Coexistence of C-type natriuretic peptide and atrial natriuretic peptide systems in the bovine cornea

Abstract

PURPOSE. To determine whether the cornea synthesizes natriuretic peptides and contains their receptors. METHODS. The synthesis of the natriuretic peptides, C-type natriuretic peptide (CNP) and atrial natriuretic peptide (ANP), in the bovine cornea was determined by high-performance liquid chromatography (HPLC) with radioimmunoassay and Southern blot analysis. The presence of natriuretic peptide receptor (NPR)-A and -B and their localizations were measured by reverse transcription-polymerase chain reaction (RT-PCR), in vitro autoradiography, and the activation of particulate guanylyl cyclase by natriuretic peptides in the corneal membrane. RESULTS. The serial dilution curves of corneal extracts were parallel to the standard curves of CNP and ANP, With reversed-phase HPLC, a major immunoreactive peak of CNP or ANP was observed at the elution time corresponding with synthetic CNP(1-53) or atriopeptin III (APIII), respectively. The presence of mRNAs of CNP and ANP was also detected in the cornea by RT-PCR and/or Southern blot analysis. Production of 3',5'-cyclic guanosine monophosphate (cGMP) by the activation of particulate guanylyl cyclase in the corneal membrane was stimulated by ANP, BNP, and CNP. More cGMP was produced by CNP than by the other natriuretic peptides. Specific I-125-Tyr(0)]-CNP(1-22) binding sites were localized in the endothelial cell layer of cornea. The apparent dissociation constant (K-d) value of the cornea was 3.06 +/- 0.73 nM and the maximum binding capacity was 3.40 +/- 0.63 femtomoles/mm(2). Both NPR-A and NPR-B mRNAs were detected by RT-PCR. CONCLUSIONS. The cornea synthesizes CNP and ANP and contains their receptors. These results suggest that the CNP and ANP systems coexist in the bovine cornea.