title: Evidence for the involvement of aquaporins in sperm motility activation of the teleost gilthead sea bream (Sparus aurata).
abstract: The expression of aquaporins in the spermatozoa of the marine teleost gilthead sea bream (Sparus aurata) and their involvement in the motility activation process were investigated. Sperm motility was activated by a hyperosmotic shock, but it was completely inhibited by 10 microM HgCl(2), such inhibition being partially recovered by beta-mercaptoethanol (ME). Conventional RT-PCR using primers specific for S. aurata aquaglyceroporin (glp) and aquaporin 1a (aqp1a) demonstrated the presence of both mRNAs in spermatozoa. Heterologous expression in Xenopus laevis oocytes showed that 10 and 100 microM HgCl(2) equally inhibited water and solute transport through S. aurata aquaporin 1a and S. aurata aquaglyceroporin, but treatment with ME only recovered aquaporin 1a-mediated water permeability. Western blot analysis using isoform-specific antisera on protein extracts from spermatozoa revealed bands that corresponded to the predicted molecular mass of S. aurata aquaglyceroporin (31 kDa) and S. aurata aquaporin 1a (28 kDa). The antisera also demonstrated that both aquaporins were localized in the head and flagellum of the spermatozoa. However, the immunoreaction at the plasma membrane of the spermatozoa head was more intense after the hyperosmotic activation, suggesting the translocation of both aquaporin 1a and aquaglyceroporin into the plasma membrane after the osmotic shock. This study therefore provides the first direct demonstration for the presence of aquaporins in fish sperm. The different sensitivities of S. aurata aquaporin 1a and S. aurata aquaglyceroporin to ME may explain the failure of this reducing agent to fully recover the mercurial inhibition of sperm motility, suggesting that these aquaporins may play different physiological roles during the activation and maintenance of sperm motility in sea bream.
authors: Laboratory of Comparative Physiology, Department of Biological and Environmental Sciences and Technologies, University of Salento, Lecce, Italy.
authors: Loredana
authors: L
authors: Zilli
authors: Roberta
authors: R
authors: Schiavone
authors: François
authors: F
authors: Chauvigné
authors: Joan
authors: J
authors: Cerdà
authors: Carlo
authors: C
authors: Storelli
authors: Sebastiano
authors: S
authors: Vilella
mesh_terms: D000818:Animals; D051397:Aquaglyceroporins / Q000378:metabolism*; D051398:Aquaporin 1 / Q000378:metabolism*; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008297:Male; D008627:Mercuric Chloride / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021541:Sea Bream / Q000502:physiology*; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa / Q000187:drug effects / Q000378:metabolism*; D014741:Video Recording
chemical_list: D051397:Aquaglyceroporins; D012333:RNA, Messenger; D051398:Aquaporin 1; D008627:Mercuric Chloride