In vitro and xenogenous capacitation-like changes of fresh, cooled, and cryopreserved stallion sperm as assessed by a chlortetracycline stain.
Abstract: Like the human female, the mare experiences reproductive tract pathology that may sometimes be circumvented by the use of assisted reproductive technologies (ARTs). One such technology, gamete intrafallopian transfer (GIFT), may be used in mares that exhibit ovulatory, oviductal, or uterine abnormalities that limit the use of common ARTs, such as embryo transfer. Homologous GIFT has been successfully performed in the horse; however, the logistics, costs, and associated risks of surgically transferring gametes to the oviducts of a recipient mare are considerably high. Use of a less costly species in a heterologous or xenogenous procedure would therefore be beneficial. This study represents the preliminary investigation into the use of sheep as recipients for xenogenous GIFT procedures using equine gametes. We investigated the capacitation response of fresh, cooled, or frozen stallion sperm after 1) in vivo incubation in the reproductive tract of estrous and anestrous ewes as well as 2) in vitro incubation in a modified Krebs/ Ringer extender at 37 degreesC with and without the addition of heparin at 10 IU/mL for up to 8 hours. A chlortetracycline (CTC) fluorescent stain was used to assess the capacitation response of sperm. Findings indicated that oviductal fluid samples recovered from estrous ewes had significantly higher numbers of sperm exhibiting capacitation-like staining patterns when compared to samples recovered from anestrous ewes (P < .05). Fresh semen yielded higher capacitation-like staining patterns after in vivo incubation than did frozen-thawed or cooled samples. A transition from majority CTC unreacted sperm to majority CTC non-acrosome intact sperm was demonstrated for both in vivo and in vitro studies. In vitro incubation of stallion sperm with heparin did not result in an increased capacitation-like staining response over time when compared with nonheparinized samples. Results from this study suggest that xenogenous capacitation of stallion sperm may occur in the estrous ewe.
Publication Date: 2000-02-12 PubMed ID: 10670518
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- Journal Article
Summary
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This research investigates the viability of using sheep as recipients for horse sperm in assisted reproductive technologies, specifically Gamete Intrafallopian Transfer. It evaluates the capacitation response of horse sperm that is fresh, cooled, or frozen after being incubated in the reproductive tracts of ewes, as well as being incubated in a lab setting.
Introduction and Objectives
- The study tests a new approach to the Assisted Reproductive technologies (ARTs) in horses, involving use of sheep as recipients through a heterologous (between different species) or xenogenous procedure.
- The main objective of the study was to investigate the “capacitation” response (a process sperm undergo to be able to fertilize an egg) of horse sperm under various conditions within a sheep’s reproductive tract, as well as in a laboratory environment.
Testing and Evaluations
- Sperm capacitation was assessed using a chlortetracycline (CTC) fluorescent stain that indicates capacitation-like changes in sperm.
- The sperm, subjected to three conditions (fresh, cooled or frozen), was incubated in the reproductive tract of both estrous (in heat) and anestrous (not in heat) ewes, and also in a lab setting using a modified Krebs/Ringer extender at 37 degrees Celsius with and without the addition of heparin.
Observations and Results
- A more successful capacitation response was observed in estrous ewes, with significantly higher numbers of sperm exhibiting capacitation-like changes.
- Fresh semen showed a higher level of capacitation-like changes than frozen-thawed or cooled sperm after in vivo incubation.
- A transition was observed from primarily unreacted sperm to non-acrosome intact sperm in both in vivo (within the body) and in vitro (in a lab) studies, indicating successful capacitation.
- Additional in vitro incubation of the sperm with heparin did not result in an increased capacitation-like staining response over time, compared to non-heparinized samples.
Implications and Conclusions
- The results suggest that horse sperm can undergo capacitation in the reproductive tract of a sheep in heat, possibly opening up new avenues for ART in horses, using sheep as more cost-effective and readily available recipients.
- However, more research is needed to fully understand the implications and potential challenges of this approach.
Cite This Article
APA
Parker NA, Bailey TL, Bowen JM, Ley WB, Purswell BJ, Dascanio JJ.
(2000).
In vitro and xenogenous capacitation-like changes of fresh, cooled, and cryopreserved stallion sperm as assessed by a chlortetracycline stain.
J Androl, 21(1), 45-52.
Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute & State University, Blacksburg 24061-0442, USA. niparker@vt.edu
MeSH Terms
- Anestrus / physiology
- Animals
- Chlortetracycline
- Cold Temperature
- Cryopreservation
- Estrus / physiology
- Female
- Heparin / pharmacology
- Horses / physiology
- Male
- Sheep
- Sperm Capacitation / drug effects
- Staining and Labeling
Citations
This article has been cited 1 times.- Benko F, Fialková V, Žiarovská J, Ďuračka M, Lukáč N, Tvrdá E. In Vitro versus Cryo-Induced Capacitation of Bovine Spermatozoa, Part 2: Changes in the Expression Patterns of Selected Transmembrane Channels and Protein Kinase A. Int J Mol Sci 2022 Nov 24;23(23).
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