Hostname: page-component-586b7cd67f-t7fkt Total loading time: 0 Render date: 2024-11-30T20:15:56.895Z Has data issue: false hasContentIssue false
  • English
  • Français

Identification of sperm morphometric subpopulations in the canine ejaculate: do they reflect different subpopulations in sperm chromatin integrity?

Published online by Cambridge University Press:  01 August 2007

I. Núñez-Martinez ,
J.M. Moran  and
F.J. Peña
I. Núñez-Martinez
Affiliation:
Veterinary Teaching Hospital, Section of Reproduction and Obstetrics, Department of Herd Health and Medicine, Faculty of Veterinary Medicine, Avd de la Universidad s/n 10071 Cáceres, Spain
J.M. Moran
Affiliation:
Veterinary Teaching Hospital, Section of Reproduction and Obstetrics, Department of Herd Health and Medicine, Faculty of Veterinary Medicine, Avd de la Universidad s/n 10071 Cáceres, Spain
F.J. Peña*
Affiliation:
Veterinary Teaching Hospital, Section of Reproduction and Obstetrics, Department of Herd Health and Medicine, Faculty of Veterinary Medicine, Avd de la Universidad s/n 10071 Cáceres, Spain
*
All correspondence to: F.J. Peña, Section of Reproduction and Obstetrics, Department of Herd Health and Medicine, Faculty of Veterinary Medicine, Avd de la Universidad s/n 10071, Cáceres, Spain. e-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

A statistical approach using sequentially principal component analysis (PCA) clustering and discriminant analysis was developed to disclose morphometric sperm subpopulations. In addition, we used a similar approach to disclose subpopulations of spermatozoa with different degrees of DNA fragmentation. It is widely accepted that sperm morphology is a strong indicator of semen quality and since the sperm head mainly comprises the sperm DNA, it has been proposed that subtle changes in sperm head morphology may be related to abnormal DNA content. Semen from four mongrel dogs (five replicates per dog) were used to investigate DNA quality by means of the sperm chromatin structure assay (SCSA), and for computerized sperm morphometry (ASMA). Each sperm head was measured for nine primary parameters: head area (A), head perimeter (P), head length (L), head width (W), acrosome area (%), midpiece width (w), midpiece area (a), distance (d) between the major axes of the head and midpiece, angle (θ) of divergence of the midpiece from the head axis; and four parameters of head shape: FUN1 (L/W), FUN2 (4π A/P2), FUN3 ((L – W)/(L + W)) and FUN 4 (π LW/4A). The data matrix consisted of 2361 observations, (morphometric analysis on individual spermatozoa) and 63 815 observations for the DNA integrity. The PCA analysis revealed five variables with Eigen values over 1, representing more than 79% of the cumulative variance. The morphometric data revealed five sperm subpopulations, while the DNA data gave six subpopulations of spermatozoa with different DNA integrity. Significant differences were found in the percentage of spermatozoa falling in each cluster among dogs (p < 0.05). Linear regression models including sperm head shape factors 2, 3 and 4 predicted the amount of denatured DNA within each individual spermatozoon (p < 0.001). We conclude that the ASMA analysis can be considered a powerful tool to improve the spermiogram.

Keywords

ASMACluster analysisDNADogFlow cytometrySCSASperm subpopulations
Type
Research Article
Information
Zygote , Volume 15 , Issue 3 , August 2007 , pp. 257 - 266
Copyright
Copyright © Cambridge University Press 2007

