The bag cells are a group of neuroendocrine cells located in the abdominal ganglion of Aplysia californica. These cells induce egg laying in the animal through the release of neurohormone(s). Previous experiments established that an extract of bag cells releases eggs from isolated ovotestis fragments in a dose-dependent manner. (F. E. Dudek and S. S. Tobe, 1978, Gen. Comp. Endocrinol. 36, 618-627.) Experiments presented here were conducted to purify and identify the component(s) of bag cell extract with the egg-releasing activity. Bag cell extracts were fractionated by means of gel-filtration chromatography followed by cation-exchange chromatography. Column eluates were assayed for egg-release activity on isolated ovotestis fragments by the method of Dudek and Tobe (1978). Only one component purified from the crude extract had egg-releasing activity. This component was identified as egg-laying hormone (ELH) based on its purification characteristics, effects on neuronal activity, and migration on thin-layer chromatography. In dose-response studies egg release increased with the concentration of ELH and had a threshold of 8 X 10(-10) M or less. ELH had the same dose-response relationship in egg release assays when present in purified form or as a component of bag cell extract. These data show that ELH acts directly on the ovotestis and that ELH is the only component of bag cell extract with egg-release activity. Taken with the results of other studies (W. D. Branton, S. Arch, T. Smock, and E. Mayeri, 1978. Proc. Nat. Acad. Sci. USA 75, 5732-5736. B. S. Rothman, P. Brownell, and E. Mayeri, 1979. Soc. Neurosci. Abstr. 5, 260. E. Mayeri and B. S. Rothman, 1982.(ABSTRACT TRUNCATED AT 250 WORDS)