Sperm Kinetics and Morphology Before and After Fractionation on Discontinuous Percoll Gradient for Sex Preselection: Computerized Analyses

Watkins, Ai-Mae; Chan, Philip J.; Patton, William C.; Jacobson, John D.; King, Alan

doi:10.3109/01485019608988494

Cited by 10 publications

(8 citation statements)

References 7 publications

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“…Motility was significantly lower in sperm from fractions 1 (10.7 Ϯ 1.3), 2 (15.9 Ϯ 1.5), and 3 (31.3 Ϯ 2.7) compared to sperm from fraction 4 (62.0 Ϯ 4.3) (P Ͻ 0.001). In contrast, the abnormal forms percent was significantly higher in sperm from fractions 1 (81.6 Ϯ 1.7), 2 (75.5 Ϯ 1.8), and 3 (51.7 Ϯ 2.5) compared to sperm from fraction 4 (20.8 Ϯ 2.1) (P Ͻ 0.001), also consistent with previous reports (Pousette et al, 1986;Le Lannou and Blanchard, 1988;Pardo et al, 1988;Menkveld et al, 1990;Van der Zwalmen et al, 1991;Mortimer and Mortimer, 1992;Saad and Guerin, 1992;Huszar et al, 1993;Hall et al, 1995;Sukcharoen, 1995;Yue et al, 1995;Watkins et al, 1996;Yao et al, 1996;Golan et al, 1997;Angelopoulos et al, 1998;Lassalle et al, 1999).…”

Section: Resultssupporting

confidence: 90%

Variation of docosahexaenoic acid content in subsets of human spermatozoa at different stages of maturation: Implications for sperm lipoperoxidative damage

2000

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Section: Resultssupporting

confidence: 90%

Variation of docosahexaenoic acid content in subsets of human spermatozoa at different stages of maturation: Implications for sperm lipoperoxidative damage

2000

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“…Kaneko et al [21] suggested that the sperm separation with Percoll gradients was due to differences in the basis of sperm velocity sedimentation, not on the basis of their buoyant density. The sperm sedimentation velocity may be influenced by differences in sperm head size or motility [15,22] but, there was no difference between the sperm head dimensions in the top and bottom fractions [22], but in contrast to a research report by Cui and Matthews [23].…”

Section: Discussioncontrasting

confidence: 46%

Assessment of Bovine X- and Y-bearing Spermatozoa in Fractions by Discontinuous Percoll Gradients with Rapid Fluorescence In Situ Hybridization

Kobayashi

Oguro

Uchida

et al. 2004

Journal of Reproduction and Development

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Abstract. This study was designed to apply the method of discontinuous Percoll gradients for sex preselection in bovine semen by using a current developed molecular technique, fluorescence in situ hybridization (FISH). In addition, we attempted to amplify the level of enrichment of X-or Y-bearing spermatozoa by treating for activating sperm motility performance with 10 mM caffeine. Bovine spermatozoa were fractionated on Percoll gradients into two major subpopulations of motile spermatozoa (bottom fraction) and weak motile spermatozoa (top fraction). The percentage of Ybearing spermatozoa in the top fraction (52.9%) slightly exceeded and that in the bottom fraction (44.3%) decreased significantly (P<0.001) compared with the theoretical ratio (50:50). Washing sperm with BO medium affected a deviation between the two sex populations, whereas semen activated with caffeine showed no difference in the percentage of X-and Y-bearing spermatozoa in both fractions compared with the theoretical ratio (50:50). These results show that the proportion of X-and Ybearing bovine spermatozoa can deviate after discontinuous Percoll gradients, although the proportion of X-and Y-bearing bovine spermatozoa was affected by sperm motility of the sample applied. Key words: Percoll separation, Semen sexing, FISH, Spermatozoa, Bovine (J. Reprod. Dev. 50: [463][464][465][466][467][468][469] 2004) ince the 1970s, several investigators have made a tt e m p t s t o s e p a r at e X -a n d Y -b e ar in g spermatozoa by means of various gradient techniques such as discontinuous albumin gradients [1, 2], Shepadex column s [2,3], discontinuous Percoll gradients [4,5] and the swim-up procedure [6]. None of these methods, however, have met with success sustainable and reproducible enough to achieve separation of Xand Y-bearing spermatozoa, and only flow cytometry has been proven to be effective [7]. One o f t h e p r o b l e m s a s s o c i a t e d w i t h r e s e a r c h attempting the separation of X-and Y-bearing spermatozoa has been the difficulty of validating the proportion of X-and Y-bearing spermatozoa within putatively separated sperm populations [8]. Early studies evaluated the efficiency of sperm separation by using quinacrine staining. Recent evidence has shown that the quinacrine method is unreliable and may not necessarily detect only the Y body at the Y chromosome in spermatozoa [9].Current development of DNA probes raised against the sex chromosome specific sequence gives

