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

Kato Y, Nagao Y - PLoS ONE (2015)

Bottom Line: Intracytoplasmic sperm injection (ICSI) has become the method of choice to treat human male infertility.One of the outstanding problems associated with this technique is our current lack of knowledge concerning the effect of sperm capacitation and motility upon the subsequent development of oocytes following ICSI.In conclusion, these findings suggest that capacitated sperm exhibiting activated motility induced chromosomal aberration during development to the blastocyst stage following ICSI.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Production Science, United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Fuchu, 183-8509, Japan; University Farm, Faculty of Agriculture, Utsunomiya University, Mohka, Tochigi 321-4415, Japan.

ABSTRACT
Intracytoplasmic sperm injection (ICSI) has become the method of choice to treat human male infertility. One of the outstanding problems associated with this technique is our current lack of knowledge concerning the effect of sperm capacitation and motility upon the subsequent development of oocytes following ICSI. In the present study, we first examined the capacitation state of sperm exhibiting normal motility, along with sperm that had been activated, and examined the effect of reactive oxygen species (ROS) produced by these sperm types upon embryogenesis following bovine in vitro fertilization (IVF) and ICSI. Data showed that activated sperm reduced the chromosomal integrity of IVF/ICSI embryos at the blastocyst stage, while capacitated sperm produced ROS in capacitation media. Secondly, we treated sperm with carbonyl cyanide m-chlorophenyl hydrazine (CCCP), a chemical known to uncouple cell respiration within the mitochondria, and investigated the effect of this treatment upon blastocyst formation and chromosomal integrity at the blastocyst stage. Activated sperm in which the mitochondria had been treated with CCCP reduced levels of chromosomal aberration at the blastocyst stage following ICSI, by reducing mitochondrial activity in activated sperm. In conclusion, these findings suggest that capacitated sperm exhibiting activated motility induced chromosomal aberration during development to the blastocyst stage following ICSI. The injection of sperm exhibiting normal motility, or activated sperm in which mitochondrial activity had been reduced, improved the quality of ICSI-derived embryos. Therefore, the selection of sperm exhibiting progressive motility may not always be better for early embryo development and fetal growth following human ICSI, and that the use of a bovine model may contribute to a deeper understanding of sperm selection for human ICSI embryo development.

No MeSH data available.


Related in: MedlinePlus

ROS production of one individual motile sperm in non-capacitation media and capacitation media.Different letters within columns indicate significant differences as determined by one-way ANOVA with Tukey-Kramer analysis (P<0.01). This experiment was repeated 3–4 times. Statistical significance compared to control: a vs b vs c.
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pone.0129285.g007: ROS production of one individual motile sperm in non-capacitation media and capacitation media.Different letters within columns indicate significant differences as determined by one-way ANOVA with Tukey-Kramer analysis (P<0.01). This experiment was repeated 3–4 times. Statistical significance compared to control: a vs b vs c.

Mentions: ROS levels were estimated using the luminol assay [16]. Luminol (5-amino-2, 3-dihydro-1,4-phthalazinedione; also, 3- aminophthalic hydrazide) is used to estimate the combined levels of total ROS. The assay was sensitized by the addition of horseradish peroxidase to the sperm suspension, so as to increase the intensity of the spontaneous luminescence levels. In brief, 2 μl of 5mM luminol was added to 75μl aliquot of sperm (15 million sperm) and the reaction initiated by adding 23 μl of 2.5 units of horseradish peroxidase at RT. Results were monitored using a luminometer (Wallac 1420 VICTOR 2 T Multilabel Counter; PerkinElmer, Inc., Wellesley, MA) for 30 minutes at RT. In Fig 6, ROS production was expressed as the % of counted photons per second (CPS). In Fig 7, ROS production (CPS) was divided by the number of motile sperm in capacitation media and non-capacitation media, then ROS production by one individual sperm was expressed as a % of CPS.


