DOCSLIB.ORG

Napierindia Gnrhr As a Molecular Target For

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Napierindia Gnrhr As a Molecular Target For Gonadotropin-releasing Hormone Receptor As A Molecular Target for Contraception in Feral Cats by India Desiree Napier A dissertation submitted to the Graduate Faculty of Auburn University in partial fulfillment of the requirements for the Degree of Doctor of Philosophy Auburn, Alabama December 12, 2015 Keywords: contraception, DNA vaccines, feline, gonadotropin-releasing hormone receptor, superparamagnetic iron oxide nanoparticles, ubiquitin Copyright 2015 by India Desiree Napier Approved by Tatiana I Samoylova, Chair, Research Professor of Scott-Ritchey Research Center Frank F Bartol, Professor of Anatomy, Physiology, and Pharmacology Timothy D Braden, Associate Professor of Anatomy, Physiology, and Pharmacology Douglas R Martin, Professor of Anatomy, Physiology, and Pharmacology Robyn R Wilborn, Associate Professor of Clinical Sciences Abstract The global overpopulation of feral cats generates concern regarding their welfare, negative impact on public health, and adverse effects on the environment. In the U.S. and other Westernized countries, the most common method for managing feral cat populations is impoundment in animal shelters, where they undergo surgical sterilization procedures as part of a shelter adoption or trap-neuter-release programs. However, such programs are labor-intensive, time-consuming, and costly. Further, several million, un-adopted cats are euthanized each year. Thus, there is a need for a permanent, nonsurgical, and low-cost method for controlling feral cat populations. Immunocontraception has the potential to provide the practical approach needed to manage stray cat populations. Gonadotropin-releasing hormone receptor (GnRHR) is an attractive target for immunocontraceptive vaccine development because it is highly expressed by anterior pituitary gonadotropic cells, important components of the hypothalamic-pituitary-gonadal axis that regulates normal mammalian reproduction. Therefore, induction of immune responses against pituitary gonadotropic cells by targeting GnRHR may impair normal reproductive function. The goal of this proof-of- concept work was to use a murine model to investigate the potentials of GnRHR as a target for immunocontraception. DNA plasmids were selected for immunocontraceptive vaccine construction because they are cost-effective, safe, and can elicit long-term ii cytotoxic and humoral immune responses in animals. In order to overcome murine self- tolerance to the GnRHR, the nucleic acid sequence of feline (f)GnRHR was cloned into a plasmid with expectations of the murine immune system to elicit a stronger response to the heterologous antigen (fGnRHR) than to the homologous protein (mouse GnRHR). As an additional strategy for enhancing the potency of the vaccine against the self- antigen GnRHR, the DNA sequence for fGnRHR fused to ubiquitin (Ub) was incorporated into a plasmid in order to enhance presentation of GnRHR epitopes from Ub-fGnRHR to circulating CD4+ T cells. This strategy may efficiently activate these T cells, producing GnRHR-specific antibodies that ultimately target GnRHR-expressing pituitary gonadotropic cells. After DNA plasmid construction, adult male mice were vaccinated once with DNA encoding fGnRHR or Ub-fGnRHR. Vaccines were administered via intramuscular (IM) needle injection; IM injection with DNA-loaded, superparamagnetic iron oxide nanoparticles followed by magnetofection; or intradermal (ID) needle injection. Mice vaccinated with DNA encoding fGnRHR or Ub-fGnRHR via IM injection or magnetofection had normal serum testosterone levels at necropsy (or 12 weeks post-immunization) compared to pretreatment values. However, mice vaccinated with plasmid encoding Ub-fGnRHR via ID needle injection had significantly low testosterone concentrations 12 weeks post-immunization. Moreover, anti-GnRHR antibodies were detected in sera from mice intradermally vaccinated with DNA encoding Ub-fGnRHR. Also, PCR analysis indicated lower GnRHR mRNA expression in these animals. In summary, ID vaccination of mice with DNA encoding Ub-fGnRHR, a heterologous and ubiquitinated protein, abated serum testosterone concentrations and pituitary expression of GnRHR mRNA, as well as induced anti-GnRHR antibody iii production in treated animals. The outcome of these parameters indicate induction of an immunological response to overcome self-tolerance to the endogenous GnRHR, resulting in impairment to anterior pituitary gonadotropes that affected testosterone production. Based on these findings, the Ub-fGnRHR DNA construct warrants further investigation, as such a vaccine profile would be beneficial as an immunocontraceptive for controlling the overpopulation of feral cats worldwide. iv Acknowledgments “It's not who I am underneath, but what I do that defines me.” Bruce Wayne, Batman Begins First, I thank my advisor, Dr. Samoylova, for providing me the opportunity to conduct my dissertation research in her