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

S.D. Johnston, C. MacCallum, D. Blyde, R. McClean, A. Lisle, W.V. Holt

Research output: Contribution to journalArticle

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Abstract

this species is unlikely to be associated with F-actin induced plasma membrane inflexibility.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 microM), 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 1410 mOsmol/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
Original languageEnglish
Pages (from-to)218-228
Number of pages11
JournalCryobiology
Volume53
Issue number2
DOIs
Publication statusPublished - 2006

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Phascolarctidae
Phascolarctos cinereus
Cryopreservation
cryopreservation
Cell membranes
Spermatozoa
Actins
spermatozoa
Cytochalasin D
Freezing
actin
Glycerol
Cell Membrane
plasma membrane
Depolymerization
Straw
Sperm Head
Citric Acid
cytochalasin D
Buffers

Cite this

@article{6bfa99d9ec9f46cf9536a4ccb08b80a3,
title = "An investigation into the similarities and differences governing the cryopreservation success of koala (Phascolarctos cinereus: goldfuss) and common wombat (Vombatus ursinus: shaw) spermatozoa.",
abstract = "this species is unlikely to be associated with F-actin induced plasma membrane inflexibility.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 microM), 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 1410 mOsmol/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",
keywords = "Cryopreservation, Cytochalasin D, F-actin, Koala, Marsupial, Osmotic tolerance, Spermatozoa, Wombat",
author = "S.D. Johnston and C. MacCallum and D. Blyde and R. McClean and A. Lisle and W.V. Holt",
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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.; Holt, W.V.

In: Cryobiology, Vol. 53, No. 2, 2006, p. 218-228.

Research output: Contribution to journalArticle

TY - JOUR

T1 - An investigation into the similarities and differences governing the cryopreservation success of koala (Phascolarctos cinereus

T2 - goldfuss) and common wombat (Vombatus ursinus: shaw) spermatozoa.

AU - Johnston, S.D.

AU - MacCallum, C.

AU - Blyde, D.

AU - McClean, R.

AU - Lisle, A.

AU - Holt, W.V.

N1 - Imported on 12 Apr 2017 - DigiTool details were: Journal title (773t) = Cryobiology. ISSNs: 0011-2240;

PY - 2006

Y1 - 2006

N2 - this species is unlikely to be associated with F-actin induced plasma membrane inflexibility.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 microM), 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 1410 mOsmol/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

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KW - Cryopreservation

KW - Cytochalasin D

KW - F-actin

KW - Koala

KW - Marsupial

KW - Osmotic tolerance

KW - Spermatozoa

KW - Wombat

U2 - 10.1016/j.cryobiol.2006.06.001

DO - 10.1016/j.cryobiol.2006.06.001

M3 - Article

VL - 53

SP - 218

EP - 228

JO - Cryobiology

JF - Cryobiology

SN - 0011-2240

IS - 2

ER -