An investigation into the similarities and differences governing the cryopreservation success of koala (Phascolarctos cinereus: goldfuss) and common wombat (Vombatus ursinus: shaw) spermatozoa

Johnston, S. D., MacCallum, C., Blyde, D., McClean, R., Lisle, A. and Holt, W. V. (2006) An investigation into the similarities and differences governing the cryopreservation success of koala (Phascolarctos cinereus: goldfuss) and common wombat (Vombatus ursinus: shaw) spermatozoa. Cryobiology, 53 2: 218-228. doi:10.1016/j.cryobiol.2006.06.001


Author Johnston, S. D.
MacCallum, C.
Blyde, D.
McClean, R.
Lisle, A.
Holt, W. V.
Title An investigation into the similarities and differences governing the cryopreservation success of koala (Phascolarctos cinereus: goldfuss) and common wombat (Vombatus ursinus: shaw) spermatozoa
Journal name Cryobiology   Check publisher's open access policy
ISSN 0011-2240
Publication date 2006
Sub-type Article (original research)
DOI 10.1016/j.cryobiol.2006.06.001
Volume 53
Issue 2
Start page 218
End page 228
Total pages 11
Editor David E Pegg
Place of publication San Diego
Publisher Academic Press Inc Elsevier Science
Collection year 2006
Language eng
Subject C1
270602 Animal Physiology - Cell
780105 Biological sciences
Abstract The aim of this study was to determine the relative cryopreservation success of koala and wombat spermatozoa and to investigate reasons for their respective post-thaw survival by examining the sperm's response to a range of osmotic media and determining the presence and distribution of F-actin. An hypothesis was proposed that F-actin may be imparting a degree of structural inflexibility to the koala sperm plasma membrane; hence, exposure of spermatozoa to cytochalasin D (5 mu M), a F-actin depolymerisation agent, should result in increased plasticisation of the membrane and greater tolerance of cell volume changes that typically occur during cryopreservation. In experiment 1, koala (n = 4) and wombat (n = 4) spermatozoa packaged in 0.25 mL straws were cryopreserved using two freezing rates (fast-3 cm above liquid N2 interface; slow-6 degrees C/min in a freezing chamber) and two glycerol concentrations (8 and 14% v/v) in a tris-citrate glucose buffer with 15% (v/v) egg yolk. Wombat spermatozoa showed better (P < 0.01) post-thaw survival (% motile, % intact plasma membranes, % decondensed sperm heads) than koala spermatozoa. When exposed to media of varying osmolality, koala spermatozoa were less tolerant (% intact plasma membrane) of hyper-osmotic conditions (920 and 1410mOsmol/kg) than wombat spermatozoa. F-actin was localised using a monoclonal antibody but only found in the wombat sperm head. When koala and wombat spermatozoa were exposed to media of varying osmolality, cytochalasin D had no beneficial effect on sperm survival (% intact plasma membranes). This study has demonstrated that wombat spermatozoa are highly tolerant of cryopreservation when compared to koala sperm but that spermatozoa from both species show greatest post-thaw survival when frozen slowly in 14% glycerol. Koala sperm are also particularly susceptible to hyper-osmotic environments but lack of detectable F-actin in the koala spermatozoan suggests that poor cryopreservation success in this species is unlikely to be associated with F-actin induced plasma membrane inflexibility. (c) 2006 Elsevier Inc. All rights reserved.
Keyword Koala
Wombat
Spermatozoa
Cryopreservation
Osmotic Tolerance
F-actin
Cytochalasin D
Marsupial
Biology
Physiology
Cold Shock
Sperm
Preservation
Morphology
Marsupials
Membranes
Calcium
Injury
Semen
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Agriculture and Food Sciences
 
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