Roles of the zona pellucida and functional exposure of the sperm‐egg fusion factor ‘<scp>IZUMO</scp>’ during <i>in vitro</i> fertilization in pigs

Tanihara, Fuminori; Nakai, Michiko; Men, Nguyen Thi; Kato, Noriko; Kaneko, Hiroyuki; Noguchi, Junko; Otoi, Takeshige; Kikuchi, Kazuhiro

doi:10.1111/asj.12164

Cited by 9 publications

(3 citation statements)

References 37 publications

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“…IZUMO 4 is a member of the IZUMO multiprotein family containing four isoforms of IZUMO, of which 1, 2 and 3 are transmembrane proteins and 4 is a soluble, intra-acrosomal protein involved in the formation of protein complexes [49]. In porcine spermatozoa, the functional exposure of IZUMO proteins is essential for gamete fusion [50]. The presence of multiple proteoforms of IZUMO 4 in our study may indicate various post-translational modifications of this protein (e.g., phosphorylation, acetylation, glycosylation, ubiquitylation) [51].…”

Section: Redox-regulated Proteins Related To Acrosome Exocytosis During the Acrosome Reactionmentioning

confidence: 99%

Bull Sperm Capacitation Is Accompanied by Redox Modifications of Proteins

Mostek

Janta

Majewska

et al. 2021

IJMS

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The ability to fertilise an egg is acquired by the mammalian sperm during the complex biochemical process called capacitation. Capacitation is accompanied by the production of reactive oxygen species (ROS), but the mechanism of redox regulation during capacitation has not been elucidated. This study aimed to verify whether capacitation coincides with reversible oxidative post-translational modifications of proteins (oxPTMs). Flow cytometry, fluorescence microscopy and Western blot analyses were used to verify the sperm capacitation process. A fluorescent gel-based redox proteomic approach allowed us to observe changes in the level of reversible oxPTMs manifested by the reduction or oxidation of susceptible cysteines in sperm proteins. Sperm capacitation was accompanied with redox modifications of 48 protein spots corresponding to 22 proteins involved in the production of ROS (SOD, DLD), playing a role in downstream redox signal transfer (GAPDHS and GST) related to the cAMP/PKA pathway (ROPN1L, SPA17), acrosome exocytosis (ACRB, sperm acrosome associated protein 9, IZUMO4), actin polymerisation (CAPZB) and hyperactivation (TUBB4B, TUB1A). The results demonstrated that sperm capacitation is accompanied by altered levels of oxPTMs of a group of redox responsive proteins, filling gaps in our knowledge concerning sperm capacitation.

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Section: Redox-regulated Proteins Related To Acrosome Exocytosis During the Acrosome Reactionmentioning

confidence: 99%

Bull Sperm Capacitation Is Accompanied by Redox Modifications of Proteins

Mostek

Janta

Majewska

et al. 2021

IJMS

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“…The porcine ovaries were collected from an abattoir and transported to the laboratory in phosphate-buffered saline (PBS) at 36-38 • C. Cumulus-oocyte complexes were obtained by aspirating 3-8 mm follicles. As described by previous studies [42][43][44][45][46], the cumulus-oocyte complexes were isolated and incubated in PBS (Solarbio, Beijing, China) supplemented with 3 mg/mL hyaluronidase at 38.5 • C for 5 min, followed by gentle pipetting for 3 to 5 min to remove cumulus cells. The zona pellucidas were prepared from the cumulus-cell-free oocytes using a microinjector (Biocompare, San Francisco, CA, USA).…”

Section: Sperm-zona Pellucida (Zp) Binding Capacitymentioning

confidence: 99%

Pyrroloquinoline Quinone Improves Ram Sperm Quality through Its Antioxidative Ability during Storage at 4 °C

