Exposure to Hg2+ and Pb2+ changes NTPDase and ecto-5'-nucleotidase activities in central nervous system of zebrafish (Danio rerio).
Neurotransmission can be affected by exposure to heavy metals, such as mercury and lead. ATP is a signaling molecule that can be metabolized by a group of enzymes called ecto-nucleotidases. Here we investigated the effects of mercury chloride (HgCl(2)) and lead acetate (Pb(CH(3)COO)(2)) on NTPDase (nucleoside triphosphate diphosphohydrolase) and ecto-5'-nucleotidase activities in zebrafish brain membranes. In vitro exposure to HgCl(2) decreased ATP and ADP hydrolysis in an uncompetitive mechanism and AMP hydrolysis in a non-competitive manner. Pb(CH(3)COO)(2) inhibited ATP hydrolysis in an uncompetitive manner, but not ADP and AMP hydrolysis. In vivo exposure of zebrafish to HgCl(2) or Pb(CH(3)COO)(2) (20mug/L, during 24, 96h and 30 days) caused differential effects on nucleotide hydrolysis. HgCl(2), during 96h, inhibited the hydrolysis of ATP, ADP and AMP. After 30 days of exposure to HgCl(2), ATP hydrolysis returned to the control levels, ADP hydrolysis was strongly increased and AMP hydrolysis remained inhibited. Exposure to Pb(CH(3)COO)(2) during 96h caused a significant decrease only on ATP hydrolysis. After 30 days, Pb(CH(3)COO)(2) promoted the inhibition of ATP, ADP and AMP hydrolysis. Semi-quantitative RT-PCR analysis showed no changes in the expression of NTPDase1 and 5'-nucleotidase, following 30 days of exposure to both metals. This study demonstrated that Hg(2+) and Pb(2+) affect the ecto-nucleotidase activities, an important enzymatic pathway for the control of purinergic signaling.
Senger
Mario Roberto
MR
Laboratório de Neuroquímica e Psicofarmacologia, Faculdade de Biociências, Pontifícia Universidade Católica do Rio Grande do Sul, Avenida Ipiranga 6681, 90619-900 Porto Alegre, RS, Brazil.
Rico
Eduardo Pacheco
EP
de Bem Arizi
Marcelo
M
Frazzon
Ana Paula Guedes
AP
Dias
Renato Dutra
RD
Bogo
Maurício Reis
MR
Bonan
Carla Denise
CD
D015720:5'-Nucleotidase / Q000096:biosynthesis / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology; D001923:Brain Chemistry / Q000187:drug effects; D002490:Central Nervous System / Q000187:drug effects* / Q000201:enzymology*; D066298:In Vitro Techniques; D007854:Lead / Q000633:toxicity*; D008566:Membranes / Q000187:drug effects / Q000378:metabolism; D008628:Mercury / Q000633:toxicity*; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000096:biosynthesis; D011983:Receptors, Purinergic / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015027:Zebrafish
D009419:Nerve Tissue Proteins; D011983:Receptors, Purinergic; D007854:Lead; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D015720:5'-Nucleotidase; D000251:Adenosine Triphosphatases; C027201:ectoATPase; D008628:Mercury