title: Spermaurin, an La1-like peptide from the venom of the scorpion Scorpio maurus palmatus, improves sperm motility and fertilization in different mammalian species.
abstract: Is it possible to identify original compounds that are able to enhance sperm motility from the venom of the scorpion Scorpio maurus palmatus?
  
  
  
  We identified a potent disulfide-rich peptide (DRP) of 73 amino acids that significantly improved the motility of fresh and frozen-thawed sperm in different mammalian species, including human, and improved fertilization outcome in mouse IVF experiments.
  
  
  
  Any disturbance of sperm motility has a strong impact on fertilization and can lead to subfertility or infertility. Significant efforts have, therefore,  been made to identify pharmacological drugs that might improve sperm motility. Such compounds are particularly useful in azoospermia to improve testicular sperm extraction and in the domain of cryopreservation because the motility of frozen-thawed sperm is reduced.
  
  
  
  This was a basic science/medical research study aimed at identifying original compounds from a library of venoms able to enhance mammalian sperm motility, including human. We first identified in the venom of a scorpion S. m. palmatus a fraction able to potently activate sperm motility. We next purified and characterized the compound by liquid chromatography, mass spectrometry and peptide synthesis. Finally, the potency and toxicity of both purified and synthetic versions of the identified compound on sperm motility were assessed using different in vitro tests in different mammalian species.
  
  
  
  For human sperm, biological samples were collected from normozoospermic donors and subfertile patients attending a reproduction department for diagnostic semen analysis. Testicular sperm was collected from cynomolgus monkeys (Macaca fascicularis) euthanized for the needs of specific authorized research projects. The peptide was also tested on bovine and mouse epidydimal sperm. We measured different sperm motility parameters with a computer-assisted sperm analysis system in the presence or absence of the peptide.
  
  
  
  Size exclusion chromatography enabled us to isolate a fraction of the venom of S. m. palmatus able to increase sperm motility. By liquid chromatography and mass spectrometry, a peptide comprising 73 amino acids with 4 disulfide bridges was identified as responsible for the biological activity and called 'spermaurin'. The identity of spermaurin was confirmed by chemical synthesis. We showed that the peptide increased the motility of fresh and frozen-thawed human sperm. We observed that the potency of the peptide was higher on fresh ejaculated spermatozoa with a low motility, achieving a 100% increase of curvilinear velocity in poorly performing sperm. We also demonstrated that peptide is effective on bovine and mouse fresh epididymal, bovine frozen-thawed ejaculated and fresh non-human primate testicular sperm. Finally, in mouse IVF, the production of 2-cell embryos was increased by 24% when sperm were treated with the peptide.
  
  
  
  This work is an in vitro evaluation of the ability of spermaurin to improve sperm motility parameters. Another limitation of this study is the small number of human sperm samples tested with the natural (n = 36) and synthetic (n = 12) peptides. Moreover, the effect of the peptide on IVF outcome was only tested in mouse and further tests with human and bovine gametes are required to confirm and extend this result in other mammalian species.
  
  
  
  This work confirms our initial study showing that venoms represent an interesting source of molecules that are able to modify sperm physiology. Moreover, this work presents the first demonstrated biological action of a venom peptide from the scorpion S. m. palmatus with sequence similarities to La1 peptide from Liocheles australasiae (Wood scorpion), a widespread family of DRPs.
  
  
  
  Not applicable.
  
  
  
  This work is part of the project 'LAB COM-14 LAB7 0004 01-LIPAV', funded by the program LabCom 2014 from the French Research Agency (ANR). Dr Arnoult reports grants from IMV Technologies during the conduct of the study. In addition, Drs Arnoult, Martinez, Ray and Schmitt have a patent EP16305642.7 pending containing some of the information presented in this manuscript.
authors.affiliation: IMV Technologies, ZI N° 1 Est, F-61300 L'Aigle, France.|Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Martinez
authors.affiliation: Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.
authors.forename: Jean-Pascal
authors.initials: JP
authors.lastname: Hograindleur
authors.affiliation: Plateforme BioPark d'Archamps, Archamps Technopole, Saint Julien en Genevois F-74160, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Voisin
authors.affiliation: Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.
authors.forename: Roland
authors.initials: R
authors.lastname: Abi Nahed
authors.affiliation: L'institut du thorax, Inserm UMR 1087/CNRS UMR 6291, Université de Nantes, NantesF44007, France.
authors.forename: Tarek M
authors.initials: TM
authors.lastname: Abd El Aziz
authors.affiliation: Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.
authors.forename: Jessica
authors.initials: J
authors.lastname: Escoffier
authors.affiliation: Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.|CHU Grenoble Alpes, UF de Biologie de la procréation, Grenoble F-38000, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Bessonnat
authors.affiliation: Molecular Imaging Research Center, MIRCen CEA/INSERM UMR1169, Fontenay-aux-Roses F-92265, France.
authors.forename: Claire-Maëlle
authors.initials: CM
authors.lastname: Fovet
authors.affiliation: L'institut du thorax, Inserm UMR 1087/CNRS UMR 6291, Université de Nantes, NantesF44007, France.
authors.forename: Michel
authors.initials: M
authors.lastname: De Waard
authors.affiliation: Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.|CHU Grenoble Alpes, UF de Biologie de la procréation, Grenoble F-38000, France.
authors.forename: Sylviane
authors.initials: S
authors.lastname: Hennebicq
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR 4301, Orléans F-45071, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Aucagne
authors.affiliation: Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.|CHU Grenoble Alpes, UF de Biochimie Génétique et Moléculaire, Grenoble F-38000, France.
authors.forename: Pierre F
authors.initials: PF
authors.lastname: Ray
authors.affiliation: IMV Technologies, ZI N° 1 Est, F-61300 L'Aigle, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Schmitt
authors.affiliation: Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.|Plateforme BioPark d'Archamps, Archamps Technopole, Saint Julien en Genevois F-74160, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Bulet
authors.affiliation: Université Grenoble Alpes, Grenoble F-38000, France.|Institute for Advanced Biosciences INSERM U1209, CNRS UMR5309, Grenoble F-38000, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Arnoult
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D015925:Cryopreservation; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects*; D004822:Epididymis / Q000166:cytology / Q000187:drug effects / Q000503:physiopathology; D005260:Female; D005299:Fertility Agents / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology*; D005307:Fertilization in Vitro; D006801:Humans; D007248:Infertility, Male / Q000188:drug therapy / Q000503:physiopathology; D008252:Macaca fascicularis; D008297:Male; D051379:Mice; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000302:isolation & purification / Q000494:pharmacology*; D012605:Scorpions; D055101:Semen Analysis; D013081:Sperm Motility / Q000187:drug effects* / Q000502:physiology; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects* / Q000473:pathology; D013111:Spider Venoms / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D013737:Testis / Q000166:cytology / Q000187:drug effects / Q000503:physiopathology
chemical_list: D005299:Fertility Agents; D019151:Peptide Library; D010455:Peptides; D013111:Spider Venoms; C000626177:spermaurin, Scorpio maurus palmatus
keywords: La1-like peptide; library screening; male infertility; peptide synthesis; pharmacology; scorpion venom; sperm motility