Head area measurements of dead, live, X- and Y-bearing bovine spermatozoa
T. Révay A E , S. Nagy B , A. Kovács B , M. E. Edvi A , A. Hidas A , W. Rens C and I. Gustavsson DA Institute for Small Animal Research, Godollo, H-2100, POB 417, Hungary.
B Research Institute for Animal Breeding and Nutrition, Department of Cell Biology, H-2053 Herceghalom, Gesztenyes u. 1, Hungary.
C University of Cambridge, Department of Clinical Veterinary Medicine, Madingley Road, Cambridge CB3 OES, UK.
D Swedish University of Agricultural Sciences, Department of Animal Breeding and Genetics, Centre of Reproductive Biology, Uppsala, Sweden.
E To whom correspondence should be addressed. email: revay@katki.hu
Reproduction, Fertility and Development 16(7) 681-687 https://doi.org/10.1071/RD04013
Submitted: 25 February 2004 Accepted: 20 September 2004 Published: 9 December 2004
Abstract
The head area of bull spermatozoa was measured after viability and acrosome staining using trypan blue and Giemsa stains, followed by X- and Y-chromosome-specific fluorescence in situ hybridisation (FISH). The former staining made possible the categorisation of cells according to morphology and membrane integrity, whereas the latter allowed distinction of spermatozoa bearing X- and Y-chromosomes. Individual spermatozoa could be followed during the consecutive steps of staining, measurement and FISH. Using a high-resolution digital imaging system and measurement software, the head area of more than 3000 cells of five bulls was determined precisely. In all bulls, morphologically normal, viable cells with intact acrosomes were significantly smaller than dead cells with damaged acrosomes. No significant difference in the head area between X- and Y-chromosome-bearing viable, acrosome-intact spermatozoa was found in individual bulls. However, significant between-bull differences were detected in all cell categories.
Bahr, G. F. , and Zeitler, E. (1964). Study of bull spermatozoa. J. Cell Biol. 21, 175–189.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Bland, J. M. , and Altman, D. G. (1986). Statistical methods for assessing agreement between two methods of clinical measurement. Lancet 1, 307–310.
| PubMed |
McEvoy, J. D. (1992). Alteration of the sex ratio. Anim. Breed. Abst. 60, 97–111.
Nagy, Sz. , Hazas, G. , Bali Papp, A. , Ivancsics, J. , Szasz, F. , Szasz, F. , Kovacs, A. , and Foote, R. H. (1999). Evaluation of sperm tail membrane integrity by light microscopy. Theriogenology 52, 1153–1159.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Rens, W. , Welch, G. R. , and Johnson, L. A. (1999). Improved flow cytometric sorting of X- and Y-chromosome bearing sperm: substantial increase in yield of sexed semen. Mol. Reprod. Dev. 52, 50–56.
| Crossref | GoogleScholarGoogle Scholar |
Rens, W. , Yang, F. , Welch, S. , Revell, S. , O’Brien, P. C. M. , Solanky, N. , Johnson, L. A. , and Ferguson Smith, M. A. (2001). An X-Y paint set and sperm FISH protocol that can be used for validation of cattle sperm separation procedures. Reproduction 121, 541–546.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Revay, T. , Kovacs, A. , Rens, W. , and Gustavsson, I. (2002). Simultaneous detection of viability and sex of bovine spermatozoa. Reprod. Fertil. Dev. 14, 373–376.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Seidel, G. E. (1999). Sexing mammalian sperm and embryos – state of the art. J. Reprod. Fertil. Suppl. 54, 475–485.
Seidel, G. E. , and Johnson, L. A. (1999). Sexing mammalian sperm – overview. Theriogenology 52, 1267–1272.
| Crossref | GoogleScholarGoogle Scholar |
van Duijn, C. (1960). Mensuration of the heads of bull spermatozoa. Mikroskopie 14, 265–276.
| PubMed |
van Duijn, C. (1975). Mensuration of spermatozoa. Bibliogr. Reprod. 25, 121–128.
van Duijn, C. , and van Voorst, C. (1971). Precision measurements of dimensions, refractive index and mass of bull spermatozoa in the living state. Mikroskopie 27, 142–167.
| PubMed |
Welch, G. R. , and Johnson, L. A. (1999). Sex preselection: laboratory validation of the sperm sex ratio of flow sorted X- and Y-sperm by sort reanalysis for DNA. Theriogenology 52, 1343–1352.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
