The mechanisms of splenic control in the Antarctic fish, Pagothenia borchgrevinki, were investigated using isolated spleen and mesenteric artery strips in vitro and perfused spleen preparations in situ. Splenosomatic index (SSI) [100 x (spleen wt/body wt)] and hematocrit were determined in animals treated with atropine and phentolamine. Atropine injection increased the SSI from 0.60 +/- 0.06 to 0.89 +/- 0.04, whereas phentolamine decreased SSI to 0.45 +/- 0.03. In atropine-injected fish, hematocrit was 18.6 +/- 1.4 before and 6.6 +/- 0.8% 3 h after injection. Electrical stimulation of the splenic nerves produced biphasic flow responses. In 11 of 12 tested preparations, atropine (3 x 10(-7) to 10(-6) NI) abolished the response, suggesting a major cholinergic component in the splenic innervation. Isolated spleen strip prepara tions contracted in response to carbachol, a response that was antagonized by atropine. The response to acetylcholine was markedly enhanced by the specific cholinesterase inhibitor BW-284c51. Catecholamine effects were somewhat irregular, and maximal contraction force with epinephrine and norepinephrine was 41 and 56%, respectively, of the carbachol response. The results suggest a mainly, if not solely, cholinergic autonomic control of the borch spleen, and a major function of the cholinergic innervation in the control of hematocrit in this species.