The prenatal development of the anterior commissure (AC) was studied in 130 hamster embryos with ages varying from E12 to E16 (E1 = day of conception and E16 = P1 = day of birth) by use of carbocyanine crystals (DiI, DiA and/or DiO) implanted into different rostrocaudal segments of the paleocortex. On E12 and E13, many AC axons were seen with tortuous trajectories pointing towards the midline (precrossing stage). On E13.5 and E14, most AC fibers abutted the midsagittal plane, led by a few pioneer axons that grew as far as 500 microns ahead into the opposite hemisphere (crossing stage). Pioneers were present in most brains at these ages irrespective of the rostrocaudal position of the carbocyanine crystal. Somata of pioneer axons could be identified by retrograde labelling. They were characteristically immature neurons, located either in the olfactory peduncle or in the superficial layers of the olfactory cortex. On E14.5 and E15, pioneers and followers were seen close to the targets and on E15.5 and E16 interstitial budding occurred, and arborization started within the olfactory peduncle and the paleocortex (postcrossing stage). If the existence of pioneer fibers represents something more than a stochastic phenomenon, their appearance in the developing AC may reflect the operation of signals at the midline and/or in the contralateral hemisphere that either accelerate the growth of pioneers, or decelerate the growth of followers.