Successful induction of pseudopregnancy using sonic vibration in mice

Wake, Yui; Endo, Marina; Tsunoda, Shigemi; Tawara, Hirosuke; Abe, Hisayuki; Nakagawa, Yuki; Kaneko, Takehito

doi:10.1038/s41598-023-30774-x

“…Unlike their method, which requires two administrations, our method requires only one administration, reducing the burden on the experimenter and pain to mice. Recently, successful mechanical mimicry of mating stimulation by sonic vibration has been reported in rats and mice (Kaneko et al, 2020; Wake et al, 2023). Although they conducted fewer experiments, the pregnancy rate of their method was high (75% to 100%); hence, so their method is effective for producing a small number of pseudo pregnant mice.…”

Section: Discussionmentioning

confidence: 99%

Intraperitoneal administration of mouse kisspeptin‐10 to mice during estrus stage induces pseudopregnancy

Mashiko,

Tokoro,

Yao

et al. 2023

Genes to Cells

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A simple method for producing pseudopregnant mice supports pup production. In this study, pregnant ICR were obtained mice without mating with vasectomized mice via administration of mouse Kisspeptin‐10 (mKp‐10) and transferring blastocysts to the uterus. Blastocyst transfer after mKp‐10 administration to mice with gapping and reddish pink vagina resulted in 65.2% (15/23) pregnancies, and 39.1% (34/87) of the transferred blastocysts showed full‐term growth. Vaginal smears were observed for accurate estrus cycle determination, and subsequent administration of mKp10 to mice during the estrus stage and blastocyst transfer resulted in 95.2% (20/21) pregnancies and 50.7% (104/205) birth rates. Regarding 2‐cell transfer after administration of mKp‐10, 100% (8/8) of the mice became pregnant, and 45.0% (36/80) of the embryos were born. Administration of mKp‐10 to mice during the estrus stage is a convenient way to generate pseudopregnant mice.

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“…For earlier-stage embryos, embryo culture to the blastocyst stage is required. However, if a surgical transfer is preferred, it is possible to adapt the CM technique to the correct timing needed for appropriate pseudopregnant recipients 2 . In general, the embryo recipients are 1 day less advanced than the embryo.…”

Section: Cytological Evaluation For Pseudopregnancy or Pregnancymentioning

confidence: 99%

“…Historically, pseudopregnant recipients have been generated by mating with sterile males, which are either surgically vasectomized or genetically infertile, and the presence of a copulation plug is assessed the following morning 1 . Recently, a protocol for sonic stimulation in mice has been developed for the surgical transfer of pronuclear or two-cell mouse embryos 2 . We have also developed a cervical manipulation (CM) protocol for use with artificial insemination and the non-surgical embryo transfer of blastocysts.…”

Section: Introductionmentioning

confidence: 99%

Inducing Pseudopregnancy in Female Mice Without the Need for Vasectomized Males Prior to Non-Surgical Embryo Transfer or Artificial Insemination

Stone

¹

,

Srodulski

²

2023

JoVE

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For successfully maintaining pregnancy with embryo transfer or artificial insemination, female recipient mice must be induced into a pseudopregnant state.Female mice are traditionally paired overnight with vasectomized males, and the following morning, the presence of a copulation plug is assessed. To increase the efficiency of producing pseudopregnant females, a cervical manipulation technique has been standardized to be used in combination with non-surgical embryo transfer or artificial insemination techniques in mice. The blunt end of a small plastic rod is inserted vaginally to contact the cervix and is vibrated for 30 s by contact with a trimmer. The procedure is quick and does not require anesthesia or analgesia. This technique increases the reliability and predictability of producing pseudopregnant females and entirely eliminates the requirement for vasectomized males. For CD1 mice, the efficiency of pseudopregnancy induction using cervical manipulation was 83% for females in estrus (N = 76) but only 38% of females in estrus were plugged by vasectomized males (N = 24). Artificial insemination in CD1 mice was performed by estrus synchronization with hormones, cervical manipulation, and the uterine transfer of sperm. Artificial insemination recipients receiving cervical manipulation (N = 76) had a pregnancy rate of 72% and an average litter size of 8.3 pups. This method can also be used to produce pseudopregnant females for non-surgical embryo transfer. Therefore, inducing pseudopregnancy by cervical manipulation is a convenient and efficient alternative to mating with a vasectomized male when performing nonsurgical assisted reproduction techniques. Using cervical manipulation provides 3Rs (replacement, reduction, and refinement) benefits for assisted reproduction techniques by reducing the number of animals required and eliminating the necessity for surgically altered males.

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“…For earlier-stage embryos, embryo culture to the blastocyst stage is required. However, if a surgical transfer is preferred, it is possible to adapt the CM technique to the correct timing needed for appropriate pseudopregnant recipients 2 . In general, the embryo recipients are 1 day less advanced than the embryo.…”

Section: Cytological Evaluation For Pseudopregnancy or Pregnancymentioning

confidence: 99%

“…Historically, pseudopregnant recipients have been generated by mating with sterile males, which are either surgically vasectomized or genetically infertile, and the presence of a copulation plug is assessed the following morning 1 . Recently, a protocol for sonic stimulation in mice has been developed for the surgical transfer of pronuclear or two-cell mouse embryos 2 . We have also developed a cervical manipulation (CM) protocol for use with artificial insemination and the non-surgical embryo transfer of blastocysts.…”

Section: Introductionmentioning

confidence: 99%

JoVE Video Dataset

0

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For successfully maintaining pregnancy with embryo transfer or artificial insemination, female recipient mice must be induced into a pseudopregnant state.Female mice are traditionally paired overnight with vasectomized males, and the following morning, the presence of a copulation plug is assessed. To increase the efficiency of producing pseudopregnant females, a cervical manipulation technique has been standardized to be used in combination with non-surgical embryo transfer or artificial insemination techniques in mice. The blunt end of a small plastic rod is inserted vaginally to contact the cervix and is vibrated for 30 s by contact with a trimmer. The procedure is quick and does not require anesthesia or analgesia. This technique increases the reliability and predictability of producing pseudopregnant females and entirely eliminates the requirement for vasectomized males. For CD1 mice, the efficiency of pseudopregnancy induction using cervical manipulation was 83% for females in estrus (N = 76) but only 38% of females in estrus were plugged by vasectomized males (N = 24). Artificial insemination in CD1 mice was performed by estrus synchronization with hormones, cervical manipulation, and the uterine transfer of sperm. Artificial insemination recipients receiving cervical manipulation (N = 76) had a pregnancy rate of 72% and an average litter size of 8.3 pups. This method can also be used to produce pseudopregnant females for non-surgical embryo transfer. Therefore, inducing pseudopregnancy by cervical manipulation is a convenient and efficient alternative to mating with a vasectomized male when performing nonsurgical assisted reproduction techniques. Using cervical manipulation provides 3Rs (replacement, reduction, and refinement) benefits for assisted reproduction techniques by reducing the number of animals required and eliminating the necessity for surgically altered males.

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Successful induction of pseudopregnancy using sonic vibration in mice

Cited by 3 publications

References 32 publications

Intraperitoneal administration of mouse kisspeptin‐10 to mice during estrus stage induces pseudopregnancy

Intraperitoneal administration of mouse kisspeptin‐10 to mice during estrus stage induces pseudopregnancy

Inducing Pseudopregnancy in Female Mice Without the Need for Vasectomized Males Prior to Non-Surgical Embryo Transfer or Artificial Insemination

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