Equine spermatozoa highly rely on oxidative phosphorylation for their energy management. The present work aimed to characterize the role of mitochondria on horse sperm motility and ROS production by incubating spermatozoa with specific inhibitors of the different mitochondrial complexes. Equine spermatozoa were incubated 1 h and 3 h at 37 °C with: complex I inhibitor rotenone (5 μM, ROT), complex II inhibitor dimethyl-malonate (10 mM, DMM), complex III inhibitor antimycin A (1.8 μM, ANTI), the uncoupling agent carbonyl cyanide m-chlorophenyl hydrazine (5 μM, CCCP), ATP synthase inhibitor oligomycin (5 μM, OLIGO), and 2 μL vehicle DMSO (control, CTL). Samples were analyzed for sperm motility and for mitochondrial membrane potential (MMP), mitochondrial integrity, mitochondrial O2•– production, and cytoplasmic H2O2. A multivariate analysis was performed on the data. CCCP caused a pronounced MMP reduction at both time points while ROT and ANTI showed the same effect at 3 h. All treatments at 3 h incubation significantly reduced the percentage of sperm with early changes in membrane permeability with active mitochondria. The H2O2 production of live cells was low at 1 h incubation in all treatments; after 3 h a slight decrease in the percentage of low-H2O2 producing cells was recorded. All treatments, except DMM, induced a significant decline in sperm motility and kinematics and modified the pattern of sperm subpopulations. The effect of DMM was evident only after 3 h, increasing the percentage of slow sperm subpopulation. In conclusion, the disruption of mitochondrial integrity induces an increase of mitochondrial ROS production that could be detrimental for cell function and survivior.

Use of specific mitochondrial complex inhibitors to investigate mitochondrial involvement on horse sperm motility and ROS production / Giaretta E.; Mislei B.; Martinez-Pastor F.; Nesci S.; Spinaci M.; Galeati G.; Nerozzi C.; Mari G.; Tamanini C.; Bucci D.. - In: RESEARCH IN VETERINARY SCIENCE. - ISSN 0034-5288. - STAMPA. - 147:(2022), pp. 12-19. [10.1016/j.rvsc.2022.03.017]

Use of specific mitochondrial complex inhibitors to investigate mitochondrial involvement on horse sperm motility and ROS production

Mislei B.;Nesci S.;Spinaci M.;Galeati G.;Nerozzi C.;Mari G.;Tamanini C.;Bucci D.
2022

Abstract

Equine spermatozoa highly rely on oxidative phosphorylation for their energy management. The present work aimed to characterize the role of mitochondria on horse sperm motility and ROS production by incubating spermatozoa with specific inhibitors of the different mitochondrial complexes. Equine spermatozoa were incubated 1 h and 3 h at 37 °C with: complex I inhibitor rotenone (5 μM, ROT), complex II inhibitor dimethyl-malonate (10 mM, DMM), complex III inhibitor antimycin A (1.8 μM, ANTI), the uncoupling agent carbonyl cyanide m-chlorophenyl hydrazine (5 μM, CCCP), ATP synthase inhibitor oligomycin (5 μM, OLIGO), and 2 μL vehicle DMSO (control, CTL). Samples were analyzed for sperm motility and for mitochondrial membrane potential (MMP), mitochondrial integrity, mitochondrial O2•– production, and cytoplasmic H2O2. A multivariate analysis was performed on the data. CCCP caused a pronounced MMP reduction at both time points while ROT and ANTI showed the same effect at 3 h. All treatments at 3 h incubation significantly reduced the percentage of sperm with early changes in membrane permeability with active mitochondria. The H2O2 production of live cells was low at 1 h incubation in all treatments; after 3 h a slight decrease in the percentage of low-H2O2 producing cells was recorded. All treatments, except DMM, induced a significant decline in sperm motility and kinematics and modified the pattern of sperm subpopulations. The effect of DMM was evident only after 3 h, increasing the percentage of slow sperm subpopulation. In conclusion, the disruption of mitochondrial integrity induces an increase of mitochondrial ROS production that could be detrimental for cell function and survivior.
2022
Use of specific mitochondrial complex inhibitors to investigate mitochondrial involvement on horse sperm motility and ROS production / Giaretta E.; Mislei B.; Martinez-Pastor F.; Nesci S.; Spinaci M.; Galeati G.; Nerozzi C.; Mari G.; Tamanini C.; Bucci D.. - In: RESEARCH IN VETERINARY SCIENCE. - ISSN 0034-5288. - STAMPA. - 147:(2022), pp. 12-19. [10.1016/j.rvsc.2022.03.017]
Giaretta E.; Mislei B.; Martinez-Pastor F.; Nesci S.; Spinaci M.; Galeati G.; Nerozzi C.; Mari G.; Tamanini C.; Bucci D.
File in questo prodotto:
File Dimensione Formato  
Postprint.docx

Open Access dal 26/03/2023

Tipo: Postprint
Licenza: Licenza per Accesso Aperto. Creative Commons Attribuzione - Non commerciale - Non opere derivate (CCBYNCND)
Dimensione 1.11 MB
Formato Microsoft Word XML
1.11 MB Microsoft Word XML Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/884094
Citazioni
  • ???jsp.display-item.citation.pmc??? 2
  • Scopus 4
  • ???jsp.display-item.citation.isi??? 4
social impact