Enriched fractions of heavily granulated (type II) and sparsely granulated (type I) somatotrophs have been prepared from male Sprague-Dawley rats by Percoll density gradient centrifugation in two steps. After 3 days of culture, basal GH release was 0.116 +/- 0.024 (n = 30) and 0.223 +/- 0.034 ng GH/microgram protein-min (n = 34) in type I and type II cells, respectively (P < 0.05). GH-releasing hormone (GHRH; 0.01-10 nM) stimulated GH release in type II cells, whereas type I cells only responded to higher doses of GHRH (1 and 10 nM). The dynamics of GH release were similar in the two cell types. Type II cells released more GH in absolute values, which may reflect the higher GH content in these cells. The somatostatin analog octreotide (100 pM) reduced basal GH release by 63% in type I cells, but by only 17% in type II cells. Octreotide also had a slightly greater effect on GHRH-induced GH release in type I cells. Both cell types responded to 100 nM GH-releasing peptide-6. We conclude that both type I and type II somatotrophs contribute to GH release, but type II cells are more sensitive to and release more GH when stimulated with GHRH. The role of type I cells may be to boost the initial secretory response at the onset of physiological pulses.