Cloning and expression pattern of peroxisome proliferator-activated receptor alpha in the thicklip grey mullet Chelon labrosus.
Aquatic organisms living in coastal and estuarine areas are exposed to diverse contaminants which can cause peroxisome proliferation. Peroxisome proliferators are agonists of peroxisome proliferator-activated receptors (PPARs), members of the nuclear receptor superfamily. We have recently demonstrated expression of the three PPAR isoforms in liver of mullet Chelon labrosus and other fish species by immunohistochemistry. The goal of the present study was first to clone PPARalpha and second to investigate its expression pattern in various tissues of mullet. PCR-based screening of mullet cDNA with PPARalpha specific degenerate primers resulted in amplification, subcloning and sequencing of a 1090 bp cDNA fragment (AY618315) that encodes mullet PPARalpha and exhibits highest amino acid identity to fish Sparus aurata PPARalpha (90%). Semi-quantitative RT-PCR was used to characterize the expression of PPARalpha in brain, muscle, liver, spleen, gill, heart and female gonad of juvenile and adult male and female mullet. For this, mullet 18S-rRNA (AY825252), beta-actin (AY836368) and elongation factor alpha (AY836369) were cloned and used as internal reference for RT-PCR. Expression of PPARalpha was detected in all tissues, was highest in liver and lowest in adult male and female muscle.
Raingeard
Damien
D
Laboratory of Cell Biology and Histology, Department of Zoology and Animal Cell Biology, University of the Basque Country, E-48080 Bilbao, Basque Country, Spain.
Cancio
Ibon
I
Cajaraville
Miren P
MP
D000199:Actins / Q000096:biosynthesis; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D015870:Gene Expression / Q000502:physiology*; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D047493:PPAR alpha / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D020648:Peptide Elongation Factor 1 / Q000096:biosynthesis; D012337:RNA, Ribosomal, 18S / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid; D023701:Smegmamorpha / Q000235:genetics / Q000502:physiology*
D000199:Actins; D017931:DNA Primers; D047493:PPAR alpha; D020648:Peptide Elongation Factor 1; D012337:RNA, Ribosomal, 18S