DOCSLIB.ORG

Characterizing Relaxin Receptor Expression and Exploring Relaxin's

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Characterizing Relaxin Receptor Expression and Exploring Relaxin's Diaz et al. BMC Urology (2020) 20:44 https://doi.org/10.1186/s12894-020-00607-4 RESEARCH ARTICLE Open Access Characterizing relaxin receptor expression and exploring relaxin’s effect on tissue remodeling/fibrosis in the human bladder Edward C. Diaz1,2*, Mason Briggs1,3, Yan Wen3, Guobing Zhuang3, Shannon L. Wallace3, Amy D. Dobberfuhl1, Chia-Sui Kao4 and Bertha C. Chen3 Abstract Background: Relaxin is an endogenous protein that has been shown to have antifibrotic properties in various organ systems. There has been no characterization of relaxin’s role in the human bladder. Our objective was to characterize relaxin receptor expression in the human bladder and assess relaxin’s effect on tissue remodeling/ fibrosis pathways in bladder smooth muscle cells. Methods: Relaxin family peptide receptor 1 (RXFP1) and RXFP2 expression was assessed using quantitative reverse transcriptase-PCR (qRT-PCR) and immunohistochemistry (IHC) on primary bladder tissue. Primary human smooth muscle bladder cells were cultured and stimulated with various concentrations of relaxin. Western blot, qRTPCR, ELISA, and zymogram assays were used to analyze fibrosis/tissue remodeling pathway proteins. Results: There was universal mRNA transcript detection and protein expression of relaxin receptors in primary bladder specimens. Immunohistochemistry demonstrated RXFP1 and RXFP2 localizing to both urothelial and smooth muscle cell layers of the bladder. 24 h of in vitro relaxin stimulation did not affect mRNA expression of selected proteins in human bladder smooth muscle cells. However, 48 h of in vitro relaxin stimulation resulted in upregulation of active (p = 0.004) and latent (p = 0.027) MMP-2 in cell lysate, and upregulation of active MMP-2 in supernatant (p = 0.04). There was a dose dependent relationship with increasing expression of MMP-2 with increasing relaxin concentration. Relaxin stimulation resulted in decreased levels of active and total TGF-β1in supernatant and extracellular matrix (p < 0.005 with 100 ng/mL relaxin stimulation). Conclusions: In the human bladder, relaxin receptors are expressed at the dome and trigone and localize to the urothelium and smooth muscle cell layers. Stimulation of human bladder SMCs with relaxin in vitro affects expression of MMP-2 and TGF-β1. Keywords: Relaxin, Fibrosis, Bladder, RXFP1, RXFP2 Background exstrophy without osteotomy in the first 48 h of life. It Relaxin 2, commonly referred to as “relaxin”, is known binds to relaxin family peptide receptor 1 (RXFP1) and in urology for its ability to facilitate closure of bladder RXFP2 [1]. In addition to its ability to increase pelvic ligament and bone laxity in pregnancy, relaxin can in- * Correspondence: [email protected] hibit fibrotic pathways [2] and modulate the extracellular 1 Department of Urology, Stanford University Medical Center, 300 Pasteur matrix [3]. This antifibrotic effect has led to clinical tri- Drive, Grant S-287, Stanford, CA 94305, USA 2Present Address: Division of Pediatric Urology, Advocate Children’s Hospital, als looking at relaxin therapy for fibrotic medical 8901 West Golf Road, Suite 301, Des Plaines, IL 60016, USA Full list of author information is available at the end of the article © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Diaz et al. BMC Urology (2020) 20:44 Page 2 of 9 conditions such as heart failure [4], scleroderma [5], and Invitrogen), Alexa