Improvement of the post-thaw qualities of Okinawan native Agu pig sperm frozen in an extender supplemented with antiapoptotic PTD-FNK protein

Shimokawa, Kazuki; Oshiro, Ryuko; Yamanaka, Kenichi; Ashizawa, Koji; Ohta, Shigeo; Tatemoto, Hideki

doi:10.1016/j.theriogenology.2012.06.005

Cited by 5 publications

(4 citation statements)

References 55 publications

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“…Recently, anti-apoptotic proteins have been used to improve sperm survival after thawing (Shimokawa et al 2012). In our experiment, AKT inhibition was not accompanied by increased ROS production and DNA fragmentation, contrary to findings in humans where inhibition of PIK3CA was accompanied by increased mitochondrial production of ROS and oxidative DNA damage without changes in mitochondrial membrane potential .…”

Section: Discussioncontrasting

confidence: 99%

Phosphorylated AKT preserves stallion sperm viability and motility by inhibiting caspases 3 and 7

Bolaños¹,

Silva²,

Muñoz³

et al. 2014

Reproduction

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AKT, also referred to as protein kinase B (PKB or RAC), plays a critical role in controlling cell survival and apoptosis. To gain insights into the mechanisms regulating sperm survival after ejaculation, the role of AKT was investigated in stallion spermatozoa using a specific inhibitor and a phosphoflow approach. Stallion spermatozoa were washed and incubated in Biggers-Whitten-Whittingham medium, supplemented with 1% polyvinyl alcohol (PVA) in the presence of 0 (vehicle), 10, 20 or 30 mM SH5, an AKT inhibitor. SH5 treatment reduced the percentage of sperm displaying AKT phosphorylation, with inhibition reaching a maximum after 1 h of incubation. This decrease in phosphorylation was attributable to either dephosphorylation or suppression of the active phosphorylation pathway. Stallion spermatozoa spontaneously dephosphorylated during in vitro incubation, resulting in a lack of a difference in AKT phosphorylation between the SH5-treated sperm and the control after 4 h of incubation. AKT inhibition decreased the proportion of motile spermatozoa (total and progressive) and the sperm velocity. Similarly, AKT inhibition reduced membrane integrity, leading to increased membrane permeability and reduced the mitochondrial membrane potential concomitantly with activation of caspases 3 and 7. However, the percentage of spermatozoa exhibiting oxidative stress, the production of mitochondrial superoxide radicals, DNA oxidation and DNA fragmentation were not affected by AKT inhibition. It is concluded that AKT maintains the membrane integrity of ejaculated stallion spermatozoa, presumably by inhibiting caspases 3 and 7, which prevents the progression of spermatozoa to an incomplete form of apoptosis.Free Spanish abstract A Spanish translation of this abstract is freely available at

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Section: Discussioncontrasting

confidence: 99%

Phosphorylated AKT preserves stallion sperm viability and motility by inhibiting caspases 3 and 7

Bolaños¹,

Silva²,

Muñoz³

et al. 2014

Reproduction

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“…The Agu pig is not only a rare pig breed in Japan but also an important resource for the pork industry in Okinawa prefecture. Considering the possibility of breed extinction due to infiltration of malignant infectious diseases, new reproductive techniques may be required for storage and utilization of germplasms from Agu pigs in addition to conventional sperm freezing as reported previously (Shimokawa et al., 2012; Yoshimoto et al., 2008). Testicular xenografting combined with testis cryopreservation could provide an opportunity to rescue genetic information from piglets that need to be eliminated after a variety of accidents that may occur in the neonatal period.…”

Section: Discussionmentioning

confidence: 99%

Embryo production by intracytoplasmic injection of sperm retrieved from neonatal testicular tissue of Agu pigs after cryopreservation and grafting into nude mice

Kaneko

Kikuchi

Men

et al. 2020

Animal Science Journal

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The Agu is the only indigenous pig breed in Japan but its population is very small. In order to estimate the efficacy of testicular xenografting for the conservation of Agu pigs, we investigated whether neonatal testicular fragments would acquire the capacity to produce sperm after they had been cryopreserved and grafted into nude mice. Although on day 180 (day 0 = xenografting), grafts showed a low proportion of seminiferous tubule cross‐sections containing sperm (0.1 ± 0.1%, mean ± SEM for four mice), the proportion reached 36.9 ± 16.7% (n = 4 mice) by day 240. When single sperm obtained on day 240 was injected into individual porcine oocytes, 28.2% of the oocytes were found to contain one male and one female pronuclei with the second polar body. Moreover, the blastocyst formation rate after injection of the xenogeneic sperm was 28.4%, whereas that in the absence of sperm injection (attributable to parthenogenesis) was 13.3%. These findings suggest that more than half of the blastocysts resulted from fertilization. Thus, testicular xenografting could assist the conservation of Agu pigs by salvaging germ cells present in neonatal testes even after cryopreservation.

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“…To improve the motility of sperm that has been frozen and then thawed, several supplements can be added to the freezing solution, including chemicals and proteins [9, 27, 31, 34]. It is known that freezing of sperm induces a lipid phase transition in the plasma membrane resulting in cholesterol depletion, which causes membrane destabilization [4].…”

Section: Introductionmentioning

confidence: 99%

Effects of Cholesterol-Loaded Cyclodextrins on the Rate and the Quality of Motility in Frozen and Thawed Rabbit Sperm

et al. 2014

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The motility of sperm after freezing and thawing is critical for effective cryopreservation. It is known that supplementation with cholesterol-loaded cyclodextrin (CLC) improves cryosurvival of sperm in various animals. To clarify the effects of supplementation with CLC on rabbit sperm motility after freezing and thawing, rabbit sperm motility was analyzed using a computer-assisted sperm analysis system. Sperm motility with CLC supplementation was 29.4 ± 9.6% (mean ± SD), which was significantly higher than that of controls (20.8 ± 7.1%, P<0.05). The curvilinear velocity of sperm with CLC exceeded that of controls, whereas the values for linearity and wobble were significantly lower in sperm with CLC compared with controls. After artificial insemination, 44.3% of recovered ova were fertilized in the CLC-supplemented group, which was higher than the percentage in the control group (36.4%). The results indicate that supplementation with CLC improves the rate and quality of motility in rabbit sperm after freezing and thawing, and would be advantageous for successful cryopreservation.

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Improvement of the post-thaw qualities of Okinawan native Agu pig sperm frozen in an extender supplemented with antiapoptotic PTD-FNK protein

Cited by 5 publications

References 55 publications

Phosphorylated AKT preserves stallion sperm viability and motility by inhibiting caspases 3 and 7

Phosphorylated AKT preserves stallion sperm viability and motility by inhibiting caspases 3 and 7

Embryo production by intracytoplasmic injection of sperm retrieved from neonatal testicular tissue of Agu pigs after cryopreservation and grafting into nude mice

Effects of Cholesterol-Loaded Cyclodextrins on the Rate and the Quality of Motility in Frozen and Thawed Rabbit Sperm

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