title: The benefits of liposomes for chilling canine sperm for 4 days at 4°C.
abstract: This study comprises 3 experiments exploring the possible benefits and mechanism of action of liposomes for chilling (4°C) canine sperm over a period of 4 days. In the first experiment, 20 ejaculates collected from 5 Beagle dogs were chilled in an extender containing 6% low density lipoproteins (LDL) (Control), or one of 7 extenders containing different concentrations (2, 4, 6, 8, 10, 15, 20%) of liposomes (LIPO). These ejaculates were chilled over 4 days and motility was assessed daily using a Hamilton Thorne analyzer (HTM-IVOS, 14.0). The 2% LIPO obtained the best results (p=0.038) after four days (72.55% motile spermatozoa and 31.4% progressive spermatozoa). In experiment 2, 10 ejaculates were collected from same 5 dogs and chilled in 6% LDL or 2% LIPO-based extenders. Sperm integrity characteristics were assessed prior to refrigeration and every 48h for four days (D0, D2, and D4). Acrosome integrity was assessed using the FITC-PSA test (Fluorescein IsoThiocyanate-Pisum Sativum Agglutinin), plasma membrane (PM) integrity using both the hypo-osmotic swelling test (HOSt) and SYBR14/Propidium Iodide test (SYBR14/PI), and DNA integrity using the Acridine-Orange test (AO). The 2% LIPO extender provided equivalent preservation of sperm integrity parameters to the reference extender (6% LDL). In experiment 3, a Langmuir-Blodgett trough was used to evaluate the mechanistic interactions between LDL, LIPO, prostatic fluid, and the canine spermatozoal membrane during chilling. Results indicate that LDL and LIPO interact differently with the biomimetic membrane. The most likely conclusion of these findings is that LDL and liposomes employ different protective mechanisms during the chilling (4°C) of canine spermatozoa.
authors.affiliation: Laboratory of Biotechnology and Pathology of Reproduction, ONIRIS: The National Veterinary, Food Agriculture, and Food Hygiene School of Loire Atlantique, BP 40706, 44307 Nantes, France; Laboratory of Biotechnology of Animal Reproduction, SAAD DAHLAB University of Blida (U.BLIDA1), BP 270, 09000 Blida, Algeria.
authors.forename: Redha
authors.initials: R
authors.lastname: Belala
authors.affiliation: Laboratory of Biotechnology and Pathology of Reproduction, ONIRIS: The National Veterinary, Food Agriculture, and Food Hygiene School of Loire Atlantique, BP 40706, 44307 Nantes, France.
authors.forename: Juliette
authors.initials: J
authors.lastname: Delay
authors.affiliation: Laboratory of Biotechnology and Pathology of Reproduction, ONIRIS: The National Veterinary, Food Agriculture, and Food Hygiene School of Loire Atlantique, BP 40706, 44307 Nantes, France.
authors.forename: Lamia
authors.initials: L
authors.lastname: Amirat
authors.affiliation: UR1268 Biopolymères Interactions Assemblages, Equipe Interfaces et Systèmes Dispersés, INRA, F-44316 Nantes Cedex 3, France.
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: Ropers
authors.affiliation: Laboratory of Biotechnology and Pathology of Reproduction, ONIRIS: The National Veterinary, Food Agriculture, and Food Hygiene School of Loire Atlantique, BP 40706, 44307 Nantes, France.
authors.forename: Jocya Le
authors.initials: JL
authors.lastname: Guillou
authors.affiliation: UR1268 Biopolymères Interactions Assemblages, Equipe Interfaces et Systèmes Dispersés, INRA, F-44316 Nantes Cedex 3, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Anton
authors.affiliation: IMV Technologies, 10 rue Clemenceau, BP 81, 61302 Aigle Cedex, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Schmitt
authors.affiliation: Department of Statistics, ONIRIS: The National Veterinary, Food Agriculture, and Food Hygiene School of Loire Atlantique, BP 40706, 44307 Nantes, France.
authors.forename: Chantal
authors.initials: C
authors.lastname: Thorin
authors.affiliation: Laboratory of Biotechnology and Pathology of Reproduction, ONIRIS: The National Veterinary, Food Agriculture, and Food Hygiene School of Loire Atlantique, BP 40706, 44307 Nantes, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Michaud
authors.affiliation: Laboratory of Biotechnology of Animal Reproduction, SAAD DAHLAB University of Blida (U.BLIDA1), BP 270, 09000 Blida, Algeria.
authors.forename: Rachid
authors.initials: R
authors.lastname: Kaidi
authors.affiliation: Laboratory of Biotechnology and Pathology of Reproduction, ONIRIS: The National Veterinary, Food Agriculture, and Food Hygiene School of Loire Atlantique, BP 40706, 44307 Nantes, France.
authors.forename: Daniel
authors.initials: D
authors.lastname: Tainturier
authors.affiliation: Laboratory of Biotechnology and Pathology of Reproduction, ONIRIS: The National Veterinary, Food Agriculture, and Food Hygiene School of Loire Atlantique, BP 40706, 44307 Nantes, France. Electronic address: djemil.bencharif@oniris-nantes.fr.
authors.forename: Djemil
authors.initials: D
authors.lastname: Bencharif
mesh_terms: D000818:Animals; D004285:Dogs; D008081:Liposomes / Q000627:therapeutic use*; D008297:Male; D012034:Refrigeration / Q000379:methods / Q000662:veterinary; D012661:Semen / Q000502:physiology; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013094:Spermatozoa / Q000502:physiology*
chemical_list: D008081:Liposomes
keywords: Canine sperm; Langmuir-Blodgett trough; Liposomes; Mechanisms of spermatozoa protection; Refrigeration