Response of retinal terminals to loss of postsynaptic target neurons in the dorsal lateral geniculate nucleus of the adult cat.

We have used the neurotoxin kainic acid to produce rapid degeneration of neurons in the dorsal lateral geniculate nucleus (dLGN) of the adult cat. This degeneration mimics the rapid loss of geniculate neurons seen after visual cortex ablation in the neonate. Subsequent anterograde transport of horseradish peroxidase injected into the eye was used to reveal the projection patterns of retinal ganglion cell axons at different survival periods after the kainic acid injection. The density of retinal projections to the degenerated regions of the geniculate was reduced considerably at 4 and 6 months survival, but at 2 months was not significantly different from normal. The laminar pattern of projections to degenerated regions of the geniculate did not change in any animals studied, even when an adjacent lamina contained surviving cells. Electron microscopic examination of degenerated dLGN revealed intact retinal (RLP) and RSD terminals at all survival times, although the density of terminals appeared much reduced when compared to controls. Some RLP terminals exhibited the "dark reaction" of degeneration and these degenerating terminals were most numerous at 2 months survival. These findings demonstrate that, in response to degeneration of their usual target cells, mature retinal ganglion cells with withdraw their axon terminals from these regions of degeneration. We conclude that mature retinal ganglion cells continue to be dependent on target integrity for the maintenance of a normal axonal arborization.     

Evidence of sub-collicular auditory projections to the medial geniculate nucleus in the cat: An autoradiographic and horseradish peroxidase study

Connections of a posteromedial region of the ventral nucleus of the lateral lemniscus were examined in the cat using the autoradiographic tracing method. This sub-collicular region previously had been shown, using retrograde transport of horseradish peroxidase, to send axons to the superior colliculus¹⁰. The autoradiographic findings revealed that many axons from the posteromedial region of the ventral nucleus of the lateral lemniscus that entered the superior colliculus continued into the midbrain reticular formation. Moreover, other axons traced rostral to the inferior colliculus into the thalamus ended in the medial geniculate nucleus, bilaterally. Experiments in which horseradish peroxidase was placed in the medial geniculate nucleus retrogradely labeled the large neurons in the posteromedial region supporting the autoradiographic observations. Other sub-collicular regions also contained labeled cells in these cases, including the main body of the ventral nucleus of the lateral lemnicus and scattered cell groups around the superior olivary complex.     

Topographical origin and ganglion cell type of the retino-pulvinar projection in the cat

In order to examine the pattern of the retino-pulvinar projection in the cat, the existence of which has been recently demonstrated using autoradiographic fiber tracing technique, a small amount of horseradish peroxidase (HRP) was injected into the lateral part of the pulvinar nucleus at various rostocaudal levels. The retrogradely labeled ganglion cells were analyzed in terms of their topographical location and cell size, as seen inretinal whole mounts. The results were compared with those obtained following injections into the lateral geniculate nucleus. Retrogradely labeled cells were found in the retina bilaterally after injections of HRP into the pulvinar nucleus. Pulvinar injections produced labeling of retinal cells in the nasal half of the retina contralaterally, and in the temporal half ipsilaterally. The labeled cells were diffusely distributed in a retinotopically organized fashion. The representation of the area centralis in the retino-pulvinar projection is displaced rostrally as compared with the retino-geniculate projection. All labeled cells after pulvinar injections were medium to small size and no large cells were encountered.     

Some aspects of the organization of the lateral geniculate nucleus inGalago senegalensis revealed by using horseradish peroxidase to label relay neurons

Following injection of horseradish peroxidase into area 17 of the prosimian Galago senegalensis, columns of labeled neurons are seen in the dorsal lateral geniculate nucleus extending through all cell layers. Individual counts of the number of labeled and unlabeled neurons reveal that from 91 to 98% of all neurons within these densely labeled columns are labeled. These results indicate that most of the neurons within the LGN of the bushbaby project to striate cortex. Average diameter measurements of labeled and unlabeled cells within the labeled columns were used to determine whether cell layers could be separated into two types (parvocellular and magnocellular) or three types (small, medium, and large) on the basis of cell body size. These measurements indicate that the LGN of the bushbaby is composed of two layers each of small (layers 4 and 5), medium (layers 3 and 6), and large (layers 1 and 2) relay neurons. These observations are consistent with the conclusion that layers 1 and 2 in the LGN of Galago are homologous with the magnocellular layers, and layers 3 and 6 homologous with the parvocellular layers, identified in the LGN of New and Old World monkeys.     

