Sexado de espermatozoides en ganado lechero (Holstein) mediante microscopia confocal y tinción DAPI
Palabras clave:
Preselección del sexo, Holstein, microscopía confocal, DAPI, K mediasResumen
La habilidad de preseleccionar el sexo de los animales se ha convertido el principal objetivo en la industria ganadera. Actualmente, el único método efectivo para separar espermatozoides en las fracciones portadoras del cromosoma –X y –Y es mediante Citometría de flujo. Sin embargo, su elevado costo y baja disponibilidad en laboratorios ha fomentado la búsqueda de nuevos métodos de sexado. En este estudio se utilizaron muestras de semen de toro Holstein y se evaluó mediante microscopía confocal, las medidas morfométricas de la cabeza espermática y la diferencia relativa del contenido de ADN a través de mediciones en el tamaño del núcleo celular teñido con DAPI en semen sin sexar y semen –X sexado por Citometría de flujo. Se realizó un pronóstico del sexo de los espermatozoides en una muestra no sexada, se compararon los datos entre las diferentes muestras por medio del estadístico conglomerado de K medias con el fin de diferenciar las subpoblaciones espermáticas. Los resultados muestran que mediante las mediciones del tamaño del núcleo es posible identificar el sexo del espermatozoide en una muestra de semen no sexado, siendo lo opuesto con las medidas morfométricas de la cabeza espermática.Descargas
Citas
Barrientos, M., Juárez, M.L., Trujillo, M.E., y Montiel, F. (2009). Alteraciones en la integridad del acrosoma y de la teca perinuclear en semen criopreservado de verraco. Zoot. Trop. 27(1): 17-24.
Betancourt,M; Fierro,R; Ambriz,D. 1993. In vitro fertilization of pig oocytes matured in vitro.Theriogenology.40:1115-1160.
Bouvet R; Paparella C;Feldman R.(2007).Efecto del tabaquismo sobre la espermatogénesis en hombres con infertilidad idiopática. Arch. Esp. Urol. 60(3).ISSN 0004-0614.
Catt,J.W; Rhodes, S.(1995).Comparative intracytoplasmic sperm injection (ICSI) in human and domestic animals. Reprod.Fértil.Dev.7:161-167.
Catt,J.W.(1996).Intracytoplasmic sperm injection (ICSI) and related technology.Anim.Reprod.Sci.42:239-250.
Chen, S.H and Seidel, G, E. (1997). Effects of oocytes activation and treatment of spermatozoa on embyonic development following intracytoplasmic sperm injection in cattle. Theriogenology.48:1265-1273.
Choi, Y.H; Love, C.C;Love, L.B; Varner, D.D; Brinsko, S; Hinrichs,K. (2002).
Developmental competence in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection with fresh of frozen-thaved spermatozoa.Reprod. 123:455-465.
Colley, A.; Buhr, M.; y Golovan, S.; (2008). Single bovine sperm sex typing by amelogenin nested PCR. Theriogenology, 70, 978–983. http://doi.org/10.1016/j.theriogenology.2008.05.060.
Council of Europe, Parliamentary Assembly. Resolution 1829: sex selection, 2011 http://assembly.coe.int/Main.asp?link=/Documents/ AdoptedText/ta11/ERES1829.htm (3 March 2013, date last accessed).
Cui, K.H. (1997). Size differences between human X and Y spermatozoa and prefertilization diagnosis. Mol Hum Reprod 3: 61–67.
Dahl E, Beutel M, Brosig B, Gru¨ssner S, Sto¨bel-Richter Y, Tinneberg HR, Bra¨hler E. Social sex selection and the balance of the sexes: empirical evidence from Germany, the UK, and the US. J Assist Reprod Genet 2006;23:311 – 318.
De Geyter C, Sterthaus O, Miny P, Wenzel F, Lapaire O, De Geyter M, Sartorius G. (2013).First successful pregnancy in Switzerland after prospective sex determination of the embryo through the separation of X-chromosome bearing spermatozoa. Swiss Med Wkly; 143:w13718.
Estrada, E; Uribe, M. (2002) Atlas de histología de vertebrados. Universidad Nacional Autónoma de México.México, D.F.227p.
Fugger, E.F., Black, S.H., Keyvanfar, K., Schulman, J.D., 1998. Births of normal daughters after MicroSort sperm separation and intra-uterine insemination, in vitro fertilization, or intracytoplasmic sperm injection. Hum. Reprod. 13 _9., 2367–2370.
Gajda,B.(2009).Factors and methods of pig oocyte and embryo quality improvement and their application in reproductive biotechnology. Reprod.Biol.9 (2):97-112.
Garcia-Roselló; Matás,C; Cánovas, S;Moreira, P.N.; Gardea,J; Coy,P.(2006).Influence of sperm pretreatment on the efficiency of intracytoplasmic sperm injection in pigs.J.Androl.27:268-275.
