example, see Calsina, et al. (2003), El-Doma (2006), Farkas, et al. (2008), and El-Doma (2008 a).

Pulsatile Flow Of Blood In A Constricted Artery With Body Acceleration, Devajyoti Biswas, Uday Shankar Chakraborty

Applications and Applied Mathematics: An International Journal (AAM)

Pulsatile flow of blood through a uniform artery in the presence of a mild stenosis has been investigated in this paper. Blood has been represented by a Newtonian fluid. This model has been used to study the influence of body acceleration and a velocity slip at wall, in blood flow through stenosed arteries. By employing a perturbation analysis, analytic expressions for the velocity profile, flow rate, wall shear stress and effective viscosity, are derived. The variations of flow variables with different parameters are shown diagrammatically and discussed. It is noticed that velocity and flow rate increase but effective viscosity decreases, …

Isoform- And Cell-Specific Function Of Tyrosine Decarboxylase In The Drosophila Malpighian Tubule, Edward M. Blumenthal

Biological Sciences Faculty Research and Publications

The biogenic amine tyramine (TA) is a potent diuretic factor when applied to the Malpighian tubule (MT) of Drosophila melanogaster, stimulating both urine production and transepithelial chloride conductance. Isolated MTs can respond not only to TA but also to its precursor, tyrosine; this observation led to the proposal that MTs are able to synthesize TA from applied tyrosine through the action of the enzyme tyrosine decarboxylase (TDC). In the current study it is shown that the non-neuronal isoform of TDC, Tdc1, is expressed in the principal cells of the MT. A mutant allele of Tdc1, Tdc1^{f03311 …}

Baseline Survey And Habitat Analysis Of Aquatic Salamanders In The Pigeon River, North Carolina, Nikki J. Maxwell

Masters Theses

The Pigeon River was severely impacted beginning in the early 1900s by a paper mill located in Canton, North Carolina. The mill discharged chemical byproducts into the Pigeon River until 1992 when the paper mill modified their processes. As a result, water quality improved but the status of salamander species in the Pigeon River was unknown. Worldwide amphibian declines over the last 20 years have drawn attention to the need for more research and a better understanding of species-specific habitat relationships. There is concern about amphibian population declines because amphibians are critical to the balance of ecosystems and are considered …