laboratory. I also thank the other members of my dissertation committee for their invaluable support during my tenure as a doctoral student. I would like to express my sincerest gratitude to my mentor, Dr. Frank Bartol, for inspiring me to be a leader in the Auburn University (AU) graduate student community and for having full confidence in my potential to be a biomedical researcher who can change the world. Dr. Timothy Braden has also been a great ally during my graduate studies at AU and taught me a valuable lesson: one must always make time for his/her family (even while enrolled in a graduate program). I also thank Drs. Douglas Martin and Robyn Wilborn for their positive attitudes, insightful comments, and encouragement throughout the dissertation process. Finally, I am thankful for the support of my family (especially my husband, Dr. James Morris-King) who encouraged me to pursue a doctoral degree and always reminded me that I am capable of achieving my educational, career, and personal goals. v Table of Contents Abstract ..........................................................................................................................ii Acknowledgments ......................................................................................................... v List of Tables ................................................................................................................ xii List of Figures .............................................................................................................. xiii List of Abbreviations .....................................................................................................xv Chapter 1: Literature Review ....................................................................................... 1 Section 1.1: The Overpopulation of Feral Cats and Its Implications ................ 1 Section 1.2: Approaches to Controlling the Overpopulation of Feral Cats ....... 6 Section 1.2.1: Shelters .......................................................................... 6 Section 1.2.2: Trap-neuter-release Programs ......................................... 7 Section 1.2.3: Surgical Sterilization ....................................................... 9 Section 1.2.4: Non-surgical Sterilization .............................................. 11 Section 1.2.4.1: Chemical Sterilants ......................................... 12 Section 1.2.4.2: Hormonal Downregulation via Sex Steroids .... 13 Section 1.2.4.3: Immunization against Zona Pellucida ............. 14 Section 1.2.4.4: Gonadotropin-releasing Hormone-based Agents ....... 16 Section 1.3: Gonadotropin-releasing Hormone (GnRH) ................................... 21 Section 1.3.1: GnRH Structure ........................................................... 21 Section 1.3:2: Hypothalamic-pituitary-gonadal Axis ............................. 23 Section 1.4: Gonadotropin-releasing Hormone Receptor (GnRHR) ............... 24 Section 1.4.1: Properties of GnRH binding to GnRHR .......................... 26 Section 1.4.2: GnRHR Signaling .......................................................... 27 Section 1.4.3: GnRHR Trafficking ........................................................ 29 Section 1.5: DNA-based Vaccines ................................................................ 33 Section 1.6: Superparamagnetic Iron Oxide Nanoparticles ........................... 37 Chapter 2: Construction and in vitro Characterization of DNA-Based Vaccines Directed against Gonadotropin-releasing Hormone Receptor ........................ 47 Section 2.1: Introduction ................................................................................ 47 Section 2.2: Materials and Methods .............................................................. 51 Section 2.2.1: Animal Samples ............................................................ 51 Section 2.2.2: RNA Isolation .................................................................. 52 Section 2.2.3: Restriction Digestion, Purification, and Ligation ............ 53 Section 2.2.4: Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and PCR ................................................................... 54 Section 2.2.5: Cloning Strategy .......................................................... 56 Section 2.2.5.1: Construction of Template Plasmid Encoding Feline (f)GnRHR ............................................................. 56 Section 2.2.5.2: Construction of Template Plasmid Encoding Ubiquitin (Ub) .................................................................. 56 Section 2.2.5.2: Construction of Template Plasmid Encoding
Load more

Recommended publications
  • Te2, Part Iii
