title: Selection of nonapoptotic sperm by magnetic-activated cell sorting in Senegalese sole (Solea senegalensis).
abstract: Senegalese sole (Solea senegalensis) is a promising species in aquaculture. However, owing to decreased sperm quality in F1 generations and the absence of courtship in those individuals born in captivity, artificial fertilization is being used to generate new progenies. The objective of this study was to implement a sperm selection method for nonapoptotic sperm subpopulation recovery before sperm cryopreservation. In particular, magnetic-activated cell sorting is used to eliminate apoptotic spermatozoa. This study represents the proof-of-concept for magnetic-activated cell sorting applicability in teleost species relevant in aquaculture. Apoptotic cell population was studied by flow cytometry using YO-PRO-1 and a caspase detection kit. Also, reactive oxygen species were measured in sperm samples. Our data demonstrated that caspase detection is more specific than YO-PRO-1 in the identification of apoptotic cells in S senegalensis seminal samples. The results showed that the percentage of apoptotic cells (caspase positive) was significantly higher (P = 0.04) in seminal samples from F1 than that from wild individuals. Magnetic-activated cell sorting removed a significant number of apoptotic cells from the samples (54% and 75% in wild and F1 individuals, respectively), decreasing the level of cells positive for reactive oxygen species (P = 0.17). In conclusion, this technique reduces the percentage of nonfunctional spermatozoa in a seminal sample before cryopreservation. This novel technique can be applied directly in the aquaculture industry.
authors.affiliation: Department of Molecular Biology, University of León, León, 24071, Spain; INDEGSAL, University of León, León, 24071, Spain.
authors.forename: D G
authors.initials: DG
authors.lastname: Valcarce
authors.affiliation: Department of Molecular Biology, University of León, León, 24071, Spain; INDEGSAL, University of León, León, 24071, Spain.
authors.forename: M P
authors.initials: MP
authors.lastname: Herráez
authors.affiliation: IEO, Spanish Institute of Oceanography, Planta de Cultivos el Bocal, Barrio Corbanera s/n Bocal, Monte, Santander 39012, Spain.
authors.forename: O
authors.initials: O
authors.lastname: Chereguini
authors.affiliation: IEO, Spanish Institute of Oceanography, Planta de Cultivos el Bocal, Barrio Corbanera s/n Bocal, Monte, Santander 39012, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Rodríguez
authors.affiliation: INDEGSAL, University of León, León, 24071, Spain; IEO, Spanish Institute of Oceanography, Planta de Cultivos el Bocal, Barrio Corbanera s/n Bocal, Monte, Santander 39012, Spain. Electronic address: robles.vanesa@gmail.com.
authors.forename: V
authors.initials: V
authors.lastname: Robles
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D015925:Cryopreservation / Q000662:veterinary; D005412:Flatfishes / Q000502:physiology*; D005434:Flow Cytometry / Q000379:methods / Q000662:veterinary*; D008280:Magnetics; D008297:Male; D017382:Reactive Oxygen Species; D013094:Spermatozoa / Q000502:physiology*
chemical_list: D017382:Reactive Oxygen Species
keywords: Caspases; MACS; Reactive oxygen species; Sole; Spermatozoa; YO-PRO-1