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“…These findings suggest the preferential processing of X‐ and Y‐chromosome‐bearing sperm by oocyte cytoplasm depending on the maturation state and support the hypothesis that X sperm show greater longevity and penetrate the majority of oocytes when they become ready for fertilization. Watkins et al (1996) revealed a tendency for X‐chromosome‐bearing sperm after 24 h of incubation to show higher longevity in motility, higher progression and hyperactivation when compared with Y‐chromosome‐bearing spermatozoa. It is not known whether this influences their longevity in vitro .…”

Section: Discussionmentioning

confidence: 98%

“…This phenomenon is not fully understood, however it may be attributed to events that occur before fertilization that favour selection of X‐ or Y‐chromosome‐bearing spermatozoa, events that occur after fertilization such as preferential development or survival of embryos of one sex or a combination of both (Kochhar et al 2001). Moreover, analysis of the X‐ and Y‐sperm populations have shown some morphological and functional differences, although it is not known if X‐ and Y‐bearing spermatozoa have the same viability in vitro (Sarkar et al 1984; Watkins et al 1996; Penfold et al 1998; Van Munster et al 1999).…”

Section: Introductionmentioning

confidence: 99%

Sperm Pre‐Incubation Prior to Insemination Affects the Sex Ratio of Bovine Embryos Produced in vitro

Lechniak¹,

Strabel²,

Bousquet

et al. 2003

Reprod Domestic Animals

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The objective of the present study was to determine whether sperm incubation prior to oocyte insemination in vitro affects the sex ratio of resulting blastocyst. Cumulus-oocyte-complexes (COCs) collected from slaughterhouse ovaries were matured in vitro and inseminated with frozen-thawed semen of three proven artificial insemination (AI) bulls pre-incubated in vitro in Sperm-Talp for 6 and 24 h. On day-9 blastocysts were collected and processed for sex determination. More than 80% of blastocyst were successfully sexed. There were no significant differences in cleavage and blastocyst rates using sperm pre-incubated for 6 h as compared with the 0-h pre-incubation control group. The cleavage and blastocyst rates were significantly lower in the 24-h pre-incubation group. The male to female ratio, when compared with the theoretical 1 : 1, differed significantly in favour of females among hatched (viable) blastocysts derived from sperm pre-incubated for 24 h prior to insemination as well as among all blastocytsts in the 6-h group. Moreover, when the sperm treatment was considered, the sex ratio was affected only among hatched blastocysts in 24-h pre-incubation group. It was concluded that prolonged sperm pre-incubation influences the rate of development and the sex ratio among hatched blastocysts.

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Sperm Kinetics and Morphology Before and After Fractionation on Discontinuous Percoll Gradient for Sex Preselection: Computerized Analyses

Cited by 10 publications

References 7 publications

Variation of docosahexaenoic acid content in subsets of human spermatozoa at different stages of maturation: Implications for sperm lipoperoxidative damage

Variation of docosahexaenoic acid content in subsets of human spermatozoa at different stages of maturation: Implications for sperm lipoperoxidative damage

Assessment of Bovine X- and Y-bearing Spermatozoa in Fractions by Discontinuous Percoll Gradients with Rapid Fluorescence In Situ Hybridization

Sperm Pre‐Incubation Prior to Insemination Affects the Sex Ratio of Bovine Embryos Produced in vitro

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