Fertilization in Dogs

Mahi‐Brown, Cherrie A.

doi:10.1007/978-1-4757-8982-9_15

Cited by 6 publications

(2 citation statements)

References 42 publications

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“…The canine oocyte contains large quantities of lipids, which impairs visualization of internal oocyte structures (Mahi‐Brown 1991). The ovulated oocyte has no or very little expanded cumulus oophorus, but retains a compact layer of corona radiata for many days after ovulation (Andersen and Simpson 1973; Renton et al.…”

Section: Introductionmentioning

confidence: 99%

Fertilizing Capacity of Frozen Epididymal Sperm Collected from Dogs

Martins

Padilha

Souza

et al. 2009

Reprod Domestic Animals

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The collection of epididymal sperm may be a valuable tool for canine reproduction especially since it can enable collection of cells after death of a valuable dog. The aim of the present study was to evaluate the viability of epididymal sperm after freeze-thawing. Epididymides were obtained from four adult dogs by elective orchiectomy. The caudal portion of the epididymides and part of the deferential ducts were squeezed by means of an anatomic clamp into a Petri dish containing either 0.9% saline solution (Group 1) or Ringer solution without lactate (Group 2). Samples were centrifuged at 800 x g for 10 min, the supernatant was removed and the pellet was diluted in one step with a Tris/citric acid/OEP (Orvus Es Paste) extender containing 7% glycerol and subjected to semen freezing. Oocytes were obtained from canine ovaries, after ovariohysterectomy. Only oocytes that were approximately 100 mum in diameter, with a dark ooplasm surrounded by three- or four-well formed cumulus cell layers were used for sperm testing. Frozen semen samples were thawed in a water bath at 70 degrees C for 8 s and analysed at room temperature for sperm motility and velocity. Oocytes were incubated with spermatozoa in humidified atmosphere containing 5% CO(2) at 38 degrees C for 18 h. Morphological and functional characteristics of spermatozoa were similar in both groups. However, the percentage of sperm cells bound to oocytes was significantly higher in Group 2 than in Group 1. This result suggests that the Ringer solution without lactate was a more suitable medium for collecting epididymal canine sperm than 0.9% saline.

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Section: Introductionmentioning

confidence: 99%

Fertilizing Capacity of Frozen Epididymal Sperm Collected from Dogs

Martins

Padilha

Souza

et al. 2009

Reprod Domestic Animals

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“…Although natural mating may be successful when performed several days before the fertilization period, the shorter duration of sperm survival of frozen-thawed semen requires that AI be made when the oocytes are ready to be fertilized. In the dog, primary oocytes are ovulated 24 to 78 h after the LH surge, and the oocytes require approximately 2 to 3 more days to mature to metaphase II (Mahi-Brown, 1991;Pheminster et al, 1973;Wildt et al, 1978). Therefore, AI using frozen-thawed semen should be performed 3 to 7 d after the LH peak.…”

Section: Introductionmentioning

confidence: 99%

A New Device for Intrauterine Artificial Insemination in the Dog

Kong¹,

Yu²,

Jeong³

et al. 2003

Asian Australas. J. Anim. Sci

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The intrauterine inseminator (IUI) was developed to provide the method of depositing dog semen into the uterine body instead of the vagina. The IUI consists of a vaginal endoscope, a balloon sheath, and injection catheter. When the endoscope is inserted into the vagina and the balloon expanded with air, the cervical os becomes visible so a injection catheter can be inserted through the cervix for deposition of the frozen-thawed semen. The efficacy of the IUI device was compared to intra-vaginal artificial insemination using semen that had been collected and frozen from pooled sperm-rich fraction of ejaculates collected from two Jindo dog donors. Aliquots of semen were extended with a Tris-egg yolk diluent, centrifuged, the seminal plasma removed, the pellet resuspended with the same diluent, and cooled to 5°C over a 2 h period. A Tris-egg yolk-glycerol extender was added at 5°C; after 1 h, semen was loaded into 0.5 ml straws, and straws were frozen in LN vapor for 5 min, and immersed in LN for storage. The final sperm concentration for freezing was approximately 100×10 6 cells/ml. The straws were thawed at 70°C for precisely 6 sec, 1.5 ml Tris-egg yolk buffer at 38°C added, and the 2 ml of thawed semen was used for a single insemination using the IUI device. Each bitch was inseminated at optimal insemination point, which was estimated by vaginal epithelial cells staining and progesterone concentration analysis. Use of the IUI device resulted in 21 of 26 females giving birth to 89 pups (4.2±1.6 pups per litter), while intra-vaginal AI resulted in 6 of 15 females whelping a total of 17 pups (2.8±1.2 pups per litter). We believe the IUI device is easier to use than previously described devices used for intrauterine insemination. In our experience the expansion of the balloon has a calming effect on the bitch that aids the inseminator. These results indicate that the IUI device was able to provide high fertility with 50 million frozen sperm per insemination and two inseminations.

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Solubilized zona pellucida proteins and progesterone induce calcium influx and the acrosome reaction in capacitated dog spermatozoa

Brewis

Morton

Moore

et al. 2001

Molecular Reproduction Devel

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Spermatozoa from the sperm-rich fractions of the semen of 6 beagle dogs were capacitated and the effect of both zona pellucida (ZP) proteins and progesterone on calcium flux and the acrosome reaction measured. Sperm calcium flux was determined using the dual wavelength calcium probe indo-1/AM (6 microM) in a flow cytometric assay (one ejaculate from each dog examined; n = 6). No calcium flux was observed in the negative control treatments (RPMI medium or DMSO). Both heat-solubilized bitch ZP proteins and progesterone caused a similar response characterized by a gradual but marked influx of calcium ions which was sustained over 2 min. Acrosomal status was assessed by indirect immunofluorescence using a specific monoclonal antibody following 1 hr incubation for each treatment (four ejaculates from each dog examined; n = 24). The level of acrosomal exocytosis was very high for samples treated with ZP proteins (70.3 +/- 2.1%) and progesterone (84.6 +/- 1.5%) and was significantly different from the respective controls (P < 0.001). Interestingly the patterns of calcium flux in response to both ZP proteins and progesterone were in contrast to the situation in other species studied to date raising the possibility that the mechanism for triggering the acrosome reaction may be different in dog spermatozoa. In addition the high degree of progesterone-induced acrosomal exocytosis compared to other species raises the probability that the majority of dog spermatozoa are already undergoing the acrosome reaction before they reach the egg ZP.

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Fertilization in Dogs

Cited by 6 publications

References 42 publications

Fertilizing Capacity of Frozen Epididymal Sperm Collected from Dogs

Fertilizing Capacity of Frozen Epididymal Sperm Collected from Dogs

A New Device for Intrauterine Artificial Insemination in the Dog

Solubilized zona pellucida proteins and progesterone induce calcium influx and the acrosome reaction in capacitated dog spermatozoa

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