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This study examined the development of individual axon terminal fields in n. laminaris (NL) of the chicken brainstem. In their mature form axons from the nucleus magnocellularis (NM), second-order auditory neurons in the chicken brainstem, project bilaterally onto the NL. Axons from the ipsilateral and contralateral NM neurons form spatially segregated, elongated arbors in the dorsal and ventral neuropil of NL, respectively. The long axes of these arbors correspond to physiologically defined isofrequency bands. To assess the development of this stereotyped arborization pattern, 6-17-day embryonic chicken brain stems were maintained in vitro while injecting horseradish peroxidase into small groups of axons. Three-dimensional reconstructions were made from serial sections and projected onto a cartesian plane for quantitative analyses. At embryonic day 6 (E6), the ventral axons already course beneath the recently migrated NL neurons. The arrival of the dorsal NM axon branches is delayed and their paths are indirect. They first loop dorsally into the the ventricular layer, where they seem to make specific connections with migrating NL neurons and use these as guides to their appropriate positions in the NL. During the period from E9 to E17 the dorsal and ventral terminal fields become similar, each adopting properties of the other's initial pattern. The dorsal terminal fields extend to form bands similar to the early ventral terminal fields, while the ventral terminal fields narrow and appear to shift position in order to achieve the tonotopic specificity characteristic of the early dorsal terminal fields. The results show that a complex, mature pattern of neuronal connections can be formed during development by the combination and reorganization of two simple patterns--each shaped, in turn, by its respective axonal trajectory.

The tonotopic and topographic organization of the bilateral projection from second-order auditory neurons of nucleus magnocellularis (NM) to nucleus laminaris (NL) was examined in young chickens. In one group of birds, the NM axons which innvervate the contralateral NL were severed by cutting the crossed dorsal cochlear tract at the midline. Heavy terminal degeneration in NL was confined to the neuropil area immediately ventral to the perikaryl lamina. Very little degeneration was seen in the dorsal neuropil region. In a second series of animals, the charactertistic frequency (CF) of cells in an area of NM was first determined by microelectrode recording techniques and then a small electrolytic lesion was made through the recording electrode. Following survival periods of 24-48 hours, the distribution of projections from the lesioned area to the ipsilateral and contralateral NL was examined using the Fink-Heimer method. As previously described in the pigeon, projections from NM terminate densely in the neuropil region immediately dorsal to the ipsilateral NL cell bodies and ventral to the perikaryl layer on the contralateral side, providing each NL neuron with segregated binaural innervation. Lesions in any area of the NM produced degeneration confined to a limited caudo-rostral and medio-lateral portion of both laminar nuclei. To investigate this topographic relationship, the cuado-rostral extents of the lesion in NM and of the resulting degeneration in both NL were determined. Linear regression and correlation analyses then related these positional values to each other and to the CF found at the center of each lesion. All correlations were highly significant and ranged from 0.78 between the position of the lesion in NM and CF to 0.91 between the caudo-rostral position of degeneration in the NL ipsilateral and contralateral to the lesion. It is concluded that neurons in NM project in a very discrete topographic, tonotopic and symmetrical fashion to NL on both sides of the brain, contributing to the binaural response properties and tonotopic organization of neurons in NL. The results also suggest that the organization of projections from NM to NL could provide a mechanism for the differential transmission delay required by a "place" model of low-frequency sound localization.

The brainstem auditory pathway of the chicken were used to examine the relationship between the maintenance of dendrites following denervation and the successful reinnervation (sprouting) by surviving afferents. In the system the third-order cells in n. laminaris receive spatially segregated binaural innervation from n. magnocellularis. Afferents from the ipsilateral n. magnocellularis innervate the dendrites on the dorsal aspect of n. laminaris cells, while afferents from contralateral magnocellular neurons innervate ventral dendrites via the crossed dorsal cochlear tract. Denervation of the ventral dendrites of n. laminaris cells by transection at the midline results in rapid and severe atrophy of the denervated dendrite. Unilateral cochlea removal induces transneuronal degeneration of 30-45% of the ipsilateral magnocellular cells, thereby partially denervating one dendrite of the n. laminaris cells on each side of the brain. In animals with long-standing transections of the crossed dorsal cochlear tract there is no evidence of sprouting the fibers from the ipsilateral n. magnocellularis when the projections of the surviving magnocellular neurons are traced with degeneration methods after a secondary cochlea removal. However, when dendrites of n. laminaris are partially denervated dendrites do not disappear. Furthermore, secondary lesions of the crossed dorsal cochlear tract or secondary cochlea removal reveal that these denervated dendrites are reinnervated by the afferents from the opposite n. magnocellularis which are normally restricted to the opposite dendrite of the n. laminaris cells.

Hair cell regeneration following acoustic trauma to the avian cochlea has been documented using DNA labeling with tritiated thymidine. The goal of this study was to identify potential precursor cell populations for regenerating hair cells. Chicks were exposed in pairs to a 1500 Hz pure tone at 120 dBSPL for 18 h. The animals received repeated injections of 3H-thymidine over a survival period of 6, 15, or 24 h, 3 days or 30 days after the completion of noise exposure. One cochlea from each animal was processed for autoradiography and the other for scanning electron microscopy. Labeled, regenerated hair cells were present by 3 days after exposure and recovery from injury was nearly complete by 30 days. Examination of animals in short survival groups suggest that two precursor populations may exist. For inferior sensory epithelial damage, cuboidal or hyaline epithelial cells appear to serve as the precursor cell population for the regeneration of both hair cells and supporting cells. With isolated superior damage, however, supporting cells may be the precursor population.

Glycine-immunoreactivity (Gly-I) was studied in the auditory brain stem of chick using a polyclonal antiserum to glycine conjugated to bovine serum albumin (Wenthold et al., 1987). During embryonic development, little Gly-I is present in nucleus magnocellularis (NM), nucleus laminaris (NL), or nucleus angularis (NA). In posthatch chicks, a few Gly-I terminals are found on nerve cell bodies in NM. Gly-labeled terminals are rare in NL and NA. When present in NL, they appear to be apposed to neuronal somata and not to dendrites. Occasionally, a Gly-labeled cell can be found in NM. After unilateral cochlea removal, Gly-labeled terminals are still present in NM. Thus, the cochlea does not appear to be the source of the glycinergic afferents to NM. The pattern of staining of Gly-I contrasts sharply with that of gamma-aminobutyric acid-immunoreactivity (GABA-I). The number of Gly-I terminals in NM, NL and NA appears to be much less than that of GABA-I. In addition, GABA-I terminals are very abundant around both the cell bodies and dendrites of neurons in NM and NL, whereas Gly-I terminals are found only on neuronal somata. Gly-I in NM also appears to be much less than that in its mammalian homologue, the anteroventral cochlear nucleus.

