Cell and Developmental Biology Commons^™

Reversal Of Cell Fate Determination In Caenorhabditis Elegans Vulval Development, Susan Euling, Victor Ambros

Dartmouth Scholarship

In Caenorhabditis elegans, the fates of the multipotent vulval precursor cells (VPCs) are specified by intercellular signals, The VPCs divide in the third larval stage (L3) of the wild type, producing progeny of determined cell types, In lin-28 mutants, vulva development is similar to wild-type vulva development except that it occurs precociously, in the second larval stage (L2), Consequently, when lin-28 hermaphrodites temporarily arrest development at the end of L2 in the dauer larva stage, these otherwise determined VPC progeny become reprogrammed back to the multipotent, signal- sensitive state of VPCs. Our results indicate that VPC fate determination by intercellular …

The Evaluation Of Succinylcholine Chloride For Harmful Effects Upon Early-Stage Mouse Embryos, Francis Warren Prescott

Biological Sciences Theses & Dissertations

Succinylcholine (SCh) chloride is a depolarizing neuromuscular blocking agent used to paralyze patients for surgical procedures. Over 1,800 clinical observations of newborn infants have demonstrated that succinylcholine does not cross the placenta in quantities detrimental to fetuses (Kolstad, et al., 1957, Bakhoum, et al., 1957, Hodges, et al., 1959, Duffield, et al., 1958, Dennis, et al., 1954, 1956, McNab, 1955, Coleman, et al., 1956, Lund, 1953). Later investigations which involved comparison of blood levels of SCh in the mother to those in fetuses bolstered the finding that SCh fails to cross the placenta in significant amounts (Moya and Kvisselgaard, 1961, …

Modulated Structures In Electrodeposited Superconducting Ba(1-X)K(X)Bio(3), Chad Bryant Huffman

Theses, Dissertations and Capstones

Compositionally modulated crystals of the superconductor Ba_1-x K_xBiO₃ have been grown electrochemically. Modulation was produced through control of the deposition potential.

Modulated crystals were imaged using optical microscopy, backscattered SEM, and STM. The expected linear relationship between the amount of time that current was applied and the modulation thickness was observed, and the growth rates were determined.

Multilayers were differentially etched using an EDTA solution. Etched and unetched samples were imaged using backscattered SEM.

The transition temperature of a modulated sample was determined by SQUID magnetometry to equal the bulk value.

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 …

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 …

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 …

Heat Shock Factor Gains Access To The Yeast Hsc82 Promoter Independently Of Other Sequence-Specific Factors And Antagonizes Nucleosomal Repression Of Basal And Induced Transcription, Alexander M. Erkine, C. C. Adams, T. Diken, D. S. Gross

Scholarship and Professional Work – COPHS

Transcription in eukaryotic cells occurs in the context of chromatin. Binding of sequence-specific regulatory factors must contend with the presence of nucleosomes for establishment of a committed preinitiation complex. Here we demonstrate that the high-affinity binding site for heat shock transcription factor (HSF) is occupied independently of other cis-regulatory elements and is critically required for preventing nucleosomal assembly over the yeast HSC82 core promoter under both noninducing (basal) and inducing conditions. Chromosomal mutation of this sequence, termed HSE1, erases the HSF footprint and abolishes both transcription and in vivo occupancy of the TATA box. Moreover, it dramatically reduces promoter chromatin …

Topographically Specific Effects Of Elf-1 On Retinal Axon Guidance In Vitro And Retinal Axon Mapping In Vivo, Masaru Nakamoto

Biology Faculty Publications

Topographic maps, which maintain the spatial order of neurons in the order of their axonal connections, are found throughout the nervous system. In the visual retinotectal projection, ELF-1, a ligand in the tectum, and its receptors in the retina show complementary gradients in expression and binding, indicating they may be positional labels for map development. Here we show that ELF-1 acts as a repellent axon guidance factor in vitro. In vivo, when the tectal ELF-1 pattern is modified by retroviral overexpression, retinal axons avoid ectopic ELF-1 patches and map to abnormally anterior positions. All these effects were seen on axons …

