Development of the olfactory epithelia of the African clawed frog, Xenopus laevis, was studied by scanning and transmission electron microscopy. Stages examined ranged from hatching through the end of metamorphosis. The larval olfactory organ consists of two chambers, the principal cavity and the vomeronasal organ (VNO). A third sensory chamber, the middle cavity, arises during metamorphosis. In larvae, the principal cavity is exposed to water-borne odorants, but after metamorphosis it is exposed to airborne odorants. The middle cavity and the VNO are always exposed to waterborne odorants. Electron microscopy reveals that in larvae, principal cavity receptor cells are of two types, ciliated and microvillar. Principal cavity supporting cells are also of two types, ciliated and secretory (with small, electron-lucent granules). After metamorphosis, the principal cavity contains only ciliated receptor cells and secretory supporting cells, and the cilia on the receptor cells are longer than in larvae. Supporting cell secretory granules are now large and electron-dense. In contrast, the middle cavity epithelium contains the same cell types seen in the larval principal cavity. The VNO has microvillar receptor cells and ciliated supporting cells throughout life. The cellular process by which the principal cavity epithelium changes during metamorphosis is not entirely clear. Morphological evidence from this study suggests that both microvillar and ciliated receptor cells die, to be replaced by newly generated cells. In addition, ciliated supporting cells also appear to die, whereas there is evidence that secretory supporting cells transdifferentiate into the adult type. In summary, significant developmental additions and neural plasticity are involved in remodeling the olfactory epithelium in Xenopus at metamorphosis.

The effects of intranasal zinc sulfate (ZnSO4) irrigation on the morphology of the olfactory epithelium and olfactory bulb were studied in mice with short survival times (as early as 1 day) and with long survival times (up to 593 days) after the irrigation procedure. As in several previous studies, the olfactory epithelium was completely destroyed within a few days after the ZnSO4 treatment. Within 2-4 days, the septum and turbinates were covered by a new, cuboidal epithelium, the cells of which differed significantly from any cells normally seen in the olfactory epithelium. Slowly, over several months, small areas of the olfactory epithelium regenerated in many of the animals. The ultrastructural changes occurring in the olfactory bulb from 1 to 25 days (the reactive stage) were characterized by degenerating olfactory axons and axon terminals, hypertrophy of astroglial cell processes, and proliferation of or extravasation by phagocytic cells. By 25 days after intranasal ZnSO4 irrigation, the number of reactive glial processes and phagocytic cells returned to normal. In some mice with survival times of 150 days or longer, there was reinnervation of small areas of the olfactory bulb by regenerated olfactory axons. These new olfactory axons innervated only superficial glomeruli or the outer portions of deeper glomeruli, but they formed synaptic contacts with mitral/tufted cells and periglomerular cells that did not differ from control animals. These findings were supported by tract-tracing experiments with 3H-amino acids and by behavioral analysis. In summary, the ultrastructural changes observed in the olfactory bulb in this study were not significantly different from those observed after surgical lesions of the olfactory epithelium or nerve.(ABSTRACT TRUNCATED AT 250 WORDS)

The relationship between olfactory axons and the cells of the olfactory bulb during normal development was analyzed to determine whether olfactory afferent axons could play a role in the induction of olfactory bulb formation. The morphology of the olfactory bulb in Xenopus larvae from stages 26 to 58 and in adult frogs was analyzed with light and electron microscopy. Axons were first observed beneath the basal lamina of the neural tube at stages 30 and 32; at stage 32, neurons in this area of the neural tube began to differentiate. Synapses of olfactory axons onto young neuronal processes were observed as early as stages 36 and 38. By stage 44, all of the layers of the olfactory bulb were present. The basic structure of the mature form of the olfactory bulb was apparent as early as stage 48/49 and remained constant throughout late larval life and even into adulthood, with only the size increasing. To determine the numerical relationship between olfactory axons from both main and vomeronasal epithelia and mitral/tufted cells in the main and accessory olfactory bulbs, a quantitative study was also performed in which the number of olfactory axons and the number of mitral/tufted cells were estimated for larval stages (stages 50 to 58) and adults. The number of axons increased with stage, with a 16-fold increase between stage 58 and adulthood. The number of mitral/tufted cells increased with stage, with only a 2.3-fold increase between stage 58 and adults. There is a correlation between axon number and mitral/tufted cell number during larval development that is consistent with the hypothesis that olfactory axons influence olfactory bulb development. The convergence ratio of olfactory axons onto mitral/tufted cells was 5:1 in larvae and increased to 34:1 in adults; this increase probably results in increased olfactory sensitivity in adult frogs.

The fine structure of synaptic terminals contacting neurons generated in the forebrain of adult male canaries was investigated by autoradiography and electron microscopy. The procedure for labeling the new neurons included pretreating adult canaries with 3H-thymidine and sacrificing them 23-45 days later. Neurons were identified as newly generated by the presence of 3H-thymidine in the cell nucleus. The new neurons in the nucleus hyperstriatum ventralis, pars caudalis (HVc) were identified by autoradiography and light microscopy and examined with electron microscopy. Several types of synaptic terminals contacted the cell body and proximal dendrites of the newly formed neurons. Synaptic junctions were formed by terminals that contained spherical, agranular vesicles, large dense-core vesicles and spherical, agranular vesicles, and pleomorphic or flattened synaptic vesicles. Terminals that contained spherical vesicles were most often associated with asymmetric synaptic densities, and terminals that contained pleomorphic or flattened vesicles formed symmetric junctions. New neurons were also contacted by small terminals that contained few vesicles and had little pre- or postsynaptic density associated with the junction; these terminals may be a special type or may be in the process of developing their synaptic contact with the new neuron. In addition, rare terminals that appeared to be degenerating or to contain debris from other degenerating neural elements contacted new neurons. In summary, these data indicate that the new neurons, which are known to be inserted into existing neural networks, receive synaptic input from at least three different sources.

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.