title: New approaches in tail-bleeding assay in mice: improving an important method for designing new anti-thrombotic agents.
abstract: This report describes a modified, simple, low-cost and more sensitive method to determine bleeding patterns and haemoglobin concentration in a tail-bleeding assay using BALB/c mice and tail tip amputation. The cut tail was immersed in Drabkin's reagent to promote erythrocyte lysis and haemoglobin release, which was monitored over 30 min. The operator was blinded to individual conditions of the mice, which were treated with either saline (NaCl 0.15m), DMSO (0.5%) or clinical anti-thrombotic drugs. Our experimental protocols showed good reproducibility and repeatability of results when using Drabkin's reagent than water. Thus, the use of Drabkin's reagent offered a simple and low-cost method to observe and quantify the bleeding and rebleeding episodes. We also observed the bleeding pattern and total haemoglobin loss using untreated animals or those under anti-coagulant therapy in order to validate the new Drabkin method and thus confirm that it is a useful protocol to quantify haemoglobin concentrations in tail-bleeding assay. This modified method provided a more accurate results for bleeding patterns in mice and for identifying new anti-thrombotic drugs.
authors.affiliation: Laboratório de Antibióticos Bioquímica Ensino e Modelagem Molecular (LABiEMol) - Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil.|Programa de Pós-Graduação em Patologia (PPG-UFF) - Hospital Universitário Antônio Pedro, Universidade Federal Fluminense, Niterói, RJ, Brazil.
authors.forename: Max Seidy
authors.initials: MS
authors.lastname: Saito
authors.affiliation: Laboratório de Antibióticos Bioquímica Ensino e Modelagem Molecular (LABiEMol) - Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil.|Programa de Pós-Graduação em Patologia (PPG-UFF) - Hospital Universitário Antônio Pedro, Universidade Federal Fluminense, Niterói, RJ, Brazil.
authors.forename: André Luiz
authors.initials: AL
authors.lastname: Lourenço
authors.affiliation: Programa de Pós-Graduação em Patologia (PPG-UFF) - Hospital Universitário Antônio Pedro, Universidade Federal Fluminense, Niterói, RJ, Brazil.
authors.forename: Hye Chung
authors.initials: HC
authors.lastname: Kang
authors.affiliation: Laboratório de Modelagem Molecular e QSAR (ModMolQSAR) - Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Carlos Rangel
authors.initials: CR
authors.lastname: Rodrigues
authors.affiliation: Laboratório de Tecnologia Industrial Farmacêutica (LabTIF) - Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Lucio Mendes
authors.initials: LM
authors.lastname: Cabral
authors.affiliation: Laboratório de Antibióticos Bioquímica Ensino e Modelagem Molecular (LABiEMol) - Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil.
authors.forename: Helena Carla
authors.initials: HC
authors.lastname: Castro
authors.affiliation: Laboratório de Antibióticos Bioquímica Ensino e Modelagem Molecular (LABiEMol) - Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil.|Laboratório de Tecnologia Industrial Farmacêutica (LabTIF) - Faculdade de Farmácia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Plínio Cunha
authors.initials: PC
authors.lastname: Satlher
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D058851:GPI-Linked Proteins; D000071020:Hemochromatosis Protein; D006470:Hemorrhage / Q000188:drug therapy*; D008297:Male; D008565:Membrane Proteins / Q000097:blood; D008807:Mice, Inbred BALB C; D015203:Reproducibility of Results; D013623:Tail / Q000293:injuries*
chemical_list: D005343:Fibrinolytic Agents; D058851:GPI-Linked Proteins; C504414:HJV protein, mouse; D000071020:Hemochromatosis Protein; D008565:Membrane Proteins
keywords: anti-thrombotic agents; bleeding time; tail-bleeding assay