Garner, D.L., Gledhill, B.L., Pinkel, D., Lake, S., Stephenson, D., Van Dilla, M.A. y Johnson, L.A. (1983). Quantification of the X- and Y-chromosome-bearing spermatozoa of domestic animals by flow cytometry. Biology of Reproduction 28 312–321. (doi:10.1095/biolreprod28.2.312).
Garner, D. L., y Seidel, G. E. (2008). History of commercializing sexed semen for cattle. Theriogenology, 69 (7), 886–895. http://doi.org/10.1016/j.theriogenology.2008.01.006
Garner DL. Hoechst 33342: The dye that enabled differentiation of living X- and Y-chromosome bearing mammalian sperm. Theriogenology 2009; 71:11–21.
Gilbert.S.(2005).Biología del desarrollo.7ma edición. Editorial Médica Panamericana. Buenos Aires, Argentina.889p.
Gledhill, B.L., Lake, S., Steinmetz, L.L., Gray, J.W., Crawford, J.R., Dean, P.N., Van Dilla, M.A., (1976). Flow microfluorometric analysis of sperm DNA content: effect of cell shape on the fluorescence distribution. J. Cell. Physiol. 87, 367–376.
Gómez J. (2005).Estudio de las alteraciones morfológicas de espermatozoides humanos con microscopía electrónica de barrido (SEM). Revista latinoamericana de fertilidad.22 (1).P.58-66.
Gómez,M.C; Catt,J.W; Evans, G; Maxwell,W.M.C.(1997).Effect of culture, incubation and acrosome reaction of fresh and frozen-thawed ram spermatozoa for in vitro fertilization and intracytoplasmic sperm injection. Reprod.Fertil. Dev.9:665-673.
Gómez, M.C; Catt, J.W; Evans, G; Maxwell,M.C.(1998).Sheep oocyte activation after intracytoplasmic sperm injection (ICSI). Reprod.Fertil.Dev.10:197-205.
González, J; Lailla, J; Fabre, E; González, E(2006) Obstetricia.5ta edición. MASSON. Barcelona, España.879p.
Goto,K; Kinoshita,A;Takuma,Y; Ogawa, K.(1990).Fertlization of bovine oocytes by the injection of inmovilized killed spermatozoa. Vet. Rec.127:517-520.
Guerino, G.; Filho, M.; Vasconcellos-Sala, R., y Ferreira-Mendanha, M. (2013). Success in Artificial Insemination - Quality of Semen and Diagnostics Employed. InTech, 39–61. http://doi.org/10.5772/45731.
Guilmoto CZ. The sex ratio transition in Asia. Popul Dev Rev 2009; 35:519 – 549
Haigo, K; Yamauchi, Y; Yazama, F; Yanagimachi, R; Horiuychi,T.( 2004). Full-term development of hámster embryos produced by injection of round spermatids into oocytes.Biol.Reprod.71:194-198.
Horiuchi, T.(2006).Application study of intracytoplasmic sperm injection for golden hasmter and cattle production.J.Reprod.Dev.52:13-21.
Hosoi,Y; Iritari,A. (1993).Rabbit microfertIlisation.Mol.Reprod.Dev.36:282-284.
Hvistendahl M. Unnatural Selection. Choosing Boys over Girls, and the Consequences of a World Full of Men. New York: Public affairs, 2011.
Johnson, L.A., 1992. Gender preselection in domestic animals using flow cytometrically sorted sperm. J. Anim. Sci. 70 _ Suppl. 2., 8–18.
Johnson, L.A., Welch, G.R., Keyvanfar, K., Dorfmann, A., Fugger, E.F., Schulman, J.D., 1993. Gender preselection in humans? Flow cytometric separation of X- and Y- spermatozoa for the prevention of X-linked diseases. Human Reprod. 8 _10., 1733–1739.
Johnson, L.A., Welch, G.R., Rens, W., 1999. The Beltsville sperm sexing technology: high-speed sorting gives improved sperm output for IVF and AI. J. Anim. Sci. 77 _Suppl. 2., 213–220.
Johnson, L.A., Guthrie, H.D., Fiser, P., Maxwell, W.M.C., Welch, G.R., Garrett, W.M., 2000. Cryopreservation of flow cytometrically sorted boar sperm: effects on in vivo embryo development. J. Anim. Sci. 78 _Suppl. 1., Abstr., in press.
Johnson, L. A.; Cran, D. G.; y Polge, C. (1994). Recent advances in sex preselection of cattle: Flow cytometric sorting of X- & Y-chromosome bearing sperm based on dna to produce progeny. Theriogenology, 41, 51–56.
