Detection of calcium ionophore induced membrane changes in dog sperm as a simple method to predict the cryopreservability of dog semen

Szász, F.; Sirivaidyapong, S.; Cheng, Fei-Fei; Voorhout, W. F.; Marks, A.; Colenbrander, B.; Solti, L.; Gadella, B.M.

doi:10.1002/(sici)1098-2795(200003)55:3<289::aid-mrd7>3.0.co;2-k

Cited by 58 publications

(38 citation statements)

References 28 publications

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“…61 This double staining for membrane integrity and acrosomal integrity is relatively reliable for fresh and in vitro capacitated sperm. 68 Problems can arise when frozen-thawed spermatozoa are analyzed. Egg yolk particles from the extender have low fluorescence and, therefore, can be assessed as live acrosome-intact sperm using the PI/FITC-PSA double-labeling method.…”

Section: Sperm Intactnessmentioning

confidence: 99%

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Hossain¹,

Johannisson²,

Wallgren³

et al. 2011

Asian J Androl

135

110

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Flow cytometry is now a recognized methodology within animal spermatology, and has moved from being a research tool to become routine in the assessment of animal semen destined to breeding. The availability of 'bench-top' flow cytometers and of newer and versatile markers for cell structure and function had allowed the instrumentation to measure more sperm parameters, from viability to reactiveness when exposed to exogenous stimuli, and to increase our capabilities to sort spermatozoa for potential fertilizing capacity, or chromosomal sex. The present review summarizes the state of the art regarding flow cytometry applied to animal andrology, albeit keeping an open comparative intent. It critically evaluates the present and future capabilities of flow cytometry for the diagnostics of potential fertility and for the development of current reproductive technologies such as sperm freezing, sperm selection and sperm sorting. The flow cytometry methods will probably further revolutionize our understanding of the sperm physiology and their functionality, and will undoubtedly extend its application in isolating many uncharacterized features of spermatozoa. However, continuous follow-up of the methods is a necessity owing to technical developments and the complexity of mapping spermatozoa.

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Section: Sperm Intactnessmentioning

confidence: 99%

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Hossain¹,

Johannisson²,

Wallgren³

et al. 2011

Asian J Androl

135

110

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“…For all other samples, membrane status and acrosome integrity were measured simultaneously using the stains PI (Sigma 287075, Sigma-Aldrich) and FITC-PNA (Sigma L-7381, Sigma-Aldrich; Graham et al 1990, Pena et al 2001. The PI/FITC-PNA staining method used herein was a combination of different dual staining protocols (Graham et al 1990, Szasz et al 2000, Pena et al 2001, as previously described for killer whales (Robeck et al 2011). Briefly, a 12.5-ml aliquot of sperm sample was transferred to a foil-covered microcentrifuge tube and mixed with 1 ml of PI (12.5 mg/ml working stock in PBS).…”

Section: Sexing and Recryopreservation Of Dolphin Spermmentioning

confidence: 99%

Evaluation of motility, membrane status and DNA integrity of frozen–thawed bottlenose dolphin (Tursiops truncatus) spermatozoa after sex-sorting and recryopreservation

Montano¹,

Kraemer²,

Love³

et al. 2012

Reproduction

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Artificial insemination (AI) with sex-sorted frozen-thawed spermatozoa has led to enhanced management of ex situ bottlenose dolphin populations. Extended distance of animals from the sorting facility can be overcome by the use of frozen-thawed, sorted and recryopreserved spermatozoa. Although one bottlenose dolphin calf had been born using sexed frozen-thawed spermatozoa derived from frozen semen, a critical evaluation of in vitro sperm quality is needed to justify the routine use of such samples in AI programs. Sperm motility parameters and plasma membrane integrity were influenced by stage of the sex-sorting process, sperm type (non-sorted and sorted) and freezing method (straw and directional) (P!0.05). After recryopreservation, sorted spermatozoa frozen with the directional freezing method maintained higher (P!0.05) motility parameters over a 24-h incubation period compared to spermatozoa frozen using straws. Quality of sperm DNA of non-sorted spermatozoa, as assessed by the sperm chromatin structure assay (SCSA), was high and remained unchanged throughout freeze-thawing and incubation processes. Though a possible interaction between Hoechst 33342 and the SCSA-derived acridine orange was observed in stained and sorted samples, the proportion of sex-sorted, recryopreserved spermatozoa exhibiting denatured DNA was low (6.6G4.1%) at 6 h after the second thawing step and remained unchanged (PO0.05) at 24 h. The viability of sorted spermatozoa was higher (P!0.05) than that of non-sorted spermatozoa across all time points after recryopreservation. Collective results indicate that bottlenose dolphin spermatozoa undergoing cryopreservation, sorting and recryopreservation are of adequate quality for use in AI.

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“…A�ention has been devoted to the high-quality evaluation of dog ejaculates in particular in relation to the results of their long-term conservation (Pena et al, 1999;Pena and Linde-Forsberg, 2000a,b;Szasz et al, 2000). Rigau et al (2001) stress the importance of sperm motility as an indicator of the quality of dog ejaculates.…”

mentioning

confidence: 99%

Functional evaluation of dog ejaculates with priority given to the aspect of acrosome integrity

Vĕzník¹,

Švecová²,

Zajicova³

et al. 2003

Vet. Med. - Czech

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38 ejaculates of dogs of various breeds from 2 to 6 years old were examined in a short-term 120 min survival test. The examination focused on the functional parameters of sperm (motility, percentage of live spermatozoa, acrosomal integrity) and their morphology. The levels of relationship between individual criteria of sperm analysis were demonstrated. Statistically highly significant correlations between motility, ratio of live spermatozoa, percentage of intact acrosomes and percentage of spermatozoa with altered superficial structures were proven. Our findings proved the sensitivity of superficial structures, primarily plasmatic membranes, and the higher resistance of structures with an organelle basis. Strict morphological sperm analysis provides important information about the rise in qualitative alterations of ejaculates caused by changes in superficial structures during the course of a short-term survival test.

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Detection of calcium ionophore induced membrane changes in dog sperm as a simple method to predict the cryopreservability of dog semen

Cited by 58 publications

References 28 publications

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Evaluation of motility, membrane status and DNA integrity of frozen–thawed bottlenose dolphin (Tursiops truncatus) spermatozoa after sex-sorting and recryopreservation

Functional evaluation of dog ejaculates with priority given to the aspect of acrosome integrity

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