SciELO - Scientific Electronic Library Online

 
vol.34 issue10Molecular characterization of DDX26, a human DEAD-box RNA helicase, located on chromosome 7p12In vitro cytotoxicity of the LDE: daunorubicin complex in acute myelogenous leukemia blast cells author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Brazilian Journal of Medical and Biological Research

On-line version ISSN 1414-431X

Abstract

CASALI, E.A. et al. Ectonucleotidase activities in Sertoli cells from immature rats. Braz J Med Biol Res [online]. 2001, vol.34, n.10, pp. 1247-1256. ISSN 1414-431X.  http://dx.doi.org/10.1590/S0100-879X2001001000003.

Sertoli 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.

Keywords : Sertoli cells; purine nucleotides; ATP diphosphohydrolase; ecto-5'-nucleotidase; ectoadenosine deaminase.

        · text in English     · pdf in English