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

Kato Y, Nagao Y - PLoS ONE (2015)

ROS production of one individual motile sperm in non-capacitation media and capacitation media.Different letters within columns indicate significant differences as determined by one-way ANOVA with Tukey-Kramer analysis (P<0.01). This experiment was repeated 3–4 times. Statistical significance compared to control: a vs b vs c.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465702&req=5

pone.0129285.g007: ROS production of one individual motile sperm in non-capacitation media and capacitation media.Different letters within columns indicate significant differences as determined by one-way ANOVA with Tukey-Kramer analysis (P<0.01). This experiment was repeated 3–4 times. Statistical significance compared to control: a vs b vs c.
Mentions: ROS levels were estimated using the luminol assay [16]. Luminol (5-amino-2, 3-dihydro-1,4-phthalazinedione; also, 3- aminophthalic hydrazide) is used to estimate the combined levels of total ROS. The assay was sensitized by the addition of horseradish peroxidase to the sperm suspension, so as to increase the intensity of the spontaneous luminescence levels. In brief, 2 μl of 5mM luminol was added to 75μl aliquot of sperm (15 million sperm) and the reaction initiated by adding 23 μl of 2.5 units of horseradish peroxidase at RT. Results were monitored using a luminometer (Wallac 1420 VICTOR 2 T Multilabel Counter; PerkinElmer, Inc., Wellesley, MA) for 30 minutes at RT. In Fig 6, ROS production was expressed as the % of counted photons per second (CPS). In Fig 7, ROS production (CPS) was divided by the number of motile sperm in capacitation media and non-capacitation media, then ROS production by one individual sperm was expressed as a % of CPS.

Bottom Line: Intracytoplasmic sperm injection (ICSI) has become the method of choice to treat human male infertility.One of the outstanding problems associated with this technique is our current lack of knowledge concerning the effect of sperm capacitation and motility upon the subsequent development of oocytes following ICSI.In conclusion, these findings suggest that capacitated sperm exhibiting activated motility induced chromosomal aberration during development to the blastocyst stage following ICSI.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Production Science, United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Fuchu, 183-8509, Japan; University Farm, Faculty of Agriculture, Utsunomiya University, Mohka, Tochigi 321-4415, Japan.

ABSTRACT
Intracytoplasmic sperm injection (ICSI) has become the method of choice to treat human male infertility. One of the outstanding problems associated with this technique is our current lack of knowledge concerning the effect of sperm capacitation and motility upon the subsequent development of oocytes following ICSI. In the present study, we first examined the capacitation state of sperm exhibiting normal motility, along with sperm that had been activated, and examined the effect of reactive oxygen species (ROS) produced by these sperm types upon embryogenesis following bovine in vitro fertilization (IVF) and ICSI. Data showed that activated sperm reduced the chromosomal integrity of IVF/ICSI embryos at the blastocyst stage, while capacitated sperm produced ROS in capacitation media. Secondly, we treated sperm with carbonyl cyanide m-chlorophenyl hydrazine (CCCP), a chemical known to uncouple cell respiration within the mitochondria, and investigated the effect of this treatment upon blastocyst formation and chromosomal integrity at the blastocyst stage. Activated sperm in which the mitochondria had been treated with CCCP reduced levels of chromosomal aberration at the blastocyst stage following ICSI, by reducing mitochondrial activity in activated sperm. In conclusion, these findings suggest that capacitated sperm exhibiting activated motility induced chromosomal aberration during development to the blastocyst stage following ICSI. The injection of sperm exhibiting normal motility, or activated sperm in which mitochondrial activity had been reduced, improved the quality of ICSI-derived embryos. Therefore, the selection of sperm exhibiting progressive motility may not always be better for early embryo development and fetal growth following human ICSI, and that the use of a bovine model may contribute to a deeper understanding of sperm selection for human ICSI embryo development.

No MeSH data available.


Related in: MedlinePlus