Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100

Zambrano, Fabiola; Águila, Luis; Arias, María Elena; Sánchez, Raúl; Felmer, Ricardo

doi:10.1016/j.theriogenology.2016.05.007

Cited by 19 publications

(28 citation statements)

References 48 publications

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“…For this reason, we analyzed embryo quality to address whether or not our sperm pretreatments impact later development. Evaluation of embryo quality—in terms of cell count, cell allocation to the inner cell mass versus trophectoderm, and the apoptotic cell index—showed no differences between the treatments and the control group, and our data were comparable to those described previously for bovine ICSI (Arias et al, ; Jo et al, ; Rho et al, ; Zambrano et al, ). Thus, pretreatment with MβCD supports sperm decondensation after injection, and neither MβCD or IBMX pretreatments affect the quality of bovine ICSI‐derived embryos.…”

Section: Discussionsupporting

confidence: 90%

“…Indeed, the highest rate of embryonic development was observed in this group—a result also reported in a mouse ICSI model using the same sperm pretreatment (Tateno, ). Similar results were recently described using glutathione (Lee et al, ) and lysolecithin (Zambrano et al, ) as sperm pretreatments, confirming the positive effect of pretreatment in promoting DNA decondensation and pronucleus formation. On the other hand, although IBMX increased the sperm membrane fluidity, this treatment showed a similar pronucleus formation rate to the control group, so disrupted membrane integrity alone is not sufficient to improve nuclear condensation following ICSI.…”

Section: Discussionsupporting

confidence: 89%

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Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Águila

Zambrano

Arias

et al. 2017

Molecular Reproduction Devel

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The efficiency of intracytoplasmic sperm injection (ICSI) in bovines is low compared to other species due in part to inadequate egg activation and sperm nucleus decondensation after injection. We hypothesized that this low efficiency is due to the lack of complete sperm capacitation, so we evaluated the effects of isobutylmethylxanthine (IBMX) and methyl-β-cyclodextrin (MβCD) on bovine sperm capacitation and on the preimplantation developmental potential of bovine embryos generated by ICSI. Treatment with IBMX and MβCD decreased sperm viability (between 13-30%); nevertheless, 0.4 mM IBMX and 1 mM MβCD increased (p < 0.05) capacitation metrics-that is, acrosome exocytosis, intracellular calcium level, plasma membrane fluidity, and tyrosine phosphorylation-compared to the control. After ICSI, embryos injected with IBMX- and MβCD-treated sperm showed similar cleavage to the untreated group (range 82-88%). Pronucleus formation rate was higher with MβCD-pretreatment (54%) compared to the control group (25%), and blastocyst rate was significantly improved with MβCD-pretreatment (24%) compared to the IBMX (18%) and control (17%) groups. Importantly, embryo quality-as assessed by the total number of cells, cell allocation, and apoptotic cell index-was not affected by the sperm treatments. In conclusion, MβCD pretreatment of sperm improved the efficiency of blastocyst production in bovine ICSI.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionsupporting

confidence: 89%

Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Águila

Zambrano

Arias

et al. 2017

Molecular Reproduction Devel

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“…This alternative protocol, designed to closer mimic physiological conditions, avoids the use of harmful chemical agents that may damage the oocyte and spermatozoa (ex. dithiothreitol, triton X-100, lysolecithin, methyl-β-cyclodextrin) [10,12,38,39,40], and improves in vitro development of ICSI bovine embryos.…”

Section: Discussionmentioning

confidence: 99%

“…In agreement with those reports, our results show that the rates of live and acrosome reacted spermatozoa increase after Co-cult treatment. Some authors have evidenced that the use of acrosome reacted sperm for ICSI improves in vitro embryo development and implantation rates in cattle and human, respectively [12,28]. For all the exposed above, we employed the Cocult treatment for ICSI assay.…”