The projections of different morphological types of ganglion cells in the cat retina.

The central projections of the retinal ganglion cells of the cat were examined using the method of retrograde transport of horseradish peroxidase. Peroxidase was injected into the lateral geniculate nucleus and into the superior colliculus by means of a recording micropipette. After injections at retinotopically homologous points in these two structures in separate animals, tha patterns of retinal ganglion cell labeling were compared. We found that there were three populations of ganglion cells: small cells, that projected predominantly to the superior colliculus; medium-sized cells, that projected predominantly to the lateral geniculate nucleus; and large cells, some of which projected to both structures, and some of which projected to the lateral geniculate nucleus alons. Quantitative studies showed that the average size of the cells in each population was smaller at the area centralis than in the periphery. These results could be directly related to physiological classifications of retinal ganglion cells proposed by other authors.     

Evidence for a sympathetic component in motor branches of the facial nerve: a horseradish peroxidase study in the cat

The retrograde transport of horseradish peroxidase (HRP) was used to examine the location of sympathetic ganglion cells with axons in facial motor branches of the cat. Large numbers of HRP-labeled neurons were observed in the rostro-anterior part of the superior cervical ganglion. In addition, some labeled neurons were found in the cervical sympathetic trunk, the accessory cervical, middle cervical and stellate ganglia.     

Localization of sympathetic postganglionic neurons of physiologically identified cardiac nerves in the dog

Cardiac nerves were identified physiologically and injected with horseradish peroxidase in 38 dogs. Retrogradely labeled neurons were present in the greatest number in the middle cervical ganglion, whereas fewer labeled neurons were present in the stellate ganglion. Only occasional neurons in the superior cervical ganglion were labeled, and no labelphysiologically and injected with horseradish peroxidase in 38 dogs. Retrogradely labeled neurons were present in the greatest number in the middle cervical ganglion, whereas fewer labeled neurons were present in the stellate ganglion. Only occasional neurons in the superior cervical ganglion were labeled, and no labelphysiologically and injected with horseradish peroxidase in 38 dogs. Retrogradely labeled neurons were present in the greatest number in the middle cervical ganglion, whereas fewer labeled neurons were present in the stellate ganglion. Only occasional neurons in the superior cervical ganglion were labeled, and no labeled cells were found in the T3 to T6 paravertebral ganglia or in the ganglia contralateral to the nerve injected. following injections into specific cardiac nerves, retrograde labeling was widespread within the middle cervical ganglion, and the distributions of labeled neurons from different nerves overlapped considerably. In the middle cervical ganglion there was little or no regional grouping of cells projecting to specific cardiac nerves. within the stellate ganglion, however, te cardiac-sympathetic cells were clustered primarily at the cranial pole near toe origin of the ventral and dorsal ansae. Mediastinal ganglia and ganglia located in cardiac nerves were frequently as heavily labeled as the ipsilateral stellate ganglion. The occurrence of heavy labeling in mediastinal and cardiac nerve ganglia indicates that these hitherto unreported ganglia play a significant role in cardiac neural regulation. These data imply that the organization of sympathetic neurons controlling the heart is much more complex than has previously been considered.     