    TERMINOLOGIA EMBRYOLOGICA Second Edition International Embryological Terminology FIPAT The Federative International Programme for Anatomical Terminology A programme of the International Federation of Associations of Anatomists (IFAA) TE2, PART III Contents Caput V: Organogenesis Chapter 5: Organogenesis (continued) Systema respiratorium Respiratory system Systema urinarium Urinary system Systemata genitalia Genital systems Coeloma Coelom Glandulae endocrinae Endocrine glands Systema cardiovasculare Cardiovascular system Systema lymphoideum Lymphoid system Bibliographic Reference Citation: FIPAT. Terminologia Embryologica. 2nd ed. FIPAT.library.dal.ca. Federative International Programme for Anatomical Terminology, February 2017 Published pending approval by the General Assembly at the next Congress of IFAA (2019) Creative Commons License: The publication of Terminologia Embryologica is under a Creative Commons Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0) license The individual terms in this terminology are within the public domain. Statements about terms being part of this international standard terminology should use the above bibliographic reference to cite this terminology. The unaltered PDF files of this terminology may be freely copied and distributed by users. IFAA member societies are authorized to publish translations of this terminology. Authors of other works that might be considered derivative should write to the Chair of FIPAT for permission to publish a derivative work. Caput V: ORGANOGENESIS Chapter 5: ORGANOGENESIS
    [Show full text]
  • Somatostatin in the Periventricular Nucleus of the Female Rat: Age Specific Effects of Estrogen and Onset of Reproductive Aging
    4 Somatostatin in the Periventricular Nucleus of the Female Rat: Age Specific Effects of Estrogen and Onset of Reproductive Aging Eline M. Van der Beek, Harmke H. Van Vugt, Annelieke N. Schepens-Franke and Bert J.M. Van de Heijning Human and Animal Physiology Group, Dept. Animal Sciences, Wageningen University & Research Centre The Netherlands 1. Introduction The functioning of the growth hormone (GH) and reproductive axis is known to be closely related: both GH overexpression and GH-deficiency are associated with dramatic decreases in fertility (Bartke, 1999; Bartke et al, 1999; 2002; Naar et al, 1991). Also, aging results in significant changes in functionality of both axes within a similar time frame. In the rat, GH secretion patterns are clearly sexually dimorphic (Clark et al, 1987; Eden et al, 1979; Gatford et al, 1998). This has been suggested to result mainly from differences in somatostatin (SOM) release patterns from the median eminence (ME) (Gillies, 1997; Muller et al, 1999; Tannenbaum et al, 1990). SOM is synthesized in the periventricular nucleus of the hypothalamus (PeVN) and controls in concert with GH-releasing hormone (GHRH) the GH release from the pituitary (Gillies, 1987; Tannenbaum et al, 1990; Terry and Martin, 1981; Zeitler et al, 1991). An altered GH status is reflected in changes in the hypothalamic SOM system. For instance, the number of SOM cells (Sasaki et al, 1997) and pre-pro SOM mRNA levels (Hurley and Phelps, 1992) in the PeVN were elevated in animals overexpressing GH. Several observations suggest that SOM may also affect reproductive function directly at the level of the hypothalamus.
    [Show full text]
  • Nomina Histologica Veterinaria, First Edition
    NOMINA HISTOLOGICA VETERINARIA Submitted by the International Committee on Veterinary Histological Nomenclature (ICVHN) to the World Association of Veterinary Anatomists Published on the website of the World Association of Veterinary Anatomists www.wava-amav.org 2017 CONTENTS Introduction i Principles of term construction in N.H.V. iii Cytologia – Cytology 1 Textus epithelialis – Epithelial tissue 10 Textus connectivus – Connective tissue 13 Sanguis et Lympha – Blood and Lymph 17 Textus muscularis – Muscle tissue 19 Textus nervosus – Nerve tissue 20 Splanchnologia – Viscera 23 Systema digestorium – Digestive system 24 Systema respiratorium – Respiratory system 32 Systema urinarium – Urinary system 35 Organa genitalia masculina – Male genital system 38 Organa genitalia feminina – Female genital system 42 Systema endocrinum – Endocrine system 45 Systema cardiovasculare et lymphaticum [Angiologia] – Cardiovascular and lymphatic system 47 Systema nervosum – Nervous system 52 Receptores sensorii et Organa sensuum – Sensory receptors and Sense organs 58 Integumentum – Integument 64 INTRODUCTION The preparations leading to the publication of the present first edition of the Nomina Histologica Veterinaria has a long history spanning more than 50 years. Under the auspices of the World Association of Veterinary Anatomists (W.A.V.A.), the International Committee on Veterinary Anatomical Nomenclature (I.C.V.A.N.) appointed in Giessen, 1965, a Subcommittee on Histology and Embryology which started a working relation with the Subcommittee on Histology of the former International Anatomical Nomenclature Committee. In Mexico City, 1971, this Subcommittee presented a document entitled Nomina Histologica Veterinaria: A Working Draft as a basis for the continued work of the newly-appointed Subcommittee on Histological Nomenclature. This resulted in the editing of the Nomina Histologica Veterinaria: A Working Draft II (Toulouse, 1974), followed by preparations for publication of a Nomina Histologica Veterinaria.
