Cochlear pathology of long term neomycin induced deafness in cats.

The central axons of Type I spiral ganglion neurons travel in the auditory nerve and terminate in the cochlear nucleus. The ascending branches of these axons innervate the anteroventral cochlear nucleus and give rise to large axosomatic endings, called the endbulbs of Held, and smaller boutons. This paper reports a study of the endbulbs of Held, stained by horseradish peroxidase and variants of the Golgi method in kittens 2, 5, 10, 20, and 45 days postnatal and adult cats. Endbulbs tend to fall into two extreme groups with some endbulbs having an intermediate appearance; consequently, we have defined three descriptive stages of endbulbs that are conceived of as representing a developmental sequence. One group of endbulbs is found mostly in kittens younger than 10 days postnatal and is similar to the classic description of endbulbs by Ramon y Cajal ('09). The other extreme group of endbulbs is found mostly in adult cats. In these cases, the parent axonal trunk divides into several thick, gnarled branches that in turn branch again, sometimes repeatedly. These branches display irregular varicosities and form a cup-shaped arborization into which the postsynaptic cell body nestles. A chronology of postnatal endbulb development has been inferred from the relative proportions of the different endbulb stages at various ages. Maturation transforms the endbulb of Held from a large, spoon-shaped swelling having many filipodia into an elaborate tree with broad trunks and many smaller branches. Some implications of the proposed developmental sequence are discussed.

Morphology of primary axosomatic endings in the anteroventral cochlear nucleus of the cat: a study of the endbulbs of Held.

During the last few decades, the structural and functional development of the cochlea has been extensively reviewed (Pujol and Hilding 1973; Rubel 1978; Romand 1983; Eggermont and Bock 1985; Pujol and Uziel 1988; Lim and Rueda 1992). This chapter does not attempt to be a general review but more specifically describes the maturation of sensory hair cells and their neural connections during in vivo development of the mammalian cochlea. An important part of this coverage concerns the ontogeny of the different types of cochlear synapses. Up until now, reviews on cochlear synapto-genesis have been either incomplete (Pujol 1986; Pujol and Sans 1986) or based on in vitro results (Sobkowicz 1992) that do not give a clear understanding of what occurs in vivo. We complement structural findings wherever possible with physiological correlations to present morphology at a functional rather than at a descriptive level.

Development of sensory and neural structures in the mammalian cochlea

The axons of physiologically characterized spiral ganglion neurons (type I) were stained throughout their arborizations in the cochlear nucleus by the intracellular injection of horseradish peroxidase (HRP). The tips of the axonal branches were marked by distinct swellings, ranging in size and shape from small boutons to large perisomatic ramifications. Electron microscopic analysis of such swellings revealed ultrastructural features characteristic of primary auditory synapses, consistent with the hypothesis that terminal swellings identifiable in the light microscope represent presynaptic endings. On the basis of light microscopic differences in size, these endings were organized into three categories. Endings of relatively small size (terminal boutons, free endings, boutons with filopodia, string endings, and small complex endings) composed 94% of all terminal endings. Within this category of small endings, there were predictable variations in relative size and regional distribution that related to the spontaneous discharge rate (SR) of the fiber. The endings of low and medium SR fibers (SR 18 spikeshecond). Furthermore, there were more endings arising from the ascending branch than from the descending branch when comparing fibers of the low and medium SR group with those of the high SR group. There were not, however, obvious morphological features of this ending category that correlated with the characteristic frequency (CF, the pure tone frequency to which the neuron is most sensitive). A second category contained medium-sized complex endings, most of which formed axosomatic contacts. This category composed 4% of the population and was found in close proximity to the perikarya of globular, octopus, and spherical cells. The endings from low and medium SR fibers were smaller on average than those from high SR fibers. These endings did not vary in their parent branch distribution with respect to fiber SR, nor did they exhibit morphological features that correlated with fiber CF. The third category contained large complex endings (endbulbs of Held) and composed 2% of the ending population. Within the anteroventral cochlear nucleus, these large, complex endings made axosomatic contact with spherical cells in the anterior division and with globular cells in the posterior division. There were no systematic variations in ending size or branch distribution that correlated with fiber SR. There was, however, a relationship between ending size and fiber CF such that fibers having CFs below 4 kHz gave rise to the largest endbulbs.

The central prod ections of intracellularly labeled auditory nerve fibers in cats: An analysis of terminal morphology

Tritiatedproline or horseradish peroxidase (HRP) was injected into the middle ectosylvian auditory cortex (AI) of one hemisphere in kittens during the first few weeks postpartum. Survival time was 24–96 hours. The tetra-methylbenzidine (TMB) (Mesulam '78) method was used to process the HRP material. By the first postnatal day (PND 1) most neurons retrogradely filled in the contralateral auditory cortex form a densely packed, continuous band that presumably represents mainly layer III. This continuous pattern is in contrast to that seen in the adult cat where callosal neurons tend to aggregate in clusters. Other retrogradely labelled cells are more sparsely distributed in deeper layers with the greatest accumulation being in layer VI. At this time callosal neurons are elongated, with their long axes parallel to radial cell columns. In autoradiographs on PND 2, silver grains are seen throughout the thickness of the cortex, indicating that callosal axons have reached the upper layers of the gray matter. Neither the alternating vertical columns nor the laminar bands of callosal afferents that characterize this projection in the adult are clearly seen in kittens of this age. By PND 8–12 the segregation of callosal afferents into vertical bands is evident in the gray matter; by PND 18 this pattern is well developed. On the other hand, the somata of callosal neurons of layer III retain their juvenile distribution well into the seventh postnatal week and show adultlike aggregations sometime around the 14th week after birth. Projections to the medial geniculate body, inferior colliculus, putamen, caudate nucleus, and pons are clearly evident early in the first postnatal week.

Postnatal development of auditory callosal connections in the kitten

Myelinated cell bodies of spiral ganglion in kittens during development can first be distinguished by light microscopic observation at the end of the first postnatal week. By mean of electron microscopic observation, the first signs of myelinated perikarya can be observed around the time of birth. Myelination is preceded by the ensheathment of the ganglion cell body by two or three layers of Schwann cell cytoplasmic processes. At birth, the first sign of myelination is visible at the basal part of the cochlea with thin thickenings which contain two or three major dense lines. At this stage, these thickenings cover only a small perikaryal area. Later on, the thickness and the number of thickenings increase and cover the major part of the cell body. The onset of myelination of the processes seems to precede the perikaryal myelination.

Ch. Mulle

Papers

Early Stages of Myelination in the Spiral Ganglion Cells of the Kitten During Development