Cell and Developmental Biology Commons^™

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 …

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 …

Assembly Of Α4Β2 Nicotinic Acetylcholine Receptors Assessed With Functional Fluorescently Labeled Subunits: Effects Of Localization, Trafficking, And Nicotine-Induced Upregulation In Clonal Mammalian Cells And In Cultured Midbrain Neurons, Raad Nashmi, Mary E. Dickinson, Sheri Mckinney, Mark Jareb, Cesar Labarca, Scott E. Fraser E. Fraser, Henry A. Lester

Biology Faculty Publications

Fura-2 recording of Ca²⁺ influx was used to show that incubation in 1 μM nicotine (2–6d) upregulates several pharmacological components of acetylcholine (ACh) responses in ventral midbrain cultures, including a MLA-resistant, DHβE-sensitive component that presumably corresponds to α4β2 receptors. To study changes in α4β2 receptor levels and assembly during this upregulation, we incorporated yellow and cyan fluorescent proteins (YFPs and CFPs) into the α4 or β2 M3–M4 intracellular loops, and these subunits were coexpressed in human embryonic kidney (HEK) 293T cells and cultured ventral midbrain neurons. The fluorescent …

Minus-End Capture Of Preformed Kinetochore Fibers Contributes To Spindle Morphogenesis, Alexey Khodjakov, Lily Copenagle, Michael B. Gordon, Duane A. Compton, Tarun M. Kapoor

Dartmouth Scholarship

Near-simultaneous three-dimensional fluorescence/differential interference contrast microscopy was used to follow the behavior of microtubules and chromosomes in living alpha-tubulin/GFP-expressing cells after inhibition of the mitotic kinesin Eg5 with monastrol. Kinetochore fibers (K-fibers) were frequently observed forming in association with chromosomes both during monastrol treatment and after monastrol removal. Surprisingly, these K-fibers were oriented away from, and not directly connected to, centrosomes and incorporated into the spindle by the sliding of their distal ends toward centrosomes via a NuMA-dependent mechanism. Similar preformed K-fibers were also observed during spindle formation in untreated cells. In addition, upon monastrol removal, centrosomes established a transient …

Asymmetry Of The Central Apparatus Defines The Location Of Active Microtubule Sliding In Chlamydomonas Flagella, Matthew J. Wargo, Elizabeth F. Smith

Dartmouth Scholarship

Regulation of ciliary and flagellar motility requires spatial control of dynein-driven microtubule sliding. However, the mechanism for regulating the location and symmetry of dynein activity is not understood. One hypothesis is that the asymmetrically organized central apparatus, through interactions with the radial spokes, transmits a signal to regulate dynein-driven microtubule sliding between subsets of doublet microtubules. Based on this model, we hypothesized that the orientation of the central apparatus defines positions of active microtubule sliding required to control bending in the axoneme. To test this, we induced microtubule sliding in axonemes isolated from wild-type and mutant Chlamydomonas cells, and then …

Exposure Of Nervous System Cells To Polychlorinated Biphenyls (Pcbs) Results In Significant Alterations In Levels Of Expression Of Neurotrophic Factors, Charles F. Ide, Jay C. Means, Trisha M. Basford, John Spitsbergen

Biological Sciences Faculty and Graduate Student Research

Toxic insult by PCBs results in learning and memory deficits in humans. Alterations in expression of neurotrophic factors and/or their receptors have been linked to changes in cognition. How PCBs affect cognition is not known. We propose that PCBs affect cognition by altering neurotrophic factor expression or effects. We exposed cultured C6 glial cells in medium containing PCB (Aroclor 1254 (10ppm)). Control cells were treated with DMSO or regular medium. Cells were incubated at 37o C for up to 72 hours. Medium samples were taken at 6hr, 24hr, 48hr, and 72hr intervals. Enzyme-linked immunosorbent assays (ELISA) were used to determine …

Manipulation Of Prenatal Hormones And Dietary Phytoestrogens During Adulthood Alter The Sexually Dimorphic Expression Of Visual Spatial Memory, Edwin D. Lephart, Trent D. Lund

Faculty Publications

In learning and memory tasks, requiring visual spatial memory (VSM), males exhibit higher performance levels compared to females (a difference attributed to sex steroid hormonal influences). Based upon the results from our companion investigation, this study examined the influence of prenatal sex steroid hormone manipulations on VSM in adulthood, as assessed in the radial arm maze. Additionally, the influence of dietary soy phytoestrogens (i.e., the presence of high or low estrogen-like compounds present in the animal's diet) on VSM was examined in combination with the prenatal hormonal manipulations. Results: Radial arm maze performance on a phytoestrogen-rich diet: 1) females treated …