Fluor 594-conjugated donkey anti- pulmonary fibrosis [6]. rabbit IgG (1:200, Invitrogen). The poorly compliant bladder demonstrates a histo- logic phenotype of fibrosis. There is an evolution of ini- Quantitative reverse transcriptase-PCR (qRT-PCR) tial hypertrophy and then loss of detrusor muscle [7], RNA was extracted with RNA Stat-60 reagent (Tel-Test muscle replacement with collagen [8], and decreased Inc.). Primers for target proteins were created using pre- elastin in the extracellular matrix [9]. Molecular path- viously described protocols [12–16]. Samples were per- ways that contribute to this process are multifactorial formed in duplicate. Brilliant SYBR Green qPCR Master [10]. Preliminary in-vivo data suggests a relationship be- Mix (Stratagene) was used. A 10 min hot start at 95 °C tween relaxin and bladder fibrosis. Using a radiated mur- with forty cycles of 30 s of denaturation (94 °C), 1 min of ine model, a group demonstrated that relaxin therapy annealing (60 °C), and extension at 72 °C for 30 s was reverses radiation-induced fibrosis and restores bladder used. Glyceraldehyde 3-phosphate dehydrogenase function [11]. This evidence supports further investiga- (GAPDH) was used as the endogenous reference. Hu- tion as to whether relaxin-related mechanisms also exist man vaginal cuff tissue was used as a positive control for in the human bladder which, to our knowledge, has not RXFP1 and RXFP2 expression. been documented. The objective of this study is to characterize relaxin receptor expression in the human Bladder smooth muscle cell culture and Relaxin treatment bladder and assess the in-vitro effect of relaxin on ex- Primary human bladder smooth muscle cell (bSMC) line pression of proteins involved in tissue remodeling and fi- was obtained commercially from Lonza. Cells were brosis in human bladder smooth muscle cells. grown in Dulbecco’s Modified Eagle Medium (DMEM) with 10% charcoal-stripped Fetal Bovine Serum (FBS) at Methods 37 °C and 5% CO2 incubator. Cells (passage 4) were – Procurement of primary bladder specimens plated in triplicate. At 80 90% confluency, cells were IRB exemption and external approval from Donor Net- then starved in serum-free DMEM with 0.2% lactalbu- work West’s (federally designated organ procurement min enzymatic hydrolysate for 24 h, and then treated – organization) Internal Research Council and Medical with various concentrations of human relaxin-2 (0 100 Advisory Board Research Subcommittee was obtained ng/mL, PeproTech) for 24 or 48 h as described previ- for collection of primary bladder tissue from brain dead ously by our group [17]. cadaveric organ transplant donors. Brain dead donation Supernatant was concentrated 100-fold using Vivaspin was selected to eliminate warm ischemia time. Bladder filtration tubes (Sartorius AG). To acquire proteins, cell tissue (Dome and trigone) was procured by a board cer- layers were washed with (PBS) and lysed by 3 successive tified urologist after removal of organs allocated for hu- 10- min treatments with Complete-Mini Protease Inhibi- man transplant and immediately cryopreserved, placed tor Cocktail (1 tab/10 mL, Sigma- Aldrich) in radioim- in formalin, and processed into cell culture. munoprecipitation assay (RIPA) buffer at 4 °C. Cell debris was removed by centrifugation. Extracellular matrix (ECM) was prepared according to Immunohistochemistry of primary tissue Pedrozo et al. [18] Cells were removed with RIPA buffer, Primary tissue was processed into paraffin blocks. 