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abaigar, T., Holt, W.W., Harrison, R.A.P. & del Barrio, G. 1999. Sperm subpopulations in boar (Sus scrofa) and gazelle (Gazela dama Mhor) semen as revealed by pattern analysis of computer assisted motility assessments. Biol. Reprod. 60, 32–4.CrossRefGoogle Scholar
Aziz, N., Fear, S., Taylor, C. & Kingsland Lewis-Jones, D.I. 1998 Human sperm head morphometric distribution and its influence on human fertility. Fertil. Steril. 70, 883–91.Google Scholar
Beatty, R.A. 1972. The genetics of size and shape of spermatozoa organelles. In Beatty, R.A., Gluecksson-Waelsch, S. eds. The Genetics of Spermatozoa. Bogtrykkerit Forum, Copenhagen Denmark. pp. 97–115.Google Scholar
Buendía, P., Soler, C., Paolicchi, F., Gago, G., Urquieta, B., Pérez Sánche, F. & Bustos Obregón, E. 2002. Morphometric characterization and classification of alpaca sperm heads using the sperm class analyzer® system. Theriogenology 57, 1207–18.Google Scholar
Burgoyne, P.S. 1975. Sperm phenotype and its relationship to somatic and germ line genotype: a study using mouse aggregation chimeras. Dev. Biol. 44, 63–76.CrossRefGoogle ScholarPubMed
Chantler, E., Abraham Peskir, J. & Roberts, C. 2004. Consistent presence of two normally distributed sperm subpopulations within normozoospermic human semen: a kinematic study . Int. J. Androl. 27, 350–9.Google Scholar
Dadoune, J.P. 2003. Expression of mammalian spermatozoal nucleoproteins. Micros. Res. Techn. 61, 56–75.CrossRefGoogle ScholarPubMed
Dalhbom, M., Anderson, M., Vierula, M. & Alanko, M. 1997. Morphometry of normal and teratospermic canine sperm heads using an image analyzer: work in progress. Theriogenology 48, 687–9.Google Scholar
den Dass, N . 1992. Laboratory assessment of semen characteristics. Anim. Reprod. Sci. 28, 87–94.Google Scholar
ESHRE Andrology special interest group 1998. Guidelines on the application of CASA technology in the analysis of spermatozoa. Human Reprod. 13, 142–5.Google Scholar
Evenson, D.P., Darzynkiewicz, Z. & Melamed, M.R. 1980 Relation of mammalian sperm chromatin heterogeneity to fertility. Science 210, 1131–3.CrossRefGoogle ScholarPubMed
Evenson, D. & Jost, L. 2000 Sperm chromatin structure assay is useful for fertility assessment. Meth. Cell. Sci. 22, 169–89.Google Scholar
Evenson, D.P., Larson, K.J. & Kost, L.K. 2002. The sperm chromatin structure assay (SCSA)™: clinical use for detecting sperm DNA fragmentation related to male infertility and comparisons with other techniques. J. Androl. 23, 25–43.CrossRefGoogle Scholar
Hair, J.F., Anderson, R.E., Tatham, R.L. & Black, W.C. 1998. Multivariate Data Analysis. 5th edn. Prentice Hall Int., New Jersey.Google Scholar
Hinst, O., Blottner, S. & Franz, C. 1995. Chromatin condensation in cat spermatozoa during epididymal transit as studied by aniline blue and acridine orange staining. Andrologia 27, 275–9.Google Scholar
Hirai, M., Boersma, A., Hoeflich, A., Wolf, E., Föll, J., Aumuller, R., Braum, J. 1997. Objectively measured sperm motility and sperm head morphometry in boars (Sus scrofa): relation to fertility and seminal plasma growth factors. J. Androl. 18, 312–23.Google Scholar
Januskauskas, A., Johannisson, A. & Rodriguez Martínez, H. 2001. Assessment of sperm quality through fluorometry and sperm chromatin structure assay in relation to field fertility of frozen–thawed semen from Swedish AI bulls. Theriogenology 55, 947–61.Google Scholar
Karabinus, D.K., Vogler, C.J., Saacke, R.G. & Evenson, D.P. 1997. Chromatin structural changes in sperm after scrotal insulation in Holstein bulls. J. Androl. 18, 549–55.CrossRefGoogle ScholarPubMed
Lewis-Jones, I., Aziz, N., Seshadri, S., Douglas, A. & Howard, P. 2003. Sperm acrosomal abnormalities are linked to sperm morphologic deformities. Fertil. Steril. 79, 212–5.CrossRefGoogle Scholar
