Attempts were made to elucidate whether thyroid hormone upregulates renal pro-epidermal growth factor (pro-EGF) gene expression, biosynthesis and release in adult rats which were rendered hypothyroid. Predominantly pro-EGF was detected in renal cortex, whereas pro-EGF and its degraded species were found in urine. We demonstrated that T3 increased pro-EGF levels in renal cortex to 2.2 +/- 0.17, 2.37 +/- 0.19, 2.73 +/- 0.25, and 3.10 +/- 0.45 fold within day 1, 2, 4 and 8, respectively following treatment. Immunoreactive EGF, assessed by immunohistochemical methods, was confined in the distal convoluted tubule and thick ascending limb of Henle. T3 markedly enhanced the density of irEGF in these nephrons. T3 augmented the concentration of urinary irEGF to 2.1, 2.2, 2.8 and 3.6 fold within day 1, 2, 4 and 8 and the abundance of urine pro-EGF to 2.53 +/- 1.39, 3.8 +/- 0.70, 3.59 +/- 1.48 fold within day 1, 2, 4, respectively. Moreover, we employed reverse transcriptase/polymerase chain reaction method to analyze relative abundance of pro-EGF mRNA in kidneys of various thyroid states and found T3 markedly increased pro-EGF mRNA levels after treatment of 1, 2 and 4 days. These results indicated that thyroid hormone augmented the gene expression, biosynthesis and excretion of pro-EGF in adult rat kidney.