dc.creatorCasali, Emerson Andre
dc.creatorSilva, Tiago Rodrigues da
dc.creatorGelain, Daniel Pens
dc.creatorKaiser, Glória Regina Rodrigues de Freitas
dc.creatorBattastini, Ana Maria Oliveira
dc.creatorSarkis, João José Freitas
dc.creatorBernard, Elena Aida
dc.date2010-04-24T04:15:28Z
dc.date2001
dc.identifier0100-879X
dc.identifierhttp://hdl.handle.net/10183/21140
dc.identifier000327088
dc.descriptionSertoli cells have been shown to be targets for extracellular purines such as ATP and adenosine. These purines evoke responses in Sertoli cells through two subtypes of purinoreceptors, P2Y2 and PA1. The signals to purinoreceptors are usually terminated by the action of ectonucleotidases. To demonstrate these enzymatic activities, we cultured rat Sertoli cells for four days and then used them for different assays. ATP, ADP and AMP hydrolysis was estimated by measuring the Pi released using a colorimetric method. Adenosine deaminase activity (EC 3.5.4.4) was determined by HPLC. The cells were not disrupted after 40 min of incubation and the enzymatic activities were considered to be ectocellularly localized. ATP and ADP hydrolysis was markedly increased by the addition of divalent cations to the reaction medium. A competition plot demonstrated that only one enzymatic site is responsible for the hydrolysis of ATP and ADP. This result indicates that the enzyme that acts on the degradation of tri- and diphosphate nucleosides on the surface of Sertoli cells is a true ATP diphosphohydrolase (EC 3.6.1.5) (specific activities of 113 ± 6 and 21 ± 2 nmol Pi mg-1 min-1 for ATP and ADP, respectively). The ecto-5'- nucleotidase (EC 3.1.3.5) and ectoadenosine deaminase activities (specific activities of 32 ± 2 nmol Pi mg-1 min-1 for AMP and 1.52 ± 0.13 nmol adenosine mg-1 min-1, respectively) were shown to be able to terminate the effects of purines and may be relevant for the physiological control of extracellular levels of nucleotides and nucleosides inside the seminiferous tubules.
dc.formatapplication/pdf
dc.languageeng
dc.relationBrazilian journal of medical and biological research = Revista brasileira de pesquisas médicas e biológicas. Ribeirão Preto, SP. Vol. 34, no. 10 (Oct. 2001), p. 1247-1256
dc.rightsOpen Access
dc.subjectSertoli cells
dc.subjectPurine nucleotides
dc.subjectATP diphosphohydrolase
dc.subjectEcto-5’-nucleotidase
dc.subjectEctoadenosine deaminase
dc.subjectBioquímica
dc.titleEctonucleotidase activities in Sertoli cells from immature rats
dc.typeArtigo de periódico
dc.typeNacional


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