    [Show full text]
  • General and Comparative Endocrinology 273 (2019) 209–217
    General and Comparative Endocrinology 273 (2019) 209–217 Contents lists available at ScienceDirect General and Comparative Endocrinology journal homepage: www.elsevier.com/locate/ygcen Effects of GnRH and the dual regulatory actions of GnIH in the pituitary explants and brain slices of Astyanax altiparanae males T Giovana Souza Brancoa,b,1, Aline Gomes Melob,1, Juliana M.B. Riccib, Melanie Digmayerb, ⁎ Lázaro W.O. de Jesusc, Hamid R. Habibid, Rafael Henrique Nóbregab, a Aquaculture Center of São Paulo State University (CAUNESP), São Paulo State University (UNESP), Jaboticabal Campus, Jaboticabal, Brazil b Reproductive and Molecular Biology Group, Department of Morphology, Institute of Biosciences, São Paulo State University (UNESP), Botucatu Campus, Botucatu, Brazil c Institute of Biological Sciences and Health, Federal University of Alagoas – A. C., Simões Campus, Maceió, Brazil d Department of Biological Sciences, University of Calgary, Calgary, Canada ARTICLE INFO ABSTRACT Keywords: The pituitary gonadotropins, Fsh (follicle-stimulating hormone) and Lh (luteinizing hormone), regulate testi- Gonadotropin-releasing hormone cular development and functions in all vertebrates. At the pituitary, different signaling systems regulate the Gonadotropin-inhibitory hormone synthesis and secretion of the gonadotropins, such as the hypothalamic neuropeptides GnRH (gonadotropin- Follicle-stimulating hormone releasing hormone) and GnIH (gonadotropin-inhibitory hormone). While GnRH exerts stimulatory roles, the Luteinizing hormone actions of GnIH remain controversial for many teleost species. Therefore, the aim of this study was to evaluate Lambari-do-rabo-amarelo the in vitro effects of chicken GnRH2 (cGnRH2) and zebrafish GnIH-3 (zGnIH-3) on the male gonadotropin and Astyanax altiparanae GnRH system expression using pituitary explants and brain slices from a neotropical species with economical and ecological relevance, Astyanax altiparanae.
    [Show full text]
  • BGD B Lecture Notes Docx
    BGD B Lecture notes Lecture 1: GIT Development Mark Hill Trilaminar contributions • Overview: o A simple tube is converted into a complex muscular, glandular and duct network that is associated with many organs • Contributions: o Endoderm – epithelium of the tract, glands, organs such as the liver/pancreas/lungs o Mesoderm (splanchnic) – muscular wall, connective tissue o Ectoderm (neural crest – muscular wall neural plexus Gastrulation • Process of cell migration from the epiblast through the primitive streak o Primitive streak forms on the bilaminar disk o Primitive streak contains the primitive groove, the primitive pit and the primitive node o Primitive streak defines the body axis, the rostral caudal ends, and left and right sides Thus forms the trilaminar embryo – ectoderm, mesoderm, endoderm • Germ cell layers: o ectoderm – forms the nervous system and the epidermis epithelia 2 main parts • midline neural plate – columnar epithelium • lateral surface ectoderm – cuboidal, containing sensory placodes and skin/hair/glands/enamel/anterior pituitary epidermis o mesoderm – forms the muscle, skeleton, and connective tissue cells migrate second migrate laterally, caudally, rostrally until week 4 o endoderm – forms the gastrointestinal tract epithelia, the respiratory tract and the endocrine system cells migrate first and overtake the hypoblast layer line the primary yolk sac to form the secondary yolk sac • Membranes: o Rostrocaudal axis Ectoderm and endoderm form ends of the gut tube, no mesoderm At each end, form the buccopharyngeal
    [Show full text]
  • Downloaded from Bioscientifica.Com at 09/29/2021 04:18:11AM Via Free Access
