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Sperm capacitation is a critical process for fertilization. This work aims to analyze the effect capacitation had on the proteome and mitochondrial parameters of bull spermatozoa. Viability, mitochondrial membrane potential (MMP), and reactive oxygen species (mROS) were assessed by flow cytometry in noncapacitated (NC) and capacitated (IVC) sperm. Proteome was evaluated using SWATH-MS. capacitation significantly induced a decrease in sperm viability, a high MMP, and an increase in mROS production. Within the group of living spermatozoa, the capacitation significantly induced a decrease in healthy mitochondrial spermatozoa, as well as an increase in mROS production, without affecting the MMP intensity. A total number of 72 differentially abundant proteins were found of which 63 were over-represented in the NC sperm group and 9 in the IVC sperm group. It was observed that many proteins associated with the sperm membrane and acrosome were lost during the capacitation process. For the IVC sperm, the functional enrichment was found in proteins related to the oxidative phosphorylation process. Our results indicate that the capacitation process induces a significant loss of seminal plasma-derived membrane proteins and a significant increase in proteins related with the oxidative phosphorylation (OXPHOS) pathway. Data are available via ProteomeXchange with identifiers PXD056424 and PXD042286.
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http://dx.doi.org/10.1021/acs.jproteome.4c00910 | DOI Listing |
Theriogenology
August 2025
Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Complutense University of Madrid, Spain. Electronic address:
In vitro embryo production (IVP) in felid species faces difficulties derived from their conservation status, challenging reproductive traits and low success rates during in vitro maturation (IVM), fertilization (IVF) and culture (IVC). This study aimed to evaluate different IVC strategies for domestic cat IVP, as a model to improve assisted reproduction techniques (ARTs) in wild felids. Three IVC media were compared: (i) synthetic oviductal fluid (SOF) supplemented with fetal bovine serum ("FBS"); (ii): SOF supplemented first with bovine serum albumin (BSA), followed by FBS addition ("BSA-FBS"); and (iii): a commercial IVC human medium (IVC-CULT®) ("COM").
View Article and Find Full Text PDFReprod Fertil
July 2025
Laboratory of Animal Germplasm Conservation, Federal Rural University of Semi-Arid, UFERSA, Mossoro, Brazil.
Abstract: The objective was to evaluate the effects of different culture systems and the addition of fibroblast growth factor (bFGF) during in vitro culture (IVC) of testicular tissue fragments from prepubertal collared peccaries. Testes from five individuals were collected, dissected, and cultured for up to 56 days (34°C and 5% CO2) in Dulbecco's Modified Eagle's Medium (DMEM), supplemented or not with FGF at 10 ng/mL, in organotypic (ORG) or 3D system culture. Samples were evaluated every 14 days for histomorphology, cell viability, DNA integrity, and proliferative activity.
View Article and Find Full Text PDFMethods Mol Biol
July 2025
Division of Animal Sciences, University of Missouri, Columbia, MO, USA.
Sperm capacitation is the final sperm maturation step that occurs in vivo in the female oviduct, triggered in a response to ovulation. Physiologically, to achieve full fertilizing ability, sperm capacitation requires a series of events to occur. This includes ion fluxes, intracellular pH increase, signaling and metabolic pathway activation, membrane reorganization, cholesterol efflux, release of decapacitating factors from the sperm surface, and membrane protein redistribution, to name a few.
View Article and Find Full Text PDFVet Med Sci
May 2025
Department of Obstetrics and Gynecology, Faculty of Veterinary Medicine, Burdur Mehmet Akif Ersoy University, Burdur, Turkey.
Background: In vitro culture media play a crucial role in supporting the growth and development of bovine embryos. Despite advancements in media formulations, there is ongoing interest in optimising these media by supplementing them with compounds like L-carnitine to improve embryo development and cryopreservation outcomes. However, the effects of L-carnitine supplementation in commercial culture media remain unclear.
View Article and Find Full Text PDFJ Proteome Res
April 2025
Department of Cellular Biology, Functional Biology and Physical Anthropology, Universitat de València, Burjassot 46100, Spain.
Sperm capacitation is a critical process for fertilization. This work aims to analyze the effect capacitation had on the proteome and mitochondrial parameters of bull spermatozoa. Viability, mitochondrial membrane potential (MMP), and reactive oxygen species (mROS) were assessed by flow cytometry in noncapacitated (NC) and capacitated (IVC) sperm.
View Article and Find Full Text PDF