Visual Spatial Memory Is Enhanced In Female Rats (But Inhibited In Males) By Dietary Soy Phytoestrogens, Edwin D. Lephart, Trent D. Lund, Timothy W. West, Lilyan Y. Tian, Lihong H. Bu, Daniel L. Simmons, Kenneth Setchell, Herman Adlercreutz

Faculty Publications

In learning and memory tasks, requiring visual spatial memory (VSM), males exhibit superior performance to females (a difference attributed to the hormonal influence of estrogen). This study examined the influence of phytoestrogens (estrogen-like plant compounds) on VSM, utilizing radial arm-maze methods to examine varying aspects of memory. Additionally, brain phytoestrogen, calbindin (CALB), and cyclooxygenase-2 (COX-2) levels were determined. Results: Female rats receiving lifelong exposure to a high-phytoestrogen containing diet (Phyto-600) acquired the maze faster than females fed a phytoestrogen-free diet (Phyto-free); in males the opposite diet effect was identified. In a separate experiment, at 80 days-of-age, animals fed the Phyto-600 …

Interactions Of Igf-Ii And Cathepsin D In Mcf-7 Breast Cancer Cells, Jesika S. Faridi

Loma Linda University Electronic Theses, Dissertations & Projects

A primary role of the IGF-II/M6P receptor is to target lysosomal enzymes from the golgi to the lysosomes. This receptor has distinct binding sites for IGF-II and M6P, however, reciprocal interactions between these ligands have been observed (Kiess et al. 1989, 1990). Since IGF-II modulates the routing of cathepsin D in MCF-7 cells by blocking the intracellular binding of cathepsin D to the IGF-II/M6P receptor (De León et al. 1996), we hypothesized that expressing a mutant form of IGF-II that does not bind the IGF-II/M6P receptor will not interfere with lysosomal enzyme trafficking.

In our present study, we report the …

An Ultrastructural Survey Of The Nuchal Glands Of Rhabdophis Tigrinus (Serpentes: Colubridae), Kathleen A. Roberts

Biological Sciences Theses & Dissertations

Rhabdophis tigrinus, an Asian natricine snake, possesses a series of integumentary glands located in the nuchal skin. Previous studies indicated that the nuchal glands are of mesodermal origin, contain cardiac steroids analogous to bufotoxins found in the parotoid glands of toads, which are included in the snake's diet. The nuchal glands are defensive in function. Preliminary histological studies revealed differences in quality of the gland tissue compared to previous studies, and difference in fixation techniques were suspected. Seven specimens of R. tigrinus were fixed by perfusion using a conservative fixation process standard for electron microscopy. The nuchal gland series …

Leukemia Inhibitory Factor (Lif): Murine Preimplantation Embryo Development, Implantation Rates, And Skeletal Development, Michael Hayes Mitchell

Theses and Dissertations in Biomedical Sciences

Leukemia inhibitory factor (LIF) is a pleiotropic cytokine which demonstrates perplexing physiological effects. It has been demonstrated that LIF is essential for implantation in mice. Little is known relating to the manner by which LIF effects pre-implantation and post-implantation development. The objectives of this project were to determine the effects LIF on pre-implantation development, to determine the effects that it may have on implantation rates, successful pregnancy rates, and resorption rates, and to determine the effects that LIF has on the skeletal development of mice. For the embryo transfer experiments, embryos were exposed to test compounds in the transfer medium …

The Role Of Glycoconjugates In Mediating Human Fertilization And Induction Of Fetomaternal Tolerance, Manish S. Patankar

Theses and Dissertations in Biomedical Sciences

Using the hemizona assay (HZA), a in vitro sperm-egg binding assay, we show that specific glycoconjugates known to inhibit immune cell interactions mediated by the selectins, potently block human sperm-egg binding. The selectin ligand sialyl Lewis^x inhibits sperm binding in the HZA by 60% at a concentration of 1 mg/ml. Our data indicates that glycodelin-A, a endometrial glycoprotein known to block sperm-egg binding in the HZA at low concentrations expresses unusual fucosylated lacdiNAc type glycans. The fucosylated lacdiNAc type sugars have been previously shown to be 15-20 fold more potent ligands of E-selectin. Glycodelin-S a seminal plasma glycoform of …

