Sperm transport in the female reproductive tract of the brushtail possum, Trichosurus vulpecula, following superovulation and artificial insemination

Assistant reproduction technologies are in constant evolution, among them the artificial insemination (AI). AI has been successfully used in pigs for decades, especially to improve boar efficienc and productivity. Lately, swine AI has taken on a new lease of life since efficient AI is essential for solving future challenges in the porcine industry and to enhance productivity. The present paper summarizes several factors concerning AI, starting with an overview of some physiological aspects including the female reproductive tract and sperm transport, as well as sperm losses during insemination and uterus sperm selection. Strategies developed to reduce the number of sperm during the AI process, are also reviewed, along with their combination with new reproductive technologies for application in pig production in the near future.

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Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.611301 ◽

2021 ◽

Vol 8 ◽

Author(s):

Eduardo G. Aisen ◽

Wilfredo Huanca López ◽

Manuel G. Pérez Durand ◽

Edita Torres Mamani ◽

Juan C. Villanueva Mori ◽

...

Keyword(s):

Seminal Plasma ◽

Artificial Insemination ◽

Reproductive Tract ◽

Vas Deferens ◽

Female Reproductive Tract ◽

Low Fertility ◽

Sperm Cells ◽

South American Camelids ◽

Thawing Process

The viscous seminal plasma (SP) is currently a major impediment to the handling of ejaculate and the development of some biotechnologies in South American camelids. The vas deferens-collected spermatozoa of alpacas is a useful technique to avoid this problem. On the other hand, SP contains a large protein component that has been implicated in the function of spermatozoa within the female reproductive tract. In this sense, the low fertility achieved using transcervical insemination with frozen-thawed spermatozoa in alpacas could be improved by adding SP. This study aimed to evaluate the effect of the whole SP on some in vitro parameters of alpaca spermatozoa after the freezing-thawing-process and the fertility after artificial insemination. It would contribute to a better understanding of the interaction between thawed sperm cells and SP. Spermatozoa were obtained by surgically diverted vas deferens. The samples were diluted with a Tris-based extender, packaged in straws, and frozen. At thawing, each straw was divided into two post-thawing conditions: with the addition of 10% of PBS (control) or with 10% SP (treatment). The sperm cells were evaluated using dynamic parameters, sperm cell morphology, and morphometry. Fertility was assessed by an artificial insemination trial. All in vitro parameters were analyzed by ANOVA. A heterogeneity test was scheduled for the fertility trial. After the freezing-thawing process, motility and plasma membrane functionality was improved when SP was added. No differences were found for post-thaw viability between the control and treatment samples. The percentage of normal cells was higher with SP at post-thawing, and a decrease of the presence of bent tailed spermatozoa with a droplet in the SP group was observed. The length of the head spermatozoa was 3.4% higher in the samples with PBS compared to those in which SP was added. Females pregnant at day 25 post-insemination were 0/12 (with SP inside the straw) and 1/10 (without SP inside the straw). In conclusion, the presence of 10% SP at post-thawing improves sperm cells' motility, functionality, and morphology, indicating that it would be beneficial to improve the frozen-thawed alpaca's physiology spermatozoa. More fertility trials must be developed to increase this knowledge.

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Preservation and evaluation of semen for artificial insemination

Reproduction Fertility and Development ◽

10.1071/rd04034 ◽

2004 ◽

Vol 16 (4) ◽

pp. 447 ◽

Cited By ~ 19

Author(s):

Lindsay Gillan ◽

W. M. Chis Maxwell ◽

Gareth Evans

Keyword(s):

Artificial Insemination ◽

Reproductive Tract ◽

Respiratory Activity ◽

Small Ruminants ◽

Functional Condition ◽

Structure And Function ◽

Female Reproductive Tract ◽

Altered State ◽

And Function ◽

Improved Methods

Many years of research have been devoted to improving the fertility of preserved semen of small ruminants. There have been few significant advances in preservation in recent times, but considerable knowledge has been gained on the effect of preservation on the structure and function of spermatozoa. It has become evident that preservation greatly affects many sperm attributes, such as motility, respiratory activity, membrane status and DNA quality. Consequently, viability is reduced, transport in the female reproductive tract is inhibited, the timing of fertilisation is altered and embryo development is affected following insemination of preserved, compared to fresh spermatozoa. A greater understanding of their functional condition may lead to the development of methods of preventing these alterations or to improved methods of using the preserved spermatozoa for artificial insemination in their altered state.

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Effects of GnRH, PGF2α and oxytocin treatments on conception rate at the time of artificial insemination in lactating dairy cows

Czech Journal of Animal Science ◽

10.17221/1283-cjas ◽

2011 ◽

Vol 56 (No. 6) ◽

pp. 279-283 ◽

Cited By ~ 1

Author(s):

A. Gümen ◽

A. Keskin ◽

G. Yilmazbas-Mecitoglu ◽

E. Karakaya ◽

S. Cevik ◽

...