Retrograde axonal transport of horseradish peroxidase by ganglion cells of the albino rat retina

After introduction of small amounts of horseradish peroxidase (HRP) into known visual centers of the brain, retinal ganglion cells projecting to these regions were detected by the accumulation of HRP-positive granules in their somata. Control experiments indicated that the HRP-positive granules had reached the ganglion cell somata by retrograde axonal transport, and did not represent blood-borne or endogenous peroxidase. Using this technique, it has been determined that axons of both large and medium-sized neurons in the ganglion cell layer of adult and immature rat retinae terminate in the superior colliculus and lateral geniculate body, and that characteristic displaced ganglion cells with axonal connections to these visual centers occur regularly in these retinae. In addition, certain small cells in the retinal ganglion cell layer are described which may represent glia or interneurons, or ganglion cells which lack the ability to transport peroxidase or which lack central connections to these visual centers of the brain.     

The afferent and sympathetic components of the lumbar spinal outflow to the colon and pelvic organs in the cat. I. The hypogastric nerve

The cell bodies of the lumbar sensory and sympathetic pre- and postganglionic neurons that project to the pelvic organs in the hypogastric nerve of the cat have been labeled retrogradely with horseradish peroxidase applied to the central end of their cut axons. The numbers, segmental distribution, location, and size of these labeled somata have been determined quantitatively. Afferent and preganglionic cell bodies were located bilaterally in dorsal root ganglia and spinal cord segments L3-L5, with the maximum numbers in L4. Very few cells lay rostral to L3. Afferent cell bodies were generally very small in cross-sectional area relative to the entire population in the dorsal root ganglia. Most of the preganglionic cell bodies lay clustered just medial to the region of the intermediolateral column and extended caudally well beyond its usual limit in the upper part of L4. These neurons were, on the average, larger than the cells of the intermediolateral column itself, with the largest cells lying in the most medial positions. Most of the post-ganglionic somata were in the ipsilateral distal lobe of the inferior mesenteric ganglion, while some (usually less than 10%) lay in accessory ganglia along the lumbar splanchnic nerves and in paravertebral ganglia L3-L5. Postganglionic somata in the inferior mesenteric ganglion were larger than both labeled and unlabeled ganglion cells in the paravertebral ganglia. From the data, it is estimated that about 1,300 afferent neurons, about 1,700 preganglionic neurons, and about 17,000 postganglionic neurons project in each hypogastric nerve in the cat.     

The retinal projection to the thalamus in the cat: a quantitative investigation and a comparison with the retinotectal pathway

The projection of cat retinal ganglion cells to the thalamus was examined using the method of retrograde axonal transport of horseradish peroxidase (HRP). After the injection site was determined physiologically, HRP was applied by one of three methods: iontophoretic injection of minimal amounts, single pressure injections and multiple pressure injections. Iontophoretic injections into single laminae of the dorsal part of the lateral geniculate nucleus (LGNd) revealed that laminae A and A1 receive almost exclusively axon terminals from alpha and beta cells. Single pressure injections elucidated the retinotopic organization of the LGNd. Multiple injections lead to HRP uptake in the whole LGNd including parts of adjacent thalamic nuclei and revealed that at least 77% of all retinal ganglion cells project to the thalamus. This pathway is made up of all alpha cells, all beta cells and almost half of the gamma cells. The thalamus receives its visual input predominantly from the ipsilateral temporal and the contralateral nasal retina; some alpha cells were also labeled in the contralateral temporal retina. The shape of the decussation line was analyzed and its width was found to be proportional to the average ganglion cell spacing along the dorsoventral axis of the retina. From a comparison of the retinothalamic and retinotectal pathways, an estimate of the number of cells with bifurcating axons could be given. The axons of all alpha cells, 10% of the beta cells, and every second gamma cell bifurcate; this amounts to 30% of the retinal ganglion cells.     

The organization of visceral sensory neurons in thoracic dorsal root gangla (DRG) of the cat studied by horseradish peroxidase (HRP) reaction using the cryostat

The organization of visceral sensory neurons in thoracic dorsal root ganglia (DRG) was studied by retrograde transport of horseradish peroxidase (HRP) from the central cut end of the left major splanchnic nerve of the cat. The majority of HRP-labeled cells were concentrated between T5 and T11. Within a DRG, labeled splanchnic neurons were found in all sectors. There was no consistent pattern of localization within the ganglion although clustering of visceral cell bodies was apparent. It may be that each clustered group of cells innervates individual viscera or reflects a degree of functional segregation.     

