Computational flow cytometry reveals that cryopreservation induces spermptosis but subpopulations of spermatozoa may experience capacitation-like changes

[EN] The reduced lifespan of cryopreserved spermatozoa in the mare reproductive tract has been attributed to both capacitative and apoptotic changes. However, there is a lack of studies investigating both phenomena simultaneously. In order to improve our knowledge in this particular point, we studied in raw and frozen-thawed samples apoptotic and capacitative markers using a wide battery of test based in flow cytometry. Apoptotic markers evaluated were caspase 3 activity, externalization of phosphatidylserine (PS), and mitochondrial membrane potential. Markers of changes resembling capacitation were membrane fluidity, tyrosine phosphorylation, and intracellular sodium. Conventional and computational flow cytometry using nonlinear dimensionally reduction techniques (t-distributed stochastic neighbor embedding (t-SNE)) and automatic classification of cellular expression by nonlinear stochastic embedding (ACCENSE) were used. Most of the changes induced by cryopreservation were apoptotic, with increase in caspase 3 activation (P < 0.01), PS translocation to the outer membrane (P < 0.001), loss of mitochondrial membrane potential (P < 0.05), and increase in intracellular Na+ (P < 0.01). Average values of markers of capacitative changes were not affected by cryopreservation; however, the analysis of the phenotype of individual spermatozoa using computational flow cytometry revealed the presence of subpopulations of spermatozoa experiencing capacitative changes. For the first time advanced computational techniques were applied to the analysis of spermatozoa, and these techniques were able to disclose relevant information of the ejaculate that remained hidden using conventional flow cytometry.SIThe authors received financial support for this study from the Ministerio de Economía y Competitividad-FEDER, Madrid, Spain, grant AGL2013-43211-R, Junta de Extremadura-FEDER (GR 15029). PMM is supported by a pre-doctoral grant from the Ministerio de Educación, Cultura y Deporte, Madrid Spain FPU13/03991. COF is supported by a post-doctoral grant from the Ministerio de Economía y Competitividad “Juan de la Cierva” IJCI-2014-21671

Cirrosis hepática canina: Estudio clínico y anatomopatológico

En el presente trabajo se describe un caso clínico de cirrosis hepática de etiología desconocida en un perro adulto. En él se realiza un estudio de los hallazgos sintomáticos, analíticos y anatomopatológicos que nos han conducido a este diagnóstico.In the present work a clinical cause of hepatic cirrhosis of etiology unknow in the adult dog we have described. In it to realised a description about sintomatics, analytics and anatomopathologics features which determinated this diagnostic.peerReviewe

Estudio del espermatozoide en el macho de la colmena de «Apis mellifera L»

La abeja melífera es una especie zootécnica de importancia creciente en la cabaña ganadera, en la cual urge llevar a cabo un programa de selección y mejora de la raza ibérica, que, ante todo, exige dominar las técnicas reproductivas utilizadas en el resto de los animales domésticos. Los espermatozoides son producidos por el elemento masculino de la colmena, denominado zángano o macho, de cuerpo grueso y pesado, ojos voluminosos.The honey bee is a zootechnical species of growing importance in the livestock herd, in which it is urgent to carry out a selection and improvement program for the Iberian breed, which, above all, requires mastering the reproductive techniques used in the rest of the animals. domestic. The spermatozoa are produced by the male element of the hive, called the drone or male, with a thick and heavy body, voluminous eyes.peerReviewe