A comparative study of the effects of freezing and frozen storage on intact and demembranated bull spermatozoa

Lindemann, Charles B.; Fisher, Mary L.; Lipton, Melissa

doi:10.1016/0011-2240(82)90121-3

Cited by 35 publications

(18 citation statements)

References 15 publications

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“…The ANOVA contrast F-values (Table 3) indicated a significance level of p < 0.001 where mitochondrial function > acrosomal integrity > plasma membrane integrity. Although it has been shown that differences existed in the effect of cryopreservation on the membranes that enclose the various sperm organelles [2][3][4][5][6][7], it is not known whether the differences in the present analyses were partly due to staining artifacts, or if they accurately reflected the hierarchy of sperm compartment functionality.…”

Section: Sybr-14amentioning

confidence: 85%

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Effect of Cryopreservation on Bovine Sperm Organelle Function and Viability As Determined by Flow Cytometry1

Thomas

Garner

DeJarnette³

et al. 1998

Biology of Reproduction

148

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Flow cytometry was used to compare the functional status of fluorescently stained sperm organelles from 12 Holstein bulls after storage for 24 h at 5 degrees C and after cryopreservation. The organelle-specific stains, SYBR-14 and LysoTracker Green DND-26, identified spermatozoa with intact plasmalemma and those with intact acrosomes, respectively. The mitochondria-specific stain, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyan ine iodide (JC-1), identified two populations of spermatozoa. One population stained red-orange because the JC-1 accumulated in the mitochondria as aggregates (characteristic of cells exhibiting a high membrane potential); a second population stained green because of JC-1 monomers within the mitochondria (characteristic of cells exhibiting a lower membrane potential). Analysis of variance revealed that within bulls, the properties of sperm viability, intact acrosomes, and mitochondrial status differed in spermatozoa stored for 24 h (p < 0.001) but not in cryopreserved spermatozoa (p > 0.11). Linear regression analyses resulted in significant models in which the proportions of stained spermatozoa stored for 24 h were indicative of those proportions observed in the cryopreserved fractions. These findings suggest that the plasmalemma, the acrosome, and the mitochondria of unfrozen spermatozoa varied as to their functional status. The cryopreservation process, however, resulted in a more uniform status of sperm organelles.

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Section: Sybr-14amentioning

confidence: 85%

“…The damage to sperm mitochondria [4][5][6] caused a decrease in ATP production and thus in sperm motility. Cryoprotective agents such as glycerol helped to increase post-thaw motility but did not protect the mitochondria from freeze-thaw injury [6].…”

Section: Introductionmentioning

confidence: 99%

Effect of Cryopreservation on Bovine Sperm Organelle Function and Viability As Determined by Flow Cytometry1

Thomas

Garner

DeJarnette³

et al. 1998

Biology of Reproduction

148

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“…Thus the freezing and thawing process produces a low motility percentage and damages membrane structures resulting in a low half-life in the female genital tract and concomitant fertility decay [18]. Furthermore, these dead and abnormal spermatozoa exert toxic [28] and lytic [39] effects on companion cells in semen, and therefore have negative effects on fertility.…”

Section: Introductionmentioning

confidence: 99%

A comparative study of Sephadex, glass wool and Percoll separation techniques on sperm quality and IVF results for cryopreserved bovine semen

Lee

Kim

et al. 2009

J Vet Sci

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The aim of this study was to compare the effects of spermatozoa separation techniques on sperm quality and in-vitro fertilization (IVF) results for cryopreserved bovine semen. Sephadex, glass wool and Percoll gradient separation techniques were used for sperm separation and sperm motility, morphology and membrane integrity were evaluated before and after separation. Also, cleavage and blastocyst developmental rate were investigated after IVF with sperm recovered by each separation technique. The motility of samples obtained by the three separation techniques were greater compared to the control samples (p < 0.05). The percentage of spermatozoa with intact plasma-membrane integrity, identified by 6-carboxyfluoresceindiacetate/propidium iodide fluorescent staining and the hypo-osmotic swelling test, was highest in the glass wool filtration samples (p < 0.05). The cleavage and blastocyst rate of total oocytes produced from glass wool filtration samples were also higher than the control and Sephadex filtration samples (p < 0.05), but were not significantly different from Percoll separation samples. However, a significantly greater number of cleaved embryos produced by glass wool filtration developed to blastocyst stage than those produced by Percoll separation (p < 0.05). These results indicate that spermatozoa with good quality can be achieved by these three separation techniques and can be used for bovine IVF. In particular, it suggests that glass wool filtration would be the most effective method of the three for improving sperm quality and embryo production for cryopreserved bovine spermatozoa.

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“…At day 36 of storage, the acrosomal membranes are deteriorated drastically and the plasmalemmae are usually absent and probably do not contribute significantly to the Mg + +-ATPase activity measured. Sperm hydrolysis of exogenous ATP correlates best with the minimal deterioration observed in the movement-related flagellar structures [9]. However, some amount of this activity may be mitochondrial in origin because some mitochondrial cristae and matricies appear degraded structurally identify the sperm head; the mitochondrial sheath (MS) distinguishes the midpiece of the sperm flagellum; and the fibrous sheath (FS) identifies the principal piece.…”

Section: Discussionmentioning

confidence: 98%

Effects of In Vitro Storage on the ATP-Phosphohydrolase Activity and Ultrastructure of Ejaculated Bull Sperm

Young

Smithwick

1983

Archives of Andrology

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Extended bull ejaculates were stored in vitro at 4 2 1°C in the dark. The pH of the ejaculates did not change significantly during storage. The percentage of sperm with intact plasmalemmae was 70% on day 2 and negligible by day 36. On day 2, acrosomes were absent in 3% of the sperm and partial in about 20%; acrosomal damage increased with storage, and by day 36 acrosomes were absent in 12% of the sperm and partially degraded in 72%. The Mg+ +-ATPase activity decreased from 2.6 to 1.9 pM Pi liberated per 2 x 10' sperm per 30 min incubation at 37" between days 2 and 9, but remained at about 65% of its initial value between days 9 and 36. Thin-sectioned sperm sampled from ejaculates at days 2, 9, and 36 of storage showed a gradual but steady deterioration, which began and was most drastic in the head. Flagellar deterioration was negligible on day 9, but by day 36 subtle evidence of deterioration appeared. The persistence of Mg+ +-ATPase at 65% of initial levels in sperm stored 36 days in vitro is probably attributable to the stability of the movement-related flagellar ATPases.

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A comparative study of the effects of freezing and frozen storage on intact and demembranated bull spermatozoa

Cited by 35 publications

References 15 publications

Effect of Cryopreservation on Bovine Sperm Organelle Function and Viability As Determined by Flow Cytometry1

Effect of Cryopreservation on Bovine Sperm Organelle Function and Viability As Determined by Flow Cytometry1

A comparative study of Sephadex, glass wool and Percoll separation techniques on sperm quality and IVF results for cryopreserved bovine semen

Effects of In Vitro Storage on the ATP-Phosphohydrolase Activity and Ultrastructure of Ejaculated Bull Sperm

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