    REPRODUCTIONRESEARCH PROOF ONLY Gonadotropin characterization, localization and expression in the European hake (Merluccius merluccius) Michela Candelma1, Romain Fontaine2, Sabrina Colella3, Alberto Santojanni3, Finn-Arne Weltzien2 and Oliana Carnevali1 1Department of Life and Environmental Sciences, Università Politecnica delle Marche, Ancona, Italy, 2Department of Basic Sciences and Aquatic Medicine, Norwegian University of Life Sciences, Oslo, Norway, and 3CNR-National Research Council of Italy, ISMAR-Marine Sciences Institute, Ancona, Italy Correspondence should be addressed to O Carnevali; Email: [email protected] Abstract In vertebrates, the regulation of gametogenesis is under the control of gonadotropins (Gth), follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh). In fish, the physiological role of Gths is not fully understood, especially in species with asynchronous ovarian development. To elucidate the role of Gths in species with asynchronous ovary, we studied European hake (Merluccius merluccius) during the reproductive season. For this aim, we first cloned and sequenced both hormones. Then, we characterized their amino acid sequence and performed phylogenetic analyses to verify the relationship to their orthologues in other species. In addition, the quantification of gene expression during their natural reproductive season was analyzed in wild-caught female hake. Our results revealed that fshb peaked during the vitellogenic phase, remaining high until spawning. This is in contrast to the situation in species with synchronous ovary. lhb, on the other hand, peaked during maturation as it is also common in species with synchronous ovarian development. Finally, combining double-labeling fluorescent in situ hybridization (FISH) for Gth mRNAs with immunofluorescence for Lh protein, we evidenced the specific expression of fshb and lhb in different cells within the proximal pars distalis (PPD) of the pituitary.
    [Show full text]
  • |||GET||| Pancreatic Islet Biology 1St Edition
    PANCREATIC ISLET BIOLOGY 1ST EDITION DOWNLOAD FREE Anandwardhan A Hardikar | 9783319453057 | | | | | The Evolution of Pancreatic Islets Advanced search. The field of regenerative medicine is rapidly evolving and offers great hope for the nearest future. Easily read eBooks on Pancreatic Islet Biology 1st edition phones, computers, or any eBook readers, including Kindle. Help Learn to edit Community portal Recent changes Upload file. Pancreatic Islet Biologypart of the Stem Cell Biology and Regenerative Medicine series, is essential reading for researchers and clinicians in stem cells or endocrinology, especially those focusing on diabetes. Because the beta cells in the pancreatic islets Pancreatic Islet Biology 1st edition selectively destroyed by an autoimmune process in type 1 diabetesclinicians and researchers are actively pursuing islet transplantation as a means of restoring physiological beta cell function, which would offer an alternative to a complete pancreas transplant or artificial pancreas. Strategies to improve islet yield from chronic pancreatitis pancreases intended for islet auto-transplantation 6. About this book This comprehensive volume discusses in vitro laboratory development of insulin-producing cells. Junghyo Jo, Deborah A. Show all. Comparative Analysis of Islet Development. Leibson A. However, type 1 diabetes is the result of the autoimmune destruction of beta cells in the pancreas. Islets can influence each other through paracrine and autocrine communication, and beta cells are coupled electrically to six to seven other beta cells but not to other cell types. Pancreatic Islet Biologypart of the Stem Cell Biology and Regenerative Medicine Pancreatic Islet Biology 1st edition, is essential reading for researchers and clinicians in stem cells or endocrinology, especially those focusing on diabetes.
