Morphology of expiratory neurons of the Bötzinger complex: An HRP study in the cat

In anesthetized and artificially ventilated cats, the physiological and morphological properties of expiratory neurons or their axons of the B?tzinger complex (BOT) were studied using intracellular recording and intracellular HRP labeling techniques. Thirteen expiratory neurons (nine cell somata and four axons) were successfully stained. Four of them were motoneurons, having relatively large cell somata in the retrofacial nucleus (RFN) and axons without any collaterals inside the brainstem. All the motoneurons showed a plateau shape of depolarization potentials during the expiratory phase. Any of the other nine expiratory neurons exhibited augmenting type firing or membrane potential changes during the expiratory phase. In five out of nine augmenting neurons, cell somata were stained and located ventral to the RFN. In four, only axons were stained. The majority of the augmenting neurons had two major axonal branches: one traveling toward the contralateral side and the other descending ipsilaterally in the brainstem. The most striking feature of the axonal trajectory was that all of the stained augmenting expiratory neurons, including the axons, had collateral branches with synaptic boutons in the BOT area, thus indicating that BOT expiratory neurons interact with some respiratory neurons in the BOT area and its vicinity.     

Morphology of augmenting inspiratory neurons of the ventral respiratory group in the cat

The present study examined, in Nembutal-anesthetized and artificially ventilated cats, the morphologic properties of the inspiratory neurons of the ventral respiratory group (VRG). Horseradish peroxidase (HRP) was injected into 21 augmenting inspiratory or late inspiratory neurons with peak firing rates in the late inspiratory phase. The majority of the stained neurons were antidromically activated by stimulation of the cervical cord. Thirteen somata, located within or around the nucleus ambiguus (AMB), between 100 microns caudally and 2,000 microns rostrally to the obex, were stained. In ten cases, the stem axons issuing from the cells of origin coursed medially to cross the midline without giving off any axonal collaterals. Three neurons gave rise to axonal collaterals on the ipsilateral side, distributing boutons in the medullary reticular formation, in the vicinity of the AMB, hypoglossal nucleus, solitary tract, and dorsal motor nucleus of the vagus. In eight neurons, only the axons were labeled; in four of these, which were antidromically activated from the spinal cord, the stem axons crossed the midline 2,000-3,000 microns rostral to the obex and descended in the reticular formation around the AMB down to the cervical cord. They issued several axonal collaterals, distributing terminal boutons at the level of the caudal end of the retrofacial nucleus and about 1,000 microns rostral and caudal from the obex. Terminals were found mainly in and around the AMB, and a few were found in the vicinity of the dorsal motor nucleus of the vagus. The remaining four nonactivated axons distributed their terminal boutons widely in the reticular formation around the AMB. Thus, the augmenting inspiratory neurons of the VRG were shown to project not only to the spinal cord, but also to the VRG, hypoglossal nucleus, and dorsal motor nucleus of the vagus.     

Morphology of single axons of tectospinal neurons in the upper cervical spinal cord

Morphology of single axons of tectospinal (TS) neurons was investigated by intraaxonal injection of horseradish peroxidase (HRP) at the upper cervical spinal cord of the cat. TS axons were electrophysiologically identified by their direct responses to stimulation of the contralateral superior colliculus (SC). None of these axons responded to thoracic stimulation at Th2. Three-dimensional reconstructions of the axonal trajectories were made from 20 well-stained TS axons at C1-C3. Cell bodies of these axons were located in the intermediate or deep layers of the caudal two-thirds of the SC. Usually, TS axons had multiple axon collaterals, and up to seven collaterals were given off per stem axon [2.7 ± 1.6 (mean ± S.D.); n = 20]. Collaterals had simple structures and ramified a few times mainly in the transverse plane. The number of terminals for each collateral was small. These collaterals terminated in the lateral parts of laminae V–IX, mainly in laminae VI, VII, and VIII. There were usually gaps free from terminal arborizations between adjacent collaterals, because the rostrocaudal spread of each collateral (mean = 700 μm) was narrower than the intercollateral interval (mean = 2,500 μm). Seven of the 19 TS axons had terminals in the lateral parts of laminae V–VIII, with little projection to lamina IX, and the other 12 axons had terminals in lamina IX besides the projection to the lateral parts of laminae V–VIII. Axon terminals in lamina IX did not appear to make contacts with the somata or proximal dendrites of retrogradely labeled motoneurons, but contacts were found with the somata of counterstained interneurons in the lateral parts of laminae V–VIII. Three spinal interneurons (two in lamina VIII and one in lamina V at C1) that received monosynaptic excitation from the SC were stained, and their axonal trajectories were reconstructed. They had multiple axon collaterals at C1-C2 and mainly projected to laminae VIII and IX, with smaller projections to lamina VII. Many axon terminals of the interneurons were found in multiple neck motor nuclei, where some of them made contacts with retrogradely labeled motoneurons. The present finding provides evidence that the direct TS projection to the spinal cord may influence activities of multiple neck muscles, mainly via spinal interneurons, and may play an important role in control of head movement in parallel with the tectoreticulospinal system. © 1996 Wiley-Liss, Inc.     