Examination Of The Cell Wall Of Micrasterias Radiosa Var Radiosa (Conjugatophyceae) By Transmission And Scanning Electron Microscopy, Jennifer A. Akin, Richard L. Meyer

Journal of the Arkansas Academy of Science

The cell wall of Micrasterias radiosa var. radiosa Ralfs 1848 (Conjugatophyceae) was examined by transmission and scanning electron microscopy. Previous electron microscopy of this taxon has not been performed; thus these are new observations. The cell wall was recognized to be of the Cosmarium type with complex pores external to the plasma membrane that penetrate the secondary wall and with ornamentation arising from the secondary wall. Subdivided regions of the pore apparatus, the pore head, pore bulb, connecting pore channel, and pore depression were detected. Pores of type 4 were located in the isthmal region and at the division of …

Modulation Of Queuine Uptake And Incorporation Into Trna By Protein Kinase C And Protein Phosphatase, Rana C. Morris, Bonnie J. Brooks, K. Lenore Hart, Mark S. Elliot

Chemistry & Biochemistry Faculty Publications

It has been suggested that the rate of queuine uptake into cultured human fibroblasts is controlled by phosphorylation levels within the cell. We show that the uptake of queuine is stimulated by activators of protein kinase C (PKC) and inhibitors of protein phosphatase; while inhibitors of PKC, and down-regulation of PKC by chronic exposure to phorbol esters inhibit the uptake of queuine into cultured human fibroblasts. Activators of cAMP- and cGMP-dependent kinases exert no effect on the uptake of queuine into fibroblast cell cultures. These studies suggest that PKC directly supports the activity of the queuine uptake mechanism, and that …

Bovine Cell Line Resistant To In Vtro Infection By Bovine Viral Darrheavirus And All Other Known Pestiviruses, Ruben O. Donis, Eduardo F. Flores

School of Veterinary and Biomedical Sciences: Faculty Publications

Bovine cell line resistant to infection by the pestiviruses Bovine Viral Diarrhea Virus (BVDV), Hog Cholera Virus (HCV) and Border Disease Virus (BDV) and all progeny and mutants thereof. A bovine cell line (CRIB) that is resistant to infection by cytopathic and non-cytopathic BVDV and by other Pestiviruses due to a stable, recessive genetic defect which blocks infection at the level of viral entry.

Características De La Osificación Craneal En Phyllomedusa Boliviana (Anura: Hylidae), Rafael O. De Sá, E. O. Lavilla

Biology Faculty Publications

RESUMEN. Dentro de la familia Hylidae disponemos de información sobre las secuencias de osificación craneal para menos del 4.0% de sus miembros. El presente trabajo describe la secuencia de osificación craneal de Phyllomedusa boliviana. El patrón general de osificación es similar a lo reportado para otras especies de la familia, pero existen algunas diferencias importantes tales como la osificación tardía de los próoticos y la osificación premetamórfica de neopalatino, vómer, cuadrado-yugal, pterigoides y columela. El lento desarrollo ulterior de la osificación resulta en juveniles con cráneos extensamente cartilaginosos.

ABSTRACT. Characteristics of the cranial ossification in Phyllomedusa …

Phase Ia/Ib Trial Of Anti-Gd2 Chimeric Monoclonal Antibody 14.18 (Ch L4.18) And Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor (Rhgm-Csf) In Metastatic Melanoma, James Murray, Eugenie Kleinerman, Shu-Fang Jia, Michael Rosenblum, Omar Eton, Antonio Buzaid, Sewa Legha, Merrick Ross, Lora Thompson, Mansoor Saleh

