Unilateral Intrauterine Horn Insemination of Frozen Semen in Cats.

Tsutsui, Toshihiko; Tanaka, Akihiro; Takagi, Yuuko; Nakagawa, Kiyoshi; Fujimoto, Youko; Murai, Mayumi; Anzai, Mizuho; Hori, Tatsuya

doi:10.1292/jvms.62.1247

Cited by 43 publications

(42 citation statements)

References 7 publications

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“…were pooled and minced in 1 mL of egg yolk tris-fructose citrate solution (EYT-FC) [20] to liberate the sperm. The resulting solution was filtered through a sterile nylon mesh (pore size, 100 μm) and sperm were recovered (~55% of sperm were progressively motile).…”

Section: Sperm Recovery and Cryopreservationmentioning

confidence: 99%

Effect of duration of in vitro maturation on nuclear maturation and fertilizability of feline oocytes

et al. 2008

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This study was conducted to improve in vitro production of embryos from domestic cats using TCM-199 as an IVM medium. The time sequence of nuclear maturation and the optimal timing of in vitro insemination were examined. Most oocytes were at the germinal vesicle stage immediately after collection; however, 8.3% had already resumed meiosis before IVM culture. After 30 h of IVM culture, the percentage of oocytes at metaphase II (MII) reached a peak (75.5%) and did not change (P>0.05) from 30 to 48 h after IVM culture. The percentage of oocytes with two pronuclei was higher (P<0.05) for oocytes matured for 30 and 36 h (38.2 and 33.0%, respectively) than for those after IVM culture for only 24 h (18.5%).Total sperm penetration rate was highest (P<0.05) for oocytes that had been matured for 30 h (46.1%). After 30 h of IVM and 18 h of IVF culture, 66.3 and 24.8% of inseminated oocytes had cleaved and developed to the blastocyst stage, respectively. We concluded that IVM of feline oocytes for 30 h in TCM-199 resulted in optimal nuclear maturation and sperm penetration.

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Section: Sperm Recovery and Cryopreservationmentioning

confidence: 99%

Effect of duration of in vitro maturation on nuclear maturation and fertilizability of feline oocytes

et al. 2008

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“…The blood vessel on the surface of the epididymis was removed and it was cut at the corpus part near the cauda epididymis and divided into the caput-corpus and caudal epididymides. Each part was minced on a plastic dish with 2 ml of extender, egg yolk tris-fructose citrate solution (EYT-FC) [16]. The solution to transmigrate sperm was filtered through a metal mesh (80 µm) to remove tissue fragments, and sperm were recovered at room temperature (22-23°C).…”

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Artificial Insemination of Frozen Epididymal Sperm in Beagle Dogs

Hori

Ichikawa

Kawakami

et al. 2004

The Journal of Veterinary Medical Science

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ABSTRACT. Freeze-storage of epididymal sperm is an important technique for the preservation of gametes in animals, including those becoming extinct. We froze canine sperm recovered from the cauda epididymis and investigated the fertility. The qualities of sperm from the cauda epididymis before freezing were: mean sperm motility, 89.4 ± 1.6 (SE) %; sperm viability, 89.1 ± 1.1%; and these were significantly higher than those of sperm from the caput-corpus epididymis (P<0.01, P<0.05). The number of sperm recovered from both cauda epididymides varied among animals: 6.3-122.3 × 10 7 , mean 61.5 ± 10.0 × 10 7 . Freezing was used only for sperm recovered from the cauda epididymis. The sperm motility and viability after thawing were 19.5 ± 2.5% and 53.1 ± 3.3%, respectively. These were slightly lower than those of frozen-thawed ejaculated sperm, but the differences were not significant. When 2 × 10 8 , 3 × 10 8 , or 4 × 10 8 sperm were inseminated in the unilateral uterus, only one animal inseminated with 3 × 10 8 sperm was fertilized (1/16, 6.3%). When 1 × 10 8 sperm were inseminated in the bilateral uterine tubes, one of six animals (16.7%) was fertilized. Therefore, although the qualities of epididymal sperm after thawing were similar to those of ejaculated sperm, the conception rate obtained with frozen-thawed epididymal sperm was low in beagle dogs. It is necessary to investigate the differences in damage between epididymal sperm after thawing and ejaculated sperm and to develop a method for improving the conception rate. KEY WORDS: canine, epididymal sperm, frozen, intratubal insemination, intrauterine insemination.J. Vet. Med. Sci. 66(1): [37][38][39][40][41] 2004 Sperm reach maturation during migration from the caput to the cauda of the epididymis and are retained in the cauda epididymis until ejaculation. It has been demonstrated in many species that sperm in the cauda epididymis exhibited fertility in in vitro fertilization and artificial insemination [1,4,[7][8][9]14]. For animals that die unexpectedly, transmigration of epididymal sperm, and its freeze-storage are important techniques for gamete preservation. These techniques are considered important for preventing species from becoming extinct. Conception obtained by artificial insemination of frozen epididymal semen has been reported in one dog [7], pigs [8], goats [1], and cats [14]. Marks et al. [7] successfully obtained delivery of one pup after intrauterine insemination despite poor semen quality after thawing. There is only one study about the quality and resistance to freezing of canine epididymal sperm, reported by Hewitt et al. [5]. Their study showed that the number of sperm that reached maturation was lower in the epididymis than in frozen ejaculated sperm, but there was no difference in oocytepenetrating ability [5]. In the method of epididymal sperm recovery, Marks et al. [7] reported that the epididymis was perfused from the seminal duct to the epididymis in boxer dogs and Hewitt et al. [5] used the mincing method. Sirivaidyapong [11] com...