5 um the remaining cell-free, non-solubilized ECM was sections were made and hematoxylin and eosin stain was washed 3 times with PBS and plates dried overnight at performed. Slides were reviewed by a genitourinary room temperature. Isolated ECM samples were digested pathologist. with plasmin (MP Biomedicals) in DMEM for 3 h at 37° to release TGFβ-118 [19]., Reaction was stopped by the Immunofluorescence for RXFP1 and RXFP2 in primary addition of aprotinin (5 μg/mL, Sigma- Aldrich). tissue Plasmin-digested ECM samples were collected and con- 5 um cryostat sections were fixed in 4% paraformalde- centrated 200-fold with Vivaspin filtration tubes. hyde, permeabilized with 0.5% Triton X-100 in phosphate-buffered saline (PBS), and blocked in 1% bo- ELISA measurement of active and latent TGF-β1 vine serum albumin. Primary antibodies: mouse anti- An ELISA kit (Promega Corp) was used. Nunc MaxiSorp human cytokeratin (AE1/AE3) IgG (5μg/ml, Chemicon 96-well plates were coated with TGF-β1 monoclonal International), mouse anti-human desmin IgG (1:50, antibody. Plates were washed and blocked and 100 μLof Sigma-Aldrich), rabbit anti-human RXFP1 IgG (10μg/ acid-treated and non-acid treated samples were then ml, Abcam), rabbit anti-human RXFP2 IgG (10μg/ml, added to the wells in duplicate. After washing, anti- Lifespan Biosciences). Secondary antibody used: Alexa TGF-β1 polyclonal antibody was applied. Horseradish Fluor 488-conjugated goat anti-mouse IgG (1:300, peroxidase HRP–conjugated antibody was used for Diaz et al. BMC Urology (2020) 20:44 Page 3 of 9 detection. Absorbance at 450 nm was measured on a SPSS Statistics 21.0 software
Load more

Recommended publications
  • The Roles Played by Highly Truncated Splice Variants of G Protein-Coupled Receptors Helen Wise
    Wise Journal of Molecular Signaling 2012, 7:13 http://www.jmolecularsignaling.com/content/7/1/13 REVIEW Open Access The roles played by highly truncated splice variants of G protein-coupled receptors Helen Wise Abstract Alternative splicing of G protein-coupled receptor (GPCR) genes greatly increases the total number of receptor isoforms which may be expressed in a cell-dependent and time-dependent manner. This increased diversity of cell signaling options caused by the generation of splice variants is further enhanced by receptor dimerization. When alternative splicing generates highly truncated GPCRs with less than seven transmembrane (TM) domains, the predominant effect in vitro is that of a dominant-negative mutation associated with the retention of the wild-type receptor in the endoplasmic reticulum (ER). For constitutively active (agonist-independent) GPCRs, their attenuated expression on the cell surface, and consequent decreased basal activity due to the dominant-negative effect of truncated splice variants, has pathological consequences. Truncated splice variants may conversely offer protection from disease when expression of co-receptors for binding of infectious agents to cells is attenuated due to ER retention of the wild-type co-receptor. In this review, we will see that GPCRs retained in the ER can still be functionally active but also that highly truncated GPCRs may also be functionally active. Although rare, some truncated splice variants still bind ligand and activate cell signaling responses. More importantly, by forming heterodimers with full-length GPCRs, some truncated splice variants also provide opportunities to generate receptor complexes with unique pharmacological properties. So, instead of assuming that highly truncated GPCRs are associated with faulty transcription processes, it is time to reassess their potential benefit to the host organism.
