title: A lectin-based cell microarray approach to analyze the mammalian granulosa cell surface glycosylation profile.
abstract: The high complexity of glycome, the repertoire of glycans expressed in a cell or in an organism, is difficult to analyze and the use of new technologies has accelerated the progress of glycomics analysis. In the last decade, the microarray approaches, and in particular glycan and lectin microarrays, have provided new insights into evaluation of cell glycosylation status. Here we present a cell microarray method based on cell printing on microarray slides for the analysis of the glycosylation pattern of the cell glycocalyx. In order to demonstrate the reliability of the developed method, the glycome profiles of equine native uncultured mural granulosa cells (uGCs) and in vitro cultured mural granulosa cells (cGCs) were determined and compared. The method consists in the isolation of GCs, cell printing into arrays on microarray slide, incubation with a panel of biotinylated lectins, reaction with fluorescent streptavidin and signal intensity detection by a microarray scanner. Cell microarray technology revealed that glycocalyx of both uGCs and cGCs contains N-glycans, sialic acid terminating glycans, N-acetylglucosamine and O-glycans. The comparison of uGCs and cGCs glycan signals indicated an increase in the expression of sialic acids, N-acetylglucosamine, and N-glycans in cGCs. Glycan profiles determined by cell microarray agreed with those revealed by lectin histochemistry. The described cell microarray method represents a simple and sensitive procedure to analyze cell surface glycome in mammalian cells.
authors.affiliation: Department of Emergency and Organ Transplantation (DETO), Veterinary Clinics and Animal Productions Unit, University of Bari Aldo Moro, SP Casamassima, km 3, 70010, Valenzano, Bari, Italy.
authors.forename: Gianluca
authors.initials: G
authors.lastname: Accogli
authors.affiliation: Department of Emergency and Organ Transplantation (DETO), Veterinary Clinics and Animal Productions Unit, University of Bari Aldo Moro, SP Casamassima, km 3, 70010, Valenzano, Bari, Italy.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Desantis
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics (DBBB), University of Bari Aldo Moro, SP Casamassima, km 3, 70010, Valenzano, Bari, Italy.|Experimental Zooprophylactic Institute of Puglia and Basilicata, Via Manfredonia 20, 71121, Foggia, Italy.
authors.forename: Nicola Antonio
authors.initials: NA
authors.lastname: Martino
authors.affiliation: Department of Biosciences, Biotechnologies and Biopharmaceutics (DBBB), University of Bari Aldo Moro, SP Casamassima, km 3, 70010, Valenzano, Bari, Italy.
authors.forename: Maria Elena
authors.initials: ME
authors.lastname: Dell'Aquila
authors.affiliation: Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-84538, Bratislava, Slovakia.
authors.forename: Peter
authors.initials: P
authors.lastname: Gemeiner
authors.affiliation: Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Sciences, Dúbravská cesta 9, SK-84538, Bratislava, Slovakia. jaroslav.katrlik@savba.sk.
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Katrlík
mesh_terms: D000818:Animals; D005260:Female; D019276:Glycocalyx / Q000378:metabolism*; D006107:Granulosa Cells / Q000166:cytology / Q000378:metabolism*; D006736:Horses; D037102:Lectins / Q000737:chemistry*; D046888:Tissue Array Analysis / Q000295:instrumentation* / Q000379:methods*
chemical_list: D037102:Lectins
keywords: Cell microarray; Cell surface glycome; Equine mural granulosa cells; Glycosylation; Lectin histochemistry; Oogenesis