Artificial Insemination with Canine Spermatozoa Frozen in a Skim Milk/Glucose-Based Extender

Abe, Yasuyuki; Lee, Dong‐Soo; Sano, Hikaru; Akiyama, Koji; Yanagimoto-Ueta, Yoshiko; Asano, Tomomi; Suwa, Yoshinori; Suzuki, Hiroshi

doi:10.1262/jrd.19148

Cited by 22 publications

(21 citation statements)

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“…Despite the fact that the dog is one of the most useful animals for various fields of research including reproduction [42][43][44], and successful cryopreservation canine embryo production in vitro is very limited, possibly because no optimal culture system for IVM of canine oocytes has yet been established. In order to clarify physiological evaluation of canine oocytes for IVM, we examined timedependent changes in MAPK and p34 cdc2 kinase activities during meiotic progression in canine oocytes, finding that both MAPK and p34 cdc2 kinase activities increased in a time-dependent manner and then reached maximal levels at 72 h of culture.…”

Section: Discussionmentioning

confidence: 99%

Kinetics of Nuclear Status and Kinase Activities During In Vitro Maturation of Canine Oocytes

Suzukamo

Hoshina

Moriya

et al. 2009

Journal of Reproduction and Development

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Abstract. In contrast to those of other mammals, canine oocytes are ovulated at the germinal vesicle (GV) stage and then progress to the metaphase II (MII) stage in the oviduct. In other species, oocytes at the MII are widely used for in vitro fertilization or as recipients in somatic cell nuclear transfer. Many researchers have tried to improve the in vitro maturation (IVM) of canine oocytes. However, the proportion of MII oocytes remains low, resulting in poor efficiency of embryogenesis in vitro. This leads us to the possibility that the in vitro cytoplasmic maturation of canine oocytes is insufficient. Furthermore, the optimal culture period for IVM of canine oocytes is controversial, and physiological evaluation is required to improve canine IVM. We show here the time-dependent changes in mitogen-activated protein kinase (MAPK) and p34 cdc2 kinase activities in canine oocytes during IVM, since it is well known that both MAPK and p34 cdc2 kinase are activated following meiotic progression and show high activities in the MII stage in other species. Immediately after collection from ovaries, most oocytes were arrested at the GV stage, which was maintained until 24 h of culture. At 48 h of culture, more than half of the oocytes had progressed beyond the MI stage. A higher proportion of MII oocytes were observed with 72 h of culture compared with other culture periods. MAPK activity was found to increase in a time-dependent manner and reached a plateau at 72 h of culture. The level of p34 cdc2 kinase activity also increased in a time-dependent manner, with its maximal level observed after 72 h of culture. Activity was decreased with 96 h of culture, although there was no significant difference in the proportion of MII oocytes between 72 and 96 h. Our data thus show that the optimal culture period for IVM of canine oocytes is 72 h because both MAPK and p34 cdc2 kinase showed high activities at that time. Key words: Canine, In vitro maturation, Kinase, Oocyte (J. Reprod. Dev. 55: [116][117][118][119][120] 2009) ocytes in most mammalian species progress to the metaphase II (MII) stage in follicles and are then ovulated into the oviduct; however, canine oocytes are ovulated as immature oocytes at the germinal vesicle (GV) stage. These ovulated canine oocytes require at least 48 h to progress to the MII stage within the oviduct [1,2]. Many researchers have reported the in vitro maturation (IVM) of canine oocytes, and the discussion in the literature now focuses on how to improve development. For example, the age of the donor bitch [3], oocyte diameter [4,5], protein [6][7][8][9][10] and hormonal supplementation of maturation media [11][12][13][14][15][16] have all been examined. However, the proportion of MII oocytes remains low (0-43.4%) [9,[15][16][17][18][19][20], and a number of essential issues have yet to be resolved.One of these issues is the optimal culture period for IVM of canine oocytes. When oocytes are recovered directly from ovarian follicles and cultured in vitro, some progress to the MII stage afte...