    [Show full text]
  • ACTH in Tetrahymena Pyriformis, 76 Adenylate Cyclase Inhibition By
    INDEX ACTH Aromatase in tetrahymena pyriformis, 76 activity, 469 Adenylate Cyclase Armatase activity inhibition by somatostatin, granulosa cells, 317 377 OS induced, 253 Administered LHRHA with testosterone enanthate, 598 Bacteria Adrenal androgens binding of TSH, 77 in prostatic cancer, 540 Blocking agents Agonist or antagonist of gonadotropin receptor, 222 binding to receptor, 242 Ca2+ Androgen insensitive cells interaction of receptor, 267 exposed to low androgen, 542 on intracellular events, 267 Androgen receptor blockers Calmodulin inhibitors cyproterone acetate, flutamide, block GnRH, 160 558 cAMP Angiotensin 11 activation of phospho­ PRL release, 359, 361 diesterase, 413 Anterior pituitary function cumulus-oocyte complex, 328 neuromodulators, 48 regulation of oocyte Antral f1uid maturation, 326 FSH, LH and prolactin, 320 cAMP accumulation steroid hormone concentrations, LH-induced, 333 319 oocyte maturation, 332 AP cells cAMP and 20a-OH-P information on membrane compart­ response to OS or FSH, 257 ments, 170 cAMP binding activity Arachidonic acid effects of FSH and LHRH, 473 action of LHRH, 6 cAMP levels Arachidonic acid (AA) pregnenolone production, 429 prostaglandins (PG), 494 cAMP phosphodieste~se Arachidonic acid conversion stimulated by Ca ,calmodulin, preovulatory follicles, 348 414 Arachidonic acid metabolites Casein secretion of LH, 6 encoded by complex split genes, Arachodonic metabolites 389 in ovulation, 344 gene structure, 389 611 612 INDEX 6 Casein mRNAs D-Arg analog sequences, 389 of LH-RH, 522 a-casein Deglycosylated
    [Show full text]
  • Pituitary P62 Deficiency Leads to Female Infertility by Impairing Luteinizing Hormone Production
    www.nature.com/emm ARTICLE OPEN Pituitary P62 deficiency leads to female infertility by impairing luteinizing hormone production 1,2,3,6 1,6 1 1 1 4 4 5 1 2,3 Xing Li , Ling Zhou✉ , Guiliang Peng✉ , Mingyu Liao , Linlin Zhang , Hua Hu , Ling Long , Xuefeng Tang , Hua Qu , Jiaqing Shao , Hongting Zheng1 and Min Long 1 © The Author(s) 2021 P62 is a protein adaptor for various metabolic processes. Mice that lack p62 develop adult-onset obesity. However, investigations on p62 in reproductive dysfunction are rare. In the present study, we explored the effect of p62 on the reproductive system. P62 deficiency-induced reproductive dysfunction occurred at a young age (8 week old). Young systemic p62 knockout (p62-/-) and pituitary-specific p62 knockout (p62flox/flox αGSUcre) mice both presented a normal metabolic state, whereas they displayed infertility phenotypes (attenuated breeding success rates, impaired folliculogenesis and ovulation, etc.) with decreased luteinizing hormone (LH) expression and production. Consistently, in an infertility model of polycystic ovary syndrome (PCOS), pituitary p62 mRNA was positively correlated with LH levels. Mechanistically, p62-/- pituitary RNA sequencing showed a significant downregulation of the mitochondrial oxidative phosphorylation (OXPHOS) pathway. In vitro experiments using the pituitary gonadotroph cell line LβT2 and siRNA/shRNA/plasmid confirmed that p62 modulated LH synthesis and secretion via mitochondrial OXPHOS function, especially Ndufa2, a component molecule of mitochondrial complex I, as verified by Seahorse and rescue tests. After screening OXPHOS markers, Ndufa2 was found to positively regulate LH production in LβT2 cells. Furthermore, the gonadotropin-releasing hormone (GnRH)-stimulating test in p62flox/flox αGSUcre mice and LβT2 cells illustrated that p62 is a modulator of the GnRH-LH axis, which is dependent on intracellular calcium and ATP.
    [Show full text]
  • Advance Publication by J-STAGE Journal of Reproduction and Development
    Advance Publication by J-STAGE Journal of Reproduction and Development Accepted for publication: February 18, 2020 Advanced Epub: February 28, 2020 Gpr120 mRNA expression in gonadotropes in the mouse pituitary gland is regulated by free fatty acids Chikaya Deura, Yusuke Kimura, Takumi Nonoyama and Ryutaro Moriyama Laboratory of Environmental Physiology, Department of Life Science, School of Science and Engineering, Kindai University, Higashiosaka 577-8502, Japan Running head: FFA regulates Gpr120 mRNA Corresponding Author: Ryutaro Moriyama, PhD Laboratory of Environmental Physiology, Department of Life Science, School of Science and Engineering, Kindai University, Higashiosaka 577-8502, Japan Phone: +81-6-4307-3435 e-mail: [email protected] Abstract GPR120 is a long-chain fatty acid (LCFA) receptor that is specifically expressed in gonadotropes in the anterior pituitary gland in mice. The aim of this study was to investigate whether GPR120 is activated by free fatty acids in the pituitary of mice and mouse immortalized gonadotrope LβT2 cells. First, the effects of palmitate on GPR120, gonadotropic hormone b-subunits, and GnRH-receptor expression in gonadotropes were investigated in vitro. We observed palmitate-induced an increase in Gpr120 mRNA expression and a decrease in Fshb expression in LβT2 cells. Furthermore, palmitate exposure caused the phosphorylation of ERK1/2 in LβT2 cells, but no significant changes were observed in the expression levels of Lhb and Gnrh-r mRNA and number of GPR120 immunoreactive cells. Next, diurnal variation in Gpr120 mRNA expression in the male mouse pituitary gland was investigated using ad libitum and night-time restricted feeding (active phase from 1900 to 0700 h) treatments.
