Cell and Developmental Biology Commons^™

Influence Of Developmental Stage, Habitat, And Captivity On Thecutaneous Bacterial Communities Of Eastern Hellbenders (Cryptobranchus Alleganiensis Alleganiensis) In West Virginia, Rachel Fern Arrick

Theses, Dissertations and Capstones

Microbes inhabit virtually all surfaces of multicellular animal hosts, with microbial cells outnumbering the hosts’ own cells 10:1. Symbiont microbes, collectively referred to as the microbiome, can have profound impacts on the metabolism, development, behavior, and disease resistance of their multicellular hosts. Because the community structure of symbiont bacteria can influence host health, the characterization of amphibian microbiomes is becoming an increasingly important tool for future conservation in the face of global amphibian declines. Eastern hellbenders are good candidates for a microbiome study because they have seen substantial declines in recent decades and learning more about the environmental and physiological …

Gene Transfer In A Newly Established Spontaneous Feline Fibrosarcoma Cell Line, Rounak Nande

Theses, Dissertations and Capstones

Fibrosarcoma is a deadly disease in cats and is most often located at classical vaccine injections sites. More rare forms of spontaneous non-vaccination site (NSV) fibrosarcomas have been described and have been found associated to genetic alterations.

Purpose of this study was to compare the efficacy of adenoviral gene therapy in NVS fibrosarcoma. The NVS fibrosarcoma cell line (Cocca-6A) was isolated from a spontaneous fibrosarcoma that occurred in a domestic calico cat. Cocca-6A fibrosarcoma cells were morphologically and cytogenetically characterized. The feline cells were karyotyped and their chromosome number was counted using a Giemsa staining. Giemsa block staining of metaphase …

Bursicon, Metamorphosis And Development Of Resilin In The Fruit Fly, David Paul Anthony Neff

Theses, Dissertations and Capstones

Hovering flight is the primary form of locomotion in the numerous and diverse forms of winged insects. A specialized elastic protein, Resilin, contributes to flight mechanics in most described insect orders. Although many studies have inferred functions of Resilin in insect flight, we report the first direct evidence that Resilin is a necessary component in flight mechanics of the fruit fly. We have successfully knocked down Resilin synthesis with RNAi and have generated a new resilin deletion mutant Df(2R)anhedral. By disturbing the formation of the Resilin rich tergopleural tendon, we have shown that Resilin functions in wing control. We also …

Comparing Digit Morphology Of An Arboreal Salamander With Potential Competitors, Eric Hugh Diefenbacher

Theses, Dissertations and Capstones

Green Salamanders (Aneides aeneus) are the only salamanders in West Virginia to exhibit an arboreal lifestyle. The focus of this study was to determine the cellular anatomy of the distal digit structures and how these structures may influence climbing ability. Comparative histology, comparative morphometrics, and comparative osteology of Green Salamanders, Cumberland Plateau Salamanders (Plethodon kentucki), and Slimy Salamanders (Plethodon glutinosus), were also used to determine if Cumberland Plateau Salamanders had the potential to compete with Green Salamanders for arboreal habitats. Histologically, Cumberland Plateau Salamanders had cell layer thickening similar to that of Green Salamanders. Morphometrically, Green Salamanders and Cumberland Plateau …

Distribution And Conservation Genetics Of The Cow Knob Salamander, Plethodon Punctatus Highton (Caudata: Plethodontidae), Matthew R. Graham

Theses, Dissertations and Capstones

Being lungless, plethodontid salamanders respire through their skin and are especially sensitive to environmental disturbances. Habitat fragmentation, low abundance, extreme habitat requirements, and a narrow distribution of less than 70 miles in length, makes one such salamander, Plethodon punctatus, a species of concern (S1) in West Virginia. To better understand this sensitive species, day and night survey hikes were conducted through ideal habitat and coordinate data as well as tail tips (10 to 20 mm in length) were collected. DNA was extracted from the tail tips and polymerase chain reaction (PCR) was used to amplify mitochondrial 16S rRNA gene fragments. …

New Microanatomical Characters For Scorpion Taxonomy (Arachnida: Scorpiones), Michael Brewer

Theses, Dissertations and Capstones

Morphological characters used in scorpion systematics are limited to a few reliable structures. The most recent high-level, comprehensive revision of scorpion phylogeny, 38 of the 105 morphological characters were based on trichobothria – socketed mechanosensory seta (Soleglad & Fet, 2003). Other microstructures such as granules, setae, spines, etc. have been found in scorpions but their use for taxonomy was limited. In an effort to better understand the complex relationships among these ancient but similar looking animals, we attempted to first locate new microstructures and then assess their reliability and variability thus determining the levels at which they are diagnostic. The …

Arsenic Toxicity In Plhc-1 Cell Line And The Distribution Of Arsenic In Central Appalachia, Yeong Nam Jeong

