Morphological and physiological characterization of individual olfactory interneurons connecting the brain and eyestalk ganglia of the crayfish.

1. In order to understand the functional organization of the crustacean olfactory system, we are using intracellular recording and staining techniques to correlate the structure and function of single, odorant-sensitive interneurons in the brain of the crayfish Procambarus clarkii. We describe here the anatomy and physiology of interneurons that connect the brain with the medullae terminales or other eyestalk ganglia. 2. All of the interneurons in our study are at least third-order olfactory neurons (second-order olfactory interneurons) because they respond to chemostimulation of the olfactory organ (the antennules) but do not branch in the olfactory lobe (the neuropil to which primary olfactory receptor cells of the antennules project). 3. Much of the central nervous system, including the three main divisions of the brain (protocerebrum, deuterocerebrum, tritocerebrum) and the medullae terminales, are involved in integrating olfactory or multimodal (including olfactory) information, since these areas contain neurites of olfactory interneurons. Previous studies have indicated that regions involved in such processing include the olfactory lobes and accessory lobes of the deuterocerebrum, and regions I, II, IV, and VII (in some species) of the medullae terminales. Our results show that also prominent among regions involved in olfactory or multimodal (including olfactory) integration are the anterior and posterior optic neuropils of the protocerebrum, the lateral and medial antennular neuropils of the deuterocerebrum, the tegumentary neuropils and the antennal neuropils of the tritocerebrum, and neuropils III, VI, XII of the medullae terminales. 4. These olfactory interneurons were sensitive to chemostimulation (unimodal), chemo- and mechanostimulation (bimodal), or chemo-, mechano-, and photostimulation (trimodal). Responses could be excitatory or inhibitory, even for a given neuron. Morphologically complex interneurons (those having bilateral branching) were more likely to have complex response characteristics (trimodal sensitivity) than were morphologically simpler interneurons (those having unilateral branching). Olfactory interneurons with a soma in the medulla terminalis showed the most complex response profiles: they were trimodal, and were exicted by odorants but were inhibited by touch and/or light. This finding suggests that these are complex, high order interneurons. 5. Our studies reveal that olfactory and other sensory information is transmitted between the brain and the medullae terminales (and possibly other eyestalk ganglia) by a coactivated, parallel array of structurally and functionally diverse neurons.