Electrophysiological activity in response to linear acceleration stimuli was recorded from Gallus Domesticus by means of subcutaneous electrodes. This investigation had two purposes: 1) to obtain normative data for our laboratory, and 2) to rule out auditory and somatosensory contributions to the Vestibular Evoked Potentials (VsEP). The stimulus consisted of a sigmoid-shaped voltage function generated by a digital-to-analog converter. This signal was amplified, attenuated, and directed to a shaker (selenoid-based, linear mechanical vibrator). The animal's head was firmly attached to a small platform which in turn was coupled to the transducer. The recorded electrophysiological activity was filtered, amplified, and averaged over 256 stimulus presentations. The VsEP are composed of a positive wave, a prominent negativity, and three to five additional positive waves which occur within the first 10 milliseconds following the stimulus. The first three elements are the most robust components of the response. Latency/acceleration and amplitude/acceleration functions were constructed for each of these three waves. Clear and replicable responses were obtained at an acceleration of 2.00 g. At this level the amplitude of the components ranged between 3 and 5 microvolts. On the average, threshold responses were recorded at 0.0935 g. VsEP were not affected by high intensity white noise. However, bilateral intralabyrinthine injection of tetrodotoxin (TTX), a voltage-gated sodium channel blocker, abolished the responses. These results suggest that the activity recorded in response to linear acceleration stimuli is of vestibular origin.

We have investigated developmental alterations in the tonotopic projection of the gerbil lateral superior olive. Single neurons were characterized in the frequency domain and the recording site marked with fast green. Transverse tissue sections from the auditory brainstem of each animal were visualized with a video-equipped microscope, and the image was digitized for subsequent alignment. The three-dimensional display indicated little variation in the rostrocaudal axis, allowing us to collapse the data into a two-dimensional tonotopic map. The tonotopic map was found to change with age such that the characteristic frequency of neurons in a given anatomical location became successively higher during development. These results are consistent with the hypothesis that the place code gradually shifts in the developing cochlea.

The influence of monaural acoustic deprivation on the size of dendrites in n. laminaris in the chick was examined. Chicks were raised in a controlled acoustic environment with one ear occluded from 2 days prior to hatching until 25 days after hatching by an earplug which provided a conductive hearing loss of approximately 40 dB across the audible frequency range. Each n. laminaris cell receives spatially segregated binaural excitatory innervation; one dendritic field received input from the plugged ear while the other received input from the normal ear. This arrangement allowed comparison of the size (length) of the "deprived" dendrites and the "nondeprived" dendrites for each cell. The tonotopic organization of n. laminaris allowed these comparisons to be made as a function of the frequency organization of the nucleus. We observed systematic changes in dendritic size which differed as a function of the tonotopic position of the neurons. In high-frequency regions the dendrites receiving information from the deprived ear were shorter than those receiving input from the normal ear. Unexpectedly, cells responsive to low frequencies showed the opposite result; the dendrites innervated from the deprived ear were longer than those responsive to the nondeprived ear. These results suggests that a relatively flat conductive hearing loss may cause nonuniform changes in activity impinging on high- and low-frequency areas of the auditory system.

The effect of unilateral basilar papilla removal on glucose uptake in the 2nd and 3rd order auditory nuclei in the chick's brain stem, nucleus magnocellularis and nucleus laminaris, respectively, was examined with [3H]2-deoxy-D-glucose (2-DG) autoradiography. The tissue was processed according to a thaw-mount technique, and the number of grains in the resulting autoradiographs was counted to assess changes in glucose uptake. It was observed that there is a greater density of grains over the neuropil regions of nucleus laminaris which receive input from the normal ear than over the corresponding regions which receive input from the operated ear. Similarly, differences in grain density are found between the normally innervated and deafferented magnocellular nuclei although these differences are not as great as those in nucleus laminaris. Differences in grain density were also apparent between the glial/fiber regions which bound the neuropil areas of nucleus laminaris; there is a greater density of grains overlying those glial/fiber regions through which fibers receiving input from the normal ear course than over those regions through which fibers which normally carry input from the operated ear travel. It is likely that this difference mainly reflects glucose uptake in the fibers although a possible contribution of glial tissue cannot be excluded. All these effects of basilar papilla removal are seen with survival times as short as 70 min and thus likely reflect the reduction of neural activity rather than the degeneration of pre- or postsynaptic elements. Finally, the same pattern of results as described above was found when using the more common [14C]2-DG procedure or when using [3H]2-DG but processing the tissue using the freeze-dried technique. The present results thus show the neuropil regions of nucleus laminaris and the adjacent glial/fiber areas to be areas of high glucose utilization. Unilateral basilar papilla removal results in the removal of an excitatory input to these regions, and this results in a reduction of glucose utilization that is specific to those neuropil regions and glial/fiber areas that receive input from the operated ear. These findings are contrasted with another study in which removal of a major excitatory input to the dentate gyrus of the rat results in a reduced glucose utilization which is not specific to the deafferented region and which largely reflects post- rather than presynatic events.

The pattern of primary auditory projections to the brain stem of young chickens was investigated using terminal degeneration methods and orthograde transport of horseradish peroxidase (HRP) or tritiated amino acid. Of particular interest was the question of whether nucleus laminaris (NL) receives primary afferents. A study of silver-stained degeneration pattersn in nucleus magnocellularis (NM) and NL at three intervals following unilateral interruption of the cochlear nerve revealed that by 48 hours after the lesion, degenerating terminals were found only in the ipsilateral nucleus angularis (NA), NM and lagenar projection areas but not in NL. Five- and eight-day survival times, however, also revealed degeneration bilaterally in NL. The appearance of terminal degeneration in NL at the longer survival times is attributed to the previously-reported severe and rapid transneuronal degeneration of neurons in NM following deafferentation and not to the presence of cochlear nerve terminals in NL. Injection of HRP or tritiated proline into the basilar papilla produced patterns of labeling similar to that seen in the 2-day degeneration material; HRP reaction product or autoradiographic label were seen only in the ipsilateral NA and NM and in the ipsilateral projection areas of the macula lagena but not in either NL. The patterns of primary auditory projections revealed by the three methods were quite similar to each other and to that previously reported for the pigeon and confirm the conslucion that the laminar nucleus of chickens does not receive primary afferents.