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 …

Cellular Mechanisms Underlying Myogenic Reactivity In Isolated Arterioles, Hui Zou

Theses and Dissertations in Biomedical Sciences

The myogenic reactivity provides one of the principal mechanisms for blood flow autoregulation. The aims of the performed studies described in this dissertation were to test the role of [Ca²⁺]_i and MLC phosphorylation in arteriolar myogenic reactivity and further examine the source(s) of activator Ca²⁺ required to initiate and maintain myogenic vasoconstriction. In addition, the possible underlying mechanism of contractile protein expression was also addressed.

These studies used male Sprague Dawley rats of 200 ~ 350 grams body weight. Experiments were carried out using rat cremaster first order arterioles and mesenteric vessels. Gel electrophoresis and immunoblotting …

Laminin Potentiates Differentiation Of Pcc4uva Embryonal Carcinoma Into Neurons, T. M. Sweeney, Roy C. Ogle, C. D. Little

School of Medical Diagnostics & Translational Sciences Publications

The embryonal carcinoma PCC4uva differentiates into neurons in response to treatment with retinoic acid and dbcAMP. We used this in vitro model system to study the effects of laminin on early neural differentiation. Laminin substrata markedly potentiate neural differentiation of retinoic acid and dbcAMP-treated cultures. Only laminin induced more rapid neural cell body clustering, neurite growth and neurite fasciculation as compared to type IV collagen, type I collagen, and fibronectin substrata. Exogenous laminin substrata promoted greater cell attachment, cellular spreading and growth to confluence than type IV collagen, type I collagen, fibronectin and glass substrata. Laminin-induced effects were inhibited by …

Embryonic Chicken Fibroblast Collagen Binding Proteins: Distribution, Role In Substratum Adhesion, And Relationship To Integrins, Roy C. Ogle, A. Jeannette Potts, Marchall Yacoe, Charles D. Little

School of Medical Diagnostics & Translational Sciences Publications

Collagen binding proteins (CBP) are hydrophobic, cell surface polypeptides, isolated by collagen affinity chromatography. Antibodies to CBPs inhibit the attachment of embryonic chicken heart fibroblasts to native type I collagen fibrils in a dose-dependent manner. The CBP antibodies also induce rounding and detachment of cells adherent to a planar substratum. This process of antibody-mediated substratum detachment resulted in a clustering of CBP and cell-associated extracellular matrix at the cell surface, and the rearrangement of filamentous actin. Other functional studies showed that cells grown within a three-dimensional gel of type I collagen cannot be immunostained at the cell surface with CBP …

Factors Affecting Thyrotropin Secretion In Superfused Rat Anterior Pituitary Cells, Frederick Scott Bartlow

Dissertations and Theses

The use of static in vitro pituitary cultures has been indispensible for examining the regulation of anterior pituitary hormone secretion. While the static cultures have shown the effects of various chemical stimulators of hormone secretion, the dynamics of such secretion has not been assessed before in vitro. The in vitro superfusion method, which partially stimulates in vivo physiologic conditions, allows for the observation of the dynamics of pituitary hormone secretion.

Sodium Flux Ratio In Voltage-Clamped Squid Giant Axons, David D. Busath, Ted Begenisich

Faculty Publications

The sodium flux ratio across the axolemma of internally perfused, voltage-clamped giant axons of Loligo pealei has been measured at various membrane potentials. The flux ratio exponent obtained from these measurements was about unity and independent of membrane voltage over the 50 mV range from about -20 to +30 mV. These results, combined with previous measurements of ion permeation through sodium channels, show that the sodium channel behaves like a multi-ion pore with two ion binding sites that are rarely simultaneously occupied by sodium.

Two-, Three-, And Four-Particle Spatial Correlations Among Tertiary Cosmic Ray Muons, David D. Busath, R O. Stevenson

Faculty Publications

Showers of muons which result from interactions of primary cosmic rays high in the atmosphere appear deep underground as bundles of nearly parallel muons. For threshold energies of 1 TeV and arrival zenith angles of 45 degrees, those events having a small mean number of muons are described by a power law distribution in shower size having an exponent of ≅ -3.7 and a radial density distribution that decreases with distance R from the shower axis a little less rapidly than exp(-R/5.4 m). Comparison of new data on four muon spatial correlations in terms of the same phenomenology with data …