Ubiquitin role in stallions with high and low freezability and application of coenzyme Q-10 as a promoter of sperm mitochondrial function

• Grant number: 16/14626-7
• Support Opportunities: Regular Research Grants
• Start date: October 01, 2016
• End date: September 30, 2018
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Rubens Paes de Arruda
• Grantee: Rubens Paes de Arruda
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Eneiva Carla Carvalho Celeghini

Abstract

Studies demonstrate that one of the major limitations to the use of equine frozen semen is observed among stallions in relation to sperm ability to survive the cryopreservation. The ubiquitin is a molecule which is involved in a route of degradation of damaged proteins or errors of synthesis. This process occurs in the passage of sperm through the epididymis, and it´s associated with infertility in males. On the other hand, the coenzyme Q-10 (CoQ-10) is a cofactor in the electron transport chain in mitochondria that is essential for the production of energy in form of ATP and can be an alternative to improve the quality of equine cryopreserved semen. Taking the above quotes into account two experiments were outlined. Both ones will make use of four stallions with high resistance to freezability (HF) and four stallions with low resistance to freezability (LF). Each animal will have five semen samples collected that will then be followed by a freezing process. Experiment 1: it will analyze the amount of ubiquitin in stallions with high and low freezability by the technique of sperm-ubiquitin tag immunoassay (SUTI) and the data obtained will be correlated with other semen parameters such as sperm motility characteristics (CASA), integrity of the plasma and acrossomal membrane and mitochondrial membrane potential (via fluorescent probes propidium iodide, hoechst 33342, JC-1 and FITC-PSA), alterations in the cytoskeleton (phalloidin-FITC) and morphology (DIC). Experiment 2: the cryopreserved semen will be treated with CoQ-10 to 50µM while the control sample is cryopreserved only with Botucrio®. Semen samples will be analyzed for the same characteristics of the experiment 1, excluding the SUTI technique and with the addition of mitochondrial function (DAB and fluorescent probe Mito Tracker Red®). (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

RENATA LANÇONI; ENEIVA CARLA CARVALHO CELEGHINI; VALDEMAR DE GIULI JÚNIOR; CARLA PATRICIA TEODORO DE CARVALHO; GABRIELA BERTAIOLLI ZOCA; LAURA NATALY GARCIA-OLIVEROS; LEONARDO BATISSACO; LETÍCIA ZOCCOLARO OLIVEIRA; RUBENS PAES DE ARRUDA. Coenzyme Q-10 improves preservation of mitochondrial functionality and actin structure of cryopreserved stallion sperm. Animal Reproduction, v. 18, n. 1, 2021-04-12. (16/14626-7)

LANCONI, RENATA; CARVALHO CELEGHINI, ENEIVA CARLA; DE GIULI JUNIOR, VALDEMAR; TEODORO DE CARVALHO, CARLA PATRICIA; ZOCA, GABRIELA BERTAIOLLI; GARCIA-OLIVEROS, LAURA NATALY; BATISSACO, LEONARDO; OLIVEIRA, LETICIA ZOCCOLARO; DE ARRUDA, RUBENS PAES. Coenzyme Q-10 improves preservation of mitochondrial functionality and actin structure of cryopreserved stallion sperm. ANIMAL REPRODUCTION, v. 18, n. 1, p. 9-pg., 2021-01-01. (16/14626-7)