Section: Discussionmentioning

confidence: 99%

“…It is still unclear whether such inconsistencies are due to the inability of bull sperm to induce complete oocyte activation, or to the poor response of bovine ooplasms to the injection stimulus, which provokes an incorrect sperm head decondensation. For this reason, several oocyte activation protocols and sperm pretreatments have been developed, though with varying results [7][8][9][10][11][12][13]. The aim of this work was to evaluate treatments that better resemble physiological processes occurring to oocytes and sperm during fertilization, in order to facilitate ICSI embryo development.…”

Section: Introductionmentioning

confidence: 99%

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Improved embryo development using high cysteamine concentration during IVM and sperm co-culture with COCs previous to ICSI in bovine

et al. 2018

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In contrast to other species, intracytoplasmic sperm injection (ICSI) in bovine remains inefficient, resulting in low embryo developmental rates. It is unclear whether such inefficiency is due to the poor response of bovine ooplasms to the injection stimulus, or to the inability of bull sperm to induce oocyte activation. In order to facilitate these events, two strategies were assessed: the use of high concentration of cysteamine [Cys] during IVM; and the selection of sperm attached to cumulus cells after incubation with COCs for ICSI. First, COCs were IVM with increasing [Cys] and subjected to IVF. Zygotes from all groups were cultured under different O tensions and development to blastocyst was evaluated. In a second experiment, sperm were co-cultured for 3 h with COCs and acrosome reaction was studied. Afterwards, the best IVM and IVC conditions determined on Experiment 1 were used for ICSI assay. COCs were matured for 21 h with 1 (Cys 1) or 0.1 mM Cys (Cys 0.1 groups, standard condition). In addition, COCs were incubated for ≥3 h with 16 × 10 sperm/ml and only sperm attached to cumulus cells were selected for ICSI (ICSI + Co-cult groups). After chemical activation, embryos were cultured in SOF medium under low O tension. Cleavage and blastocyst rates were evaluated at days 2 and 7 of IVC, respectively. Finally, the relative expression of eight genes indicators of embryo quality was compared between ICSI and IVF control blastocysts by qPCR. Cleavage rates were higher for Cys 0.1 ICSI + Co-cult and Cys 1 ICSI + Co-cult groups (n = 117, 92% and n = 116, 79%, respectively) compared to their controls (n = 132, 60% for Cys 0.1 ICSI and n = 108, 52% for Cys 1 ICSI) (p ≤ 0.05). Interestingly, the combined treatment (Cys 1 ICSI + Co-cult) showed higher blastocyst rates than all other ICSI groups (23 vs. 11, 18 and 14% for Cys 0.1 ICSI + Co-cult, Cys 1 ICSI, and Cys 0.1 ICSI, respectively) (p ≤ 0.05). Moreover, incubation with COCs increased the rates of live acrosome reacted sperm (p ≤ 0.05). The relative abundance of mRNAs coding for INFτ, CAT, DNMT1, OCT4, and HDAC3 did not differ between treatments (p ≤ 0.05). SOD2, HADC1 and HADC2 expression was higher for Cys 0.1 ICSI than for IVF embryos (p ≤ 0.05). Group Cys 1 ICSI did not differ from IVF for those three genes, neither did Cys 1 ICSI + Co-cult, except for HDAC1 (p ≤ 0.05). In conclusion, the use of 1 mM Cys during IVM and of sperm incubated with mature COCs might be a good strategy to improve ICSI outcomes in cattle.

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Lower blastocyst quality after conventional vs. Piezo ICSI in the horse reflects delayed sperm component remodeling and oocyte activation

Salgado

Brom-de-Luna

Resende

et al. 2018

J Assist Reprod Genet

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Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100

Cited by 19 publications

References 48 publications

Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Improved embryo development using high cysteamine concentration during IVM and sperm co-culture with COCs previous to ICSI in bovine

Lower blastocyst quality after conventional vs. Piezo ICSI in the horse reflects delayed sperm component remodeling and oocyte activation

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