Spatial and Temporal Patterns of Evoked Neural Activity from Auditory Nuclei in Chick Brainstem Detected by Optical Recording

Abstract

In order to detect the spatial patterning of the auditory projection of the embryonic chick brainstem, anatomical methods such as orthograde transport of horseradish peroxidase have been used. However, these methods do not provide the continuous information required about the absolute value and time-course of varying neural excitement. Furthermore, the use of conventional electrophysiological methods makes it difficult or impossible to detect the transmembrane voltage change because of the small size and fragility of the cells of the young chick brainstem. We thus believe that optical measurement of membrane potential might be beneficial in circumstances where electrodes are difficult to use for reasons of cell size, complexity, or membrane topology. In the present work, we therefore examined the feasibility of an optical method for delineating the synaptic transmission of afferent input in the auditory nuclei in the chick brainstem. We used embryonic chick brainstem slice preparations featuring an intact eighth nerve, and loaded depolarizing square current pulses from tungsten microelectrodes into the auditory nerve for stimulation of these preparations. In this approach, we used a multiple-site optical recording system comprising a 16 x 16-element photodiode array and a voltage sensitive dye (NK-2761). Neural excitation evoked by stimulation to the left auditory nerve was propagated to the dorsal side of the brainstem. This area in which the optical signal was detected is located on the auditory nuclei. Since the physiological spatial patterning of the auditory nerve projection could be roughly estimated by the optical technique, the technique is considered useful for examining the electrical activity generated from auditory nuclei in the brainstem. This is the first report of spatial patterning of auditory neurons in the embryonic chick brainstem generated through optical recording.