Zhu,

Li,

Yang

et al. 2024

Antioxidants

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Sperm motility is an important factor in the migration of sperm from the uterus to the oviduct. During sperm preservation in vitro, sperm generates excessive ROS that damages its function. This study aims to investigate whether the addition of pyrroloquinoline quinone (PQQ) to the diluted medium could improve chilled ram sperm quality, and then elucidates the mechanism. Ram semen was diluted with Tris-citric acid-glucose (TCG) medium containing different doses of PQQ (0 nM, 10 nM, 100 nM, 1000 nM, 10,000 nM), and stored at 4 °C. Sperm motility patterns, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential, reactive oxygen species (ROS) levels, malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity, and ATP levels were measured after preservation. Furthermore, the expressions of NADH dehydrogenase 1 (MT-ND1) and NADH dehydrogenase 6 (MT-ND6) in sperm were also detected by western blotting. In addition, sperm capacitation and the ability of sperm to bind to the zona pellucina were also evaluated. It was observed that the addition of PQQ significantly (p < 0.05) improved ram sperm motility, membrane integrity, and acrosome integrity during preservation. The percentage of sperm with high mitochondrial membrane potential in the PQQ treatment group was much higher than that in the control. In addition, supplementation of PQQ also decreased the sperm MDA and ROS levels, while increasing ATP levels. Interestingly, the levels of MT-ND1 and MT-ND6 protein in sperm treated with PQQ were also higher than that of the control. Furthermore, the addition of 100 nM PQQ to the medium decreased ROS damage in MT-ND1 and MT-ND6 proteins. The addition of 100 nM PQQ significantly (p < 0.05) increased protein tyrosine phosphorylation in ram sperm after induced capacitation. Furthermore, the value of the sperm–zona pellucida binding capacity in the 100 nM PQQ treatment group was also much higher than that of the control. Overall, during chilled ram- sperm preservation, PQQ protected ram sperm quality by quenching the ROS levels to reduce ROS damage and maintain sperm mitochondrial function, and preserved the sperm’s high ability of fertilization.

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“…However, the specific role of ZP glycoproteins in other mammalian species remains controversial. Moreover, there is a dearth of knowledge for livestock species like pigs, where the incidence of polyspermy after in vitro fertilisation remains a substantial problem (Tanihara et al 2014). In the pig, the ZP is comprised of the ZP2, ZP3, and ZP4 glycoproteins (Hedrick & Wardrip 1987), and ZP4 has been shown to orchestrate the sperm-ZP binding prior fertilisation (Yonezawa et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Sperm binding to ZP2-coated beads improve the efficiency of porcine in vitro fertilisation

2020

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The role of specific zona pellucida (ZP) glycoproteins in gamete interaction has not yet been elucidated in many species. A recently developed 3D model based on magnetic sepharose beads (B) conjugated to recombinant ZP glycoproteins (BZP) and cumulus cells (CBZP) allows the study of isolated ZP proteins in gamete recognition studies. The objective of this work was to study the role of porcine ZP2, ZP3 and ZP4 proteins in sperm binding, cumulus cell adhesion and acrosome reaction triggering. ZP protein-bound beads were incubated with fresh ejaculated boar spermatozoa and isolated cumulus cells for 24 h. The number of sperm bound to the beads, the acrosomal shrouds (presence of acrosomal content) on the bead’s surface, and the acrosome integrity (by means of PNA-FITC lectin) in bound and unbound sperm were studied. Finally, in vitro matured porcine oocytes mixed with BZP2were inseminated in vitro using fresh sperm and fertilization results evaluated. Over 60 % of beads had at least one sperm bound after 2 hours of co-incubation. ZP2-beads (BZP2) and cumulus-ZP2-bead complexes (CBZP2) reached the highest number of sperm per bead, whereas BZP3 and BZP4 models showed the highest number of unbound reacted sperm cells and acrosomal shrouds. Fertilization efficiency and monospermy rate increased when oocytes were fertilized in the presence of BZP2. We, therefore, conclude that in pigs it is mainly ZP2 that is involved in sperm-ZP binding whereas ZP3 and ZP4 induce acrosome reaction. Using magnetic sepharose ZP2-bound beads might be a valuable tool to improve the fertilization rate in pigs.

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Roles of the zona pellucida and functional exposure of the sperm‐egg fusion factor ‘IZUMO’ during in vitro fertilization in pigs

Cited by 9 publications

References 37 publications

Bull Sperm Capacitation Is Accompanied by Redox Modifications of Proteins

Bull Sperm Capacitation Is Accompanied by Redox Modifications of Proteins

Pyrroloquinoline Quinone Improves Ram Sperm Quality through Its Antioxidative Ability during Storage at 4 °C

Sperm binding to ZP2-coated beads improve the efficiency of porcine in vitro fertilisation

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