Keyword(s):

Dairy Cows ◽

Artificial Insemination ◽

Reproductive Tract ◽

Female Reproductive Tract ◽

Conception Rate ◽

Beneficial Effects ◽

Lactating Dairy Cows ◽

Conception Rates ◽

Prostaglandin F ◽

And Control

In several studies, hormones such as gonadotropin-releasing hormone (GnRH), prostaglandins and oxytocin were used to increase pregnancy rate by inducing ovulation and improving the sperm transport in the female reproductive tract in lactating dairy cattle. The objective of this study was to compare the effects of GnRH, prostaglandin F2α (PGF2α) and oxytocin treatments at the time of artificial insemination (AI) after spontaneous oestrus on the conception rate (CR) of lactating dairy cows. Oestrus was detected by visual observations by experienced personnel. All cows (n = 430, 308 Holstein-Frisian and 122 Swedish-Red dairy cows) were inseminated based on the am/pm rule by veterinarians of the farm. After AI, cows were alternately assigned to one of the four treatment groups: (1) GnRH (n = 113); (2) PGF2α (n = 106); (3) oxytocin (n = 106) and (4) non-treated control (n = 105). Pregnancy diagnosis was performed 28–34 and 58–64 days post-insemination by transrectal ultrasonography. Conception rates on days 28–34 and 58–64 were not different among GnRH (46.0%; 52/113 and 44.3%; 50/113), PGF2α (37.7%; 40/106 and 35.9%; 38/106) and control (49.5%; 52/105 and 47.6%; 50/105) groups. However, conception rates were lower (P = 0.02) in oxytocin (31.1%; 33/106 and 30.2%; 32/106) than in GnRH and control groups on days 28–34 and 58–64. Other covariant factors, such as milk production, days in milk (DIM), breed, parity, service number did not affect the conception rate. Thus, there were no beneficial effects of treatments with GnRH and PGF2α at the time of AI, and oxytocin had an adverse effect on CR in lactating dairy cows in this study.

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A glimpse at sperm function in vivo: sperm transport and epithelial interaction in the female reproductive tract

Animal Reproduction Science ◽

10.1016/s0378-4320(00)00130-5 ◽

2000 ◽

Vol 60-61 ◽

pp. 337-348 ◽

Cited By ~ 57

Author(s):

Mary A Scott

Keyword(s):

Reproductive Tract ◽

Female Reproductive Tract ◽

Sperm Function ◽

Sperm Transport

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Ultrastructural observations on in vivo fertilisation in the brushtail possum, Trichosurus vulpecula, following PMSG/LH superovulation and artificial insemination

Zygote ◽

10.1017/s0967199499000714 ◽

1999 ◽

Vol 7 (4) ◽

pp. 307-320 ◽

Cited By ~ 9

Author(s):

M.K. Jungnickel ◽

A.J. Harman ◽

J.C. Rodger

Keyword(s):

Artificial Insemination ◽

Acrosome Reaction ◽

Brushtail Possum ◽

Trichosurus Vulpecula ◽

Chromatin Decondensation ◽

Large Hole ◽

Enzymatic Action ◽

Acrosomal Membrane ◽

Sperm Penetration

Information on the dynamics of gamete interaction in marsupials is very limited and not available for any species from the major Australian Order Diprotodontia which includes most of the more familiar animals such as kangaroos, possums and the koala. This study addressed this deficiency by examining the ultrastructure of in vivo fertilised eggs from common brushtail possums (Trichosurus vulpecula). Females were superovulated by treatment with 15 IU PMSG and then 4 mg porcine LH 3 days later, and inseminations were performed 910-13 h after LH) using epididymal spermatozoa. Between 33 and 39 h after LH injection females were killed, reproductive tracts excised and the oviduct ampulla segment flushed for eggs. Three of the six eggs examined were fertilised as judged by the presence of sperm remnants in the cytoplasm. On the basis of these eggs it was found that sperm penetration left a large hole in the zona pellucida (ZP), suggesting that sperm zona penetration occurs primarily by the enzymatic action of acrosomal enzymes. Sperm lying within the perivitelline space were lacking both an outer acrosomal membrane and the associated acrosomal contents, while both these structures were found on sperm embedded within the mucoid layer, which is consistent with induction of the acrosome reaction by binding to the ZP. Once inside the egg cytoplasm, the sperm head travelled only a short distance before chromatin decondensation occurred. Fertilised eggs showed signs of cytoplasmic activation including cytoskeleton association with apparently dividing mitochondria and prominent rough endoplasmic reticulum. Unfertilised eggs appeared to be undergoing degenerative changes and lacked any evidence of activation. This study has demonstrated that superovulation and laparoscopic intravaginal artificial insemination provide a system through which perifertilisation events in the possum and other monovular Australian marsupials can be examined experimentally.