Johnson, L.A., Flook, J.P., Look, M.V., 1987. Flow cytometry of X- and Y-chromosome-bearing sperm for DNA using an improved preparation method and staining with Hoechst 33342. Gamete Res. 17, 203–212.
Karabinus DS.(2009). Flow cytometric sorting of human sperm: MicroSort clinical trial update. Theriogenology; 71:74 – 79.
Katayama,M;Koshida,M;Miyake,M.(2002).Fate of the acrosome in ooplasm in pigs after IVF and ICSI.Hum.Reprod.17:2657-2664.
Keskintepe, L; Brackett, B.G.(2000).Cryopreservation of bovine blastocysts obtained by intracytoplasmic sperm injection. Theriogenology.53:1041-1052.
Kimura, Y; Yanagimachi,R.(1995).Intracytoplasmic sperm injection in the mouse.Biol.Reprod.52:709-720.
Kolbe, T; Holtz,W.(2000).Birth of a piglet derived from an oocyte fertilizad by intracytoplasmic sperm injection (ICSI).Anim.Reprod.Sci.64:97-101.
Kwon,I.K; Park,K.E; Niwa,K.(2004).Activation pronuclear formation and development in vitro of pig oocytes following intracytoplasmic injection of freeze-dried spermatozoa.Biol.Reprod.71:1430-1436.
Kyung-Bon, L; Koji, N.(2006). Fertilization and development in vitro of bovine oocytes following intracytoplasmic injection of head-dried sperm heads. Biol.Reprod.74:146-152.
Lee,J.W; Tian, X.C; Yang, X.(2003).Failure of male pronucleus formation is the major cause of lack of fertilization and embryo development in pig oocytes subjected to intracytoplasmic injection. Biol.Reprod.68:1341-1347.
Lee.J.W; Niwa,K.(2006).Fertilization and development in vitro of bovine oocytes following intracytoplasmic injection of head-dried sperm heads. Biol.Reprod.74:146-152.
Martin,M.J.(2000).Development of in vivo-matured porcine oocytes following intracytoplasmic sperm injection.Biol.Reprod.63:109-112.
Moruzzi, J.F.,(1979). Selecting a mamalian species for the separation of X- and Y-chromosome-bearing spermatozoa. J. Reprod. Fertil. 57, 319–323.
Nakai,M; Kashiwazaki,N; Takizawa, A;Maedomari,N; Ozawa, M; Noguchi,J; Kaneko, H; Shino, M; Kikuchi,K.(2007).Effects of chelating agents during freeze-drying of boar spermatozoa on DNA fragmentation and developmental ability in vitro and in vivo after intracytoplasmic sperm head injection. Zygote.15:15-24.
Oguna, A; Ogunuki, N; Inoue, K; Mochida, K.(2003).New microinsemination techniques for laboratory animals. Theriogenology.59:87-94.
Oikawa, T; Takada, N; Kikuchi,T;Numabe, T;Takenaka,M; Horiuchi,T. (2005).Evaluation of activation treatments for blastocyst production and birth of viable calves following bovine intracytoplasmic sperm injection. Anim. Reprod.Sci.86:187-194.
Palermo,G; Hubert, J; Devroey,P; Van Steirteghem,A.(1992).Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte. Lancet.340:17-18.
Patricio,P; Tucker,M.J.;Guelman,V.(2003).Atlas de reproducción asistida. Ed.Mc Graw Hill. México, Distrito Federal.pp.81
Pennings G. Personal desires of patients and social obligations of geneticists:
applying preimplantation genetic diagnosis for non-medical sex selection. Prenat Diagn 2002;22: 1123 – 1129.
Pinkel, D., Gledhill, B.L., Van Dilla, M.A., Stephenson, D., Watchmaker, G., (1982). High resolution DNA measurements of mammalian sperm. Cytometry 3, 1–9.
Prasad, S.; Rangasamy, S.; Y Satheshkumar, S. (2010). Sex Preselection in Domestic Animals - Current status and Future prospects, 3(7), 346–348.
Pope,C.E; Johnston,C.A; McRae,M; Keller, G.L.(1998). Development of embryos produced by intracytoplasmic sperm injection of cat oocytes. Anim.Reprod.Sci.53:221-236.
Rath, D., Long, C.R., Dobrinsky, J.R., Welch, G.R., Schreier, L.L., Johnson, L.A., 1999. In vitro production of sexed embryos for gender preselection: high speed sorting of X-chromosome-bearing sperm to produce piglets after embryo transfer. J. Anim. Sci. 77, 3346–3352.
Rath, D., Barcikowski, S., Graaf, S. De, Garrels, W., Grossfeld, R., Klein, S., y Washausen, S. (2013). Sex selection of sperm in farm animals: Status report and developmental prospects. Reproduction, 145(1). http://doi.org/10.1530/REP-12-0151.
