Cryopreservation of macropodid spermatozoa: New insights from the cryomicroscope

Holt, W. V., Penfold, L. M., Johnston, S. D., Temple-Smith, P., Shaw, J., McCallum, C., Lindemans, W. and Blyde, D. (1999) Cryopreservation of macropodid spermatozoa: New insights from the cryomicroscope. Reproduction Fertility and Development, 11 6: 345-353. doi:10.1071/RD99076

Author Holt, W. V.
Penfold, L. M.
Johnston, S. D.
Temple-Smith, P.
Shaw, J.
McCallum, C.
Lindemans, W.
Blyde, D.
Title Cryopreservation of macropodid spermatozoa: New insights from the cryomicroscope
Journal name Reproduction Fertility and Development   Check publisher's open access policy
ISSN 1031-3613
Publication date 1999-01-01
Sub-type Article (original research)
DOI 10.1071/RD99076
Open Access Status
Volume 11
Issue 6
Start page 345
End page 353
Total pages 9
Place of publication Canberra, Australia
Publisher CSIRO Publishing
Language eng
Subject C1
300402 Animal Reproduction
780105 Biological sciences
060602 Animal Physiology - Cell
Abstract This study examined the effects of cooling and cryopreservation upon macropod spermatozoa (eastern grey kangaroo, Macropus giganteus and red-necked wallaby, Macropus rufogriseus). Sperm survival during and after freezing to -30 degrees C or -70 degrees C in minimum essential medium (MEM) + 5, 10, 20 or 30% (v/v) glycerol, MEM + 10 or 20% (v/v) ethylene glycol and MEM containing a mixture of 7.5% (v/v) glycerol + 10% (v/v) dimethylsulphoxide was examined by cryomicroscopy. The MEM/glycerol mixtures permitted better post-thaw sperm recovery than the other cryoprotectants. After freezing to -30 degrees C at 10 degrees C min(-1) in 20% glycerol, then rewarming at 20 degrees C min(-1), flagellar activity resumed in more than 50% of spermatozoa when the temperature increased into the range 5-10 degrees C. However, as the temperature increased into the range 20-25 degrees C, motility declined rapidly so that less than 5% motile cells were seen at 35 degrees C. Spermatozoa in MEM without cryoprotectant were also examined by cryomicroscopy to evaluate changes in flagellar configuration, swimming behaviour and viability during cooling from 35 degrees C to approximately -7 degrees C, and rewarming to 35 degrees C. Cooling from 35 to 28 degrees C induced kangaroo spermatozoa to exhibit rigid principal-piece bending and non-linear motility, which was reversed by further cooling and the spermatozoa resumed their normal linear movement. Rewarming induced principal-piece bending in the range of 20-30 degrees C, but this effect was reversed by further warming. Although red-necked wallaby spermatozoa showed these effects, they also exhibited a tendency to form rosette-like clusters during rewarming, especially when the temperature reached approximately 14 degrees C. The clusters were induced when the flagellar end-pieces became anteriorly reflected, producing hook-like flagellar conformations, which then became interlinked. © CSIRO 1999
Keyword Developmental Biology
Reproductive Biology
Genetic Resource Banks
Plasma Membrane
Lipid Phase-transitions
Sperm Plasma-membrane
Brushtail Possum
Mammalian Sperm
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Agriculture and Food Sciences
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Citation counts: TR Web of Science Citation Count  Cited 29 times in Thomson Reuters Web of Science Article | Citations
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Created: Tue, 10 Jun 2008, 22:25:42 EST