Changes in Sperm Motility and Capacitation Induce Chromosomal Aberration of the Bovine Embryo following Intracytoplasmic Sperm Injection

Kato, Yoku; Nagao, Yoshikazu

doi:10.1371/journal.pone.0129285

Cited by 25 publications

(23 citation statements)

References 47 publications

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“…Kato and Nagao suggested that capacitated sperm exhibiting hyperactivated motility induce chromosomal aberration in bovine ICSI‐embryos owing to a high mitochondrial activity and increased production of reactive oxygen species (Kato & Nagao, ). This was likely not influencing our present study because the spermatozoa were not selected based on motility.…”

Section: Discussionmentioning

confidence: 99%

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: Discussionmentioning

confidence: 99%

Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Águila

Zambrano

Arias

et al. 2017

Molecular Reproduction Devel

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“…After an injection it is possible to activate oocytes, in order to provide extrusion of the second polar body, by influence of 7 % etanol for 3-5 minutes [17,25]. However, outer activation of an oocyte can cause virginal reproduction [18].…”

Section: биология и биотехнологииmentioning

confidence: 99%

“…Oocytes demonstrating extrusion of the second PB after 3-4 hours after activation are considered to be insemintated [17,25].…”

Section: биология и биотехнологииmentioning

confidence: 99%

Features of the preparation of biological material for genome editing in cattle

Баркова

Makutina²,

Modorov³

et al. 2019

Agrarian Bulletin of The

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Abstract. To carry out genome editing in cattle, an effective and well-functioning system for obtaining gametes, fertilizing eggs and their cryopreservation is necessary. Aim of the work: review and research of present-day existing methods of obtaining, insemination and cryopreservation of donor material, in order to provide genome editing in cows. Methods and materials. The work is completed according to the theme No. 0532-2019-0001 “Development of complex technology of marker-based genome selection of agricultural animals” within State Order of Ministry of Education and Science of the Russian Federation. The analysis of open scientific literature on the issues of in vitro fertilization in animals, cryopreservation of oocytes and embryons, sperm preparation and methods of insemination of cows’ oocytes, and cryopreservation of oocytes and embryons of animals is done. Features of the preparation of biological material of cattle for genome editing by microinjection into ooplasm are described. Results of research and duscussion. At present time there are two ways to obtain donor material from cattle: from live animals and taking ovaries after slaughtering cows. Material transportation is carried out at a temperature of 30–37 °C depending on the distance to the laboratory and expected time period of transportation. Oocyte-cumulus complexes can be removed by ovarian dissection and aspiration of visible follicles. In both cases, immature eggs are predominantly obtained. Subsequent ripening is carried out in vitro using special media in a CO2 incubator. The culture medium for oocyte maturation should contain hormones that mimic the peak of LH (luteinizing hormone), which occurs in vivo during the maturation of oocytes before ovulation. To accumulate a certain number of eggs at the stage of MII, it is recommended to carry out their cryopreservation by the method of vitrification, having previously released the oocyte from the cumulus cells. After thawing, oocytes need to be incubated for 2–3 hours 38.5 °C in 5–6.5% CO2 to restore the spindle. In order to make editing more effective, the introduction of genetic material is recommended to be carried out in parallel with the fertilization method “icsi”. In humans, mice and rabbits, an injection of sperm into the cytoplasm is sufficient to activate the oocyte, however, in cattle, just micro-injection of the sperm is not enough and often the male pronucleus does not form. To solve the problem, various methods are used, including freezing-thawing of sperm, resulting in damage of a membrane, or addition of heparin-glutathione into the medium that increases decondensation of the sperm DNA.

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“…The evaluation of chromatin damage is an important aspect of sperm analysis because some sperm with chromatin abnormalities can fertilize the oocyte, but the embryo development is impaired, which leads to a high rate of embryonic death (ELLINGTON et al, 1998;KHALIFA et al, 2008;KATO;NAGAO, 2015).…”

Section: Chromatin Integritymentioning

confidence: 99%

Techniques for sperm evaluation using fluorescent probes

2015

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A variety of laboratory tests were developed to obtain more reliable results of sperm evaluation and increase the accuracy of sperm fertility predictions. These tests detected damage of sperm specific compartments or organelles, which cannot be detected in routine sperm analysis. The use of fluorescent probes and detection using fluorescent microscopy or flow cytometry is an important tool but a more precise and accurate laboratory test is needed. Propidium iodide and 6-carboxyfluorescein diacetate are used for evaluations of plasmatic membrane integrity. Fluorescein isothiocyanate, associated with conjugated lecithin Psium sativum or Arachis hypogaea, are used for evaluations of acrosome integrity. Two probes, MitoTracker or Rhodamine123, are generally used to measure the absence or presence of mitochondrial potential. However, a better option is 5,5'; 6,6' -tetrachloro -1,1'; 3,3' -tetraetilbenzimidazolil-carbocyanine (JC-1) dye, which assesses not only the presence of mitochondrial potential and distinguished spermatozoa with poorly and highly functional mitochondria. Two techniques, TUNEL or COMETA, and the Acridine Orange Test (AOT) dye are used to evaluate chromatin integrity. A fluorescence technique based on chlortetracycline (CTC) or Merocyanine 540 is used to estimate whether sperm pass by or through the capacitation process. This review focuses on the fluorescent probes that are most widely used to evaluate plasma membrane integrity, capacitation, acrosome integrity, chromatin integrity and mitochondrial potential. Key words: Fluorescence microscopy, sperm, cryopreservation ResumoDiversos testes laboratoriais têm sido desenvolvidos com o intuito de obter resultados confiáveis nas avaliações espermáticas, aumentando a qualidade e a confiabilidade do sêmen utilizado em técnicas de reprodução assistida. Esses testes têm permitido detectar danos em compartimentos e organelas específicas da célula espermática, que não são detectados nas análises de rotina. Dentre esses testes, o uso de sondas fluorescentes e sua detecção em microscópio de epifluorescência ou citometria de fluxo se tornaram ferramenta importante quando uma avaliação mais acurada é necessária. Para a avaliação da integridade de membrana plasmática, pode ser utilizado o iodeto de propídio, associado ao diacetato de 6-carboxifluoresceína. As avaliações de integridade acrossomal podem ser feitas através do isotiocianato de fluoresceína, associado às lecitinas conjugadas, como a Psium sativum ou Arachis hypogaea. O potencial mitocondrial pode ser avaliado quanto à ausência ou presença através da sonda Mitotracker ou Rodamina 123. Outra opção ainda melhor pode ser observada utilizando o corante iodeto de 5,5'; 6,6' -tetracloro -1,1'; 3,3' -tetraetilbenzimidazolil-carbocianina (JC-1), este corante avalia a presença do potencial mitocondrial e avalia quanto à classificação do grau. Para avaliar a integridade

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Changes in Sperm Motility and Capacitation Induce Chromosomal Aberration of the Bovine Embryo following Intracytoplasmic Sperm Injection

Cited by 25 publications

References 47 publications

Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Sperm capacitation pretreatment positively impacts bovine intracytoplasmic sperm injection

Features of the preparation of biological material for genome editing in cattle

Techniques for sperm evaluation using fluorescent probes

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