Martínez Pastor, F., García Macías, V., Alvarez, M., Herráez, P., Anel, L. & de Paz, P. 2005. Sperm subpopulations in Iberian red deer epididymal sperm and their changes through the conservation process. Biol. Reprod. 72, 316–27.CrossRefGoogle Scholar
McLay, D.W. & Clarke, H.J. 2003. Remodelling the paternal chromatin at fertilization in mammals. Reproduction 125, 625–33.CrossRefGoogle ScholarPubMed
Mujica, A., Navarro García, F., Hernández González, E.O. & Juarez-Mosqueda, M.L. 2003. Perinuclear theca during spermatozoa maturation leading to fertilization. Microscopy Research and Technique 61, 76–87.Google Scholar
Osteremeier, G.C., Sargeant, G.A., Yandell, B.S., Evenson, D.P. & Parrish, J.J. 2001. Relationship of bull fertility to sperm nuclear shape. J. Androl. 22, 595–603.CrossRefGoogle Scholar
Pena, A., Johannisson, A. & Linde Forsberg, C. 1999. Post-thaw evaluation of dog spermatozoa using a new triple fluorescent staining and flow cytometry. Theriogenology 52, 965–80.Google Scholar
Peña, A.I., Johannisson, A. & Linde Forsberg, C. 2001 Validation of flow cytometry for assessment of viability and acrosomal integrity of dog spermatozoa and for evaluation of different methods of cryopreservation. J. Reprod. Fertil. 57 (Suppl.), 371–6.Google ScholarPubMed
Peña Martinez, A.I. 2004, Canine fresh and cryopreserved semen evaluation, Anim. Reprod. Sci. 82–83, 209–24.CrossRefGoogle Scholar
Peña, F.J., Saravia, F., García Herreros, M., Núñez Martínez, I., Tapia, J.A., Johanisson, A., Wallgren, M. & Rodríguez Martínez, H. 2005. Identification of sperm morphometric subpopulations in two different portions of the ejaculate and its relation to post thaw quality. J. Androl. 26, 716–23.CrossRefGoogle Scholar
Phillips, N.J., MacGowan, M.R., Johnston, S.D. & Mayer, D.G. 2004. Relationship between thirty post-thaw spermatozoal characteristics and the field fertility of 11 high use Australian dairy AI sires. Anim. Reprod. Sci. 81, 47–61.Google Scholar
Quintero-Moreno, A., Miró, J., Rigau, T. & Rodríguez Gil, J.E. 2003. Identification of sperm subpopulations with specific motility characteristics in stallion ejaculates. Theriogenology 59, 1973–90.CrossRefGoogle ScholarPubMed
Rathi, R., Colenbrander, B., Bevers, B.B. & Gadella, B.M. 2001. Evaluation of in vitro capacitation of stallion spermatozoa. Biol. Reprod. 65, 462–70.CrossRefGoogle ScholarPubMed
Rijsselaere, T., Van Soom, A., Hoflack, G., Maes, D. & de Kruif, A. 2004. Automated sperm morphometry and morphology analysis of canine semen by the Hamilton Thorne Analyser. Theriogenology 62, 1292–1306.Google Scholar
Rodriguez Martinez, H. 2003. Laboratory semen assessment and prediction of fertility: still utopia? Reprod. Dom. Anim. 38, 312–8.Google Scholar
Roldan, E.R.S., Cassinello, J., Abaigar, T. & Gomendio, M. 1998. Inbreeding, fluctuating asymmetry and ejaculate quality in an endangered ungulate. Proc. R. Soc. Lond. B. Biol. Sci. 265, 243–8.CrossRefGoogle Scholar
Said, T.M., Aziz, N., Sharma, R.K., Lewis-Jones, I., Thomas, A. & Agarwal, A. 2001. Novel association between the sperm deformity index and oxidative stress induced DNA damage in infertile male patients. Asian J. Androl. 7, 121–6.CrossRefGoogle Scholar
Soler, C., de Monserrat, J.J., Gutierrez, R., Nuñez, J., Sancho, M., Perez-Sánchez, F. & Cooper, T.G. 2003. Use of the sperm class analyser for objective assessment of human sperm morphology. Int. J. Androl. 26, 262–70.CrossRefGoogle ScholarPubMed
Thurston, L.M., Watson, P.F., Mileham, A.J. & Holt, W.V. 2001 Morphological sperm subpopulations defined fourier shape descriptors in fresh ejaculates correlate with variation in boar semen quality following cryopreservation. J. Androl. 22, 382–94.Google Scholar