Theses, Dissertations and Capstones

Our ecosystem has geobiophysical relationships. Environmental pollution can lead to immediate consequences like organism death or lack of reproduction. It can also lead to long-range consequences through mutations and DNA damage. Fish can be affected very directly by environmental contamination in water but the study of fish is not performed well. We are developing a quantitative assay to assess DNA damage in vitro due to heavy metal contamination using a tissue culture line of Poeciliopsis lucida, the desert pupfish, which is used for other toxicology assays. To detect DNA damage, Single Cell Gel Electrophoresis (Comet Assay) is very useful. …

Analysis Of Genotypic And Phenotypic Differences In Desmognathus Quadramaculatus Across The Southern Appalachians, Deborah Susan Merritt

Theses, Dissertations and Capstones

Populations of Desmognathus quadramaculatus were compared for genotypic and phenotypic differences across 5 states in the Southern Appalachians. Series consisting of 10 salamanders each were collected from West Virginia, Virginia, Tennessee, North Carolina and Georgia. Salamanders were assessed for genotypic differences by using allozyme (proteins with polymorphic loci) electrophoresis staining for 13 loci of the muscle, liver and stomach tissue. Nei’s genetic distance was used to score genetic variation between populations. Phenotypic differences were assessed by comparing 10 external morphological measurements between populations. Mahalanobis’ distance was used to score morphological differences between populations. Both genetic and morphological differences were correlated …

Aging Affects Stretch-Induced P70s6k And 4e-Bp1 Phosphorylation In Fast- And Slow-Twitch Muscle, Sreevani Uddemarri

Theses, Dissertations and Capstones

In the present investigation we compare the expression, basal activation and the ability of muscle stretch to activate the p70^S6k pathway in the fast-twitch extensor digitorum longus (EDL) and slow-twitch soleus of adult (6 mo. old), aged (30 mo. old) and very aged (36 mo. old) Fischer 344 x Brown Norway rats. Immunoblotting demonstrated that the tissue content of mTOR, p70^S6k, 4E-BP1 and GSK-3β decreased in the EDL and soleus in aged rats, while SHP-2 increased in the EDL and decreased in the soleus when compared to adult rats. Basal phosphorylation of 4E-BP1 increased in both the …

Response Properties Of Tibial Campaniform Sensilla On The Cockroach Leg In Restrained Preparations And Freely Moving Animals, Angela L. Ridgel

Theses, Dissertations and Capstones

The ability to detect changes in forces is important for effective use of a leg in posture and locomotion. This thesis examines how forces are detected in the legs of cockroaches by tibial campaniform sensilla. Campaniform sensilla are mechanoreceptors that encode forces through ovoid cuticular caps embedded in the exoskeleton. The tibial sensilla are unique in that they consist of two subgroups with mutually perpendicular cap orientations.

We characterized the responses of the tibial receptors in restrained preparations by applying forces to the leg at controlled magnitudes and rates. The tibial sensilla, as a group, were sensitive to increasing and …

Interaction Of Laminin Beta2 With Eps8 And Desmin In Muscle Cells, Luke Xueliang Cui

Theses, Dissertations and Capstones

Laminin β2 is localized to the synaptic basal lamina (BL) in muscle and glomerular BL in kidney. To find interacting proteins, a mouse kidney cDNA library was screened with domain I of rat laminin β2 using the yeast two-hybrid system. Fifteen positive clones were found. The DNA sequence of three each of these clones corresponded to the phosphoprotein Eps8 (epidermal growth factor receptor pathway substrate clone 8, which is tyrosine phosphorylated after EGF stimulation) and the intermediate filament (IF) protein keratin. To determine if this interaction occurred in muscle cells, C2 cells transfected with the rat …

The Role Of Domain I In Laminin Chain Assembly, George D. Kamphaus

Theses, Dissertations and Capstones

Laminin, a major component of basement membrane, is a trimeric glycoprotein comprised of three chains - α, β and γ (Burgeson et al., 1994). An order for trimer assembly has been deduced: first, the β and γ chains bind to form a dimer and subsequently α is added to complete the trimer (I. Hunter et al., 1990 & 1992; Utani et al., 1994 & 1995). The C-terminal portions, found within the protein structural domain I of the p and y chains, are implicated in dimer and trimer formation by biochemical studies performed extracellularly (Utani, et al., 1994 & 1995; Nomizu …

In Vivo Incorporation Of Tritium To Measure Lipogenesis In Red Skeletal Muscle: Significance Of A Nonlipogenic Diet In La/N-Fa(K)("Corpulent") And Zucker Fa Rat Strains, Karin Traci Mann

Theses, Dissertations and Capstones

Skeletal muscle lipogenesis in rodents may equal that of liver but its importance in the obese animals is not known. We evaluated the rate of fatty acid and triglyceride synthesis and storage content in red skeletal muscles, liver, inguinal fat, and retroperitoneal fat of the LA/N-fa^k (“corpulent”) and Zucker fa obese rat strains. Obese-lean pairs of both rat strains, weaned at 32 days, fed a nonlipogenic diet (54% starch for 21 days) were either dosed with 2 µCi 3H-H² O per gram of body weight and tested after 60 minutes for whole body 3H-incorporation into fatty acids and …