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Oxytocin Receptors in the Mammary Gland and Reproductive Tract of a Marsupial, the Brushtail Possum (Trichosurus Vulpecula)1

Biology of Reproduction ◽

10.1095/biolreprod45.5.673 ◽

1991 ◽

Vol 45 (5) ◽

pp. 673-679 ◽

Cited By ~ 8

Author(s):

Conrad Sernia ◽

Walter G. Thomas ◽

Robert T. Gemmell

Keyword(s):

Mammary Gland ◽

Reproductive Tract ◽

Brushtail Possum ◽

Trichosurus Vulpecula ◽

Oxytocin Receptors

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Secretory proteins from the female reproductive tract of the brushtail possum Trichosurus vulpecula: binding to sperm and effects on sperm survival in vitro

Reproduction Fertility and Development ◽

10.1071/rd00010 ◽

1999 ◽

Vol 11 (6) ◽

pp. 329 ◽

Cited By ~ 6

Author(s):

K. S. Sidhu ◽

K. E. Mate ◽

F. C. Molinia ◽

A. M. Glazier ◽

J. C. Rodger

Keyword(s):

Molecular Mass ◽

Reproductive Tract ◽

Serum Proteins ◽

Female Reproductive Tract ◽

Conditioned Media ◽

Secretory Proteins ◽

Brushtail Possum ◽

Sperm Survival ◽

Uterine Proteins

Previous studies have demonstrated that co-culture of brushtail possum epididymal spermatozoa with oviduct epithelial cell monolayers prolongs sperm survival and results in the re-orientation of the sperm head and tail to the T-shape (thumbtack) configuration. Transformation of sperm to thumbtack orientation is believed to be associated with marsupial sperm capacitation. Here we report that incubation in oviduct-conditioned media also significantly prolongs sperm survival and results in the transformation of sperm to the thumbtack orientation. The major objective of the current study was to examine the proteins present in the conditioned media, to determine whether any of these proteins specifically bound to sperm and the relationship between these proteins and sperm survival and thumbtack orientation. Co-culturing brushtail possum sperm with biotin-labeled proteins in conditioned media (CM) from ampulla, isthmus and uterine explants demonstrated strong binding of two proteins of molecular mass 230 and 61 kD and weak binding of nine proteins of molecular mass 200, 180, 120, 140, 55, 52, 48, 34, 30 kD to sperm within 30 min. The binding of the 61-kD protein from the conditioned media appeared specific as increasing concentrations of non-labeled oviduct proteins, but not serum proteins, inhibited the binding of labeled proteins. The binding of oviduct and uterine proteins in the conditioned media significantly prolonged sperm survival and percentage motility and also transformed a large number of sperm to a thumbtack orientation. The implication of binding of these proteins is discussed in the context of sperm survival and capacitation in this species.

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Glycosaminoglycans of the uterine and vaginal cul-de-sac tissue in the brushtail possum (Trichosurus vulpecula)

Reproduction Fertility and Development ◽

10.1071/rd9960819 ◽

1996 ◽

Vol 8 (4) ◽

pp. 819 ◽

Cited By ~ 4

Author(s):

M Legge ◽

BL Hill ◽

GH Shackell ◽

BJ McLeod

Keyword(s):

Reproductive Tract ◽

Keratan Sulfate ◽

Progressive Increase ◽

Implant Removal ◽

Brushtail Possum ◽

Similar Response ◽

Trichosurus Vulpecula ◽

Uterine Tissue ◽

Hormonal Stimulation ◽

Sterile Saline

Live-captured feral brushtail possums (Trichosurus vulpecula) received slow-release progestagen implants for 16 days after removal of pouch young. At the time of implant removal one group was injected intramuscularly with pregnant mare serum gonadotrophin (PMSG, 2 I.U. kg-1 in 0.2 mL saline) and another group received an equivalent volume of sterile saline. Animals from each group were killed on Days 0, 3, 4, 5 or 6 after implant removal (PMSG, n = 4 per day; saline controls, n = 3 per day). The uterus and vaginal cul-de-sac were excised and tissue was analysed for glycosaminoglycans (GAGs) and keratan sulfate content. GAG concentration increased progressively over the six-day period following implant removal. Mean uterine GAG concentrations were significantly higher than those in vaginal cul-de-sac from Day 3 to Day 6 in PMSG-treated and saline-treated animals following progestagen implant removal (P < 0.05). Keratan sulfate concentrations showed a similar response--concentrations were higher in uterine tissue than in vaginal cul-de-sac tissue and PMSG-treated animals showed a greater response than did saline controls in the uterine tissue, but the same response in cul-de-sac tissue (P < 0.05 for all groups). The progressive increase in GAG concentrations of reproductive tract tissue following removal of the progesterone implant is indicative of hormonal stimulation and suggests that GAGs may have a supportive role in gamete and embryo nutrition and growth in the brushtail possum.

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