Five groups of 10-day-old chicks were continuously exposed to either 500 or 1500 Hz pure tone at 125 dB for 4 or 12 h and killed 10 days later. The basilar papillae were fixed, embedded in plastic, sectioned, and hair cells were counted according to type: tall or short. Short hair cells were found to be more susceptible to acoustic overstimulation than tall hair cells. Further, the position of maximum short hair cell loss varied along the length of the basilar papilla as a function of the exposure frequency while the position of tall hair cell loss did not. Similarities between morphological response of short hair cells in avians after acoustic trauma and outer hair cells in mammals are discussed.

Large spherical cells of the mammalian anteroventral cochlear nucleus (AVCN) receive direct excitatory input from auditory nerve axons. Trans-synaptic regulation of neuronal cell size and cell number after cochlear ablation has been previously demonstrated in neonates of several vertebrate species, including the gerbil. Such changes may be related to loss of spontaneous or evoked auditory nerve electrical activity or to loss of activity-independent factors. We have developed a method to chronically, yet reversibly, block auditory nerve electrical activity without violating the integrity of the inner ear. Tetrodotoxin (TTX) was embedded in an ethylene-vinyl acetate copolymer resin (Elvax). A small piece of Elvax containing TTX was placed next to the round window membrane, which allowed TTX to diffuse into the inner ear. As a measure of the effectiveness of manipulation, the onset, duration, and magnitude of the auditory threshold shift were measured by the auditory brainstem response. The sound-evoked response was abolished within 10 minutes of placement of TTX on the round window membrane. The duration of threshold shift was dose-dependent and lasted 24-46 hours. Implants of Elvax without TTX did not produce a significant threshold shift. TTX, which blocks voltage-gated sodium channels, did not abolish the potassium-based cochlear microphonic response. The consequence of blocking afferent electrical activity on gerbil AVCN large spherical cells was examined by measuring their cross-sectional area after each of four manipulations: unilateral auditory nerve action potential blockade with TTX; unilateral surgical cochlear ablation; ipsilateral TTX exposure/contralateral cochlear ablation; and unilateral sham operation (Elvax without TTX).(ABSTRACT TRUNCATED AT 250 WORDS)

Nucleus magnocellularis is the avian homologue of the spherical cell region of the mammalian anteroventral cochlear nucleus. Its primary excitatory synaptic input is from large end bulbs of Held from the eighth nerve ganglion cells. We have examined the effects of three peripheral manipulations--middle ear ossicle (columella) removal (monaural and binaural), columella removal and oval window puncture (monaural), and monaural earplug--on cross-sectional cell area ("cell size") of second-order auditory neurons in n. magnocellularis of the chicken. Manipulations were performed between embryonic day 19 and posthatch day 4. Survival time was varied from 2 to 60 days. Air conduction and bone conduction thresholds were determined to assess for conductive and sensorineural hearing loss associated with each of these manipulations. Hair cell counts were made from basilar papillae of each experimental group. We found that a columella removal alone, which produced a 50-55-dB purely conductive hearing loss, was not associated with changes in cell size of n. magnocellularis neurons. Similarly, chronic monaural earplugging did not affect the cross-sectional area of these second-order auditory neurons. Conversely, a combined columella removal and oval window puncture, which produced a mixed hearing loss with a 15-40-dB sensorineural component was associated with an 18-20% reduction in n. magnocellularis cell area. Hair cell counts for experimental ears were not significantly different from control ears. These results, in conjunction with measurements of multiunit activity recorded in n. magnocellularis, suggest that manipulations which markedly attenuate extrinsic auditory stimulation, but do not result in chronic change in the average activity levels, also do not influence the size of n. magnocellularis cell bodies. On the other hand, a manipulation which influences overall activity levels, but does not result in degeneration of receptor cells, resulted in marked changes in n. magnocellularis cell size.

Ten-day-old chicks were exposed to a pure tone (1.5 kHz) or white noise at 125 dB SPL avg. RMS for 4 to 24 hours, and were sacrificed either 10, 30 or 60 days after exposure. The basilar papillae were embedded in plastic, sectioned, and hair cells were counted at 100-micrometer intervals throughout the length of the papilla. As sound exposure duration increased, both the maximum number of hair cells lost, and the extent of the damaged area along the basilar membrane increased. Short hair cells situated on the free area of the basilar papilla were more susceptible to damage than were tall hair cells. The location of hair cell loss varied as a function of frequency band of exposure; the pure tone produced a well localized basal lesion, while wide-band noise produced a more general lesion which extending toward the apex. Degeneration continued with increased survival time up to 30 days. It is concluded that avians respond to acoustic overstimulation in a manner very similar to mammals. The convenience of this preparation along with the relative simplicity of its cochlea may render it useful for future investigations of the mechanisms of acoustic trauma.

The length of the cochlear duct was measured in chicks aged embryonic day 14 to post-hatch day 469. Chicks were anesthetized, decapitated and their cochlear ducts exposed under an operating microscope. Because of the very thin bone and cartilage surrounding the relatively straight tube of the papilla the entire cochlear duct could rapidly be exposed and measured without fixation or removal from the head. The length of the duct was measured using a computer based Zeiss Videoplan Image Analysis System. A total 41% increase in length was seen from embryonic day 14 to post-hatch day 469; 20% of this increase occurred after hatching. It is suggested that this increase in cochlear duct length could influence basilar membrane properties important to frequency coding mechanisms during development.

Three electrophysiological functions of the chick basilar papilla were studied during development by recording the compound actin potential (AP) at the round window. The auditory thresholds showed a continuous maturation between the fifteenth day of incubation (E15) and the first post-hatching day (P1), when they attained adult values. Responses matured first to low frequencies and later to high frequency stimuli. The input-output (intensity-amplitude) functions matured regularly and never demonstrated the classical two slopes seen in mammals. The tuning properties, studied by tone-on-tone masking of the AP, achieved mature values before the thresholds; the Q10s reached adult values at E17 for a 500-Hz probe tone and at E19 for a 1000-Hz probe tone. The fact that a low-to-high frequency development trend was found with the embryonic middle ear cleared of fluids further suggests that this property of auditory ontogeny may be a function of changes in the transduction properties of the cochlea.