    [Show full text]
  • Extracellular Matrix Grafts: from Preparation to Application (Review)
    INTERNATIONAL JOURNAL OF MOleCular meDICine 47: 463-474, 2021 Extracellular matrix grafts: From preparation to application (Review) YONGSHENG JIANG1*, RUI LI1,2*, CHUNCHAN HAN1 and LIJIANG HUANG1 1Science and Education Management Center, The Affiliated Xiangshan Hospital of Wenzhou Medical University, Ningbo, Zhejiang 315700; 2School of Chemistry, Sun Yat-sen University, Guangzhou, Guangdong 510275, P.R. China Received July 30, 2020; Accepted December 3, 2020 DOI: 10.3892/ijmm.2020.4818 Abstract. Recently, the increasing emergency of traffic acci- Contents dents and the unsatisfactory outcome of surgical intervention are driving research to seek a novel technology to repair trau- 1. Introduction matic soft tissue injury. From this perspective, decellularized 2. ECM-G characterization matrix grafts (ECM-G) including natural ECM materials, and 3. Methods of decellularization treatments their prepared hydrogels and bioscaffolds, have emerged as 4. Removal of residual cellular components and chemicals possible alternatives for tissue engineering and regenerative 5. Application of ECM-P in regenerative medicine medicine. Over the past decades, several physical and chemical 6. Challenges and future outlook on ECM-P decellularization methods have been used extensively to deal 7. Conclusions with different tissues/organs in an attempt to carefully remove cellular antigens while maintaining the non-immunogenic ECM components. It is anticipated that when the decellular- 1. Introduction ized biomaterials are seeded with cells in vitro or incorporated into irregularly shaped defects in vivo, they can provide the The extracellular matrix (ECM) derived from organs/tissues is appropriate biomechanical and biochemical conditions for a complex, highly organized assembly of macromolecules with directing cell behavior and tissue remodeling.
    [Show full text]
  • Extracellular Matrix Degradation and Remodeling in Development and Disease
    Downloaded from http://cshperspectives.cshlp.org/ on September 30, 2021 - Published by Cold Spring Harbor Laboratory Press Extracellular Matrix Degradation and Remodeling in Development and Disease Pengfei Lu1,2, Ken Takai2, Valerie M. Weaver3, and Zena Werb2 1Breakthrough Breast Cancer Research Unit, Paterson Institute for Cancer Research and Wellcome Trust Centre for Cell Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester M20 4BX, United Kingdom 2Department of Anatomy and Program in Developmental Biology, University of California, San Francisco, California 94143-0452 3Department of Surgery and Center for Bioengineering and Tissue Regeneration, University of California, San Francisco, California 94143 Correspondence: [email protected] The extracellular matrix (ECM) serves diverse functions and is a major component of the cellular microenvironment. The ECM is a highly dynamic structure, constantly undergoing a remodeling process where ECM components are deposited, degraded, or otherwise modified. ECM dynamics are indispensible during restructuring of tissue architecture. ECM remodeling is an important mechanism whereby cell differentiation can be regulated, including processes such as the establishment and maintenance of stem cell niches, branch- ing morphogenesis, angiogenesis, bone remodeling, and wound repair. In contrast, abnor- mal ECM dynamics lead to deregulated cell proliferation and invasion, failure of cell death, and loss of cell differentiation, resulting in congenital defects and pathological processes including tissue fibrosis and cancer. Understanding the mechanisms of ECM remodeling and its regulation, therefore, is essential for developing new therapeutic inter- ventions for diseases and novel strategies for tissue engineering and regenerative medicine. he extracellular matrix (ECM) forms a milieu versatile and performs many functions in addi- Tsurrounding cells that reciprocally influ- tion to its structural role.
    [Show full text]
  • Immune Clearance of Senescent Cells to Combat Ageing and Chronic Diseases
    cells Review Immune Clearance of Senescent Cells to Combat Ageing and Chronic Diseases Ping Song * , Junqing An and Ming-Hui Zou Center for Molecular and Translational Medicine, Georgia State University, 157 Decatur Street SE, Atlanta, GA 30303, USA; [email protected] (J.A.); [email protected] (M.-H.Z.) * Correspondence: [email protected]; Tel.: +1-404-413-6636 Received: 29 January 2020; Accepted: 5 March 2020; Published: 10 March 2020 Abstract: Senescent cells are generally characterized by permanent cell cycle arrest, metabolic alteration and activation, and apoptotic resistance in multiple organs due to various stressors. Excessive accumulation of senescent cells in numerous tissues leads to multiple chronic diseases, tissue dysfunction, age-related diseases and organ ageing. Immune cells can remove senescent cells. Immunaging or impaired innate and adaptive immune responses by senescent cells result in persistent accumulation of various senescent cells. Although senolytics—drugs that selectively remove senescent cells by inducing their apoptosis—are recent hot topics and are making significant research progress, senescence immunotherapies using immune cell-mediated clearance of senescent cells are emerging and promising strategies to fight ageing and multiple chronic diseases. This short review provides an overview of the research progress to date concerning senescent cell-caused chronic diseases and tissue ageing, as well as the regulation of senescence by small-molecule drugs in clinical trials and different roles and regulation of immune cells in the elimination of senescent cells. Mounting evidence indicates that immunotherapy targeting senescent cells combats ageing and chronic diseases and subsequently extends the healthy lifespan. Keywords: cellular senescence; senescence immunotherapy; ageing; chronic disease; ageing markers 1.
