Modification of equine sperm chromatin decondensation method to use fluorescence in situ hybridization (FISH).

The aim of this study was a cytogenetic analysis of stallions semen to find sex chromosome aberrations and to determine if there was an association between stallion's age and aberration frequency for the sex chromosomes. Sperm samples were collected from 22 stallions of various age from 3 to 23 years. Multicolour FISH was performed on each sample, using probes for the sex chromosomes and EGFR gene, localized on 4p12 in domestic horse. A total of 26199 sperm cells were analysed (from 1 070 to 1 532 per animal). Among the analysed cells, there were 50.318% with X chromosome, 48.543% with Y chromosome and 1.139% with aberrant chromosomes. The frequency of aberrations was: sex chromosomes nullisomy (0.466%), XY aneuploidy (0.454%), XX disomy (0.146%), YY disomy (0.041%), diploidy (0.024%) and trisomy XXY (0.008%). Additionally there was a correlation between the age of an animal and the frequency of sex chromosome aberration and a significant positive correlation between age and disomy of XY, XX, YY, trisomy of XXY, autosomal disomy was seen. A Correlation between the age of a stallion and the level of nullisomy was negative. The present study demonstrated that FISH technique is a powerful method to identify sex chromosome aberrations in equine spermatozoa and might be very helpful for a breeder during a selection for the best stallion.

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“…The spermatozoa were decondensed according to Blottner et al. (1990) with some modifications (Bugno‐Poniewierska et al. 2009).…”

Section: Methodsmentioning

confidence: 99%

“…The spermatozoa were decondensed according to Blottner et al (1990) with some modifications (Bugno-Poniewierska et al 2009). After thawing, the sperm cells were washed in 0.9% NaCl, spun down and diluted in 0.5 ml of 0.9% NaCl.…”

Section: Methodsmentioning

confidence: 99%

Determination of the Correlation Between Stallion’s Age and Number of Sex Chromosome Aberrations in Spermatozoa

Bugno‐Poniewierska

Kozub

Pawlina

et al. 2011

Reprod Domestic Animals

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“…The first ECAY-specific whole-chromosome painting probes were generated by chromosome microdissection (Raudsepp and Chowdhary, 1999;Kubickova et al, 2002) and used for quick identification of ECAY in cytogenetic preparations (Kubickova et al, 2002), for sperm FISH to detect ECAY aneuploidy (Bugno-Poniewierska et al, 2009), and for horse-donkey Zoo-FISH (Raudsepp and Chowdhary, 1999). Typically, only 10-20 copies of ECAY are microdissected and amplified by DOP-PCR (Telenius et al, 1992).…”

Section: Molecular Probesmentioning

confidence: 99%

The Y‐Chromosome

2013

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“…Sperm‐FISH has also been optimized for stallions (Bugno‐Poniewierska et al. ) and used for the study of sex chromosome aneuploidies in the sperm of reproductively normal stallions (Bugno et al. ) and for evaluating correlation between stallion age and the rate of sperm chromosome abnormalities (Bugno‐Poniewierska et al.…”

Section: Introductionmentioning

confidence: 99%

Fertility Assessment in Sorraia Stallions by Sperm‐Fish and Fkbp6 Genotyping

Kjöllerström

Oom

Chowdhary

et al. 2016

Reprod Domestic Animals

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The Sorraia, a critically endangered indigenous Iberian horse breed, is characterized by low genetic variability, high rate of inbreeding, bad sperm quality and subfertility. Here, we studied 11 phenotypically normal but subfertile Sorraia stallions by karyotyping, sex chromosome sperm-FISH and molecular analysis of FKBP6 - a susceptibility locus for impaired acrosome reaction (IAR). The stallions had normal sperm concentration (>300 million cells/ml), but the numbers of progressively motile sperm (21%) and morphologically normal sperm (28%) were invariably low. All stallions had a normal 64,XY karyotype. The majority of sperm (89%) had normal haploid sex chromosome content, although 11% of sperm carried various sex chromosome aneuploidies. No correlation was found between the percentage of sperm sex chromosome abnormalities and inbreeding, sperm morphology or stallion age. Direct sequencing of FKBP6 exon 4 for SNPs g.11040315G>A and g.11040379C>A revealed that none of the stallions had the susceptibility genotype (A/A-A/A) for IAR. Instead, all animals had a G/G-A/A genotype - a testimony of low genetic variability. The findings ruled out chromosomal abnormalities and genetic predisposition for IAR as contributing factors for subfertility. However, low fertility of the Sorraia stallions could be partly attributed to relatively higher rate of sex chromosome aneuploidies in the sperm.

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Modification of equine sperm chromatin decondensation method to use fluorescence in situ hybridization (FISH).

Cited by 11 publications

References 15 publications

Determination of the Correlation Between Stallion’s Age and Number of Sex Chromosome Aberrations in Spermatozoa

Determination of the Correlation Between Stallion’s Age and Number of Sex Chromosome Aberrations in Spermatozoa

The Y‐Chromosome

Fertility Assessment in Sorraia Stallions by Sperm‐Fish and Fkbp6 Genotyping

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