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“…The epididymis was cut at the corpus into two equal parts. Each part was minced in 1 ml of egg yolk Tris-fructose citrate solution (EYT-FC) [23] to transmigrate the sperm. The solution was filtered through a stainless steel mesh (80 µm) and sperm were recovered.…”

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“…Sperm from 6 animals were supplemented with 1% OEP. When there were no major differences in sperm quality between the right and left epididymides, sperm from bilateral epididymides were combined and frozen.Artificial insemination: Ovulation was induced by subcutaneous injection of 100 IU hCG on the third or fourth day of estrus [23]. Semen was inseminated in a unilateral uterine horn with more mature ovarian follicles 20 hr after hCG administration before ovulation [23].…”

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Artificial Insemination with Frozen Epididymal Sperm in Cats.

Tsutsui

Wada

Anzai

et al. 2003

The Journal of Veterinary Medical Science

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ABSTRACT. Artificial insemination with frozen cauda epididymal sperm was performed in cats. Sperm were transmigrated from the epididymides in 10 male cats. The mean sperm motility and viability were 67% and 82.5%, respectively, and 11.6 × 10 7 sperm were recovered. The mean sperm motility after thawing was 24.0%. Eleven female cats received unilateral intrauterine insemination of 5 × 10 7 sperm, and the conception rate was 27.3% (3/11). This was the first case of conception obtained with frozen epididymal sperm in cats. KEY WORDS: artificial insemination, epididymal sperm, feline.J. Vet. Med. Sci. 65(3): 397-399, 2003 It has been demonstrated in many species that epididymal sperm have fertility in in vitro fertilization and artificial insemination (AI) [4,8,[11][12][13][14][15]25], and fertilization with frozen epididymal sperm was confirmed in pigs [11] and dogs [9].Freeze-storage of epididymal sperm is an important technique for gamete preservation of species, particularly for preservation of animals becoming extinct. As wild animals in the cat family are becoming extinct, we selected cats as an animal model.We have performed studies of reproductive techniques such as in vitro fertilization [5,6], embryo transfer [6,24], and AI [18,[20][21][22][23] in cats.With regard to feline epididymal sperm, studies of the qualities [1,2], state of fertility acquisition [2,3,12], and freeze-storage method [4,8,17] are progressing, and fertility of fresh epididymal sperm [12] and frozen semen [4,8] has been shown, but no in vivo study with frozen epididymal sperm has been performed. With frozen feline ejaculated semen, Platz et al.[16] and we [23] have succeeded in obtaining newborns by intravaginal and intrauterine insemination, respectively. Therefore, we froze epididymal sperm by the same method as for freezing ejaculated sperm and investigated the possibility of inducing conception by intrauterine insemination. It has been shown that frozen semen supplemented with Orvus ES Paste (OEP, Nova Chemical Sales Scituate, Inc., MA, U.S.A.) has superior properties after thawing semen without OEP in dogs [9] and pigs [7]. In this way, the usefulness of OEP was also investigated in this study.Animals: Ten male crossbred cats aged 1.0-3.3 years (mean: 1.9 ± 0.3, SE) were used. For AI, 11 female cats aged 2-4 years were used. The cats were kept in groups in an animal room measuring 4 × 7 m with temperature controlled to 23 ± 2°C. The animals were given commercial dry cat food (Hill's Feline Maintenance Dry, U.S.A.).Transmigration of sperm from epididymis: Surgically excised testis and epididymis were weighed. The epididymis was cut at the corpus into two equal parts. Each part was minced in 1 ml of egg yolk Tris-fructose citrate solution (EYT-FC) [23] to transmigrate the sperm. The solution was filtered through a stainless steel mesh (80 µm) and sperm were recovered. The time from excision of the epididymis to transmigration of the sperm was within 1 hr.Sperm quality test and freezing: The recovered sperm were examined by the gen...

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Unilateral Intrauterine Horn Insemination of Frozen Semen in Cats.

Cited by 43 publications

References 7 publications

Effect of duration of in vitro maturation on nuclear maturation and fertilizability of feline oocytes

Effect of duration of in vitro maturation on nuclear maturation and fertilizability of feline oocytes

Artificial Insemination of Frozen Epididymal Sperm in Beagle Dogs

Artificial Insemination with Frozen Epididymal Sperm in Cats.

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