Intracellular calcium concentration in equine spermatozoa attached to oviductal epithelial cells in vitro.
Abstract: Interaction of spermatozoa with oviductal epithelial cells (OEC) in the oviductal isthmus prolongs the life span of spermatozoa. The hypothesis that the interaction of equine spermatozoa with OEC affects their intracellular calcium concentration ([Ca2+]i) was tested in a sperm-OEC coculture model. Changes in [Ca2+]i in spermatozoa loaded with the fluorescent calcium indicator indo-1 acetoxymethylester (AM) were determined for spermatozoa attached to OEC or to Matrigel, as well as for free-swimming spermatozoa incubated without oviductal epithelium. [Ca2+]i was determined before incubation and at 0.5, 2, 4, and 6 h of incubation by ratio image analysis of fluorescent images captured at 405 nm and 490 nm. At each time point, [Ca2+]i was lower in motile spermatozoa attached to OEC than in free-swimming spermatozoa. [Ca2+]i in spermatozoa attached to Matrigel was lower than in free-swimming spermatozoa and was comparable to [Ca2+]i in spermatozoa attached to OEC only at 0.5 h incubation. Beyond 0.5 h of incubation, [Ca2+]i was higher in spermatozoa attached to Matrigel than in spermatozoa attached to OEC. These results indicate that spermatozoa with low [Ca2+]i might preferentially attach to OEC and to Matrigel, but that [Ca2+]i is maintained at this basal level only in spermatozoa attached to OEC. The reduced [Ca2+]i in spermatozoa associated with OEC may prevent premature capacitation and acrosomal exocytosis of spermatozoa stored in the isthmic sperm reservoir.
Publication Date: 1996-04-01 PubMed ID: 8924497DOI: 10.1095/biolreprod54.4.783Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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This study investigated if the attachment of equine spermatozoa to oviductal epithelial cells (OEC) in horses affects the intracellular calcium concentration. Results showed that the calcium levels were lowest in spermatozoa that had attached to OEC, suggesting this may help to prolong the life span of the sperm by avoiding premature capacitation.
Study Objective and Hypothesis
- The study aimed to evaluate the potential effects of equine spermatozoa’s interaction with oviductal epithelial cells (OEC), found in the oviductal isthmus of horses, on the sperm’s intracellular calcium concentration.
- The scientists hypothesized that this interaction between OEC and sperm cells might have an impact on calcium ion concentration within the sperm cells.
Research Design and Methodology
- The researchers set up a sperm-oviductal epithelial cell co-culture model to test their hypothesis.
- Changes in calcium concentration within the sperm cells, which were loaded with a fluorescent calcium indicator, were determined for those sperm cells attached to either OEC or Matrigel.
- Calcium concentration was also measured for free-swimming sperm cells that were incubated without oviductal epithelium.
- Measurements were taken before the incubation process as well as at 0.5, 2, 4, and 6 hours into incubation.
Results and Findings
- It was found that the intracellular calcium concentration was lower in spermatozoa that were attached to OEC compared to those free-swimming at all the time frames of incubation.
- The attached spermatozoa to the Matrigel also had lower calcium concentration compared to the free-swimming ones, and this level was usually similar to those attached to the OEC at 0.5-hour incubation.
- However, after 0.5 hours of incubation, the spermatozoa attached to Matrigel showed higher calcium concentration compared to those attached to OEC.
- Therefore, sperm cells with lower calcium ions concentration might be more likely to attach to OEC and Matrigel, but this lower level is only maintained with OEC attachment.
Implications of Findings
- The findings suggest that the reduced intracellular calcium concentration in sperm cells attached to OEC might help prevent premature capacitation and acrosomal exocytosis. This mechanism could therefore help prolong the lifespan of spermatozoa stored in the isthmic sperm reservoir.
Cite This Article
APA
Dobrinski I, Suarez SS, Ball BA.
(1996).
Intracellular calcium concentration in equine spermatozoa attached to oviductal epithelial cells in vitro.
Biol Reprod, 54(4), 783-788.
https://doi.org/10.1095/biolreprod54.4.783 Publication
Researcher Affiliations
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
MeSH Terms
- Acrosome / physiology
- Acrosome / ultrastructure
- Animals
- Calcium / metabolism
- Cells, Cultured
- Coculture Techniques
- Epithelial Attachment / physiology
- Fallopian Tubes / cytology
- Fallopian Tubes / metabolism
- Female
- Fluorescent Dyes
- Horses / physiology
- Indoles
- Male
- Sperm Motility / physiology
- Spermatozoa / metabolism
Grant Funding
- HD19584 / NICHD NIH HHS
Citations
This article has been cited 10 times.- Soto-Heras S, Sakkas D, Miller DJ. Sperm selection by the oviduct: perspectives for male fertility and assisted reproductive technologies†. Biol Reprod 2023 Apr 11;108(4):538-552.
- Kölle S. Sperm-oviduct interactions: Key factors for sperm survival and maintenance of sperm fertilizing capacity. Andrology 2022 Jul;10(5):837-843.
- Gimeno BF, Bariani MV, Laiz-Quiroga L, Martínez-León E, Von-Meyeren M, Rey O, Mutto AÁ, Osycka-Salut CE. Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen-Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status. Animals (Basel) 2021 Jan 3;11(1).
- Machado SA, Sharif M, Wang H, Bovin N, Miller DJ. Release of Porcine Sperm from Oviduct Cells is Stimulated by Progesterone and Requires CatSper. Sci Rep 2019 Dec 20;9(1):19546.
- Oh-Oka T, Saxena DK, Tanii I, Yoshinaga K, Toshimori K. Changes in the activity of sperm nitric oxide synthase in the oviductal reservoir during ovulation. Reprod Med Biol 2003 Jun;2(2):75-81.
- Firman RC, Bentley B, Bowman F, Marchant FG, Parthenay J, Sawyer J, Stewart T, O'Shea JE. No evidence of sperm conjugate formation in an Australian mouse bearing sperm with three hooks. Ecol Evol 2013 Jul;3(7):1856-63.
- Kadirvel G, Machado SA, Korneli C, Collins E, Miller P, Bess KN, Aoki K, Tiemeyer M, Bovin N, Miller DJ. Porcine sperm bind to specific 6-sialylated biantennary glycans to form the oviduct reservoir. Biol Reprod 2012 Jun;87(6):147.
- Mugnier S, Kervella M, Douet C, Canepa S, Pascal G, Deleuze S, Duchamp G, Monget P, Goudet G. The secretions of oviduct epithelial cells increase the equine in vitro fertilization rate: are osteopontin, atrial natriuretic peptide A and oviductin involved?. Reprod Biol Endocrinol 2009 Nov 19;7:129.
- Zelinger E, Brumfeld V, Rechav K, Waiger D, Kossovsky T, Heifetz Y. Three-dimensional correlative microscopy of the Drosophila female reproductive tract reveals modes of communication in seminal receptacle sperm storage. Commun Biol 2024 Feb 6;7(1):155.
- Miller DJ. Sperm in the Mammalian Female Reproductive Tract: Surfing Through the Tract to Try to Beat the Odds. Annu Rev Anim Biosci 2024 Feb 15;12:301-319.
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