title: An inducible nitric oxide synthase (NOS) is expressed in hemocytes of the spiny lobster Panulirus argus: cloning, characterization and expression analysis.
abstract: Nitric oxide (NO) is a free radical gas involved in a variety of physiological processes in invertebrates, such as neuromodulation, muscle contraction and host defense. Surprisingly, little is known about the involvement of NO synthase (NOS) in the immune system of crustaceans. This work is focused on the study of the NOS gene of the spiny lobster Panulirus argus, a crustacean with commercial interest, and its relationship with the immune response to a microbial elicitor. A NOS full-length DNA was isolated from hemocytes by reverse transcription-polymerase chain reaction (RT-PCR) using degenerated primers. The open reading frame (ORF) encodes a protein of 1200 amino acids, with an estimated molecular mass of 135.9 kDa, which contains the conserved domains and binding motifs of NOS found in a variety of organisms. NOS gene expression in lobster gills, heart, stomach, digestive gland, abdominal muscle, gut and hemocytes was studied by Real Time quantitative PCR (Real Time qPCR). The expression was higher in hemocytes, heart and gills. In addition, when lobster hemocytes were exposed in vitro to Escherichia coli O55:B5 lipopolysaccharide (LPS), an increase in the NOS activity and also in the NOS gene expression evaluated by Real Time qPCR was observed, thus demonstrating the presence of an inducible crustacean NOS by a microbial elicitor of the immune response. The information is relevant in providing basic knowledge for further studies of crustacean defense mechanisms.
authors.affiliation: Center for Marine Research, University of Havana (CIM-UH), Cuba.
authors.forename: Tania
authors.initials: T
authors.lastname: Rodríguez-Ramos
authors.forename: Yamila
authors.initials: Y
authors.lastname: Carpio
authors.forename: Jorge
authors.initials: J
authors.lastname: Bolívar
authors.forename: Georgina
authors.initials: G
authors.lastname: Espinosa
authors.forename: Jorge
authors.initials: J
authors.lastname: Hernández-López
authors.forename: Teresa
authors.initials: T
authors.lastname: Gollas-Galván
authors.forename: Laida
authors.initials: L
authors.lastname: Ramos
authors.forename: Carlos
authors.initials: C
authors.lastname: Pendón
authors.forename: Mario Pablo
authors.initials: MP
authors.lastname: Estrada
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006434:Hemocytes / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D019001:Nitric Oxide Synthase / Q000235:genetics* / Q000276:immunology*; D033521:Palinuridae / Q000145:classification / Q000201:enzymology* / Q000235:genetics* / Q000276:immunology; D016415:Sequence Alignment
chemical_list: D000276:Adjuvants, Immunologic; D008070:Lipopolysaccharides; D019001:Nitric Oxide Synthase