Cholesterol-loaded-cyclodextrins and fertility potential of stallions spermatozoa.
Abstract: Irreversible damage occurs to spermatozoal membranes, during the phase transition, when spermatozoa are cooled from room temperature to 5 degrees C. Some of this damage can be ameliorated by adding cholesterol to the membrane, thereby altering membrane lipid composition. Adding cholesterol-loaded cyclodextrins (CLCs) to stallion spermatozoa prior to freezing, increases cell cryosurvival. However, the fertilizing potential of CLC-treated stallion spermatozoa is unknown. To address this, experiments were conducted which evaluated the ability of CLC-treated stallion spermatozoa to capacitate, acrosome react, and bind to the zona pellucida in vitro, and to fertilize oocytes in vivo. When CLC-treated cryopreserved stallion spermatozoa were treated with various agents to induce capacitation and the acrosome reaction (AR), dilauroylphosphatidylcholine (PC-12) and lysophosphatidylcholine (LPC) induced the AR in control cells (62% and 55%, respectively) but not in CLC-treated cells (17% and 14%, respectively, P0.05). Control- and CLC-treated stallion spermatozoa bound to ZP of cattle oocytes equally well (0.44+/-0.16 vs. 0.25+/-0.09, respectively; P>0.05). In addition, the fertility rates of mares inseminated with control- and CLC-treated sperm were similar (P>0.05).
Copyright 2009 Elsevier B.V. All rights reserved.
Publication Date: 2009-08-15 PubMed ID: 19762177DOI: 10.1016/j.anireprosci.2009.08.001Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research is about the impact of adding cholesterol-loaded cyclodextrins (CLCs) to stallion sperm cells on their survival during freezing, their interaction with female egg cells, and ultimately their potential for successful reproduction.
Background
- The researchers have noted that when sperm cells of horses are cooled, damage to their membrane structure can occur. This damage is reportedly less severe if cholesterol is added to the membrane, which alters its lipid makeup.
- A substance called cholesterol-loaded cyclodextrins (CLCs) had a positive effect on the survival of these sperm cells during freezing, so the researchers wanted to understand more about how CLCs affected the fertility capacity of these preserved cells.
Methodology
- To gather needed data, the team exposed CLC-treated stallion sperm cells to various agents known to stimulate capacitation (the sperm’s final stages of maturation) and the acrosome reaction (important for the sperm to penetrate an egg).
- The agents used included dilauroylphosphatidylcholine (PC-12) and lysophosphatidylcholine (LPC), both of which failed to induce the acrosome reaction in CLC-treated cells.
- However, another agent, the calcium ionophore A23187, was able to trigger the acrosome reaction irrespective of whether the cells were CLC-treated or not.
- Additionally, researchers examined the ability of both CLC-treated and untreated stallion sperm cells to bind to the zona pellucida (outer layer of the egg cell) of cattle oocytes, which was found to be similar.
- Lastly, they compared fertility rates of female horses inseminated with CLC-treated and untreated sperm cells.
Outcomes
- The results showed that although PC-12 and LPC could not induce the acrosome reaction in CLC-treated cells, this reaction could be provoked uniformly by A23187, regardless of whether the cells had been treated with CLCs.
- The binding ability of CLC-treated and untreated stallion sperm cells to cattle oocytes was found to be similar, with no statistical significance between the two groups.
- Furthermore, no significant variation was noticed between the fertility rates of mares inseminated with CLC-treated and untreated sperm cells.
- These findings indicate that adding CLCs to stallion spermatozoa before freezing does not significantly alter their reproductive capacity.
Cite This Article
APA
Spizziri BE, Fox MH, Bruemmer JE, Squires EL, Graham JK.
(2009).
Cholesterol-loaded-cyclodextrins and fertility potential of stallions spermatozoa.
Anim Reprod Sci, 118(2-4), 255-264.
https://doi.org/10.1016/j.anireprosci.2009.08.001 Publication
Researcher Affiliations
- Animal Reproduction and Biotechnology Laboratory, Colorado State University, Campus Delivery 1680, Fort Collins, CO 80523, USA.