    [Show full text]
  • Review Genetic Dissection of Mammalian Fertility Pathways Martin M
    fertility supplement review Genetic dissection of mammalian fertility pathways Martin M. Matzuk*†‡# and Dolores J. Lamb†§ Departments of *Pathology, †Molecular and Cellular Biology and ‡Molecular and Human Genetics, and §Scott Department of Urology, Baylor College of Medicine, Houston, TX 77030, USA #e-mail: [email protected] The world’s population is increasing at an alarming rate and is projected to reach nine billion by 2050. Despite this, 15% of couples world-wide remain childless because of infertility. Few genetic causes of infertility have been identified in humans; nevertheless, genetic aetiologies are thought to underlie many cases of idiopathic infertility. Mouse models with reproductive defects as a major phenotype are being rapidly created and discovered and now total over 200. These models are helping to define mechanisms of reproductive function, as well as identify potential new contracep- tive targets and genes involved in the pathophysiology of reproductive disorders. With this new information, men and women will continue to be confronted with difficult decisions on whether or not to use state-of-the-art technology and hormonal treatments to propagate their germline, despite the risks of transmitting mutant genes to their offspring. espite advances in assisted reproductive have been produced by spontaneous muta- Where it all begins technologies, infertility is a major health tions, fortuitous transgene integration, Reproductive development and physiology problem worldwide. Approximately 15% of retroviral infection of embryonic stem are evolutionarily conserved processes couples are unable to conceive within one cells, ethylnitrosurea (ENU) mutagenesis across eutherian mammalian species and year of unprotected intercourse. The fertil- and gene targeting technologies3,7,8.
    [Show full text]
  • A Narrative Review on Plasminogen Activator Inhibitor-1 and Its (Patho)Physiological Role: to Target Or Not to Target?
    International Journal of Molecular Sciences Review A Narrative Review on Plasminogen Activator Inhibitor-1 and Its (Patho)Physiological Role: To Target or Not to Target? Machteld Sillen and Paul J. Declerck * Laboratory for Therapeutic and Diagnostic Antibodies, Department of Pharmaceutical and Pharmacological Sciences, KU Leuven, B-3000 Leuven, Belgium; [email protected] * Correspondence: [email protected] Abstract: Plasminogen activator inhibitor-1 (PAI-1) is the main physiological inhibitor of plasminogen activators (PAs) and is therefore an important inhibitor of the plasminogen/plasmin system. Being the fast-acting inhibitor of tissue-type PA (tPA), PAI-1 primarily attenuates fibrinolysis. Through inhibition of urokinase-type PA (uPA) and interaction with biological ligands such as vitronectin and cell-surface receptors, the function of PAI-1 extends to pericellular proteolysis, tissue remodeling and other processes including cell migration. This review aims at providing a general overview of the properties of PAI-1 and the role it plays in many biological processes and touches upon the possible use of PAI-1 inhibitors as therapeutics. Keywords: plasminogen activator inhibitor-1; PAI-1; fibrinolysis; cardiovascular disease; cancer; inflammation; fibrosis; aging Citation: Sillen, M.; Declerck, P.J. 1. Introduction A Narrative Review on Plasminogen Plasminogen activator inhibitor-1 (PAI-1) belongs to the family of serine protease Activator Inhibitor-1 and Its inhibitors (serpins) and is an important regulator of the plasminogen/plasmin system (Patho)Physiological Role: To Target (Figure1)[ 1]. This system revolves around the conversion of the zymogen plasmino- or Not to Target?. Int. J. Mol. Sci. 2021, gen into the active enzyme plasmin through proteolytic cleavage that is mediated by 22, 2721.
