Lacking expression of paternally-expressed gene confirms the failure of syngamy after intracytoplasmic sperm injection in swamp buffalo (Bubalus bubalis)

Chankitisakul, Vibuntita; Tharasanit, Theerawat; Phutikanit, Nawapen; Tasripoo, K.; Nagai, Takashi; Techakumphu, Mongkol

doi:10.1016/j.theriogenology.2011.11.007

Cited by 5 publications

(4 citation statements)

References 59 publications

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“…In the present study, we were able to successfully collect sperm from collared peccary testis xenografts and, also for the first time, harvest peccary oocytes, which subsequently after standard ICSI procedures (Pereyra-Bonnet et al 2008) resulted in diploid embryos. Using the PCR technique, we observed that twothirds of the four-cell embryos obtained expressed the paternally imprinted gene NNAT (Park et al 2011, Chankitisakul et al 2012. The low ICSI efficiency reported in the present study probably occurred due to the absence of standardized protocols for this species.…”

Section: Experimental Groupcontrasting

confidence: 52%

Derivation of sperm from xenografted testis cells and tissues of the peccary (Tayassu tajacu)

Campos-Júnior¹,

Costa²,

Avelar³

et al. 2014

Reproduction

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Because the collared peccary (Tayassu tajacu) has a peculiar Leydig cell cytoarchitecture, this species represents a unique mammalian model for investigating testis function. Taking advantage of the well-established and very useful testis xenograft technique, in the present study, testis tissue and testis cell suspensions from immature collared peccaries (nZ4; 3 months old) were xenografted in SCID mice (nZ48) and evaluated at 2, 4, 6, and 8 months after grafting. Complete spermatogenesis was observed at 6 and 8 months after testis tissue xenografting. However, probably due to de novo testis morphogenesis and low androgen secretion, functionally evaluated by the seminal vesicle weight, a delay in spermatogenesis progression was observed in the testis cell suspension xenografts, with the production of fertile sperm only at 8 months after grafting. Importantly, demonstrating that the peculiar testicular cytoarchitecture of the collared peccary is intrinsically programmed, the unique Leydig cell arrangement observed in this species was re-established after de novo testis morphogenesis. The sperm collected from the xenografts resulted in diploid embryos that expressed the paternally imprinted gene NNAT after ICSI. The present study is the first to demonstrate complete spermatogenesis with the production of fertile sperm from testis cell suspension xenografts in a wild mammalian species. Therefore, due to its unique testicular cytoarchitecture, xenograft techniques, particularly testis cell suspensions, may represent a new and very promising approach to evaluate testis morphogenesis and to investigate spermatogonial stem cell physiology and niche in the collared peccary.

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Section: Experimental Groupcontrasting

confidence: 52%

Derivation of sperm from xenografted testis cells and tissues of the peccary (Tayassu tajacu)

Campos-Júnior¹,

Costa²,

Avelar³

et al. 2014

Reproduction

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“…Our previous study demonstrated a failure of male pronucleus formation after ICSI in swamp buffalo oocytes [1]. In the present study, we investigated the effects of sperm pretreatments on male pronucleus formation following ICSI.…”

Section: Discussionmentioning

confidence: 94%

“…The treated sperm were diluted using 10% polyvinylpyrrolidone in H-SOF. Sperm were injected as described previously [1]. Injection occurred within one hour of each sperm treatment.…”

Section: Methodsmentioning

confidence: 99%

“…We have previously reported that an injected sperm itself poorly stimulates the activation process in buffalo oocytes. Although additional chemical activation following ICSI can promote the development of buffalo ICSI oocytes to the blastocyst stage, a failure of male pronucleus formation has been largely observed [1]. A possible reason for this failure may be the lack of an important signaling molecule when a sperm head is injected into an oocyte.…”

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confidence: 99%

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Sperm Pretreatment with Dithiothreitol Increases Male Pronucleus Formation Rates After Intracytoplasmic Sperm Injection (ICSI) in Swamp Buffalo Oocytes

Chankitisakul

Am-In

Tharasanit

et al. 2013

Journal of Reproduction and Development

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Failure of male pronucleus formation has hampered the success of intracytoplasmic sperm injection (ICSI) in swamp buffalo. The aim of the present study was to improve male pronucleus formation by pretreating sperm with various chemicals before ICSI. In Experiments1 and 2, sperm were treated according to one of the following protocols: (1) 0.1% Triton-X 100 (TX) for 1 min, (2) 10 µM calcium ionophore (CaI) for 20 min, (3) freezing and thawing (FT) without any cryoprotectant, or (4) no treatment (control). These sperm treatment groups then either did or did not receive additional sperm treatment with 5 mM dithiothreitol (DTT) for 20 min. Acrosomal integrity (Experiment 1) and DNA fragmentation (Experiment 2) were evaluated in the sperm before ICSI. In Experiment 3, oocytes matured in vitro were subjected to ICSI using pretreated sperm as described above and then were cultured either with or without activation. The TX- and CaI-treated sperm caused an increase in the number of acrosome-loss sperm, whereas the FT treatment and control increased the proportion of acrosome-reacted sperm (P<0.05). The DNA fragmentation did not differ among treatments (P>0.05). At 18 h post-ICSI, pronucleus (PN) formation was found only in activated oocytes. The majority of the activated ICSI oocytes contained intact sperm heads. Normal fertilization was observed in the CaI and FT treatment groups and control group when sperm were treated with DTT before ICSI. In conclusion, DTT treatment of sperm with reacted acrosomes before ICSI together with activation of the ICSI oocytes is important for successful male pronucleus formation.

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Past, present and future of ICSI in livestock species

Briski

Salamone

2022

Animal Reproduction Science

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Lacking expression of paternally-expressed gene confirms the failure of syngamy after intracytoplasmic sperm injection in swamp buffalo (Bubalus bubalis)

Cited by 5 publications

References 59 publications

Derivation of sperm from xenografted testis cells and tissues of the peccary (Tayassu tajacu)

Derivation of sperm from xenografted testis cells and tissues of the peccary (Tayassu tajacu)

Sperm Pretreatment with Dithiothreitol Increases Male Pronucleus Formation Rates After Intracytoplasmic Sperm Injection (ICSI) in Swamp Buffalo Oocytes

Past, present and future of ICSI in livestock species

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