The study shows that the growth‑hormone‑releasing peptide GHRP‑2 makes pituitary cells from sheep fire more by letting extra calcium flow into the cells. This calcium influx is needed for the peptide’s effect and can be blocked by calcium‑channel blockers.

The newly synthesized GH-releasing peptide, GHRP-2 (D-Ala-D-beta Nal-Ala-Trp-D-Phe-Lys-NH2), was studied in somatotroph-enriched populations of ovine pituitary cells in primary culture. Nystatin-perforated whole-cell recordings were made on identified somatotrophs after 4-14 days of culture. Using a standard bath solution (containing Na+, Ca2+) and an electrode solution containing K+ in current-clamp recordings, GHRP-2 (10 nM) depolarized the membrane potential of the cells triggering a burst of action potentials. Voltage-clamp recordings indicated that GHRP-2 produced a slowly inactivated inward current with a slight reduction in outward current. The inward current was blocked by the Ca2+ channel blocker, Co2+ (0.5 mM). Ca2+ currents were then isolated using tetraethylammonium bath solution and an electrode solution containing Cs+. Ovine somatotrophs possess transient (T type) and long lasting (L type) Ca2+ currents. The L type current was abolished by addition of nifedipine (3 microM) to the bath solution and T type current was isolated on this basis. Current-voltage relationships indicated an increase in both T and L type Ca2+ currents in response to GHRP-2. The voltage-dependent inactivation curve for T type Ca2+ current was shifted towards a less negative level by the peptide. Intracellular free Ca2+ concentration ([Ca2+]i) in somatotroph-enriched populations was specifically increased by GHRP-2 but this effect was totally abolished by blockade of membrane Ca2+ channels. These data show that GHRP-2 causes an influx of Ca2+ leading to an increase in [Ca2+]i in ovine somatotrophs.(ABSTRACT TRUNCATED AT 250 WORDS)