Characterization of two channel catfish, Ictalurus punctatus, glucocorticoid receptors and expression following an acute stressor

Two channel catfish glucocorticoid receptor genes, ipGR1 (NR3C1_1) and ipGR2 (NR3C1_2) were partially characterized. Identification and analysis of the genomic organization of two channel catfish glucocorticoid (GC) receptors (GRs) revealed differences in the lengths of exons 1 and 2 and the addition of an extra 27-bp exon inserted after exon 2 in the GR1 gene, yielding a 9-aa insert in the receptor protein. Sequence of the 9-aa insert in ipGR1 (WRARQNTHG) is unique compared to other teleost fish GRs. Amino acid sequence alignment of the two channel catfish GRs, revealed 55% sequence identity between them, with a high degree of sequence conservation (82%) in the DNA binding and ligand binding domains. Real-time PCR indicated that ipGR1 and ipGR2 were expressed in all tissues evaluated. Channel catfish GR1 was predominantly expressed in the gills, nearly 25-fold higher than in the liver. GR1 expression was higher than GR2 expression in gills, intestine, head kidney and heart (P < 0.05). Channel catfish hepatic GR1 mRNA expression was significantly (P < 0.05) increased from prestress expression 30 min following removal of the acute stressor. After 30 min of stress and during the 2 h recovery period, ipGR1 mRNA expression was higher relative to ipGR2 expression. Hepatic ipGR2 expression was not affected (P > 0.05) by the acute stress event. The present study adds to the growing body of information on GR evolution and function and further demonstrates the unique regulation of the GC/GR system in teleost fish.