Improvement of the post-thaw qualities of Okinawan native pig spermatozoa frozen in an extender supplemented with ascorbic acid 2-O-α-glucoside

Yoshimoto, Teppei; Yamauchi, Shogo; Muto, Norio; Nakada, Tadashi; Ashizawa, Koji; Tatemoto, Hideki

doi:10.1016/j.cryobiol.2008.05.002

Cited by 30 publications

(25 citation statements)

References 32 publications

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“…Many investigators have focused on the use of antioxidants in the freezing media to reduce the negative effects of ROS on spermatozoa (Aitken 1995;Amidi et al 2011;Bilodeau et al 2000;Gadea et al 2011;Taylor et al 2009;Yoshimoto et al 2008;Kirilova et al 2015). Indeed, the major objective of this review is to discuss the findings of different studies in favor of the role of different antioxidants on sperm cryoinjury during the cryopreservation/thaw process regarding mechanisms which improve resistance to cryodamage of spermatozoa.…”

Section: Introductionmentioning

confidence: 98%

“…Moreover, BSA (bovine serum albumin) increases motility and viability of spermatozoa after long term storage in low temperatures (Matsuoka et al 2006;Yoshimoto et al 2008) and cryopreservation (Blesbois and Caffin 1992). Also, a good membrane protection in terms of resistance to hypoosmotic shock was also attained when BSA and egg yolk were added to the extender (Amidi et al 2010).…”

Section: Introductionmentioning

confidence: 99%

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The role of antioxidants in sperm freezing: a review

et al. 2016

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Cryopreservation of spermatozoa is becoming more important because of new clinical requirements and current clinical practice. Despite the success of sperm cryopreservation this routinely used procedure induces serious detrimental changes in sperm function. Some researchers believe that cryopreservation is associated with DNA fragmentation and DNA single strand breaks in sperm. Mechanisms of cryodamage to human spermatozoa are thought to be multifactorial including: cold shock, osmotic stress, intracellular ice crystal formation, oxidative stress, and combinations of these conditions. Additives showing antioxidative properties reported to reduce the impact of ROS-induced and cold shock damages. Many studies exist as regards the effects of antioxidants on the cryopreservation aimed at improving the quality of post-thaw semen. Hence, this review will clarify results of recent applications of various antioxidants used in numerous research efforts to improve cryopreservation of spermatozoa. This review is to increase the understanding of the roles of these antioxidants concerning mechanisms which enhance resistance to cryodamage of spermatozoa.

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Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

The role of antioxidants in sperm freezing: a review

et al. 2016

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“…The plasmalemma integrity of frozen-thawed sperm was assessed using dual-color fluorescent staining with 6-carboxyfluorescein diacetate (CFDA, 10 μg) and propidium iodide (PI, 10 μg) as previously described [21,22]. At least 270 sperm per treatment condition were observed (× 400) under a fluorescence microscope (Nikon, Tokyo, Japan) equipped with blue illumination (excitation at 450-490 nm, emission at 520 nm).…”

Section: Sperm Plasmalemma Integritymentioning

confidence: 99%

“…Acrosomal proteolytic activity in single sperm was assessed using the gelatin plate assay as described previously [22][23][24]. Frozen-thawed sperm were washed three times by centrifugation at 700 × g for 4 min and resuspended in PBS, giving a concentration of 4 × 10 6 sperm/ml.…”

Section: Acrosomal Proteolytic Activity Assaymentioning

confidence: 99%

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Characteristics of Okinawan Native Agu Pig Spermatozoa After Addition of Low-Density Lipoprotein to Freezing Extender

Yamauchi

Lay

Azuma

et al. 2009

Journal of Reproduction and Development

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Abstract. Technical refinement of boar sperm cryopreservation is indispensable for effective breeding of the rare Okinawan native pig, the Agu. The objective of the present study was to determine whether addition of low-density lipoprotein (LDL) extracted from hen egg yolk to the freezing extender improves the characteristics of cryopreserved Agu spermatozoa. Ejaculated Agu sperm frozen in extender supplemented with 2, 4, 6, 8 or 10% LDL instead of egg yolk was thawed, and the post-thaw sperm characteristics were evaluated. Treatment with 4-8% LDL during cooling and freezing significantly increased the intracellular cholesterol content, as compared to that of sperm frozen in extender containing 20% egg yolk (P<0.05). Higher potential resistance to cell damage from cryoinjury was also observed in sperm frozen in extender supplemented with LDL: the integrities of plasmalemma and DNA, mitochondrial activity and proteolytic activity of the acrosomal content in the post-thaw sperm were superior to those of sperm that were not treated with LDL. Moreover, the percentages of total motile sperm and the extent of rapid progressive motility at 1 and 3 h after incubation were markedly higher in sperm treated with 4 or 6% LDL, and these sperm also had more ATP. However, LDL did not inhibit in vitro sperm penetrability, even though the cholesterol content of post-thaw sperm was higher after treatment with LDL. These findings indicate that addition of 4-6% LDL instead of egg yolk to the freezing extender improves the post-thaw characteristics of Agu sperm by protecting sperm against cold shock damage during cryopreservation. Key words: Boar spermatozoa, Cell damage, Cryopreservation, Low-density lipoproteins (LDL) (J. Reprod. Dev. 55: [558][559][560][561][562][563][564][565] 2009) ecently, the male reproductive ability (i.e., semen volume, sperm quality and sperm concentration) of the Okinawan native pig, the Agu, has markedly decreased because of repeated inbreeding within a minority of the closed population for the past 20 years, which has resulted in a marked reduction in reproductive efficiency under natural mating conditions. Consequently, longterm preservation and storage of Agu sperm for artificial insemination (AI) has become a subject of interest because it permits insemination of a large number of females in a short period of time, avoids transmission of disease and conserves valuable germplasm stocks. However, the poor post-thaw survival of boar sperm has limited the success of AI with frozen sperm and has prevented its widespread acceptance in commercial breeding [1,2].Boar sperm are extremely sensitive to the damaging effects of cold shock [3]. The susceptibility of sperm to cryopreservation differs across species: rooster, human, monkey, rabbit and dog sperm are more resistant to cold shock than bull, ram, horse and boar sperm [3][4][5][6]. According to previous reports [4,6], the ratio of proteins to phospholipids in the plasma membrane is lowest in rooster sperm, at 0.46, moderate in bull and stallion sperm ...

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