Acrosin is essential for sperm penetration through the zona pellucida in hamsters

Acrosomal exocytosis (AR) is a critical process that sperm need to undergo to fertilize an egg. The evaluation of the presence or absence of the acrosome is usually performed by using lectins or dyes in fixed cells. With this approach, it is neither possible to monitor the dynamic process of exocytosis and related molecular events while discriminating between live and dead cells, nor to evaluate the acrosomal status while sperm reside in the female reproductive tract. However, over the last two decades, several new methodologies have been used to assess the occurrence of AR in living cells allowing different groups to obtain information that was not possible in the past. These techniques have revolutionized the whole study of this process. This review summarizes current methods available to analyze AR in living cells as well as the important information that emerged from studies using these approaches.

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“…In contrast, acrosin is essential for sperm penetration through the ZP in hamsters (Hirose et al, 2020).…”

Section: Transgenic Sperm Containing Egfp In Their Acrosomes Andmentioning

confidence: 92%

Seeing is believing: Current methods to observe sperm acrosomal exocytosis in real time

Balestrini

Jabloñski

Schiavi‐Ehrenhaus

et al. 2020

Molecular Reproduction Devel

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“…A caveat here is that much of this information about specific proteins' functions, as well as these recent data about polyspermy in vivo has come from the mouse model, and some differences in fertilization biology between species have been reported, for example, for the dependence of fertilization on the sperm protease acrosin (Baba, Azuma, Kashiwabara, & Toyoda, 1994; Adham, Nayernia, & Engel, 1997; Isotani et al, 2017; Hirose, et al, 2020). As noted above, data from numerous studies provide evidence that there likely are differences in how mammalian species prevent polyspermic fertilization.…”

Section: Further Concepts and Future Perspectives For Polyspermy Prevmentioning

confidence: 99%

Preventing polyspermy in mammalian eggs—Contributions of the membrane block and other mechanisms

Evans

2020

Molecular Reproduction Devel

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The egg's blocks to polyspermy (fertilization of an egg by more than one sperm) were originally identified in marine and aquatic species with external fertilization, but polyspermy matters in mammalian reproduction too. Embryonic triploidy is a noteworthy event associated with pregnancy complications and loss. Polyspermy is a major cause of triploidy with up to 80% of triploid conceptuses being the result of dispermic fertilization. The mammalian female reproductive tract regulates the number of sperm that reach the site of fertilization, but mammals also utilize egg-based blocks to polyspermy. The egg-based blocks occur on the mammalian egg coat (the zona pellucida) and the egg plasma membrane, with apparent variation between different mammalian species regarding the extent to which one or both are used. The zona pellucida block to polyspermy has some similarities to the slow block in water-dwelling species, but the mammalian membrane block to polyspermy differs substantially from the fast electrical block that has been characterized in marine and aquatic species. This review discusses what is known about the incidence of polyspermy in mammals and about the mammalian membrane block to polyspermy, as well as notes some lesser-characterized potential mechanisms contributing to polyspermy prevention in mammals. K E Y W O R D S embryo, fertilization, oocyte, plasma membrane, polyspermy How to cite this article: Evans JP. Preventing polyspermy in mammalian eggs-Contributions of the membrane block and other mechanisms.

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“…We developed a new method, called genome editing via oviductal nucleic acid delivery (GONAD), which was subsequently renamed "improved GONAD (i-GONAD)", for the production of genome-edited mice [16][17][18], rats [19,20], and hamsters [21]. This technology is based on the injection of a solution (1-1.5 µL) containing genome editing reagents into the lumen of an oviduct via the oviductal wall of pregnant female animals at the late zygote to two-cell stage following in vivo EP of the entire oviduct using tweezer-type electrodes under a dissecting microscope [22,23].…”

Section: Introductionmentioning

confidence: 99%

“…According to Gurumurthy et al [22], the optimal electric conditions for B6 involve in vivo EP being carried out under a constant current of 100 mA. To our knowledge, two electroporators, NEPA21 (NEPA GENE Co. Ltd., Chiba, Japan) and CUY21EditII, are those that have been most frequently used for successful GONAD/i-GONAD [16][17][18][19][20][21]. CUY21EditII can provide a constant current, whereas NEPA21 cannot.…”

Section: Introductionmentioning

confidence: 99%

Modification of i-GONAD Suitable for Production of Genome-Edited C57BL/6 Inbred Mouse Strain

Kobayashi

Aoshima

Ito

et al. 2020

Cells

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Improved genome editing via oviductal nucleic acid delivery (i-GONAD) is a novel method for producing genome-edited mice in the absence of ex vivo handling of zygotes. i-GONAD involves the intraoviductal injection of clustered regularly interspaced short palindromic repeats (CRISPR) ribonucleoproteins via the oviductal wall of pregnant females at 0.7 days post-coitum, followed by in vivo electroporation (EP). Unlike outbred Institute of Cancer Research (ICR) and hybrid mouse strains, genome editing of the most widely used C57BL/6J (B6) strain with i-GONAD has been considered difficult but, recently, setting a constant current of 100 mA upon EP enabled successful i-GONAD in this strain. Unfortunately, the most widely used electroporators employ a constant voltage, and thus we explored conditions allowing the generation of a 100 mA current using two electroporators: NEPA21 (Nepa Gene Co., Ltd.) and GEB15 (BEX Co., Ltd.). When the current and resistance were set to 40 V and 350–400 Ω, respectively, the current was fixed to 100 mA. Another problem in using B6 mice for i-GONAD is the difficulty in obtaining pregnant B6 females consistently because estrous females often fail to be found. A single intraperitoneal injection of low-dose pregnant mare’s serum gonadotrophin (PMSG) led to synchronization of the estrous cycle of these mice. Consequently, approximately 51% of B6 females had plugs upon mating with males 2 days after PMSG administration, which contrasts with the case (≈26%) when B6 females were subjected to natural mating. i-GONAD performed on PMSG-treated pregnant B6 females under conditions of average resistance of 367 Ω and average voltage of 116 mA resulted in the production of pregnant females at a rate of 56% (5/9 mice), from which 23 fetuses were successfully delivered. Nine (39%) of these fetuses exhibited successful genome editing at the target locus.

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Acrosin is essential for sperm penetration through the zona pellucida in hamsters

Cited by 83 publications

References 31 publications

Seeing is believing: Current methods to observe sperm acrosomal exocytosis in real time

Seeing is believing: Current methods to observe sperm acrosomal exocytosis in real time

Preventing polyspermy in mammalian eggs—Contributions of the membrane block and other mechanisms

Modification of i-GONAD Suitable for Production of Genome-Edited C57BL/6 Inbred Mouse Strain

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