Morphological study of long axonal projections of ventral medullary inspiratory neurons in the rat

The aim of this study was to examine medullary and spinal axonal projections of inspiratory bulbospinal neurons of the rostral ventral respiratory group (VRG) in the rat. A direct visualization of long (9.8–33 mm) axonal branches, including those projecting to the contralateral side of the medulla oblongata and the spinal cord, was possible due to intracellular labeling with neurobiotin and long survival times (up to 22 h) after injections. Seven of the nine labeled neurons had bilateral descending axons, which were located in discrete regions of the spinal white matter; ipsilateral axons in the lateral and dorsolateral funiculus, contralateral in the ventral and ventromedial funiculus. The collaterals issued by these axons at the mid-cervical level formed close appositions with dendrites of phrenic motoneurons, which had also been labeled with neurobiotin. None of these collaterals crossed the midline. The significance of this finding is discussed in relation to the crossed-phrenic phenomenon. Additional spinal collaterals were identified in the C₁ and T₁ segments. Within the medulla, collaterals with multiple varicosities were identified in the lateral tegmental field and in the dorsomedial medulla (in the hypoglossal nucleus and in the nucleus of the solitary tract). These results demonstrate that inspiratory VRG neurons in the rat have some features which have not been previously described in the cat, including frequent bilateral spinal projection and projection to the nucleus of the solitary tract. In addition, this study shows that intracellular labeling with neurobiotin offers an effective way of tracing long axonal projections, supplementing results previously obtainable only with antidromic mapping, and providing morphological details which could not be observed in previous studies using labeling with horseradish peroxidase.     

Efferent projections of inspiratory neurons of the ventral respiratory group. A dual labeling study in the rat

The efferent projections of the medullary respiratory neurons of the rat were studied using an anterograde tracer, Phaseolus vulgaris leucoagglutinin (PHA-L). In Nembutal-anesthetized rats, PHA-L was iontophoretically applied to (1) the area of inspiratory neurons of the ventral respiratory group (VRG) around the nucleus ambiguus, or (2) the area ventrolateral to the solitary tract. In addition, a fluorescence retrograde tracer, Fast blue (FB), was injected into the cervical phrenic nerve several days after the PHA-L injection. When PHA-L was injected into the area of predominantly inspiratory neurons of VRG, dense PHA-L-labeled axons were observed bilaterally in the spinal cord: the ipsilateral projections were noticeably denser than the contralateral ones. Fine axonal branches were distributed around a column of the phrenic motoneurons and boutons were observed on the somata of the FB-labeled motoneurons, suggesting monosynaptic connections between VRG inspiratory neurons and phrenic motoneurons. On the other hand, when PHA-L was injected into the area ventrolateral to the solitary tract, only a few descending axons to the spinal cord were seen bilaterally. No contacts between the PHA-L-labeled axons and the FB-labeled phrenic motoneurons were observed. The brainstem projections of the VRG were found bilaterally in the nuclei ambigui, Cajal's interstitial nuclei of the solitary nucleus, the solitary nuclei, the hypoglossal nuclei, the K?lliker-Fuse's nuclei, and the subcoeruleus areas.     

Morphological and electrophysiological identification of gigantocellular tegmental field neurons with descending projections in the cat: II. Bulb