Depletion of cortical target induced by prenatal ionizing irradiation: effects on the lateral geniculate nucleus and on the retinofugal pathways.

Studies using neonatal surgical lesions to reduce the target area of the retina have supported the idea that developing axons show only a limited specificity in their targeting. This investigation tested whether retinogeniculate axons adjust for partial target depletion by repositioning of axons. We used adult Swiss mice exposed to gamma rays at the time when layer IV cells are generated in the ventricular zone (16 days of gestation). Nissl-stained brain sections were used for histological analyses in thalamus and cortex. Retinal ganglion cells were backfilled from the optic tract with horseradish peroxidase. Intraocular injections of horseradish peroxidase were used to study the retinal projections. In the posterior cortex there was a nearly complete absence of layer IV. The irradiated animals showed a 75% reduction of the dorsal lateral geniculate nucleus. The ventral division, superior colliculus, and other visually related nuclei were not affected. The loss in the ganglion cells (15.7%) was significant but clearly smaller than that observed in the dorsal lateral geniculate nucleus (75%). Therefore, the shrinkage of the dorsal lateral geniculate nucleus led to a reduction in the area available for retinal projections. Despite partial target loss, pattern of retinal projections did not differ from that of the controls. The effect on the dorsal lateral geniculate nucleus is discussed in the light of differences between prenatal and neonatal damage of the presumptive visual cortex. The absence of aberrant retinal projections suggests that repositioning of axons is not the first mechanism employed by retinal axons to match connections in numerically disparate populations.     

Electrical activity regulates dendritic reorganization in ganglion cells after neonatal retinal lesion in the cat

During the first month of postnatal life, the dendritic arborizations of cat retinal ganglion cells continue to develop and undergo a substantial remodeling. Mechanical and pharmacological interferences with the normal development induce, during this period of time, substantial modifications in ganglion cell morphology. Specifically, the degeneration of those neurons whose axons were severed by a neonatal retinal lesion leads to a zone depleted of ganglion cells. Neurons at the border of the depleted area develop an abnormal elongation of the dendritic trees toward the empty space. In the present paper, we report data showing that this dendritic reorganization can be prevented by blocking the electrical activity with repeated tetrodotoxin injections into the eye during the whole critical period. Our analysis was performed on neurons filled with horseradish peroxidase.     

Direct vagal input to neurons in the area postrema which project to the parabrachial nucleus: An electron microscopic-HRP study in the cat

This study in cat examines the synaptic relationship of vagal afferents to parabrachial projecting neurons in the area postrema (AP) using anterograde and retrograde transport of horseradish peroxidase (HRP). Wheat germ agglutinin-HRP injected into the parabrachial nucleus (PBN) produced retrograde neuronal labeling in the AP and in the nucleus of the tractus solitarius bilaterally, but with an ipsilateral predominance. Labeled neurons were confined mainly to the caudal 2/3's of the AP. Following injection of WGA-HRP into the PBN and HRP into the nodose ganglion in the same animal, examination of sections of the AP with the electron microscope revealed anterogradely labeled axon terminals in apposition to retrogradely labeled somata and dendrites. In some instances, labeled terminals were observed to form synaptic contacts with retrogradely labeled neurons. We conclude that in the cat a vagal input to neurons in the AP is monosynaptically relayed to the PBN.     

The distribution of the cat''s carotid sinus nerve afferent and efferent cell bodies using the horseradish peroxidase technique

The location of both afferent and efferent carotid sinus nerve (CSN) cell bodies in the cat has been determined using the horseradish peroxidase (HRP) technique. Following a limited exposure of the central cut end of the CSN to HRP, labeled sensory ganglion cells were found in both the petrosal and superior ganglia of the IXth cranial nerve. An average of 387 in the former and 16 cells in the latter ganglion were labeled.