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Section: Discussionmentioning

confidence: 99%

Kinetics of Nuclear Status and Kinase Activities During In Vitro Maturation of Canine Oocytes

Suzukamo

Hoshina

Moriya

et al. 2009

Journal of Reproduction and Development

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“…The collected semen was cryopreserved using the SM‐based extender described in the previous report with some modifications (Abe et al., ). To prepare the SM‐based extender, 30 mg/ml of skim milk (232100; Difco, Le Pont de Claix, France) and 119 mg/ml raffinose pentahydrate (R7630) were dissolved in Milli‐Q water at 60°C, and then, the solution was centrifuged at 10,000 g for 15 min at room temperature.…”

Section: Methodsmentioning

confidence: 99%

“…As an alternative to EY, skim milk (SM) seems to be a suitable semen extender in canine species, because it is the most commonly used cryoprotectant for mouse (Nakagata, ) and goat spermatozoa (Dorado, Rodriguez, & Hidalgo, ). We previously reported that SM is an effective cryoprotectant for cryopreservation of canine spermatozoa (Abe et al., ). Motility and other motion parameters of canine spermatozoa frozen with a simple extender composed of SM, glucose and glycerol were equivalent or much better than the EY‐based extender.…”

Section: Introductionmentioning

confidence: 99%

Fertilizing ability of canine spermatozoa cryopreserved with skim milk‐based extender in a retrospective study

Abe

Yokozawa

Umemiya‐Shirafuji

et al. 2017

Reprod Domestic Animals

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We previously reported that skim milk (SM) is an effective cryoprotectant for cryopreservation of canine spermatozoa instead of egg yolk (EY), which is the conventional cryoprotectant. In this study, the fertilizing ability and practical use of frozen canine spermatozoa prepared with SM were evaluated by transcervical insemination. Frozen-thawed spermatozoa were inseminated one to four times on days 2-9 after the LH surge. In SM group, a single transcervical insemination (TCI) on Day 5 led to higher delivery rate (83%) than any other days (33%-50%) post-LH surge. In EY group, delivery rate in double TCI on days 5 and 6 (71%) was higher compared to any other experimental groups (0%-44%). Regardless of single or double, TCI on Day 5 or Day 6 led to higher litter sizes in SM or EY groups, respectively. The breeding efficiency and litter size of single TCI on Day 5 (4.2) and double TCI on Day 5 and Day 6 (3.7) were significantly higher than in the other experimental groups in SM and EY groups, respectively (p < .05). These findings suggest that skim milk is a suitable alternative to egg yolk for cryopreservation of canine spermatozoa, and the suitable timing for insemination might be on Day 5 post-LH surge.

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“…Egg yolk the most common component used in canine semen extenders to protect spermatozoa from cold shock during chilled process (Mason, 2016). However, the presence of an avian influenza outbreak (Abe et al, 2008) and egg yolk affected by microbial contamination (Tarig et al, 2017) causes the desire to replace the egg yolk in extenders. Thus, it is necessary to develop extenders without use egg yolk.…”

Section: Advances In Animal and Veterinary Sciencesmentioning

confidence: 99%

A Comparative Study on the Effects of Coconut Water Based Extenders on the Quality of Kintamani Dog Semen Preserved at 4oC

Puja¹,

Sawitri²,

Maharani³

et al. 2018

Adv. Anim. Vet. Sci.

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| The objective of this study was to determined the effects of different extenders on chilled kintamani dog semen quality parameters. In this study, the impact of extenders on motility, viability and DNA integrity of kintamani dog semen were investigated. A total of 12-second fraction of ejaculates were collected from four kintamani dogs using manual stimulation. Sperms were diluted in each of the five extenders at room temperature and then cooled to 4 0 C. Samples were then evaluated every day until five days. Chilled semen samples were assessed for motility, viability and DNA integrity. Results showed that the progressive motility of the sperm cells was significantly higher in extender A and B compared to other extenders. The extender containing 20% egg yolk and 20 % tender coconut water preserved the motility of more than 60% of the spermatozoa up to the day 5 post-sampling. Percentage of live sperm decreased slightly from day 0 to day 5. There was no significant difference in live spermatozoa due to the extender after five day. The DNA was not unaltered by different of extender and storage refrigeration process. It concluded that kintamani dog semen qualities could be maintained for up five days when semen extended with coconut water-based extender with addition fructose and store at 4 0 C. Keywords

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Artificial Insemination with Canine Spermatozoa Frozen in a Skim Milk/Glucose-Based Extender

Cited by 22 publications

References 10 publications

Kinetics of Nuclear Status and Kinase Activities During In Vitro Maturation of Canine Oocytes

Kinetics of Nuclear Status and Kinase Activities During In Vitro Maturation of Canine Oocytes

Fertilizing ability of canine spermatozoa cryopreserved with skim milk‐based extender in a retrospective study

A Comparative Study on the Effects of Coconut Water Based Extenders on the Quality of Kintamani Dog Semen Preserved at 4oC

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