    [Show full text]
  • The Biological and Clinical Relevance of G Protein-Coupled Receptors to the Outcomes of Hematopoietic Stem Cell Transplantation: a Systematized Review
    International Journal of Molecular Sciences Review The Biological and Clinical Relevance of G Protein-Coupled Receptors to the Outcomes of Hematopoietic Stem Cell Transplantation: A Systematized Review Hadrien Golay 1 , Simona Jurkovic Mlakar 1, Vid Mlakar 1, Tiago Nava 1,2 and Marc Ansari 1,2,* 1 Platform of Pediatric Onco-Hematology research (CANSEARCH Laboratory), Department of Pediatrics, Gynecology, and Obstetrics, University of Geneva, Bâtiment La Tulipe, Avenue de la Roseraie 64, 1205 Geneva, Switzerland 2 Department of Women-Children-Adolescents, Division of General Pediatrics, Pediatric Onco-Hematology Unit, Geneva University Hospitals (HUG), Avenue de la Roseraie 64, 1205 Geneva, Switzerland * Correspondence: [email protected] Received: 14 June 2019; Accepted: 7 August 2019; Published: 9 August 2019 Abstract: Hematopoietic stem cell transplantation (HSCT) remains the only curative treatment for several malignant and non-malignant diseases at the cost of serious treatment-related toxicities (TRTs). Recent research on extending the benefits of HSCT to more patients and indications has focused on limiting TRTs and improving immunological effects following proper mobilization and engraftment. Increasing numbers of studies report associations between HSCT outcomes and the expression or the manipulation of G protein-coupled receptors (GPCRs). This large family of cell surface receptors is involved in various human diseases. With ever-better knowledge of their crystal structures and signaling dynamics, GPCRs are already the targets for one third of the current therapeutic arsenal. The present paper assesses the current status of animal and human research on GPCRs in the context of selected HSCT outcomes via a systematized survey and analysis of the literature.
    [Show full text]
  • 2011: Macrophage Phenotype As a Determinant of Remodeling Outcome with the Use of Biologic Scaffolds
    Macrophage Phenotype as a Determinant of Remodeling Outcome with the use of Biologic Scaffolds Stephen F. Badylak1, Kathryn A. Kukla1,3, Li Zhang1, Neill J. Turner1, Bryan N. Brown1,2 Corresponding Author: Stephen F. Badylak 1McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA; 2Department of Biomedical Engineering, Cornell University, Ithaca, NY; and 3Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA Statement of Purpose: The ultimate determination of deposition of dense and disorganized connective tissue clinical success for an implanted biomaterial is the consistent with encapsulation. Remodeling of the response of the host tissue following implantation. The autograft was characterized by necrosis of the muscular innate immune mechanisms that participate in and component of the tissue and deposition of dense mature modulate the host response include effector cells such as connective tissue and adipose tissue consistent with macrophages. Macrophages have been classified as scarring. having either an M1 or an M2 phenotype, depending upon gene expression, effector molecule production, and cell Despite differences in the tissue remodeling outcome, a function. To date, the causes and the effects of morphologically indistinguishable population of macrophage polarization towards an M1 or M2 phenotype macrophages was observed following implantation of have been studied largely in the context of the host each scaffold material. However, immunolabeling response to pathogens and in cancer biology. Recently, techniques showed that scaffolds which resulted in M1 and M2 macrophages have been shown to play constructive remodeling (i.e., UBM) were associated with distinct roles in the remodeling process following tissue a predominant M2 (regulatory, pro-wound healing) injury and the host response to biologic scaffold materials macrophage population, while scaffolds which resulted in (1).