MeSH Terms
- Acrosome Reaction / drug effects
- Animals
- Calcimycin / pharmacology
- Calcium / analysis
- Cell Membrane / chemistry
- Cell Membrane / physiology
- Cholesterol / administration & dosage
- Cholesterol / analysis
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cyclodextrins / administration & dosage
- Female
- Fertility
- Horses / physiology
- Insemination, Artificial / veterinary
- Ionophores / pharmacology
- Male
- Pregnancy
- Semen Preservation / adverse effects
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sperm Capacitation / drug effects
- Sperm-Ovum Interactions / drug effects
- Spermatozoa / chemistry
- Spermatozoa / physiology
- Spermatozoa / ultrastructure
- Zona Pellucida / metabolism
Citations
This article has been cited 11 times.- Sharafi M, Borghei-Rad SM, Hezavehei M, Shahverdi A, Benson JD. Cryopreservation of Semen in Domestic Animals: A Review of Current Challenges, Applications, and Prospective Strategies. Animals (Basel) 2022 Nov 24;12(23).
- Gobato MLM, Segabinazzi LGTM, Scheeren VFC, Bandeira RS, Freitas-Dell'Aqua CP, Dell'Aqua JA Jr, Papa FO. Ability of donkey sperm to tolerate cooling: Effect of extender base and removal of seminal plasma on sperm parameters and fertility rates in mares. Front Vet Sci 2022;9:1011899.
- Asano A, Priyadarshana C. Membrane-Mediated Regulation of Sperm Fertilization Potential in Poultry. J Poult Sci 2022 Apr 25;59(2):114-120.
- Carro MLM, Ramírez-Vasquez RRA, Peñalva DA, Buschiazzo J, Hozbor FA. Desmosterol Incorporation Into Ram Sperm Membrane Before Cryopreservation Improves in vitro and in vivo Fertility. Front Cell Dev Biol 2021;9:660165.
- Rivera-Egea R, Garrido N, Sota N, Meseguer M, Remohí J, Dominguez F. Sperm lipidic profiles differ significantly between ejaculates resulting in pregnancy or not following intracytoplasmic sperm injection. J Assist Reprod Genet 2018 Nov;35(11):1973-1985.
- Buschiazzo J, Ríos GL, Canizo JR, Antollini SS, Alberio RH. Free cholesterol and cholesterol esters in bovine oocytes: Implications in survival and membrane raft organization after cryopreservation. PLoS One 2017;12(7):e0180451.
- Kiso WK, Asano A, Travis AJ, Schmitt DL, Brown JL, Pukazhenthi BS. Pretreatment of Asian elephant (Elephas maximus) spermatozoa with cholesterol-loaded cyclodextrins and glycerol addition at 4°C improves cryosurvival. Reprod Fertil Dev 2012;24(8):1134-42.
- Del Llano E, Rasschaert M, Arnoult C, Robert P, Ray PF, Loeuillet C. Administration of ethiodized poppy seed oil-based contrast agent into the uterus enhances fertilization rate in mice inseminated with low sperm numbers. Hum Reprod 2026 Jan 1;41(1):69-77.
- de Zutter BM, de Paula Freitas-Dell'Aqua C, Dell'Aqua-Junior JA, Monteiro GA, Troncarelli T, Papa FO. Optimising Stallion Semen Cryopreservation: Preliminary Insights Into Pre-Centrifugation Extender Effects. Reprod Domest Anim 2025 Oct;60(10):e70135.
- Aly AS, Rozeboom KJ, Parrish JJ. Using Cholesterol-Loaded Cyclodextrin to Improve Cryo-Survivability and Reduce Capacitation-Like Changes in Gender-Ablated Jersey Semen. Animals (Basel) 2025 Jul 11;15(14).
- Ahmet E, Ramazan A, Selin Y, İzem SA, Nur E, Kamber D, Mithat E, Kemal A. The Effect of Cholesterol-Loaded Cyclodextrin and Resveratrol Compounds on Post-Thawing Quality of Ram Semen. Vet Med Sci 2025 Jan;11(1):e70172.
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