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Tissue distribution of a novel aquaporin in a teleost fish (Sparus auratus)

dc.contributor.authorEstêvão, Dulce
dc.contributor.authorPower, Deborah
dc.date.accessioned2020-01-29T14:04:26Z
dc.date.available2020-01-29T14:04:26Z
dc.date.issued2003
dc.description.abstractThe aquaporin family consists of two principal groups of transmembrane proteins which function as channels for non-ionic compounds. The CHIP (CHannel forming Integral Protein) group functions only as water channels (aquaporins) and the GLP (GLycerol intrinsic Protein) group transports primarily glicerol but also water and other small solutes. A sea bream AQP-3 cDNA encoding a protein of 298 amino acids was isolated from a kidney cDNA library. The tissue distribution of this GLP family member was investigated by in situ hybridisation. Tissues, liver, gastrointestinal tract, kidney, head kidney, gonads and gills, were fixed in 4% PFA, dehydrated, embedded in wax and serial sections (8µm) mounted on poly-L-lysine coated slides. The AQP-3 probe was generated from the full length cDNA after digestion with BamHI followed by purification. In vitro transcription was carried out using T7 RNA Polymerase and Digoxigenin-RNA labeling mix to synthetize a specific riboprobe. Tissues were hybridised overnight at 54ºC followed by stringency washes in 2xSSC. Detection of hybridised probe was carried out using anti-digoxigenin-AP Fab fragments. The chromagens for colour detection were NBT (4-Nitroblue tetrazolium chloride) and BCIP (5-Bromo-4-Chloro 3-indolylphosphate). Sections were analysed using a microscope coupled to a digital image analysis systems. Results showed that the expression of AQP-3 was abundant in cells scattered in the lamina propria of the hindgut and in occasional cells localised at the interface of the circular and longitudinal muscle layer in the hindgut. Expression was also abundant in some tubules of the kidney and in putative chloride cells in the gills. Some cells stained intensely in the head kidney and in gonads signal was intense in the cytoplasm of larger oocytes, in the head of some sperm and in unidentified cells scattered in testicular tissue. The signal obtained in the mid gut and duodenum was much weaker and no signal was detected in the liver.pt_PT
dc.description.sponsorshipFCT pluriannual grant to CCMAR and M.D. Estêvão was funded by FCT Grant 2/5.3/Prodep/2001.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationEstêvão, M. D.; Power, D. M. – Tissue distribution of a novel aquaporin in a teleost fish (Sparus auratus). 4º Congreso de la Asociación Ibérica de Endocrinologia Comparada, 8-10/9/2003, Córdoba.pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.1/13435
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.titleTissue distribution of a novel aquaporin in a teleost fish (Sparus auratus)pt_PT
dc.typeconference object
dspace.entity.typePublication
oaire.citation.conferencePlaceCórdobapt_PT
oaire.citation.title4º Congreso de la Asociación Ibérica de Endocrinologia Comparadapt_PT
person.familyNameEstêvão
person.familyNamePower
person.givenNameMaria Dulce da Mota Antunes de Oliveira
person.givenNameDeborah Mary
person.identifierI-4695-2014
person.identifier.ciencia-id881B-53E8-0DB1
person.identifier.ciencia-id891A-8A44-3CAE
person.identifier.orcid0000-0002-7151-8363
person.identifier.orcid0000-0003-1366-0246
person.identifier.scopus-author-id8381590400
person.identifier.scopus-author-id7101806760
rcaap.rightsopenAccesspt_PT
rcaap.typeconferenceObjectpt_PT
relation.isAuthorOfPublication67887883-2695-48cc-8221-aa46780ac143
relation.isAuthorOfPublicationc68f5ffb-63f6-4c70-8957-29e464fb59c0
relation.isAuthorOfPublication.latestForDiscoveryc68f5ffb-63f6-4c70-8957-29e464fb59c0

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