Previous studies of the avian auditory system have revealed that removal of the peripheral receptor (the cochlea) leads to a transneuronal degeneration of auditory relay neurons in nucleus magnocellularis (NM) of the brain stem. An early manifestation of the degeneration which can be observed within 12 hours is a decrease of histochemical staining for RNA (Nissl staining); such a decrease could reflect an alteration in protein synthetic activity within the NM neurons. The present study evaluates this possibility by determining whether the cochlea removal led to an alteration incorporation of protein precursors in the target neurons which exhibit transneuronal degeneration and if so, how early the changes appeared. The cochlea was removed unilaterally in seventeen 10-day-old chicks and two 66-week-old mature chickens, and incorporation of protein precursors was evaluated in the neurons of NM at 0.5, 1.5, 3, 6, 12, and 24 hours following the cochlea removal. Each chick received an intravenous injection of 3H leucine, and was allowed to survive for 30 minutes after the injection of precursor. The brains were then prepared for autoradiography. The extent of incorporation by neurons in NM was determined by counting grains overlying each cell body and determining grain density/micrometers2 of neuron cross-sectional area. We found that auditory relay neurons whose synaptic inputs have been silenced exhibit dramatic decreases in protein synthesis within 30 minutes after removal of the cochlea; leucine incorporation was reduced by about 50%. In chicks sacrificed 3 to 24 hours after removal of the cochlea, some neurons (about 1/3) were entirely unlabeled despite heavy labeling of their neighbors and heavy labeling of all NM neurons on the opposite side of the brain. The remaining neurons exhibited about a 15% reduction in incorporation in comparison with the cells in the contralateral (control) NM. While the decreases in incorporation were apparent at all survival intervals, there was no consistent decrease in Nissl staining until 6 hours after cochlea removal. There were no changes in protein precursor incorporation following removal of the cochlea in adult birds, a result which is in keeping with the relative absence of transneuronal degeneration following removal of the cochlea at maturity. The results suggest a very rapid transneuronal regulation of protein metabolism within target neurons in young animals, perhaps by activity-related events.

Afferent regulation of neurons in the cochlear nucleus as a function of age was investigated at the light microscope level. Unilateral cochlea removal was performed on Mongolian gerbils of three age groups: 1, 8, and 20 weeks postnatal. Animals survived for either 2 days or 2 weeks. An additional group of neonatally operated animals had a prolonged survival of 9 weeks. The number of neurons in anteroventral cochlear nucleus (AVCN) was counted, and cross-sectional area measurements of large spherical cells in AVCN were made. In animals 1 week old at the time of surgery, there was a 35% reduction in neuron number in AVCN after 2 days, a 58% reduction after 2 weeks, and a 59% reduction 9 weeks after inner ear destruction. However, in animals 20 weeks old at the time of surgery, there was no cell loss in AVCN either 2 days or 2 weeks after surgery. Animals in each age group showed a reduction in cross-sectional area of large spherical cells in AVCN after cochlea ablation. The gerbils that underwent cochlea removal at 8 and 20 weeks showed an average decrease of 14-18%. This effect was seen as early as 2 days after cochlea removal. Animals that underwent cochlea removal at 1 week exhibited the greatest change; a 25% decrease at 2 days progressed to 38% at 2 weeks following cochlea removal. No appreciable further changes were seen at 9 weeks after neonatal cochlea removal. The results indicate greater susceptibility of 1-week-old gerbil cochlear nucleus neurons to peripheral loss than found in older animals.(ABSTRACT TRUNCATED AT 250 WORDS)

The distribution and time course of gamma-aminobutyric acid (GABA) immunoreactivity was investigated in the cranium of the chick embryo from 2 to 16 days of incubation (E2-16). A fraction of nerve fibers transiently stains GABA-positive in all cranial motor nerves and in the vestibular nerve. Cranial motor nerves stain GABA-positive from E4 to E11, including neuromuscular junctions at E8-11; labeled fibers are most frequent in the motor trigeminal root (E6-9.5). Substantial GABA staining is present from E4 to E10 in a subpopulation (1-2%) of vestibular ganglion cells. Their peripheral processes are labeled in the vestibular endorgan, predominantly in the posterior crista. Some GABA-positive fibers are present in the olfactory nerve (after E5) and in the optic nerve (after E9.5); their immunoreactivity persists throughout the period investigated. Transient GABA immunoreactivity follows "pioneer" fiber outgrowth and coincides with the formation of early synaptic contacts. GABA-containing neurons may change their neuronal phenotype (loss of GABA expression) or they may be eliminated by embryological cell death. Periods of cell death were determined in cranial ganglia and motor nuclei by aggregations of pycnotic cells in the same embryonic material. The periods of embryonic cell death partly coincide with transient GABA immunoreactivity. The function(s) of transient GABA expression is unknown. Some lines of evidence suggest that GABA has neurotrophic functions in developing cranial nerves or their target tissue. In the developing neuromuscular junction, GABA may be involved in the regulation of acetylcholine receptors.

Nucleus laminaris (NL) is a third-order auditory nucleus in the avian brain stem which receives spatially-segregated binaural inputs from the second-order magnocellular nuclei. The organization of dendritic structure in NL was examined in Golgi-impregnated brains from hatchling chickens. Quantitative analyses of dendritic size and number were made from camera lucida drawings of 135 neurons sampled from throughout the nucleus. The most significant results of this study may be summarized as follows: (1) The preponderant neuron in n. laminaris may be characterized as having a cylindrical-to-ovoid cell body, about 20 micrometer in diameter. The neurons comprising NL were found to be nearly completely homogeneous in issuing their dendrites in a bipolar fashion: one group of dendrites is clustered on the dorsal surface of the cells, the other group on the ventral. The dendrites of NL are contained within the glia-free neuropil surrounding the nucleus. From the rostromedial to the caudolateral poles of NL there is a gradient of increasing extension of the dendrites, increasing number of tertiary and higher-order dendrites, and increasing distance from the somata of the occurrence of branching. (2) The total dendritic size (sum of the dorsal) and ventral dendritic lengths of the cells) increases 3-fold from the rostromedial to the caudolateral poles of NL. About 50% of the variance in dendritic size is accounted for by the position of the cells in NL, and the gradient of dendritic size increase has the same orientation across NL as the tonotopic gradient of decreasing characteristic frequency in NL. (3) From the rostromedial pole to the caudolateral pole of NL there is an 11-fold decrease in the number of primary dendrites along a gradient coinciding with the length and frequency gradients. Sixty-six percent of the variance in dendrite number is accounted for by position in the nucleus. (4) The correlation of dorsal and ventral dendritic size on a cell-by-cell basis is not high (r = 0.47), indicating a fair amount of variability on the single-cell level. On the other hand, the average dorsal dendritic length within an isofrequency band in NL correlates very highly with the average ventral dendritic length. Thus, on an areal basis, the amount of dendritic surface area offered to the dorsal and ventral afferents is tightly regulated. (5) The dorsal and ventral dendrites have separate gradients of increasing length and number across NL. The dorsal gradients are skewed toward the rostrocaudal axis, while the ventral dendritic gradients are skewed mediolaterally. (6) There was no correlation between either dendritic size or number of primary dendrites and the size of the somata in NL, which remains relatively constant throughout the nucleus. Several hypotheses about the ontogenetic control of dendritic structure are examined in light of the above data. Of these, the hypotheses that the ontogeny of dendritic size and number is largely under afferent control receives a great deal of circumstantial support.