Haematology and Oncology, East Africa

We performed a phase Ia/Ib trial of chimeric anti-GD2 monoclonal antibody 14.18 (ch14.18) in combination with recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) to determine the maximum tolerated dose as well as immunologic and biologic responses to the regimen. Sixteen patients with metastatic malignant melanoma received escalating doses of ch14.18 (15-60 mg/m2) administered intravenously for 4 h on day 1. Twenty-four hours later, subcutaneous injections of rhGM-CSF were administered daily for a total of 14 days. Significant side effects were related to ch14.18 infusion and consisted of moderate to severe abdominal and/or extremity pain, blood pressure changes, headache, nausea, diarrhea, peripheral …

Development And Lipid Composition Of The Harpacticoid Copepod Nitocra Spinipes Reared On Different Diets, Gregory M. Weiss, George B. Mcmanus, H. Rodger Harvey

OES Faculty Publications

We reared the harpacticoid copepod Nitocra spinipes on diets of bacteria, a diatom, or a macroalga, evaluating survivorship and growth in short-term (≤ 1 generation) experiments. Lipid content of the copepods and their diets was measured and used as an index of nutrition. Although growth, survivorship and lipid content of N. spinipes were significantly greater when fed the diatom, which had the highest lipid content of the 3 diets, the copepod was able to develop from egg to adult when fed a lipid- poor bacterial diet. Furthermore, this species was able to go through developmental molts without the addition of …

Connexin32 Is A Myelin-Related Protein In The Pns And Cns, Steven S. Scherer, Suzanne M. Deschênes, Yi-Tian Xu, Judith B. Grinspan, Kenneth H. Fischbeck, David L. Paul

Biology Faculty Publications

We have examined the expression of a gap junction protein, connexin32 (Cx32), in Schwann cells and oligodendrocytes. In peripheral nerve, Cx32 is found in the paranodal myelin loops and Schmidt-Lanterman incisures of myelinating Schwann cells, and the levels of Cx32 protein and mRNA change in parallel with those of other myelin-related genes during development, Wallerian degeneration, and axonal regeneration. In the central nervous system, Cx32 is found in oligodendrocytes and their processes, but not in compact myelin, and the levels of Cx32 protein and mRNA increase during development in parallel with those of the other myelin genes. Thus, Cx32 is …

Mutations In Rad27 Define A Potential Link Between G₁ Cyclins And Dna Replication, Elizabeth Ann Vallen, Frederick R. Cross , '78

Biology Faculty Works

The yeast Saccharomyces cerevisiae has three G1 cyclin (CLN) genes with overlapping functions. To analyze the functions of the various CLN genes, we examined mutations that result in lethality in conjunction with loss of cln1 and cln2. We have isolated alleles of RAD27/ERC11/YKL510, the yeast homolog of the gene encoding flap endonuclease 1, FEN-1.cln1 cln2 rad27/erc11 cells arrest in S phase; this cell cycle arrest is suppressed by the expression of CLN1 or CLN2 but not by that of CLN3 or the hyperactive CLN3-2. rad27/erc11 mutants are also defective in DNA damage repair, as determined by their increased sensitivity to …

Trophic Transfer Of Fatty Acids, Sterols, And A Triterpenoid Alcohol Between Bacteria, A Ciliate, And The Copepod Acartia Tonsa, Melissa C. Ederington, George B. Mcmanus, H. Rodger Harvey

OES Faculty Publications

The incorporation of lipids into the copepod Acartia tonsa and its eggs was measured when it was fed either a bacterivorous ciliate (Pleuronema sp.) or a diatom (Thalassiosira weissflogii). Egg production was 10-fold higher on the diatom diet, whereas hatch success of eggs was the same for algal and ciliate diets. Adult copepods fed diatoms contained more total fatty acid and sterols than copepods fed the ciliate diet, and individual lipids reflected the dietary source. Eggs from diatom-fed copepods had fewer fatty acids but more sterols than eggs from copepods on a ciliate diet. Ciliate-fed copepods and …

In Vitro Expression Of Human Zona Pellucida Protein 3, Chih-Wei Chen

Theses and Dissertations in Biomedical Sciences

Human zona pellucida protein 3 (hZP3) is the putative receptor on the zona pellucida of the mature oocyte that recognizes and binds sperm, and therefore plays a critical role in fertilization.

