Use of ¹²⁵I-[Y³⁹]exendin-4 to characterize exendin receptors on dispersed pancreatic acini and gastric chief cells from guinea pig

We synthesized and iodinated an exendin-4 analogue, [Y³⁹]exendin-4 (700 Ci/mmol), for use as a radioligand to characterize exendin receptors on dispersed pancreatic acini and gastric chief cells from guinea pig. Binding of this bioactive radioligand was rapid, temperature-dependent and specific (not inhibited by other pancreatic or gastric secretagogues). Measurement of the ability of exendin-4 to inhibit the binding of ¹²⁵I-[Y³⁹]exendin-4 indicated the presence of two classes of receptors. Pancreatic acini had 12.5·10¹⁰ binding sites/mg acinar protein of which 6% were high affinity (K_d = 0.5 nM) and 94% were low affinity (K_d = 0.1 μM). Chief cells had 3370 binding sites/cell of which 9% were high affinity (K_d = 0.3 nM) and 91% were low affinity (K_d = 0.2 μM). Washing with 0.2 M acetic acid (pH 2.5), 0.2 M glycine (pH 10.5), or trypsin (100 μg/ml) after 30 min incubation at 37°C, indicated that 63 and 49% of radioligand was internalized in acini and chief cells, respectively. Truncated glucagon-like peptide-1 (tGLP-1), a mammalian peptide sharing 53% homology with exendin-4, inhibited radioligand binding at the same concentrations that altered secretion from acini and chief cells. Glucagon, GLP-1 and GLP-2 inhibited ¹²⁵I-[Y³⁹]exendin-4 binding only at concentrations ≥ 100 nM. Exendin(9-39)NH₂, a specific exendin-receptor antagonist, potently inhibited ¹²⁵I-[Y³⁹] exendin-4 binding (IC₅₀ = 6.1 and 3.5 nM in acini and chief cells, respectively). In pancreatic acini and gastric chief cells from guinea pig, exendin-3, exendin-4 and tGLP-1 increase cellular cAMP and modulate enzyme secretion by interacting with high-affinity exendin receptors. ¹²⁵I-[Y³⁹] exendin-4 is a useful radioligand for studying exendin receptors.