    [Show full text]
  • Gonadotropin-Releasing Hormone Signaling in Behavioral Plasticity Hans a Hofmann
    Gonadotropin-releasing hormone signaling in behavioral plasticity Hans A Hofmann Sex and reproduction sculpt brain and behavior throughout life Studies in songbirds and cichlid fishes have highlighted and evolution. In vertebrates, gonadotropin-releasing hormone the surprising plasticity exhibited by the GnRH system in (GnRH) is essential to these processes. Recent advances have adult animals in response to seasonal and social cues. uncovered novel regulatory mechanisms in GnRH signaling, such as the initiation of sexual maturation by kisspeptins. Yet GnRH genes and GnRH receptor genes despite our increasing molecular knowledge, we know very GnRH is a decapeptide (see Box 1), the structure of which little about environmental influences on GnRH signaling and has been highly conserved throughout evolution, espe- reproductive behavior. Alternative model systems have been cially at the N- and C- termini, which are involved in crucial for understanding the plasticity of GnRH effects within receptor binding and activation [2,3]. GnRH genes encode an organismal context. For instance, GnRH signaling is under a preprohormone precursor (see glossary) that needs to be the control of seasonal cues in songbirds, whereas social cleaved and correctly folded before the peptide can signals regulate GnRH in cichlid fishes, with crucial become physiologically active [4]. Although peptide consequences for reproduction and behavior. Analyzing maturation has mostly been studied in cell lines, it is cellular signaling cascades within an organismic context is assumed that it is highly conserved across vertebrates. essential for an integrative understanding of GnRH function. Similar to other neuropeptide systems [5], a signal pep- tidase first removes the N-terminal sequence. The result- Addresses ing prohormone is then further processed at specific sites Harvard University, Bauer Center for Genomics Research, 7 Divinity Avenue, Cambridge, MA 02138, USA by prohormone convertases in conjunction with specific regulators, such as proSAAS and 7B2.
    [Show full text]
  • Evidence of Regeneration of Testicular and Epididymal Tissue Structure and Function Following Withdrawal from Sub-Chronic Khat E
    Journal of Applied Life Sciences International 23(9): 10-23, 2020; Article no.JALSI.61095 ISSN: 2394-1103 Evidence of Regeneration of Testicular and Epididymal Tissue Structure and Function Following Withdrawal from Sub-chronic Khat Exposure: Studies in the Rabbit Animal Model Albert W. Nyongesa1*, Esther M. Maluki2 and Jemimah A. Simbauni2 1Department of Veterinary Anatomy and Physiology, University of Nairobi, P.O.Box 30197-00100, Nairobi, Kenya. 2Department of Zoological Sciences, Kenyatta University, P.O.Box 43844-00100, Nairobi, Kenya. Authors’ contributions This work was carried out in collaboration among all authors. Author AWN conceived the idea and designed the experiment, interpreted hormonal and histological data and results reporting and provided critical analysis to paper writing and formatting. Author EMM designed the experiment, contributed reagents and materials, performed the experiments. Author JAS contributed to the experimental design, performed critical analysis of data and paper writing. All authors read and approved the final manuscript. Article Information DOI: 10.9734/JALSI/2020/v23i930183 Editor(s): (1) Dr. Muhammad Kasib Khan, University of Agriculture Faisalabad, Pakistan. Reviewers: (1) Kasula Siddartha, Krishna Institute of Medical Sciences, India. (2) Mehmet İrfan Karadede, İzmir Katip Çelebi University, Turkey. (3) Thyagaraju Kedam, Sri Venkateswara University, India. Complete Peer review History: http://www.sdiarticle4.com/review-history/61095 Received 10 July 2020 Original Research Article Accepted 16 September 2020 Published 29 September 2020 ABSTRACT Khat, Catha edulis, use is rampant in Eastern Africa and Middle East countries with associated reports of reproductive function impairment in the body of the user. Reports on recovery post long- term khat exposure are obscure.
    [Show full text]