Flow cytometry of X and Y chromosome‐bearing sperm for DNA using an improved preparation method and staining with Hoechst 33342

Johnson, L.A.; Flook, J. P.; Look, M. V.

doi:10.1002/mrd.1120170303

Cited by 123 publications

(74 citation statements)

References 9 publications

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“…Sexed semen is now widely available in dairy industries around the world, and is nearing commercial application in several other species (Seidel, 2012). The process distinguishes X-and Y-chromosome bearing sperm by measuring differences in fluorescence following staining the sperm with a non-toxic, DNA-binding dye (Hoechst 33342) (Johnson et al, 1987). Despite reliably producing~90% gender bias (Cerchiaro et al, 2007;Borchersen and Peacock, 2009;DeJarnette et al, 2009), the fertility of the sexed semen product is compromised (Seidel, 2012).…”

Section: Introductionmentioning

confidence: 99%

Applications and cost benefits of sexed semen in pasture-based dairy production systems

Butler

Hutchinson

Cromie

et al. 2014

Animal

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Sexed semen technology is now commercially available in many countries around the world, and is primarily used in dairy cattle breeding. Sperm are sorted by flow cytometry on the basis of a 4% difference in DNA content between sperm containing X and Y chromosomes. Despite reliably producing a 90% gender bias, the fertility of the sexed semen product is compromised compared with conventional semen. The negative implications of the reduced fertility of sexed semen are amplified in seasonal systems of dairy production, as the importance of fertility is greater in these systems compared with year-round calving systems. A review of the literature indicates that conception rates (CR) to 1st service with frozen-thawed sexed semen are~75% to 80% of those achieved with conventional frozen-thawed semen. Preliminary results from a large-scale field trial carried out in Ireland in 2013 suggest that significant improvements in the performance of sexed semen have been made, with CR of 87% of those achieved with conventional semen. The improved fertility of a sexed semen product that delivers a 90% gender bias has considerable implications for the future of breeding management in pasture-based dairy production systems. Sexed semen may facilitate faster, more profitable dairy herd expansion by increasing the number of dairy heifer replacements born. Biosecurity can be improved by maintaining a closed herd during the period of herd expansion. In a non-expansion scenario, sexed semen may be used to increase the value of beef output from the dairy herd. The replacement heifer requirements for a herd could be met by using sexed semen in the 1st 3 weeks of the breeding season, with the remaining animals bred to beef sires, increasing the sale value over that of a dairy bull calf. Alternatively, very short gestation sires could be used to shorten the calving interval. Market prices have a considerable effect on the economics of sexed semen use, and widespread use of sexed semen should be restricted to well managed herds that already achieve acceptable herd fertility performance.

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Section: Introductionmentioning

confidence: 99%

Applications and cost benefits of sexed semen in pasture-based dairy production systems

Butler

Hutchinson

Cromie

et al. 2014

Animal

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“…After it was demonstrated that it is not possible to differentiate sex-specific markers by physical methods, Gledhill et al (1982) published the first experiments on flow cytometry on spermatozoa, which were followed by further technical adaptations to these specific cells (Fulwyler 1977, Dean et al 1978, Stovel et al 1978, Garner et al 1983, Johnson & Pinkel 1986, Johnson et al 1987a, 1987b, Johnson & Clarke 1988. The first pre-sexed offspring using this technique were born in 1988 (Morrell et al 1988).…”

Section: Sex Sorting Of Spermatozoa By Flow Cytometrymentioning

confidence: 99%

Sex selection of sperm in farm animals: status report and developmental prospects

Rath¹,

Barcikowski²,

Graaf³

et al. 2013

Reproduction

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Pre-selection of spermatozoa based on the relative DNA difference between X-and Y-chromosome bearing populations by flow cytometry is an established method that has been introduced into commercial cattle production. Although several important improvements have increased the sort efficiency, the fertilising ability of sexed spermatozoa based on offspring per insemination is still behind farmers' expectations. The main stress factors, especially on mitochondria, that reduce the lifespan of spermatozoa are described, and new technical as well as biological solutions to maintain the natural sperm integrity and to increase the sorting efficiency are discussed. Among these methods are the identification of Y-chromosome bearing spermatozoa by bi-functionalised gold nanoparticles and triplex hybridisation in vivo as well as new laser-controlled deflection system that replaces the deflection of spermatozoa in the electrostatic field. Additionally, as well as a new nonsurgical transfer system of spermatozoa into the oviduct of cows has been developed and allows a significant reduction of spermatozoa per transfer. Altogether, the improvements made in the recent years will allow a broader use of sex-sorted spermatozoa even in those species that require more cells than cows and sheep.Reproduction (2013) 145 R15-R30

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“…Based on the excitation, spermatozoa are separated into discrete populations. In domestic animals the differences in DNA content between X and Y bearing spermatozoa ranges from 3 -4.5% (Johnson et al, 1987;Johnson, 2000). Success rate in this method has been reported to be 85 -95% (Pinkel et al, 1982;Johnson et al, 1989Johnson et al, , 2000.…”

Section: Flow Cytometrymentioning

confidence: 93%

Sexing of Spermatozoa in Farm Animals: a Mini Review

Prakash¹

2014

Adv. Anim. Vet. Sci.

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Livestock farmers always have a wish for producing young ones of desired sex. Among the several techniques available, use of sexed semen for artificial insemination is recognized as more pragmatic and easy way to pre -select the sex of the offspring. Selective use of sexed semen in breeding will not only increase the genetic progress from the daughter -dam path but would also help in producing good male germplasm from elite bulls for future breeding. Several attempts have been made, elsewhere in the globe, to develop methods that efficiently separate bovine semen into fractions containing higher concentrations of X -or Y -bearing sperm. These technologies include sex specific antibodies, centrifugation and flow cytometry. Of these attempts, the only method proven to be commercially viable is flow cytometry. However, sorting pressure, speed, electrical deviation, laser radiation all leads to membrane alteration and pre -capacitation like changes in the sorted sperm leading to reduced fertility. Despite these limitations, production of sexed semen usually followed by cryopreservation is being used commercially for cattle production. Development of the instrument for increasing the sorting rate and also purity of sorting without affecting the viability and fertility is still an active area of research. The aim of this review is to update the readers with the recent developments in sexing of spermatozoa with special reference to farm animals.All copyrights reserved to Nexus® academic publishers

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Flow cytometry of X and Y chromosome‐bearing sperm for DNA using an improved preparation method and staining with Hoechst 33342

Cited by 123 publications

References 9 publications

Applications and cost benefits of sexed semen in pasture-based dairy production systems

Applications and cost benefits of sexed semen in pasture-based dairy production systems

Sex selection of sperm in farm animals: status report and developmental prospects

Sexing of Spermatozoa in Farm Animals: a Mini Review

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