Receptor activated C kinase is down-regulated in the male gonad of the marine bivalve mollusc Mya arenaria exposed to tributyltin (TBT).
This study was aimed at investigating the molecular mechanisms by which tributyltin (TBT) impairs the reproductive processes in the marine bivalve Mya arenaria. The suppression polymerase chain reaction subtractive hybridization (SSH) method was used to identify differentially expressed transcripts in the gonads of adult M. arenaria 72 h after a single injection of 160 ng TBT in the adductor muscle. Subtractive cDNA libraries comprising 322 clones were obtained. These clones were sequenced and corresponded to 55 female and 26 single male non-redundant cDNAs. Following similarity searches in genome databases, some of the transcripts could be assigned to cellular functions including mitochondrial respiration, structural proteins, structure of cytoskeleton, nucleic acid regulation, general metabolism and signal transduction. Among the potentially differentially regulated transcripts, Receptor for activated C kinase 1 (RACK1) represented 6% of the total down-regulated clones in males and the corresponding protein exhibited a high degree of similarity (80%) with the human polypeptide. The RACK1 cDNA from M. arenaria consists of 1085 bp, encoding a 318 deduced polypeptide which contains five internal tryptophan-aspartate (WD) repeats, six putative PKC phosphorylation sites, one tyrosine kinase site, four putative N-myristoylation sites as well as a transmembrane segment spanning amino acid 228-251. A significant down-regulation (by approximately 30% (p<0.05)) of RACK1 expression in male gonads exposed to TBT was confirmed by quantitative real-time RT-PCR. Transcript levels of RACK1 were higher in the female gonads than in the mantle, gills and male gonads. Gene expression as detected by in situ hybridization was strong in mature oocytes comparatively to primary germ cells. RACK1 may be a useful biomarker for TBT exposure in the reproductive system of bivalve molluscs.
Siah
A
A
Laboratory of Ecotoxicology, University of Le Havrse, 25 rue P. Lebon, BP 540, 76058 Le Havre Cedex, France.
Masson
R
R
Loup
B
B
Bultelle
F
F
Pellerin
J
J
Leboulenger
F
F
Danger
J M
JM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry; D015536:Down-Regulation; D004781:Environmental Exposure; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D006066:Gonads / Q000187:drug effects; D057927:Hydrophobic and Hydrophilic Interactions; D008297:Male; D008969:Molecular Sequence Data; D049892:Mya / Q000145:classification / Q000187:drug effects*; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D012337:RNA, Ribosomal, 18S / Q000032:analysis; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000032:analysis / Q000187:drug effects* / Q000235:genetics; D012098:Reproduction / Q000187:drug effects; D016415:Sequence Alignment; D014220:Trialkyltin Compounds / Q000633:toxicity*
D018076:DNA, Complementary; D012333:RNA, Messenger; D012337:RNA, Ribosomal, 18S; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D014220:Trialkyltin Compounds; C011559:tributyltin