Retrogradely labeled neurons were found only within the ipsilateral medulla. These cells were both round and spindle shaped and had an average somal diameter of 19 μm. The number of these CSN efferent cell bodies ranged from 1 to a maximum of 20 in a given animal. They were found in both the nucleus parvocellularis and the retrofacial nucleus. In 8 cases axonal labeling was observed. Axons generally projected dorsomedially from the ventrolateral medulla.     

The retinohypothalamic tract in the cat: retinal ganglion cell morphology and pattern of projection

The pattern of retinal projection to the hypothalamus and the morphological properties of the retinal ganglion cells that comprise the retinohypothalamic tract have been examined in the cat. Intraocular injections of horseradish peroxidase revealed a dense retinal projection to the ventral suprachiasmatic nucleus; however, lighter projections were seen in the dorsal suprachiasmatic nucleus, and in hypothalamic regions both dorsal and lateral to the suprachiasmatic nucleus. Intrasuprachiasmatic nucleus injections of horseradish peroxidase retrogradely labelled retinal ganglion cells that were small to medium in soma size. The labelled ganglion cells exhibited long thin dendrites that were sparsely branched. The labelled retinal ganglion cells exhibited a significant change in soma size associated with retinal eccentricity. The morphological characteristics of the ganglion cells that project to the suprachiasmatic nucleus are similar to those of gamma cells.     

Displaced ganglion cells in the retina of the rat

The distribution, laterality of projection, and perikaryal sizes of displaced ganglion cells (DGCs) were examined in whole-mounted retinae after massive unilateral injections of horseradish peroxidase along the optic tract in pigmented rats. The DGCs were found predominantly in the lower temporal periphery of the retina. Nearly all DGCs labeled had contralaterally projecting axons. The sizes of the labeled DGCs spanned the range of ordinary ganglion cells, but few middle-sized DGCs were labeled. The results support the hypothesis that displaced ganglion cells are late-developing neurons that do not complete their migration toward the ganglion cell layer during retinal histogenesis.     

Patterns of synaptic input to layer 4 of cat striate cortex

Although cells in layer 4 of cat striate cortex represent the first stage in the cortical processing of visual information, they have considerably more complicated receptive field properties than the afferents to the layer from the lateral geniculate nucleus. In considering how these properties are generated, we have focused on the intrinsic cortical circuitry, and particularly on the projection to layer 4 from layer 6. Layer 6 pyramidal cells were injected with horseradish peroxidase and examined at the light and electron microscopic level. The labeled axon terminals were found to form asymmetric synapses and to show a strong preference for contacting dendritic shafts. Serial reconstruction of dendrites postsynaptic to labeled layer 6 cell axon terminals showed that a large proportion of the postsynaptic dendrites belonged to smooth and sparsely spiny stellate cells, suggesting a selective innervation of these cell types. In contrast, the geniculate projection to layer 4 made synapses primarily with dendritic spines and, as a result, the large majority of terminals ended on spiny cells. Since smooth and sparsely spiny stellate cells are thought to mediate inhibition within the cortex, we suggest that one effect of the layer 6 to layer 4 projection could be to contribute to inhibitory features of the receptive fields of layer 4 cells.     

Retinal ganglion cell projections to individual layers of the lateral geniculate body in Galago crassicaudatus

The genus Galago provides an unique opportunity to study the relation between layers of the lateral geniculate body and classes of retinal ganglion cells. In the present experiments HRP was restricted to individual layers of the lateral geniculate body with the following results: After injections of the magnocellular layers, layers 1 and 2, labeled retinal ganglion cells ranged in size from 8 to 20 μm. After injections of the parvocellular layers, layers 3 and 6, labeled retinal ganglion cells ranged in size from 6 to 12 μm. After injections involving layers 4 and 5, which layers contain only very small, pale cells, labeled retinal ganglion cells ranged in size from 5 to 14 μm. Thus, the very largest ganglion cells were labeled only after injections of magnocellular layers 1 and 2, while small and medium retinal ganglion cells were labeled after HRP injections in every layer of the lateral geniculate body. Because the magnocellular layers actually contain a mixture of large, medium, and small-sized cells, we suggest that retinal ganglion cells of different size-classes project to geniculate relay cells of the corresponding size-class.