title: Aquaporin-Mediated Water and Hydrogen Peroxide Transport Is Involved in Normal Human Spermatozoa Functioning.
abstract: Different aquaporins (AQPs) are expressed in human sperm cells and with a different localization. Their function has been related to cell volume control in response to the osmotic changes encountered passing from the epididymal fluid to the cervical mucus or involved in the end stage of cytoplasm removal during sperm maturation. Recently, AQPs have also shown hydrogen peroxide (H₂O₂) permeability properties. Here, we investigate the expression, localization and functioning of AQPs in human sperm cells with particular attention to their role as peroxiporins in reactive oxygen species (ROS) scavenging in both normospermic and sub-fertile human subjects. Western blotting and immunocytochemistry were used to confirm and clarify the AQPs expression and localization. Water and H₂O₂ permeability was tested by stopped flow light scattering method and by the CM-H2DCFDA (5-(and-6)-chloromethyl-2',7'-dichlorodihydro-fluorescein diacetate, acetyl ester) H₂O₂ fluorescence probe, respectively. AQP3, -7, -8, and -11 proteins were found in human sperm cells and localized in the head (AQP7), in the middle piece (AQP8) and in the tail (AQP3 and -11) in both the plasma membrane and in intracellular structures. Sperm cells showed water and H₂O₂ permeability which was reversibly inhibited by H₂O₂, heat stress and the AQP inhibitor HgCl₂. Reduced functionality was observed in patients with compromised basal semen parameters. Present findings suggest that AQPs are involved in both volume regulation and ROS elimination. The relationship between sperm number and motility and AQP functioning was also demonstrated.
authors.affiliation: Department of Molecular Medicine, University of Pavia, I-27100 Pavia, Italy. lumberto@unipv.it.
authors.forename: Umberto
authors.initials: U
authors.lastname: Laforenza
authors.affiliation: Department of Molecular Medicine, University of Pavia, I-27100 Pavia, Italy. giorgia.pellavio@gmail.com.
authors.forename: Giorgia
authors.initials: G
authors.lastname: Pellavio
authors.affiliation: Scientific Direction, Fondazione IRCCS Policlinico San Matteo, I-27100 Pavia, Italy. a.marchetti@smatteo.pv.it.
authors.forename: Anna Lisa
authors.initials: AL
authors.lastname: Marchetti
authors.affiliation: Center for Reproductive Medicine, Obstetrics and Ginecology Unit, Fondazione IRCCS Policlinico San Matteo, I-27100 Pavia, Italy. claudia.omes@unipv.it.
authors.forename: Claudia
authors.initials: C
authors.lastname: Omes
authors.affiliation: Center for Reproductive Medicine, Obstetrics and Ginecology Unit, Fondazione IRCCS Policlinico San Matteo, I-27100 Pavia, Italy. todarofederica@gmail.com.
authors.forename: Federica
authors.initials: F
authors.lastname: Todaro
authors.affiliation: Department of Molecular Medicine, University of Pavia, I-27100 Pavia, Italy. gastaldi@unipv.it.
authors.forename: Giulia
authors.initials: G
authors.lastname: Gastaldi
mesh_terms: D051400:Aquaporin 3 / Q000378:metabolism; D020346:Aquaporins / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism* / Q000494:pharmacology; D007150:Immunohistochemistry; D007248:Infertility, Male / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D008627:Mercuric Chloride / Q000494:pharmacology; D016877:Oxidants / Q000378:metabolism / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D013077:Sperm Head / Q000378:metabolism; D013094:Spermatozoa / Q000378:metabolism / Q000502:physiology*; D013696:Temperature; D014867:Water / Q000378:metabolism*
chemical_list: C511251:AQP11 protein, human; C496027:AQP3 protein, human; C114141:AQP7 protein, human; D020346:Aquaporins; D016877:Oxidants; D017382:Reactive Oxygen Species; C108330:aquaporin 8; D014867:Water; D051400:Aquaporin 3; D008627:Mercuric Chloride; D006861:Hydrogen Peroxide
keywords: aquaporins-7; aquaporins-8; oxidative stress; sperm motility; sterility; water channel