title: Molecular characterization of exosomes and their microRNA cargo in human follicular fluid: bioinformatic analysis reveals that exosomal microRNAs control pathways involved in follicular maturation.
abstract: OBJECTIVE
  To characterize well-represented microRNAs in human follicular fluid (FF) and to ascertain whether they are cargo of FF exosomes and whether they are involved in the regulation of follicle maturation.
  
  
  METHODS
  FF exosomes were characterized by nanosight, flow cytometry, and exosome-specific surface markers. Expression microRNA profiles from total and exosomal FF were compared with those from plasma of the same women.
  
  
  METHODS
  University laboratory and an IVF center.
  
  
  METHODS
  Fifteen healthy women who had undergone intracytoplasmic sperm injection.
  
  
  METHODS
  None.
  
  
  METHODS
  TaqMan low-density array to investigate the expression profile of 384 microRNAs; DataAssist and geNorm for endogenous control identification; significance analysis of microarrays to identify differentially expressed microRNAs; nanosight, flow-cytometry, and bioanalyzer for exosome characterization; bioinformatic tools for microRNAs target prediction, gene ontology, and pathway analysis.
  
  
  RESULTS
  We identified 37 microRNAs upregulated in FF as compared with plasma from the same women. Thirty-two were carried by microvesicles that showed the well-characterized exosomal markers CD63 and CD81. These FF microRNAs are involved in critically important pathways for follicle growth and oocyte maturation. Specifically, nine of them target and negatively regulate mRNAs expressed in the follicular microenvironment encoding inhibitors of follicle maturation and meiosis resumption.
  
  
  CONCLUSIONS
  This study identified a series of exosomal microRNAs that are highly represented in human FF and are involved in follicular maturation. They could represent noninvasive biomarkers of oocyte quality in assisted reproductive technology.
authors.affiliation: Department Gian Filippo Ingrassia, Biologia Cellulare e Molecolare, Genetica, Genomica Giovanni Sichel, Università di Catania, Catania, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Santonocito
authors.affiliation: Servizio di PMA/Azienda Ospedaliera Cannizzaro, Catania, Italy.
authors.forename: Marilena
authors.initials: M
authors.lastname: Vento
authors.affiliation: Department Gian Filippo Ingrassia, Biologia Cellulare e Molecolare, Genetica, Genomica Giovanni Sichel, Università di Catania, Catania, Italy.
authors.forename: Maria Rosa
authors.initials: MR
authors.lastname: Guglielmino
authors.affiliation: Department Gian Filippo Ingrassia, Biologia Cellulare e Molecolare, Genetica, Genomica Giovanni Sichel, Università di Catania, Catania, Italy.
authors.forename: Rosalia
authors.initials: R
authors.lastname: Battaglia
authors.affiliation: Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Jessica
authors.initials: J
authors.lastname: Wahlgren
authors.affiliation: Department Gian Filippo Ingrassia, Biologia Cellulare e Molecolare, Genetica, Genomica Giovanni Sichel, Università di Catania, Catania, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Ragusa
authors.affiliation: Department Gian Filippo Ingrassia, Biologia Cellulare e Molecolare, Genetica, Genomica Giovanni Sichel, Università di Catania, Catania, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Barbagallo
authors.affiliation: Servizio di PMA/Azienda Ospedaliera Cannizzaro, Catania, Italy.
authors.forename: Placido
authors.initials: P
authors.lastname: Borzì
authors.affiliation: Servizio di PMA/Azienda Ospedaliera Cannizzaro, Catania, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Rizzari
authors.affiliation: Department Gian Filippo Ingrassia, Biologia Cellulare e Molecolare, Genetica, Genomica Giovanni Sichel, Università di Catania, Catania, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Maugeri
authors.affiliation: Servizio di PMA/Azienda Ospedaliera Cannizzaro, Catania, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Scollo
authors.affiliation: Department of Life, Health and Environmental Sciences, University of L'Aquila, L'Aquila, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Tatone
authors.affiliation: Department of Rheumatology and Inflammation Research, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Hadi
authors.initials: H
authors.lastname: Valadi
authors.affiliation: Department Gian Filippo Ingrassia, Biologia Cellulare e Molecolare, Genetica, Genomica Giovanni Sichel, Università di Catania, Catania, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Purrello
authors.affiliation: Department Gian Filippo Ingrassia, Biologia Cellulare e Molecolare, Genetica, Genomica Giovanni Sichel, Università di Catania, Catania, Italy. Electronic address: dipietro@unict.it.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Di Pietro
mesh_terms: D000328:Adult; D019295:Computational Biology; D055354:Exosomes / Q000502:physiology*; D005260:Female; D015571:Follicular Fluid / Q000378:metabolism*; D063990:Gene Ontology; D006801:Humans; D035683:MicroRNAs / Q000097:blood / Q000378:metabolism*; D006080:Ovarian Follicle / Q000378:metabolism / Q000502:physiology*; D020554:Sperm Injections, Intracytoplasmic; D060225:Tetraspanin 28 / Q000378:metabolism; D060149:Tetraspanin 30 / Q000378:metabolism; D015854:Up-Regulation
chemical_list: C554586:CD63 protein, human; C554798:CD81 protein, human; D035683:MicroRNAs; D060225:Tetraspanin 28; D060149:Tetraspanin 30
keywords: Follicular fluid; exosomes; intrafollicle molecular communication; microRNA profile