Two experiments using embryonic and hatchling chickens examined how the representation of frequency along the basilar membrane changed during hearing development. In experiment 1, chicks were exposed to high intensity pure tones (500, 1,500, or 3,000 Hz) at one of three different ages. Analysis of hair cell degeneration indicated a discrete region of damage which systematically changed as a function of exposure frequency and age. With maturation, each frequency produced damage at progressively more apical locations. In experiment 2, the representation of frequency in the brain stem auditory nuclei was compared in embryonic, hatchling, and adult chickens. Microelectrode recordings indicated a systematic shift in the frequency representation. Neurons, which are activated by high frequencies in the adult, initially respond to only low frequencies. These experiments indicate how the mature pattern of frequency representation along the basilar membrane gradually emerges during the stages of hearing development.

A simple circuit to detect avian vocalizations is described. Adjustments of five different controls (frequency, bandwidth, amplitude, duration and spacing) allow the circuit to accurately detect the vocalizations of different ages and species of birds. Analyses of over 4000 peeps and 500 inter-peep intervals from 40 chicks and 16 ducklings showed that the circuit and an experienced observer agreed closely in the timing and counting of vocalizations.

Developmental changes in an unconditioned response to acoustic stimulation were observed in young chickens. Specifically, durations of distress call (peep) suppression were measured after the onsets of tones that differed in intensity and frequency in 384 newly hatched and 4-day-old chicks. Resuppression was also measured after a 6% change in the frequency of these tones, once the animals had habituated to the original tone. The data showed that the suppression varied systematically as a function of age, intensity, and frequency: (a) the duration of suppression increased with increasing stimulus intensity, as expected; (b) responsiveness to high frequencies grew more rapidly over the first 4 days than responsiveness to low frequencies, an effect indicating a developmental gradient across frequencies with age; (c) resuppression to the 6% change in frequency increased in duration with age; and (d) young birds suppressed vocalizations longer to loud tones in the range of their species' maternal assembly call than to other frequency-intensity combinations. These developmental trends indicate rapid changes in "perceived loudness" and "perceptual sharpening" over the first few days of postnatal life.

The immotile cilia syndrome appears to be a congenital defect in the ultrastructure of cilia that renders them incapable of movement. Respiratory tract cilia and sperm are predominantly affected. Bronchiectasis, sinusitis and male sterility are the main clinical findings. Situs inversus may be found. To these findings can be added otitis media. The defect appears to be a complete or partial absence of dynein arms which are believed to be essential for generating movement of cilia or sperm tails. Six patients suspected of having immotile cilia were compared to six patients in a control group. In affected patients, no cilia movement in the middle ear or nasopharynx was observed using the operating microscope. Electron microscopy of cilia from the mucosa of the middle ear and nasopharynx appeared to confirm the ultrastructural defect in two of six patients suspected of having the syndrome.

The consequences of cochlea removal on neuron number and soma cross-sectional area were examined in the second order auditory nucleus (n. magnocellularis) of chickens. Both the age of the subjects at the time of cochlea (basilar papilla) removal (1-66 weeks) and the survival period (1-45 days) were varied. Neuron number and soma cross-sectional area were determined from Nissl stained sections. Additional material was processed to examine the relationship of ganglion cell loss to changes in n. magnocellularis. Neuron number decreased by 25-30% and soma cross-sectional area decreased by 10-20% ipsilateral to the cochlea removal in chickens operated on during the first 6 weeks after hatching. In contrast, in chickens operated on at 66 weeks posthatch neuron number decreased less than 10% and there was no change in soma area. The changes were rapid, being nearly complete 2 days after cochlea removal. An initial change (1 and 2 days after surgery) observed in animals operated on up to 6 weeks posthatch was the presence of a large number of neurons in which no Nissl substance could be detected. These results demonstrate an age-dependent change in the susceptibility of NM neurons to deafferentation. This change is not temporally related to other measures of functional maturation of the auditory system.

4- to 8-day-old chicks were used to examine the ultrastructure of end bulbs and neurons of nucleus magnocellularis by freeze-fracture and thin section techniques. Both normal animals and animals deafferented by removal of the basilar papilla were used to describe the pre- and postsynaptic membranes of these neurons. Postsynaptic membrane specializations similar to those described in rodents were found in normal and deafferented chicks. In the presynaptic end bulb there was an immediate cessation of endocytotic vesicle formation after deafferentation. There was also a shift in the preferential fracture plane from the presynaptic to postsynaptic membranes 48-72 h after basilar papilla removal. These results indicate that the ultrastructure of this synaptic complex is comparable to that seen in mammals.

N. laminaris dendrites begin to atrophy almost immediately after they are deafferented. Accompanying this rapid change in shape is a loss of microtubules and neurofilaments at the base of the dendrite, and a decrease in the density of the dendritic cytoplasm. However, degenerative changes in the dendritic plasma membrane were not evident until 2 days after deafferentation. Thus it was unknown what happened to the volume and membrane lost from the atrophying dendrites before this time. The soma was investigated in this study as a possible recipient of the volume of the atrophying dendrite. Soma size increased significantly by 2 hours after deafferentation and continued to increase for 1-8 days after deafferentation. The nucleus, which is normally concentric with the soma, moved continuously to the dorsal pole of the soma, toward the innervated side of the cell. The cytoplasm on the ventral side of the soma showed a decrease in density and loss of cytoskeleton similar to what was found in the initial portion of the ventral primary dendrites in the accompanying paper. These changes are interpreted as indicative of a rapid resorption of the ventral dendrite back into the soma following deafferentation.

The time course and specificity of the changes in dendritic morphology following deafferentation were examined in nucleus laminaris of young chickens. The dendrites of nucleus laminaris neurons are segregated into dorsal and ventral domains, which are innervated separately from the ipsilateral and contralateral nucleus magnocellularis, respectively. Transection of the crossed dorsal cochlear tract deafferents the ventral dendrites of nucleus laminaris bilaterally without interrupting the matching input to the dorsal dendrites. In 10-day-old chicks, atrophy of the ventral dendrites began immediately after transecting the tract; the ventral dendrites were 10% shorter by 1 hour and 16% shorter by 2 hours after deafferentation. The length of the ventral dendrites progressively decreased over the next 2 weeks, resulting in at least a 60% loss of ventral dendrite 16 days after surgery. The dorsal dendrites of the same cells, whose afferents remained intact, did not change in length during the time course of this study. However, 16 days after the lesion, spines appeared on the normally smooth dorsal and ventral dendrites. The time course of dendritic atrophy and its restriction to the deafferented postsynaptic surface are related to possible mechanisms by which afferents regulate and maintain their target neurons.