An Immunomodulating Mycotoxin Interferes With The Development Of Autoimmune Diabetes In Diabetes-Prone Bb/Wor Rats, Honggang Liu

Theses, Dissertations and Capstones

Various fungal products have immunomodulating activity and some have been studied regarding prevention of transplantation rejection. Prior to this investigation, the mycotoxin, gliotoxin (GT), has never been investigated as an immunotherapeutic drug for autoimmune disease. GT is a fungal secondary metabolite and a member of the epipolythiodioxopiperazine (ETP) family which has been shown to inhibit phagocytosis, induction of cytolytic T cells and the proliferation of T cells following mitogen stimulation. GT also induces in vitro apoptosis in certain immune cell types. More importantly, GT exhibits selective activity towards cells of hemopoietic origin.

Autoimmune diseases are disorders caused by immune responses …

Use Of The (3)H-Tetracycline Rat Model In The Study Of Skeletal Compartmentalization And Metabolism Of Calcium In The Maturing Male And Female Rat And The Role Of The Skeleton In Calcium Homeostasis In The Adolescent Male Rat, Darrin Lee Demoss

Theses, Dissertations and Capstones

The (3)H-tetracycline bone labeling procedure was employed to monitor bone resorption from urinary loss curves in male and female rats of various ages. In addition, whole body dry skeletal mass and the loss of (3)H-tetracycline from individual bones was determined. It was found that the dry skeletal mass/body mass ratio of 24-week-old females was 30-40% greater than that for males. The urinary loss of (3)H-tetracycline was described by a double exponential equation indicating the presence of two distinct and independent exchangeable bone compartments. Both compartments decrease in size with age, but their label loss activities were different. The label loss …

Regulation Of Melanogenesis In B16 Mouse Melanoma Cells By Protein Kinase C (Pkc), Harish Mahalingam

Theses, Dissertations and Capstones

The pigment cell-specific expression of tyrosinase and TRP1 has been shown to be important for the production of melanin in pigmented cells. Using a pigmented cell line, B16 mouse melanoma, we obtained evidence that PKC plays a major role in regulating melanogenesis. Chronic treatment with phorbol dibutyrate (PDBu) leads to downregulation of PKC activity and protein levels. This is accompanied by a loss of pigmentation which is correlated with a 50% reduction and a complete loss of TRP1 and tyrosinase respectively. Similar results were obtained with Northern and Western blotting indicating that PKC may regulate the steady state levels of …

Postnatal Development Of The Neural Retina In A South American Opossum: Monodelphis Domestica, Tracy L. Soltesz

Theses, Dissertations and Capstones

Postnatal retinal development was studied in a marsupial opossum, Monodelphis domestica using light microscopy and 3H-thymidine autoradiography. For the light microscopic study, opossum neonates at 1, 3, 7, 14, 21, 25 and 28 days of age were euthanized, fixed and processed into paraffin sections for hematoxylin and eosin staining. The distinct ganglion cell layer, first observed on postnatal day (P) 7, was separated from the outer neuroblasts by the inner plexiform layer. The neuroblast layer was divided into inner and outer nuclear layers on P25 by the presumptive outer plexiform layer, indicated by discrete intercellular spaces located between the nuclear …

Genetic Analysis Of Rhinichthys Atratulus (Pisces: Cyprinidae) In North Central West Virginia, Alan Harper Tennant

Theses, Dissertations and Capstones

The blacknose dace, Rhinichthys atratulus, is a cyprinid minnow common to shallow streams throughout North America. It has been shown to be sensitive to anthropogenic stress and a study of the genetic nature of R. atratulus populations may yield information about their habitats. In West Virginia, two subspecies of R. atratulus are present, R. a. atratulus and R. a. obtusus, which are separated by the Allegheny divide. In north central West Virginia, R. a. atratulus was observed in the Cheat River drainage, an area previously thought be within the range of R. a. obtusus. Fish of the R. a. atratulus …

Geographic And Nongeographic Variation In The Genus Microtus (Mammalia: Rodentia) In West Virginia, Tina M. Savage

Theses, Dissertations and Capstones

Geographic and nongeographic variation was assessed on 12 cranial and 4 external characters in approximately 400 specimens of Microtus (M. ochrogaster, M. chrotorrhinus, M. pinetorum, M. pennsylvanicus) in West Virginia. Univariate and multivariate statistical analyses were used to examine age-related variation, sexual dimorphism, elevation and geographic variation and litter size for each species of Microtus in the state. Cranial characters were more beneficial than external in detecting variation. Examination of data for each species of Microtus studied demonstrated that values and trends in variation is a specific rather than a generic trait. Age-related variation was present in some, …