title: Adenine Nucleotides Control Proliferation In Vivo of Rat Retinal Progenitors by P2Y1Receptor.
abstract: Previous studies demonstrated that exogenous ATP is able to regulate proliferation of retinal progenitor cells (RPCs) in vitro possibly via P2Y1receptor, a G protein-coupled receptor. Here, we evaluated the function of adenine nucleotides in vivo during retinal development of newborn rats. Intravitreal injection of apyrase, an enzyme that hydrolyzes nucleotides, reduced cell proliferation in retinas at postnatal day 2 (P2). This decrease was reversed when retinas were treated together with ATPγ-S or ADPβ-S, two hydrolysis-resistant analogs of ATP and ADP, respectively. During early postnatal days (P0 to P5), an increase in ectonucleotidase (E-NTPDase) activity was observed in the retina, suggesting a decrease in the availability of adenine nucleotides, coinciding with the end of proliferation. Interestingly, intravitreal injection of the E-NTPDase inhibitor ARL67156 increased proliferation by around 60 % at P5 rats. Furthermore, immunolabeling against P2Y1receptor was observed overall in retina layers from P2 rats, including proliferating Ki-67-positive cells in the neuroblastic layer (NBL), suggesting that this receptor could be responsible for the action of adenine nucleotides upon proliferation of RPCs. Accordingly, intravitreal injection of MRS2179, a selective antagonist of P2Y1receptors, reduced cell proliferation by approximately 20 % in P2 rats. Moreover, treatment with MRS 2179 caused an increase in p57KIP2and cyclin D1 expression, a reduction in cyclin E and Rb phosphorylated expression and in BrdU-positive cell number. These data suggest that the adenine nucleotides modulate the proliferation of rat RPCs via activation of P2Y1receptors regulating transition from G1 to S phase of the cell cycle.
authors: Department of Neurobiology, Institute of Biology, Fluminense Federal University, Niterói, Brazil.
authors: Luana
authors: L
authors: de Almeida-Pereira
authors: Department of Neurobiology, Institute of Biology, Fluminense Federal University, Niterói, Brazil.
authors: Camila Feitosa
authors: CF
authors: Magalhães
authors: Department of Neurobiology, Institute of Biology, Fluminense Federal University, Niterói, Brazil.
authors: Marinna Garcia
authors: MG
authors: Repossi
authors: Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors: Maria Luiza Prates
authors: MLP
authors: Thorstenberg
authors: Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors: Alfred
authors: A
authors: Sholl-Franco
authors: Institute of Biophysics Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors: Robson
authors: R
authors: Coutinho-Silva
authors: Department of Neurobiology, Institute of Biology, Fluminense Federal University, Niterói, Brazil.
authors: Ana Lucia Marques
authors: ALM
authors: Ventura
authors: Department of Neurobiology, Institute of Biology, Fluminense Federal University, Niterói, Brazil. lfragel@id.uff.br.|Laboratório de Desenvolvimento e Regeneração Neural, Departmento de Neurobiologia, Universidade Federal Fluminense, Cx. Postal 100180, Niterói, RJ, 24020-141, Brazil. lfragel@id.uff.br.
authors: Lucianne
authors: http://orcid.org/0000-0001-6747-2828
authors: L
authors: Fragel-Madeira
mesh_terms: D000244:Adenosine Diphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D051381:Rats; D058472:Receptors, Purinergic P2Y1 / Q000378:metabolism*; D012160:Retina / Q000187:drug effects* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects*
chemical_list: C111914:N(6)-methyl-2'-deoxyadenosine 3',5'-diphosphate; D058472:Receptors, Purinergic P2Y1; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C027201:ectoATPase; D001973:Bromodeoxyuridine
keywords: Development; P2Y1 receptor; Proliferation; Purines; Retina