We have examined one of the metabolic consequences of unilateral cochlea (basilar papilla) removal in the chick brain stem auditory system. We assessed changes in succinate dehydrogenase (SDH), a mitochondrial enzyme involved in energy metabolism, in neurons of second-order n. magnocellularis (NM) and third-order n. laminaris (NL). Chickens undergoing surgery at 10 days of age were perfused 4 hours to 35 days postlesion. Chickens 6 or 66 weeks of age at cochlea removal were examined 1 or 8 days after surgery. In all groups, cryostat sections were prepared for SDH histochemistry or Nissl staining. In normal chickens, NM cell bodies and NL neuropil contain SDH reaction product. In young birds, the density of SDH reaction product in NM shows a rapid biphasic response to cochlea removal. From 8 to 60 hours postlesion, density increases ipsilateral to cochlea removal; for survival times of 3-35 days, SDH density decreases in ipsilateral NM. In NL, no changes were observed until 3 days after cochlea removal. Then we observed a long-lasting decrease in density of SDH reaction product in the neuropil regions receiving input from the deafferented NM. All of these changes are age-dependent in that they were observed only following cochlea removal on or before 6 weeks of age.

The role of auditory experience in the ontogeny of perceptual coding was investigated in hatching chickens. In Experiment 1, auditory frequency generalization gradients were determined by using a habituation-generalization paradigm and an orienting response. One-day and 3-4 day chicks were habituated to 800-Hz tones and then tested at five frequencies ranging from 800 to 1,000 Hz. One-day chicks displayed reliably flatter generalization gradients than 3-4-day chicks. In Experiment 2 an auditory deprivation method is described in which an ear impression compound was injected into the external ear. This method (a) is fully calibrated and provides approximately 40 dB of attenuation across a .125- to 4.0-kHz range, (b) appears to be fully reversible, and (c) is applicable prior to normal auditory experience as well as at subsequent stages. In Experiment 3, this method was used to investigate the effects of auditory deprivation on the "perceptual sharpening" described in Experimental 1. Chicks were deprived from Embryonic Day 18 1/2 to 3-4 days posthatch, at which time normal, deprived, and sham-operated chicks were tested as in Experiment 1. The generalization gradients of the deprived chicks were significantly flatter than those of the 3-4 day normal and sham-operated chicks and were similar to those of 1-day chicks. This effect cannot be attributed to (a) operative procedures, (b) changes in auditory thresholds, or (c) changes in response rate with age or condition. Thus the perceptual sharpening normally occurring between 1 and 3-4 days posthatch fails to occur during that time in the absence of a normal acoustic environment.

Recovery of hair cells was studied at various times after acoustic trauma in adult quail. An initial loss of hair cells recovered to within 5 percent of the original number of cells. Tritium-labeled thymidine was injected after this acoustic trauma to determine if mitosis played a role in recovery of hair cells. Within 10 days of acoustic trauma, incorporation of [3H]thymidine was seen over the nuclei of hair cells and supporting cells in the region of initial hair cell loss. Thus, hair cell regeneration can occur after embryonic terminal mitosis.

Deafness mutant mice show no stimulus-related cochlear potentials as well as abnormal electrically-evoked responses recorded from the inferior colliculus. Abnormal spontaneous activity in the auditory periphery could result in abnormal development and/or maintenance of the central auditory pathways. We therefore assessed spontaneous activity of cochlear origin in the central nuclei of the mutants by ablating one cochlea and subsequently using the 2-deoxyglucose (2DG) technique to study metabolic activity. Any asymmetries in labeling in a given nucleus should be due to spontaneous activity in the cochlear nerve on the unoperated side. In control animals (+/dn mice undergoing unilateral cochlea ablation), statistically significant decreased 2DG labeling was observed in the ipsilateral PVCN and AVCN, and contralateral MNTB and IC; all receive primary excitatory input from the ablated ear. No significant differences in labeling between right and left sides were observed in any of the nuclei studied in the mutant animals. These findings suggest that there is no spontaneous activity of cochlear origin in these mutants, even though many cochlear nerve fibers and spiral ganglion cells survive.

The development of gamma-aminobutyric acid-immunoreactivity (GABA-I) in nucleus magnocellularis (NM) and nucleus laminaris (NL) of the chick was studied by using an antiserum to GABA. In posthatch chicks, GABA-I is localized to small, round punctate structures in the neuropil and surrounding nerve cell bodies. Electron microscopic immunocytochemistry demonstrates that these puncta make synaptic contact with neuronal cell bodies in NM; thus, they are believed to be axon terminals. GABAergic terminals are distributed in a gradient of increasing density from the rostromedial to the caudolateral regions of NM. The distribution of GABA-I was studied during embryonic development. At embryonic days (E) 9-11, there is little GABA-I staining in either NM or NL. Around E12-14, a few fibers are immunopositive but no gradient is seen. More GABA-I structures are present at E14-15. They are reminiscent of axons with varicosities along their length, preterminal axonal thickenings and fiber plexuses. At E15, terminals become apparent circumscribing neuronal somata and are also discernible in the neuropil of both nuclei. In E16-17 embryos, terminals are the predominantly labeled GABA-I structures and they are uniformly distributed throughout NM. The density of GABAergic terminals increases in caudolateral regions of NM such that by E17-19, there is a gradient of increasing density of GABA-I terminals from the rostromedial to caudolateral regions of NM. The steepness of this gradient increases during development and is the greatest in posthatch (P) chicks. Cell bodies labeled with the GABA antiserum are located around the borders of both NM and NL and in the neuropil between these two nuclei. Occasionally, GABA-I neurons can be found within these auditory brainstem nuclei in both embryonic and posthatch chicks. Nucleus angularis (NA) contains some GABAergic cells. The appearance of GABA-I terminals around E15 is correlated in time with the formation of end-bulbs of Held on NM neurons. Thus, the ontogeny of presumed inhibitory inputs to chick auditory brainstem nuclei temporally correlates with, and could modulate the development of, excitatory auditory afferent structure and function.

The possible synergism between noise and aspirin for causing cochlear damage was examined histologically. Six chicks fed aspirin for five days and five chicks fed a normal diet only were paired and placed in sound chambers. They were exposed to a 1500-Hz tone at 115 dB sound pressure level for eight hours. The mean serum salicylate level just before noise exposure was 24 mg/dL (1.74 mmol/L). Ten days later they were killed, and the temporal bones were processed. Hair cell counts were made at 100-microns intervals throughout the length of the basilar papilla (cochlea). The noise produced a discrete cochlear lesion centered about 30% of the distance from the base to apex. The addition of aspirin did not significantly alter the extent or location of this lesion. One aspirin-fed chick had a unilateral middle ear effusion, and a striking shift in the center of damage toward the apex was noted in this cochlea.

