title: Investigating sperm cryopreservation in a model tunicate, Ciona intestinalis sp. A.
abstract: In cryopreservation procedures, the capacity to protect the cells from freezing and thawing processes is sensitive to the choice of the cryoprotective agent (CPA) and to its optimal concentration. The advancement of research on Tunicate model species has raised interest in liquid nitrogen cryopreservation for the storage and distribution of genetic resources. Ciona intestinalis (Linnè, 1767) consists of a complex of cryptic taxa that are central to several areas of investigation, from comparative genomics to invasive biology. Here we investigated how five CPAs, three chilling rates and two freezing rates influence semen cryopreservation in C. intestinalis sp. A. By using larval morphology and motility as endpoints, we estimated that long term semen storage requires 10% dimethyl sulfoxide as a protective agent, -1°C/min chilling rate (18°C to 5°C) and -13°C/min freezing rate (5°C to -80°C), followed by immersion in liquid nitrogen.
authors.affiliation: Stazione Zoologica Anton Dohrn, Villa Comunale, I-80121 Naples, Italy.
authors.forename: Gerarda
authors.initials: G
authors.lastname: Sorrenti
authors.affiliation: Laboratory of Cryobiology, Department of Biological Sciences, University of Naples "Federico II", Via Mezzocannone 16, I-80134 Naples, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Bagnoli
authors.affiliation: Stazione Zoologica Anton Dohrn, Villa Comunale, I-80121 Naples, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Miraglia
authors.affiliation: Stazione Zoologica Anton Dohrn, Villa Comunale, I-80121 Naples, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Crocetta
authors.affiliation: Laboratory of Cryobiology, Department of Biological Sciences, University of Naples "Federico II", Via Mezzocannone 16, I-80134 Naples, Italy; CRIAcq, Interdepartmental Research Center for Acquaculture, University of Naples "Federico II", Via Università 100, Portici, I-80055 Naples, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Vitiello
authors.affiliation: Stazione Zoologica Anton Dohrn, Villa Comunale, I-80121 Naples, Italy.
authors.forename: Filomena
authors.initials: F
authors.lastname: Ristoratore
authors.affiliation: Stazione Zoologica Anton Dohrn, Villa Comunale, I-80121 Naples, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Cirino
authors.affiliation: Laboratory of Cryobiology, Department of Biological Sciences, University of Naples "Federico II", Via Mezzocannone 16, I-80134 Naples, Italy; CRIAcq, Interdepartmental Research Center for Acquaculture, University of Naples "Federico II", Via Università 100, Portici, I-80055 Naples, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Sansone
authors.affiliation: Stazione Zoologica Anton Dohrn, Villa Comunale, I-80121 Naples, Italy; CNR ISAFOM - Institute for Agricultural and Forest Systems in the Mediterranean, National Research Council, Str. le Lancia, Blocco Palma I, Zona Industriale, I-95121 Catania, Italy. Electronic address: paolo.sordino@szn.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Sordino
mesh_terms: D000818:Animals; D002938:Ciona intestinalis / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D003247:Conservation of Natural Resources; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro; D005615:Freezing; D007814:Larva / Q000254:growth & development; D008297:Male; D009865:Oocytes / Q000166:cytology / Q000502:physiology; D012662:Semen Preservation / Q000379:methods*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects* / Q000502:physiology
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide
keywords: Ciona intestinalis; Cryopreservation; Dimethyl sulfoxide; Fertilization success; Spermatozoon