    [Show full text]
  • Relaxin Receptor RXFP1 and RXFP2 Expression in Ligament, Tendon, and Shoulder Joint Capsule of Rats
    ORIGINAL ARTICLE Rehabilitation & Sports Medicine http://dx.doi.org/10.3346/jkms.2016.31.6.983 • J Korean Med Sci 2016; 31: 983-988 Relaxin Receptor RXFP1 and RXFP2 Expression in Ligament, Tendon, and Shoulder Joint Capsule of Rats Jae Hyung Kim,1,2 Sang Kwang Lee,3 Numerous musculoskeletal disorders are caused by thickened ligament, tendon stiffness, or Seong Kyu Lee,4 Joo Heon Kim,5 fibrosis of joint capsule. Relaxin, a peptide hormone, can exert collagenolytic effect on and Michael Fredericson1 ligamentous and fibrotic tissues. We hypothesized that local injection of relaxin could be used to treat entrapment neuropathy and adhesive capsulitis. Because hormonal effect 1Division of Physical Medicine and Rehabilitation, Department of Orthopaedic Surgery, Stanford depends on the receptor of the hormone on the target cell, it is important to confirm the University, Stanford, CA, USA; 2Department of presence of such hormonal receptor at the target tissue before the hormone therapy is Physical Medicine & Rehabilitation, Eulji University initiated. The aim of this study was to determine whether there were relaxin receptors in Hospital and Eulji University School of Medicine, the ligament, tendon, and joint capsular tissues of rats and to identify the distribution of Daejeon, Korea; 3Eulji Medi-Bio Research Institute, Daejeon, Korea; 4Department of Biochemistry, Eulji relaxin receptors in these tissues. Transverse carpal ligaments (TCLs), inguinal ligaments, University School of Medicine, Daejeon, Korea; anterior cruciate ligaments (ACLs), Archilles tendons, and shoulder joint capsules were 5Department of Pathology, Eulji University School of obtained from male Wistar rats. Western blot analysis was used to identify relaxin receptor Medicine, Daejeon, Korea isoforms RXFP1 and RXFP2.
    [Show full text]
  • Apoptosis During Embryonic Tissue Remodeling Is Accompanied by Cell Senescence
    www.impactaging.com AGING, November 2015, Vol 7 N 11 Research Paper Apoptosis during embryonic tissue remodeling is accompanied by cell senescence Carlos I. Lorda‐Diez1, Beatriz Garcia‐Riart1, Juan A. Montero 1, Joaquín Rodriguez‐León2, Juan A 1 1 Garcia‐Porrero , and Juan M. Hurle 1 Departamento de Anatomía y Biología Celular and IDIVAL, Universidad de Cantabria, Santander 39011, Spain; 2 Departamento de Anatomía y Biología Celular,Universidad de Extremadura, Badajoz 07006, Spain. , s , v l p n , , n l , AS , Key words: programmed cell death sene cence limb de e o me t β‐galactosidase sy dacty y S P INZ Received: 07/01/15; Accepted: 11/02/15; Published: 11/14/15 Correspondence to: Juan M. Hurlé, PhD; E‐mail: [email protected] Copyright: Lorda‐Diez et al. This is an open‐access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Abstract This study re‐examined the dying process in the interdigital tissue during the formation of free digits in the developing limbs. We demonstrated that the interdigital dying process was associated with cell senescence, as deduced by induction of β‐gal activity, mitotic arrest, and transcriptional up‐regulation of p21 together with many components of the senescence ‐associated secretory phenotype. We also found overlapping domains of expression of members of the Btg/Tob gene family of antiproliferative factors in the regressing interdigits. Notably, Btg2 was up‐regulated during interdigit remodeling in species with free digits but not in the webbed foot of the duck.
