Centrifugation Force and Time Alter CASA Parameters and Oxidative Status of Cryopreserved Stallion Sperm
Giuseppina Marzano,
Natalina Moscatelli,
Mariangela Di Giacomo,
Nicola Antonio Martino,
Giovanni Michele Lacalandra,
Maria Elena Dell’Aquila,
Giuseppe Maruccio,
Elisabetta Primiceri,
Maria Serena Chiriacò,
Vincenzo Zara,
Alessandra Ferramosca
Affiliations
Giuseppina Marzano
Department of Mathematics and Physics E. de Giorgi, University of Salento, 73100 Via per Arnesano, Lecce, Italy
Natalina Moscatelli
Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Via per Monteroni, Lecce, Italy
Mariangela Di Giacomo
Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Via per Monteroni, Lecce, Italy
Nicola Antonio Martino
Department of Biosciences, Biotechnologies & Biopharmaceutics, University of Bari Aldo Moro, km3 Strada per Casamassima, 70100 Valenzano, Bari, Italy
Giovanni Michele Lacalandra
Department of Veterinary Medicine, University of Bari Aldo Moro, km3 Strada per Casamassima, 70100 Valenzano, Bari, Italy
Maria Elena Dell’Aquila
Department of Biosciences, Biotechnologies & Biopharmaceutics, University of Bari Aldo Moro, km3 Strada per Casamassima, 70100 Valenzano, Bari, Italy
Giuseppe Maruccio
Department of Mathematics and Physics E. de Giorgi, University of Salento, 73100 Via per Arnesano, Lecce, Italy
Elisabetta Primiceri
Institute of Nanotechnology, CNR NANOTEC, 73100 Via per Monteroni, Lecce, Italy
Maria Serena Chiriacò
Institute of Nanotechnology, CNR NANOTEC, 73100 Via per Monteroni, Lecce, Italy
Vincenzo Zara
Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Via per Monteroni, Lecce, Italy
Alessandra Ferramosca
Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Via per Monteroni, Lecce, Italy
Conventional sperm selection techniques used in ARTs rely on centrifugation steps. To date, the different studies reported on the effects of centrifugation on stallion sperm motility provided contrasting results and do not include effects on mitochondrial functionality and different oxidative parameters. The effects of different centrifugation protocols (300× g for 5′, 300× g for 10′, 1500× g for 5′ and 1500× g for 10′ vs. no centrifugation) on motility and oxidative status in cryopreserved stallion sperm, were analyzed. After centrifugation, almost all motility parameters were significantly altered, as observed by computer-assisted sperm analysis. A polarographic assay of oxygen consumption showed a progressive decrease in mitochondria respiration from the gentlest to the strongest protocol. By laser scanning confocal microscopy, significant reduction of mitochondrial membrane potential, at any tested protocol, and time-dependent effects, at the same centrifugal force, were found. Increased DNA fragmentation index at any tested protocol and time-dependent effects at the same centrifugal force were found, whereas increased protein carbonylation was observed only at the strongest centrifugal force. These results provide more comprehensive understandings on centrifugation-induced effects on cryopreserved stallion sperm and suggest that, even at a weak force for a short time, centrifugation impairs different aspects of equine sperm metabolism and functionality.