A full length cDNA of hZP3 (1278 bp) was amplified from the human ovary mRNA by reverse transcription-polymerase chain reaction (RT-PCR). The hZP3 cDNA was subcloned into PSK and pREP4 expression vectors. The cDNA of hZP3 was further characterized by restriction mapping, PCR, auto-sequencing and Southern blot analysis by using an internal oligonucleotide probe, and found to be identical to the one reported by J. Dean. Using autosequencing, 289 …

Investigation Of The Substrate Recognition Characteristics And Kinetics Of Mammalian Mitochondrial Dna Topoisomerase I, Zeki Topcu

Theses and Dissertations in Biomedical Sciences

Topoisomerases are DNA-modifying enzymes found in prokaryotes, eukaryotes, viruses and organelles such as chloroplast and mitochondria. Information about these enzymes in eukaryotic systems is mostly limited to nuclear enzymes, although our laboratory has been characterizing the biochemical and biophysical properties of the mammalian mitochondrial topoisomerases. We have determined the polarity of the attachment of mitochondrial topoisomerase I to its substrate DNA. To study the substrate preference and kinetic parameters of mitochondrial topoisomerase I, selected regions of mammalian mitochondrial DNA (mtDNA) were inserted into pGEM plasmid vectors following a series of modification and optimization experiments of currently available methods for PCR-cloning. …

Heterozygosity Mapping Of Partially Congenic Lines: Mapping Of A Semidominant Neurological Mutation, Wheels ( Whl), On Mouse Chromosome 4, Patrick M. Nolan, Patricia J. Sollars, Barbara A. Bohne, Warren J. Ewens, Gary E. Pickard, Maja Bucan

School of Veterinary and Biomedical Sciences: Faculty Publications

We identified a semidominant, chemically induced, mouse use mutation with a complex array of abnormal behaviors including bidirectional circling and hyperactivity, abnormal circadian rhythmicity and abnormal responses to light. In this report, we genetically and phenotypically characterized the circling/ waltzing component of the abnormal behavior. We mapped the locus controlling this trait by heterozygosity mapping of partially congenic lines carrying the mutagenized chromosome outcrossed to different inbred strains for three generations. Analysis of 68 PCR-based markers in 13 affected individuals indicated that the mutant locus, named Wheels (Whl), resides in the subcentromeric portion of mouse chromosome 4. The …

A Kinetic Investigation Of The Mechanism Of Muscle Contraction With A Series Of Nucleotides, Wei Jiang

Theses and Dissertations in Biomedical Sciences

Muscle contraction is thought to be accomplished by sliding of myosin filaments along actin filaments. Although actomyosin naturally uses ATP as an energy source, it can also use other nucleoside triphosphates (NTP) as substrates. In this work, the dependence of rate and equilibrium constants of the various steps in the muscle contraction mechanism upon nucleotide structure was investigated to unravel the dependence of the mechanical properties of the muscle upon changes in different biochemical steps of the mechanism.

The experiments measuring the dissociation of actomyosin by NTP (or mant-NTP) shows that both the rate constant of actomyosin-S1 dissociation and second …

Enhancer Trap Technique: A Novel Tool For Identification And Developmental Characterization Of Genes Of Drosophila, Amit Singh

Biology Faculty Publications

The classical technique of mutational screen for identification of genes controlling early development has now approached saturation. A new era in genetic identification and developmental characterization of genes in Drosophila has commenced with the advent of the enhancer trap technique. This technique involves mobilization of a P-lacZ vector to diverse chromosomal locations in the fruit fly genome to bring it under the regulation of developmentally expressed genes or their enhancer elements. The technique offers a strikingly elegant method of gaining entry into fruit fly genes.