    [Show full text]
  • The Myofibroblast in Wound Healing and Fibrosis
    F1000Research 2016, 5(F1000 Faculty Rev):752 Last updated: 17 JUL 2019 REVIEW The myofibroblast in wound healing and fibrosis: answered and unanswered questions [version 1; peer review: 2 approved] Marie-Luce Bochaton-Piallat1, Giulio Gabbiani1, Boris Hinz2 1Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, Geneva, Switzerland 2Laboratory of Tissue Repair and Regeneration, Matrix Dynamics Group, Faculty of Dentistry, University of Toronto, Toronto, Canada First published: 26 Apr 2016, 5(F1000 Faculty Rev):752 ( Open Peer Review v1 https://doi.org/10.12688/f1000research.8190.1) Latest published: 26 Apr 2016, 5(F1000 Faculty Rev):752 ( https://doi.org/10.12688/f1000research.8190.1) Reviewer Status Abstract Invited Reviewers The discovery of the myofibroblast has allowed definition of the cell 1 2 responsible for wound contraction and for the development of fibrotic changes. This review summarizes the main features of the myofibroblast version 1 and the mechanisms of myofibroblast generation. Myofibroblasts originate published from a variety of cells according to the organ and the type of lesion. The 26 Apr 2016 mechanisms of myofibroblast contraction, which appear clearly different to those of smooth muscle cell contraction, are described. Finally, we summarize the possible strategies in order to reduce myofibroblast F1000 Faculty Reviews are written by members of activities and thus influence several pathologies, such as hypertrophic the prestigious F1000 Faculty. They are scars and organ fibrosis. commissioned and are peer reviewed before Keywords publication to ensure that the final, published version Myofibroblast , mechanotransduction , myofibroblast generation , is comprehensive and accessible. The reviewers myofibroblast contraction , hypertrophic scars , organ fibroses who approved the final version are listed with their names and affiliations.
    [Show full text]
  • The Role of the Relaxin Receptor RXFP1 in Brain Cancer
    The role of the relaxin receptor RXFP1 in brain cancer By Usakorn Kunanuvat A Thesis submitted to the Faculty of Graduate Studies of The University of Manitoba in partial fulfilment for the requirements of the degree of MASTER OF SCIENCE Department of Human Anatomy and Cell Science University of Manitoba Winnipeg Copyright © 2012 by Usakorn Kunanuvat ABSTRACT Relaxin (RLN2) promotes cell migration/invasion, cell growth, and neoangiogenesis through binding to the relaxin receptor RXFP1 in many types of cancers. However, there have been no studies to determine the role of this system in brain tumors, especially in Glioblastoma Multiforme (GB), the most lethal primary brain tumor in adults. GB is a systemic brain disease and aggressively invades brain tissue. In this study, we have identified RXFP1 receptor, but not RLN2, in GB cell lines and primary GB cells from patients. RLN2 treatment resulted in a significant increase in migration of GB cell line and primary GB cells. To determine molecular mechanisms that facilitate RXFP1-mediated migration in GB cells, we employed a pseudopodia assay and 2D LC-MS/MS to investigate the protein composition at cell protrusions (pseudopodia) during GB cell migration. We also observed the expression of known mediators promoting tissue invasion upon RLN2 treatment. We identified PGRMC1, a candidate protein from 2D LC-MS/MS as a novel relaxin target protein in RXFP1-expressing brain tumor cells. RLN2 treatment also caused an increase in cathepsin (cath)-B and -L and enhanced production of as the small Rho-GTPases Rac1 and Cdc42 in GB cells. Collectively, these findings indicate that RXFP1-induced cell migration is mediated by the upregulation and intracellular actions of Rac1, Cdc42 and by cath-B and cath–L who serve as matrix modulating factors to facilitate brain tumor cells migration.
    [Show full text]