The technique of measuring soma cross-sectional area at the plane of the nucleolus leads to systematic errors that depend on how far the nucleolus is displaced from the center of the soma. A set of correction factors was produced based on calculations from a geometric model of the measurement process. Applying the correction factors to measurements of second-order auditory neurons led to substantial changes in the estimated soma area.

Early acoustic experience affects the structure of neurons in the brainstem of young chickens. In the binaurally innervated cells of the nucleus laminaris the symmetry of the dorsal and ventral dendritic trees normally increases during the embryonic and early postnatal period. Unilateral ear plugs disrupt that development. This study shows that balanced stimulation plays an important role in the development of symmetrical neuronal structures in the central auditory pathway.

The second- and third-order auditory nuclei in the brainstem of the chicken, nucleus magnocellularis (NM) and nucleus laminaris (NL), receive afferents that are immunoreactive to gamma-aminobutyric acid (GABA). In order to investigate the source(s) of these GABAergic afferents, we examined the distribution, morphology, and connectivity of GABAergic neurons in and adjacent to NM and NL in chicks from 7 days of incubation to 12 days posthatch. Immunocytochemical techniques revealed the presence of approximately 150 GABA-labeled neurons within the neuropil surrounding NM and NL on each side of the brainstem. Most of these neurons are located between NM and NL and along the lateral border of NM. GABAergic neurons are multipolar; their thick dendritic processes branch extensively and give rise to several thin, secondary processes. Frequently, the GABA-labeled processes arborize within NM or NL. The morphology of these non-NM/NL neurons was investigated further with Golgi impregnation and specific neuronal markers (antisera to microtubule-associated protein). Our observations suggest that a considerable portion of GABAergic input to NM and NL originates from local GABAergic neurons. In order to determine other possible sources of GABAergic input to NM and NL, we injected tracers unilaterally into NM/NL. A small number (20-30) of neurons were retrogradely labeled in the trapezoid body, almost exclusively ipsilaterally. No labeled cells were found in other regions of the brainstem, except for the contralateral NM. Unilateral injections of horseradish-peroxidase-labeled wheat germ agglutinin into the paraflocculus revealed only minor terminal labeling in the lateral region of NL bilaterally. The number and distribution of GABAergic terminals in NM and NL appeared normal after transection of the crossed dorsal cochlear tract.

Developmental changes in the site of receptor damage following pure-tone acoustic overstimulation were examined in the basilar papillae of embryonic and hatchling chickens. During development, a systematic shift in the position of damage toward the apex of the cochlea was produced by each of three frequencies, suggesting that the transduction properties of the sensory epithelium systematically shift with age. These results imply that neurons in the central nervous system may be maximally stimulated by different sounds during development.

The morphological development of the cochlea begins in the base or midbasal region and spreads toward the apex. In adults, the base responds maximally to high-frequency sounds and lower frequencies are represented progressively toward the apex. This predicts that responses to sound should occur initially to high frequencies and gradually change to include lower frequencies. Paradoxically, animals respond first to relatively low frequencies and last to high frequencies. We have previously proposed that this discrepancy results from an ontogenetic change in spatial coding of frequency along the cochlea (Rubel et al., '76). According to this model, only the basal end of the cochlea transduces sound early in development but it responds to low frequencies. During maturation the representation of low and midrange frequencies shifts apically and the base becomes responsive to high frequencies. This hypothesis predicts that the tonotopic organization within the central nervous system should change during development; neurons at any given location within an auditory nucleus should become maximally responsive to successively higher frequency sounds during development. In the present study this prediction was tested by using microelectrode recording procedures to map the tonotopic organization of nucleus magnocellullaris (NM) and nucleus laminaris (NL), first- and second-order auditory nuclei, in chickens at three ages: embryonic day 17, 1 day posthatch, and 2-4 weeks posthatch. The characteristic frequencies of neurons having the same anatomical location were quantitatively compared across ages. The tonotopic order in NM and NL was similar at all ages; responses to high-frequency sounds were recorded anteromedially and lower frequencies were located progressively more caudolaterally. However, there was a striking quantitative change in tonotopic organization. Neurons at a given location in both nuclei became maximally responsive to progressively higher frequencies during development. The characteristic frequencies of neurons in embryos and newly hatched chicks averaged, respectively, 1.00 (+/- 0.06, S.E.M.) and 0.34 (+/- 0.04) octaves lower than their predicted adult values. All regions in both nuclei showed a statistically significant increase in characteristic frequency during development except the most posterolateral (low-frequency) sector. Too few neurons were recorded from this region to be able to reliably estimate characteristic frequency. These results support the hypothesis that the spatial coding of frequency along the cochlea shifts during development.(ABSTRACT TRUNCATED AT 400 WORDS)

Ten-day-old chicks were continuously exposed to pure tones (500, 1 500, or 3 000 Hz) at 125 dB SPL for 12 hours and sacrificed 10 days after exposure. The basilar papillae were embedded in plastic, sectioned and hair cells were counted at 100-mum intervals throughout the length of the papilla. The position of hair cell loss along the basilar membrane varied systematically as a log-linear function with the frequency of stimulation. This systematic relationship was used to predict the frequency organization of the chick basilar papilla. It is concluded that although the avian basilar papilla differs greatly in morphological detail from the mammalian cochlea, its response to intense acoustic stimulation is quite similar.

This study describes qualitative and quantitative changes in dendritic ultrastructure during the rapid atrophy of nucleus laminaris (NL) dendrites following deafferentation. The dendrites of n. laminaris neurons in the chick auditory system are segregated into dorsal and ventral dendritic tufts, which receive spatially separated innervation from the ipsilateral and contralateral nucleus magnocellularis, respectively. We have previously shown that removing the input to the ventral side of NL results in the rapid atrophy of the ventral dendrites, whereas the nondeafferented dorsal dendrites of the same cells do not change in length. The ultrastructure of NL was examined in normal animals and after deafferentation. Changes in dendritic ultrastructure were not qualitatively apparent 4 hours after deafferentation. Between 12 and 48 hours the cytoplasm of the ventral dendrites became progressively more lucent, and a gap formed in the transition between the soma and ventral dendritic cytoplasm. Many of the dendrite tips, however, appeared normal even 2 days after deafferentation. Degeneration of dendrite plasma membrane was not visible until 2 days after deafferentation. On the other hand, quantitative measurements revealed a 30% decrease in microtubule density in the initial portion of the ventral dendrite by 4 hours, and a 50-60% decrease from 12 to 48 hours after deafferentation. Neurofilament density in the initial ventral dendrites decreased 50% by 12 hours, and 70% by 2 days after deafferentation. Many of the terminals of the severed afferents remained attached to the atrophying dendrite until 2 days after surgery, when they were in advanced stages of degeneration. Glia apparently were not involved in dendrite loss. The implications of these results on the role of cytoskeleton in the production and maintenance of dendritic shape are discussed.

