Cluster analysis reveals a binary effect of storage on boar sperm motility function1
Heiko Henning A , Anna M. Petrunkina B C E , Robin A. P. Harrison D and Dagmar Waberski A
+ Author Affiliations
- Author Affiliations
A Unit for Reproductive Medicine of Clinics, Clinic for Pigs and Small Ruminants, University of Veterinary Medicine, Bünteweg 15, 30559 Hannover, Germany.
B Department of Medicine, Box 157, University of Cambridge, Hills Road, CB2 0QQ, UK.
C Unit for Reproductive Medicine of Clinics, Clinic for Horses, University of Veterinary Medicine Hannover, Bünteweg 15, 30559 Hannover, Germany.
D 11 London Road, Great Shelford, Cambridge CB22 5DB, UK.
E Corresponding author. Email: anna.petrunkina@gmx.de
Reproduction, Fertility and Development 26(5) 623-632 https://doi.org/10.1071/RD13113
Submitted: 18 February 2013 Accepted: 9 April 2013 Published: 7 June 2013
Abstract
Storage of liquid-preserved boar spermatozoa is associated with a loss of fertilising ability of the preserved spermatozoa, which standard semen parameters barely reflect. Monitoring responses to molecular effectors of sperm function (e.g. bicarbonate) has proven to be a more sensitive approach to investigating storage effects. Bicarbonate not only initiates capacitation in spermatozoa, but also induces motility activation. This occurs at ejaculation, but also happens throughout passage through the oviduct. In the present study we tested whether the specific response of boar sperm subpopulations to bicarbonate, as assessed by motility activation, is altered with the duration of storage in vitro. Three ejaculates from each of seven boars were diluted in Beltsville thawing solution and stored at 17°C. Only minor changes in the parameters of diluted semen were revealed over a period of 72 h storage. For assessment of bicarbonate responses, subsamples of diluted spermatozoa were centrifuged through a discontinuous Percoll gradient after 12, 24 and 72 h storage. Subsequently, spermatozoa were incubated in two Ca2+-free variants of Tyrode’s medium either without (TyrControl) or with (TyrBic) 15 mM bicarbonate, and computer-aided sperm analysis motility measurements were made. Cluster analysis of imaging data from motile spermatozoa revealed the presence of five major sperm subpopulations with distinct motility characteristics, differing between TyrBic and TyrControl at any given time (P < 0.001). Although there was an increasing loss of motility function in both media, bicarbonate induced an increase in a ‘fast linear’ cohort of spermatozoa in TyrBic regardless of storage (66.4% at 12 h and 63.9% at 72 h). These results imply a binary pattern in response of sperm motility function descriptors to storage: although the quantitative descriptor (percentage of motile spermatozoa) declines in washed semen samples, the qualitative descriptor (percentage of spermatozoa stimulated into fast linear motion by bicarbonate) is sustained independent of the duration of storage.
Additional keyword: bicarbonate.
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