Research lines
The main line of research aims to study the signalling mechanisms in sperm in vitro systems: training and cryopreservation, in addition to analyze its effect on the genital tract of the female and, finally, the definition of fertility biomarkers in the male.
In animals with internal fertilization, in various taxa/classes of animals, a male transfers to the female not only sperm but also a rather complex fluid known as seminal fluid or seminal plasma, which contains proteins, peptides, cytokines and RNA, among other components. Increasing data suggest that seminal fluid influences the survival of fertilizing sperm over various intervals by modulating the female's immune system to tolerate paternal antigens and, in mammals, toward tolerance of fetuses and placenta until term. The main objective of the research group is to determine the signaling functions of exosomes from seminal fluid/plasma and their load of coding and non-coding RNAs (with special attention to small RNAs) in Sus scrofa (pig), of proven fertility. For this purpose, the components of seminal plasma are analyzed and their possible application in the improvement of two in vitro processes, training and cryopreservation. Furthermore, the detailed study of the physiology of the changes that occur after training and cryopreservation, as well as the differential effect of the treatment of sperm groups in the epithelia of the female sperm storage sites (utero-tubal junction), are two of the secondary objectives of the research group. Furthermore, defining the effect of such changes in variations in the expression of genes related to the immune process could represent a basic mechanism that governs internal fertilization, thus guaranteeing successful reproductive performance. The knowledge generated will potentially help improve artificial reproduction techniques, the design of new semen extenders and considerations to modulate immunological processes in reproduction.
Methodologies and techniques
- Analysis of sperm motility and kinetic parameters, using the CASA system.
- Analysis by epifluorescence and/or flow cytometry of: membrane integrity, state of the acrosomal membrane, early changes of membrane destabilization, mitochondrial potential, oxidative stress indicators (ROS), lipid peroxidation, capacitation starus, etc.
- Protein analysis by Western Blotting or massive proteomics (LC-MS/MS).
- Analysis of coding and non-coding RNA, using RT-qPCR or RNA sequencing.
- Extraction, quantification and characterization of extracellular vesicles.
- Cryopreservation and in vitro handling of sperm.