To determine if there is an ontogenetic change in stimulus coding, chickens between the day of hatching and 9-10 days old were tested using a habituation-generalization paradigm. Experiment 1 indicated that 1 day-old and 3--4-day-old chicks show similar habituation of an eye-opening response to auditory stimuli in the 800--1,200-Hz range. In Experiment 2 the eye-opening response to a 1,000-Hz stimulus was habituated and then immediately tested using stimuli which varied between 800 and 1,200 Hz. Each age-group (1 day, 3--4 days, and 9--10 days) showed a symmetrical stimulus generalization gradient around the 1,000-Hz stimulus and the 1-day-old chicks displayed a reliably flatter gradient than either of the older groups, which did not differ. In a third experiment, the position of the gradients relative to the baseline was shifted without altering the relative shapes. These results allow general arousal, general auditory responsiveness, overall error rate, and metric characteristics of the independent and dependent variables to be eliminated as possible sources of the age differences in gradient shape. The changes in stimulus generalization, therefore, support the view that during normal development there is a sharpening of perceptual coding processes.

The effects of a conductive or mixed (conductive and sensorineural) hearing loss on anatomical and physiological properties of the chicken auditory system were examined. Animals used in the anatomical studies underwent either a columella (ossicle) removal, which produced a moderate conductive hearing loss, or an oval window puncture, which produced a severe mixed hearing loss, at 4 days posthatch. In a companion study, multiunit spike counts were obtained from 3-week-old chickens before, during, and after consecutive tympanic membrane puncture, columella removal, and oval window puncture. Tympanic membrane puncture and columella removal (conductive hearing loss) are not associated with either cell area changes in the nucleus magnocellularis or changes in spontaneous neuronal activity. Conversely, an oval window puncture (sensorineural damage) is associated with a cell area reduction of 20%, as well as a marked decline in activity within auditory nuclei.

Absolute auditory thresholds were estimated in chickens at 0 and 4 days after hatching. Momentary suppressions of the chicks' regular peeping, following the onset of a tone, were used as indications of stimulus detection. In the first experiment a staircase procedure was used to estimate thresholds. The absolute thresholds of both ages were the same at low frequencies (250-500 Hz), but at higher frequencies (1-2 kHz) 4-day-old chicks had lower thresholds than the 0-day-old chicks. The estimates of thresholds at 1 kHz were corroborated in the second experiment with a method of constant stimuli. A more efficient modified method of limits was used to replicate the age by frequency interaction in the third experiment. These changing thresholds are likely to reflect a developmental process somewhere in the auditory system and not some nonsensory artifact for two reasons: similar thresholds at low frequencies show that developmental differences are not due to differences in the sensitivity of the testing procedure at the two ages and thresholds obtained from the 4-day-old birds are similar to estimates from mature birds. In conclusion, responsiveness to low frequencies develops before responsiveness to higher frequencies, showing that the development of absolute thresholds is correlated with other measures of functional maturation in the auditory system.

The onset of responsiveness to eighth nerve stimulation was examined in n. magnocellularis and n. laminaris, (second- and third-order neurons) of the chick brainstem auditory system. Extracellular microelectrode mapping techniques were used to examine postsynaptic responses in in vitro brainstem preparations. Two specific questions were addressed. First, what is the earliest time at which postsynaptic action potentials can be evoked in n. magnocellularis and n. laminaris by eighth nerve stimulation? Second, does responsiveness to eighth nerve stimulation develop along a spatial gradient in n. magnocellularis and, if so, how does this gradient compare with other developmental events observed in the chick auditory system? Postsynaptic responses in n. magnocellularis were first recorded at 11 days of incubation. Nucleus laminaris responses to direct stimulation of n. magnocellularis were also first recorded at 11 days, although n. laminaris responses to eight nerve stimulation were not seen until 12 days of incubation. A gradient of response development within n. magnocellularis was indicated by mapping of responsive sites on days 11-13. At 11 days, responses to eighth nerve stimulation were restricted to the most anteromedial portion of n. magnocellularis. Between 11 and 13 days, cells in increasingly more posterolateral portions of n. magnocellularis became responsive. This anteromedial-to-posterolateral gradient in n. magnocellularis is correlated with the basal-to-apical gradient of morphogenesis observed in the basilar papilla and morphogenetic gradients previously observed in n. magnocellularis and n. laminaris.

Nucleus magnocellularis (NM) and nucleus laminaris (NL) are, respectively, second- and third-order auditory nuclei in the chicken brain stem. In this report the morphogenesis of these nuclei is examined. The times of origin of the cells of these nuclei were studied with 3H-thymidine autoradiography. The number of cells in each nucleus and their rostro-caudal distribution within each nucleus was determined in Nissl-stained sections at 9, 11, 13, 15, 17, 19, and 21 days of incubation. For the above ages the volumes of NM and NL were also calculated planimetrically and the rostro-caudal distribution of volume within each nucleus was studied.

The nucleus laminaris (NL), a third-order brain stem auditory nucleus in birds, receives afferents to its dorsal dendrites from the ipsilateral nucleus magnocellularis (NM), while the ventral dendrites of NL neurons are innervated by axons from the contralateral NM via the crossed dorsal cochlear tract (CTrX). The CTrx was transected in young chickens and, 96 h later, NL was examined for cytological changes. A morphometric analysis of electron micrographs from lesioned and shamd axon terminals were localized almost entirely to the ventral neuropil region of NL. (2) The volume density of dendrite in the ventral region of NL of lesioned animals was reduced by 85% compared to both the dorsal dendritic region of the same animals and the ventral dendritic region of sham-operated control animals. (3) The frequency with which primary dendrites were encountered in the ventral neuropil of lesioned animals was 81% lower than sham-operated controls. (4) Frequently, there was an apparent reduction in the amount of rough endoplasmic reticulum, Golgi apparatus and cytoplasmic granularity in the deafferented part of the NL neurons. (5) The data indicate that the cytological integrity of dendritic processes cna be specifically, profoundly and very rapidly compromised by removing a significant proportion of their afferents, suggesting that a tonic influence may be exerted by the presynaptic terminals.