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The CatSper channel modulates boar sperm motility during capacitation.
Linköping University, Department of Clinical and Experimental Medicine, Division of Clinical Sciences. Linköping University, Faculty of Medicine and Health Sciences.ORCID iD: 0000-0002-0256-1958
Linköping University, Faculty of Medicine and Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Division of Children's and Women's health.ORCID iD: 0000-0003-0120-354X
Linköping University, Faculty of Medicine and Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Division of Children's and Women's health.ORCID iD: 0000-0002-5194-2124
2017 (English)In: Reproductive biology, ISSN 2300-732X, Vol. 17, no 1, 69-78 p.Article in journal (Refereed) Published
Abstract [en]

The cation channel of sperm (CatSper) comprises four transmembrane subunits specifically expressed in human, equine, murine and ovine spermatozoa, apparently implicated in capacitation, hyperactivation and acrosome exocytosis. Western blotting and immunocytochemistry showed hereby that CatSper subunits are also present in boar spermatozoa, primarily over the sperm neck, tail and cytoplasmic droplets; albeit CatSper -1 presented in addition some distribution over the membrane of the acrosome and CatSper -2 and -4 over the membrane of the post-acrosome. The role of the Catsper channel in boar spermatozoa was investigated by extending the spermatozoa in media containing different calcium (Ca(2+)) availability and exposure to the capacitation-trigger bicarbonate, to progesterone or CatSper inhibitors (Mibefradil and NNC 55-0396), separately or sequentially, at physiological and toxicological doses. Extracellular Ca(2+) availability, combined with bicarbonate exposure (capacitation-inducing conditions) decreased sperm motility, similarly to when spermatozoa incubated in capacitation-inducing conditions was exposed to Mibefradil and NNC 55-0396. Exposure of these spermatozoa to progesterone did not cause significant changes in sperm motility and nor did it revert its decrease induced by CatSper antagonists. In conclusion, the CatSper channel regulates sperm motility during porcine capacitation-related events in vitro.

Place, publisher, year, edition, pages
Elsevier, 2017. Vol. 17, no 1, 69-78 p.
Keyword [en]
CatSper, Spermatozoa, Capacitation, Sperm motility, Boar
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:liu:diva-134398DOI: 10.1016/j.repbio.2017.01.001ISI: 000397370900009PubMedID: 28077244Scopus ID: 2-s2.0-85009212156OAI: oai:DiVA.org:liu-134398DiVA: diva2:1076654
Note

Funding agencies: The Swedish Research council VR, Stockholm [521-2011-6553]; Research Council FORMAS, Stockholm [221-2011-512]; FORSS (Forskningsradet i Sydostra Sverige, Sweden [473121]

Available from: 2017-02-23 Created: 2017-02-23 Last updated: 2017-05-29Bibliographically approved
In thesis
1. Sperm Membrane Channels, Receptors and Kinematics: Using boar spermatozoa for drug toxicity screening
Open this publication in new window or tab >>Sperm Membrane Channels, Receptors and Kinematics: Using boar spermatozoa for drug toxicity screening
2016 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Internal fertilization usually implies that a spermatozoon, with intact attributes for zygote formation, passes all hurdles during its transport through the female genitalia and reaches the oocyte. During this journey, millions to billions of other spermatozoa perish. Spermatozoa are highly differentiated motile cells without synthetic capabilities. They generate energy via glycolysis and oxidative phosphorylation to sustain motility and to maintain the stability and functionality of their plasma membrane. In vivo, they spend their short lifespan bathing in female genital tract fluids of different origins, or are in vitro exposed to defined media during diverse sperm handling i.e. extension, cryopreservation, in vitro fertilization, etc. Being excitable cells, spermatozoa respond in vivo to various stimuli during pre-fertilization (capacitation, hyperactivation, oocyte location) and fertilization (acrosome reaction, interaction with the oocyte) events, mediated via diverse membrane ion-conducting channels and ligand-gated receptors. The present Thesis has mapped the presence and reactivity (sperm intactness and kinematics) of selected receptors, water and ion channels in ejaculated boar spermatozoa. The final aim was to find a relevant alternative cell type for in vitro bioassays that could ease the early scrutiny of candidate drugs as well as decreasing our needs for experimental animals according to the 3R principles. Spermatozoa are often extended, cooled and thawed to warrant their availability as fertile gametes for breeding or in vitro testing. Such manipulations stress the cells via osmotic variations and hence spermatozoa need to maintain membrane intactness by controlling the exchange of water and the common cryoprotectant glycerol, via aquaporins (AQPs). Both AQPs-7 and -9 were studied for membrane domain changes in cauda- and ejaculated spermatozoa (un-processed, extended, chilled or frozen-thawed). While AQP-9 maintained location through source and handling, thawing of ejaculated spermatozoa clearly relocated the labelling of AQP-7, thus appearing as a relevant marker for non-empirical studies of sperm cryopreservation. Alongside water, spermatozoa interact with calcium (Ca2+) via the main Ca2+ sperm channel CatSper. Increments in intracellular Ca2+ initiate motility hyperactivation and the acrosome reaction. The four subunits of the CatSper channel were present in boar spermatozoa, mediating changes in sperm motility under in vitro capacitation-inducing conditions (increased extracellular Ca2+ availability and bicarbonate) or challenge by the CatSper antagonists mibefradil and NNC 55-0396. Uterine and oviduct fluids are richest in endogenous opioids as β-endorphins during mating and ovulation. Both μ- and δ- opioid receptors were present in boar spermatozoa modulating sperm motility, as in vitro challenge with known agonists (μ: morphine; δ: DPDPE and κ: U 50488) and antagonists (μ: naloxone; δ: naltrindole and κ: nor-binaltrorphimine) showed that the μ-opioid receptor maintained or increased motility while the δ-opioid receptor mediated decreased motility over time. Finally, boar spermatozoa depicted dose-response effects on sperm kinematics and mitochondrial potential following in vitro challenge with 130 pharmacological drugs and toxic compounds as well as with eight known mito-toxic compounds. In conclusion, boar spermatozoa expressing functional water (AQPs-7 and -9) and ion (CatSper 1-4) channels as well as μ- and δ-opioid receptors are able to adapt to stressful environmental variations, capacitation and pharmacological compounds and drug components. Ejaculated sperm suspensions are easily and painlessly obtained from breeding boars, and are suitable biosensors for in vitro drug-induced testing, complying with the 3R principles of reduction and replacement of experimental animals, during early toxicology screening.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2016. 65 p.
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 1526
Keyword
Plasma membrane, membrane channels, membrane receptors, kinematics, 3Rprinciples, spermatozoa, boar.
National Category
Cell and Molecular Biology Pharmacology and Toxicology Cell Biology Pharmaceutical Sciences Biochemistry and Molecular Biology Veterinary Science
Identifiers
urn:nbn:se:liu:diva-131862 (URN)10.3384/diss.diva-131862 (DOI)9789176857267 (ISBN)
Public defence
2016-11-11, Berzeliussalen, Campus US, Linköping, 09:00 (English)
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Supervisors
Available from: 2016-10-11 Created: 2016-10-11 Last updated: 2017-05-29Bibliographically approved

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The full text will be freely available from 2018-01-08 14:02
Available from 2018-01-08 14:02

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