There are three different descending projections from the bulbar gigantocellular tegmental field (BFTG) in the cat, as defined by intracellular recording and intracellular horseradish peroxidase (HRP) techniques. The first pathway arises from neurons which send axons to the contralateral medial longitudinal fasciculus (cMLF neurons); cMLF neurons show excitatory postsynaptic potentials (EPSPs) after stimulation of the ipsilateral pontine gigantocellular tegmental field (PFTG). Most cMLF neurons have large ellipsoid-polygonal somata (mean, 56.8 micron), thick axons (average diameter, 3.09 microns), mostly non-spiny dendrites and dendritic fields flattened in the anteroposterior direction. No cMLF neurons with axon collaterals in the BFTG are present in the data of this study. The second pathway arises from neurons which send axons to the ipsilateral MLF (iMLF neurons); iMLF neurons show EPSPs after stimulation of the ipsilateral PFTG. Most iMLF neurons have large ellipsoid-polygonal somata (mean, 60.2 microns), thick axons (average diameter, 3.00 microns), mostly non-spiny dendrites and dendritic fields that are only slightly flattened in the anteroposterior direction. As with cMLF neurons, no iMLF neurons with axon collaterals in the BFTG are present in the data of this study. The third pathway arises from neurons that send axons directly into the ipsilateral caudal bulbar reticular formation (iBRF neurons). Most iBRF neurons have smaller ellipsoid-polygonal somata (mean, 38.6 microns), thinner axons (average diameter, 1.84 microns), mostly nonspiny dendrites, and dendritic fields that are flattened in the anteroposterior direction. In contrast to cMLF and iMLF neurons, axon collaterals are present in 73% of iBRF neurons. About half of iBRF neurons have bifurcated axon collaterals with both anterior and posterior projections, and in these neurons antidromic spike potentials are elicited by stimulation of the ipsilateral PFTG.     

Morphological and electrophysiological characteristics of projection neurons in the nucleus interpositus of the cat cerebellum

The populations of neurons in the nucleus interpositus (IP) of the cat cerebellum which project to the ventral lateral nucleus of the thalamus (VL), the red nucleus (RN), the nucleus reticularis tegmenti pontis (NRTP), the pontine nuclei (PN), the inferior olive (IO), and the cerebellar cortex were identified by intracellular and extracellular injections of HRP and studied electrophysiologically. When HRP was simultaneously injected into the VL, RN, and IO, over 95% of the neurons in the IP nuclei were labeled; indicating that there are few, if any, local circuit neurons. The vast majority (86%) of the larger IP neurons (soma length ≥ 20 μm) project rostrally to the RN and thalamus. These neurons typically have long, relatively spine free dendrites and axons which in a few cases gave rise to recurrent collaterals. Two intracellularly stained projection neurons which had exceptionally long spiny dendrites had axons which gave rise to nucleocortical collaterals in addition to several local collaterals. IP neurons projecting to the NRTP and PN were located primarily in the lateral aspect of the nucleus interpositus anterior. Electrophysiological experiments established that neurons projecting to the NRTP also project to the VL. The IP neurons projecting to the IO have small fusiform or multipolar somata, long thin dendrites, and receive excitatory inputs from the IO. At least 73% of the small neurons in the IP project to the IO, and some of these, in addition, project to the VL. There are at least three morphologically distinguishable populations of projection neurons in the IP, large VL projection neurons, small IO projection neurons, and neurons with nucleocortical collaterals. The projection of the IP to diverse regions of the brain is accomplished mainly by axon collateralization, but regional and morphological specialization also play a role in the organization of the output of the IP.     

Axonal branching patterns of nucleus accumbens neurons in the rat

The patterns of axonal collateralization of nucleus accumbens (Acb) projection neurons were investigated in the rat by means of single-axon tracing techniques using the anterograde tracer biotinylated dextran amine. Seventy-three axons were fully traced, originating from either the core (AcbC) or shell (AcbSh) compartment, as assessed by differential calbindin D28k-immunoreactivity. Axons from AcbC and AcbSh showed a substantial segregation in their targets; target areas were either exclusively or preferentially innervated from AcbC or AcbSh. Axon collaterals in the subthalamic nucleus were found at higher than expected frequencies; moreover, these originated exclusively in the dorsal AcbC. Intercompartmental collaterals were observed from ventral AcbC axons into AcbSh, and likewise, interconnections at pallidal and mesencephalic levels were also observed, although mostly from AcbC axons toward AcbSh targets, possibly supporting crosstalk between the two subcircuits at several levels. Cell somata giving rise to short-range accumbal axons, projecting to the ventral pallidum (VP), were spatially intermingled with others, giving rise to long-range axons that innervated VP and more caudal targets. This anatomical organization parallels that of the dorsal striatum and provides the basis for possible dual direct and indirect actions from a single axon on either individual or small sets of neurons.     