Restoration Of Circadian Behavior By Anterior Hypothalamic Heterografts, Patricia J. Sollars, Daniel P. Kimble, Gary E. Pickard

School of Veterinary and Biomedical Sciences: Faculty Publications

The suprachiasmatic nucleus (SCN) of the anterior hypothalamus (AH) is a circadian oscillator and an important component of the mammalian circadian system. To determine whether the SCN is the dominant circadian pacemaker responsible for generating a species-typical characteristic of circadian rhythms [i.e., period length (T)], neural transplantation was conducted using fetal AH donors of different species and SCN-lesioned (SCNx) hosts. The circadian behavior of each of the three donor species is clearly distinguishable by its species-typical T. The extent of SCN pacemaker autonomy was assessed by noting whether the period of the restored circadian rhythm following heterograft transplantation was characteristic …

Isolation Of A Mouse Cdna Encoding Mtj1, A New Murine Member Of The Dnaj Family Of Proteins, Shannon E. Brightman, Gregory L. Blatch, Bruce R. Zetter

Biology Faculty Publications

We report the isolation and sequencing of MTJ1, a 1792-bp cDNA from an M27 murine lung carcinoma cell line. The largest ORF within MTJ1 encodes a 63,869-Da protein, containing a 73-amino-acid (aa) sequence (the J domain) that is conserved in proteins of the DnaJ family of chaperonins. The J domain of MTJ1 is bracketed by potential transmembrane domains in a similar configuration to the J domain of the yeast DnaJ-like protein, SEC63. Polyclonal antibodies raised against deduced aa sequences within MTJ1 recognized antigens of 62, 42 and 41 kDa that were enriched in the nuclear and heavy microsome subcellular fractions …

An Improved Method For Chemical Devitellinization Of X-Gal Stained Drosophila Embryos, Amit Singh, Madhuri Kango-Singh, P. Sinha

Biology Faculty Publications

In Drosophila developmental biological studies, X-gal staining is commonly employed to study the spatio-temporal expression of the lacZ reporter gene in the transformed flies or their embryos. Study of the lacZ pattern in embryos often suffers from the lack of an efficient and high yieldirrg technique for devitellinization of X-gal stained embryos. Devitellinization techniques employed during antibody staining, in situ hybridization or embryonic cuticular preparations generally do not give satisfactory results when used for similar purpose in X-gal stained embryos. This results in the flaky appearance of the blue stain. We present here an improved chemical devitellinization technique which gives …

Increased Stem Cell Factor Release By Hemangioma-Derived Endothelial Cells, C. J. Meininger, Shannon Brightman, K. A. Kelly, B. R. Zetter

Biology Faculty Publications

Background: Capillary hemangiomas, the most common tumors in young children, consist of proliferating capillary vessels and endothelial cells. These tumors also contain large numbers of mast cells, compared with the normal surrounding skin or tissue. We have recently shown that stem cell factor (SCF), the gene product of the murine steel locus, can act as a chemoattractant for mast cells. In this study, we investigated whether SCF might be involved in the recruitment and maintenance of mast cells in hemangiomas. Experimental Design: Cultured endothelial cells derived from a murine hemangioma were compared with normal vascular endothelial cells for the ability …

Complementary Gradients In Expression And Binding Of Elf-1 And Mek4 In Development Of The Topographic Retinotectal Projection Map, Masaru Nakamoto

Biology Faculty Publications

Topographic maps with a defined spatial ordering of neuronal connections are a key feature of brain organization. Such maps are believed to develop in response to complementary position-specific labels in presynaptic and postsynaptic fields. However, the complementary labeling molecules are not known. In the well studied visual map of retinal axons projecting to the tectum, the labels are hypothesized to be in gradients, without needing large numbers of cell-specific molecules. We recently cloned ELF-1 as a ligand for Eph family receptors. Here, RNA hybridization shows matching expression gradients for ELF-1 in the tectum and its receptor Mek4 in the retina. …

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, …

Preparation Of A Site-Specific Lymphotoxin- Mutant To Be Used In Protein Characterization And Receptor Binding Studies, Derek Andrew Knight

Theses Digitization Project

No abstract provided.