Lignosulfonic acid blocks in vitro fertilization of macaque oocytes when sperm are treated either before or after capacitation

Theodore L Tollner, James W. Overstreet, Ming W. Li, Stuart A Meyers, Ashley I. Yudin, Edward R. Salinas, Gary N. Cherr

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Lignin-derived macromolecules (LDMs) are biologically active compounds that affect a variety of cell-to-cell interactions including the inhibition of fertilization and embryo development in a number of nonmammalian species. The effect of lignosulfonic acid (LSA), a highly sulfonated LDM, on cynomolgus macaque sperm-oocyte interaction was evaluated with a zona pellucida binding assay and by in vitro fertilization (IVF). Sperm were treated with LSA (1.5 mg/mL) either before washing or after capacitation. Capacitation included centrifugation through 80% Percoll followed by 2 consecutive washes with medium, overnight incubation, and activation with dibutyryl cyclic adenosine monophosphate and caffeine. The zona binding assay was performed using immature oocytes that had adhered to the center of glass "binding chambers." The number of capacitated sperm that attached to the zona over a 3-minute period was recorded. Sperm attachment was significantly inhibited by LSA as compared to controls whether treatment occurred after capacitation (92.5%; P < .001) or before washing (82.5%; P < .001). When sperm were treated similarly with fucoidin, a sulfated polysaccharide known to inhibit sperm-oocyte interaction, sperm-zona binding was significantly inhibited by postcapacitation treatment but not by prewash treatment. Treatment of sperm with LSA consistently blocked fertilization over 4 IVF cycles both before washing and after capacitation. Fertilization rate for controls was 65% ± 17%. No LSA-treated sperm were observed on the surface of lightly rinsed oocytes after 4 hours of coincubation. Localization of biotinylated LSA showed labeling over the entire sperm surface with the greatest intensity observed over the head and midpiece. LSA treatment had no effect on the percentage of motile sperm or quality of sperm motility. Due to the antifertility properties of this nontoxic molecule, LSA appears to have potential as a vaginal contraceptive.

Original languageEnglish (US)
Pages (from-to)889-898
Number of pages10
JournalJournal of Andrology
Volume23
Issue number6
StatePublished - Nov 2002

Fingerprint

Macaca
Fertilization in Vitro
Oocytes
Spermatozoa
Acids
Herpes Zoster
Sperm-Ovum Interactions
Lignin
Fertilization
Therapeutics
Zona Pellucida
Sperm Count
Sperm Motility
Contraceptive Agents
Caffeine
Contraception
Centrifugation
Cell Communication
Cyclic AMP
Embryonic Development

Keywords

  • Contraception
  • Fucoidin
  • Microbicide
  • Polysulfonation
  • Zona pellucida

ASJC Scopus subject areas

  • Endocrinology

Cite this

Lignosulfonic acid blocks in vitro fertilization of macaque oocytes when sperm are treated either before or after capacitation. / Tollner, Theodore L; Overstreet, James W.; Li, Ming W.; Meyers, Stuart A; Yudin, Ashley I.; Salinas, Edward R.; Cherr, Gary N.

In: Journal of Andrology, Vol. 23, No. 6, 11.2002, p. 889-898.

Research output: Contribution to journalArticle

Tollner, Theodore L ; Overstreet, James W. ; Li, Ming W. ; Meyers, Stuart A ; Yudin, Ashley I. ; Salinas, Edward R. ; Cherr, Gary N. / Lignosulfonic acid blocks in vitro fertilization of macaque oocytes when sperm are treated either before or after capacitation. In: Journal of Andrology. 2002 ; Vol. 23, No. 6. pp. 889-898.
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AU - Yudin, Ashley I.

AU - Salinas, Edward R.

AU - Cherr, Gary N.

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AB - Lignin-derived macromolecules (LDMs) are biologically active compounds that affect a variety of cell-to-cell interactions including the inhibition of fertilization and embryo development in a number of nonmammalian species. The effect of lignosulfonic acid (LSA), a highly sulfonated LDM, on cynomolgus macaque sperm-oocyte interaction was evaluated with a zona pellucida binding assay and by in vitro fertilization (IVF). Sperm were treated with LSA (1.5 mg/mL) either before washing or after capacitation. Capacitation included centrifugation through 80% Percoll followed by 2 consecutive washes with medium, overnight incubation, and activation with dibutyryl cyclic adenosine monophosphate and caffeine. The zona binding assay was performed using immature oocytes that had adhered to the center of glass "binding chambers." The number of capacitated sperm that attached to the zona over a 3-minute period was recorded. Sperm attachment was significantly inhibited by LSA as compared to controls whether treatment occurred after capacitation (92.5%; P < .001) or before washing (82.5%; P < .001). When sperm were treated similarly with fucoidin, a sulfated polysaccharide known to inhibit sperm-oocyte interaction, sperm-zona binding was significantly inhibited by postcapacitation treatment but not by prewash treatment. Treatment of sperm with LSA consistently blocked fertilization over 4 IVF cycles both before washing and after capacitation. Fertilization rate for controls was 65% ± 17%. No LSA-treated sperm were observed on the surface of lightly rinsed oocytes after 4 hours of coincubation. Localization of biotinylated LSA showed labeling over the entire sperm surface with the greatest intensity observed over the head and midpiece. LSA treatment had no effect on the percentage of motile sperm or quality of sperm motility. Due to the antifertility properties of this nontoxic molecule, LSA appears to have potential as a vaginal contraceptive.

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