Rho,G.J; Kawarsky, S; Jhonson, W.H.; Kochlar,K; Betteridge,K.J.(1998). Sperm and oocyte treatments to improve the formation of male and female pronuclei and subsequent development following intracytoplasmic sperm injection bovine oocytes. Biol.Reprod.59:918-924.
Santamaría, E. (2013) Entender y combatir la progresión de la distrofia muscular de Duchenne en modelos animales. Universitat Pompeu Fabra, Barcelona, España.
Seidel, G.E. Jr., Schenk, J.L., Herickoff, L.A., Doyle, S.P., Brink, Z., Green, R.D., Cran, D.G., 1999. Insemination of heifers with sexed sperm. Theriogenology 52, 1407–1420.
Seidel, G. E. (2007). Overview of sexing sperm. Theriogenology, 68(3), 443–446. http://doi.org/10.1016/j.theriogenology.2007.04.005.
Sharpe, J. C., y Evans, K. M. (2009). Advances in flow cytometry for sperm sexing. Theriogenology, 71, 4–10. http://doi.org/10.1016/j.theriogenology.2008.09.021.
Shenfield F, Pennin.gs G, Devroey P, Sureau C, Tarlatzis B, Cohen J. (2003).ESHRE Ethics Task Force. Taskforce 5: preimplantation genetic diagnosis. Hum Reprod 18:649–651.
Shirazi, A; Ostad-Hosseini,S; Ahmadi,E; Heidari,B; Shams-Esfandabadi, N. (2009). In vitro developmental competence of ICSI-derived activated ovine embryos.Theriogenology.71:342-348.
Strasinger,S; Schaub, M.(2010). Análisis de orina y líquidos corporales.5ta edición. Editorial Medica Panamericana. Buenos Aires, Argentina.301p.
Squires,E.L; Wilson,J.M; Kato,H;Blaszczyk,A.(1996).A pregnancy after intracytoplasmic sperm injection into equine oocytes matured in vitro. Theriogenology.45:306.
Urbina M; Lerner J.(2008).Fertilidad y reproducción asistida, Editorial Médica Panamericana.Caracas,Venezuela.625p.
Universidad de Navarra (2012) Enfermedades ligadas al sexo. Facultades de ciencias. Departamento de ciencias .Madrid, España.
Van Balen F. Attitudes towards sex selection in the Western world. Prenat Diagn 2006;26:614 – 618.
Van Munster, E. B., Stap, J., Hoebe, R. Te Meerman, G. J., y Aten, J. (1999). Difference in volume of X- and Y-chromosome-bearing bovine sperm heads matches difference in DNA content. Cytometry, 35, 125–128.http://doi.org/10.1002/(SICI)1097-0320(19990201)35:2<125::AID-CYTO3>3.0.CO;2-H
Vázquez, C; Martin, A; Siloniz, I; Serrano, S (2010) Técnicas básicas de Microbiología observación de bacterias. Universidad Complutense, Madrid, España.
Wang, B; Baldessare,H; Pierson, J; Cote, F; Rao,K.M; Karatzas,C.N. (2003).The in vitro and in vivo development of goat embryos produced by intracytoplasmic sperm injection using tail-cut spermatozoa. Zigote.11:219-227.
Wani,N.A.(2002).In vitro maturation and in vitro fertilization of sheep oocytes, Small Rum.Res.44:89-95.
Welch, G. R., y Johnson, L. (1999). Sex preselection: Laboratory validation of the sperm sex ratio of flow sorted X- and Y-sperm by sort reanalysis for DNA. Theriogenology, 52(8), 1343–1352. http://doi.org/10.1016/S0093-691X(99)00221-6.
WHO. Preventing gender-biased sex selection. Geneva: World Health Organization,2011.http://whqlibdoc.who.int/publications/2011/9789241501460_eng.pdf (3 March 2013, date last accessed).
Yamauchi, Y; Yanagimachi, R; Horiuchi, T.(2002).Full term development of golden hámster oocytes following intracytoplasmic sperm head injection.Biol. Reprod.67:534-539.
Publicado
Cómo citar
Número
Sección
Licencia
Todos los contenidos de CULCYT se distribuyen bajo una licencia de uso y distribución “Creative Commons Reconocimiento-No Comercial 4.0 Internacional” (CC-BY-NC). Puede consultar desde aquí la versión informativa de la licencia.
Los autores/as que soliciten publicar en esta revista, aceptan los términos siguientes: a) los/las autores/as conservarán sus derechos de autor y garantizarán a la revista el derecho de primera publicación de su obra; y b) se permite y recomienda a los/las autores/as agregar enlaces de sus artículos en CULCYT en la página web de su institución o en la personal, debido a que ello puede generar intercambios interesantes y aumentar las citas de su obra publicada.