The morphology of single lateral vestibulospinal tract axons in the lower cervical spinal cord of the cat

The intraspinal morphology of single lateral vestibulospinal tract (LVST) axons was investigated with the method of intra-axonal staining with horseradish peroxidase (HRP) and three-dimensional reconstruction of the axonal trajectory. Axons penetrated in the ventral funiculus at C5-C8 were identified as LVST axons by their monosynaptic responses to stimulation of the ipsilateral vestibular nerve and by their direct responses to stimulation of the ipsilateral Deiters' nucleus and LVST. Reconstructions were made from 34 well-stained LVST axons. Of these, 23 terminated in the brachial segments (C5-Th1) and the other 11 projected below Th2. These axons were traced over distances of 2.9-16.3 mm rostrocaudally. Within these lengths, one to seven axon collaterals (mean +/- S.D., 3.2 +/- 2.0, N = 19) were given off at right angles from the stem axons of LVST axons terminating in the brachial segments. The mean diameters of stem axons and primary collaterals were 4.5 microns and 1.6 micron, respectively. In the gray matter, collaterals ramified successively, pursued a delta-like path, and terminated mainly in laminae VII and VIII or lamina IX. The rostrocaudal extension of a single collateral was very restricted (mean +/- S.D., 760 +/- 220 microns, N = 16), in contrast to the extensive dorsoventral and mediolateral extent of the terminal arborization. There were usually gaps between adjacent collateral arborizations from the same stem axons, since the intercollateral distances ranged from 400 to 4,300 microns (mean = 1,490 microns). LVST axons terminating in brachial segments were divided into two groups--a medial group and a lateral group--on the basis of their projection sites in the transverse plane of the gray matter. The axons of the medial type had their main projection to laminae VII and VIII of Rexed, while those of the lateral type terminated in lamina IX. The terminal arborizations of the medial type LVST axons were mainly distributed over lamina VIII, where synaptic boutons appeared to make contact with proximal dendrites or somata of medium-sized and large neurons in the ventromedial nucleus and also in the medial portion of lamina VII adjacent to the central canal and dorsal to lamina VIII. Five out of 15 medial type axons had a bilateral projection. One or two collaterals of each of these axons crossed the midline through the anterior commissure and terminated in lamina VII or VIII. It was concluded that the contralateral projection was sparse.(ABSTRACT TRUNCATED AT 400 WORDS)     

Projections and terminations of single respiratory axons in the cervical spinal cord of cat

Position, divergence, branching, and termination patterns of single, respiratory axons were studied in cat cervical spinal cord by injecting horseradish peroxidase (HRP) intra-axonally. We stained 12 axons which were characterized by their firing patterns and by electrical stimulation. Five axons discharged during inspiration (I); the remaining 7 discharged during expiration (E). No injected axon was evoked by stimulating ipsilateral phrenic nerve roots while 7 (4 I, 3 E) of 12 were excited at a short latency from stimulating at a medullary site (on the midline, 1-2 mm rostral to the obex, approximately 3 mm below the dorsal medullary surface) where many bulbospinal respiratory axons decussate. All injected stem axons were located in the ventral and ventrolateral funiculi, traversed in a rostrocaudal direction, and were stained for lengths ranging from 3.6 to 12.4 mm. Mean axonal diameter was 2.9 microns. In 6 axons (4 I, 2 E), 14 collaterals were stained: 1 on each E axon, 2 on one I axon, 3 each on 2 others and 4 on another I axon. Collaterals emerged perpendicularly from the descending stem axon and projected directly to the ventral horn. The average distance between neighboring collaterals was 1.0 mm (n = 7). Collaterals did not arborize until they were near or within the ventral horn. Both en passant and terminaux types of presynaptic boutons were found primarily within the rostrocaudal cylinder that defined the phrenic motor column. In addition, some boutons were located dorsomedial to the phrenic motor column. We conclude that I axons, presumably of medullary origin, have multiple collaterals which terminate primarily in the phrenic motor column. However, the same axon can have terminals in different regions of the ventral horn, which are known to contain dendrites of phrenic motoneurons.     

Morphological and electrophysiological identification of gigantocellular tegmental field neurons with descending projections in the cat: I. Pons

Two different descending projections from the pontine gigantocellular tegmental field (PFTG) were defined by the use of intracellular recording and intracellular horseradish peroxidase (HRP) techniques in the cat. Type I neurons (reticulospinal neurons) had antidromic spike potentials produced by stimulation of the ipsilateral medial longitudinal fasciculus (MLF) and sent axons to the ipsilateral MLF. Most type I neurons had large ellipsoidpolygonal somata (mean, 59.7 microns), thick axons (average diameter, 3.33 microns), and slightly oblate large dendritic fields. The mean anteroposterior extent of the dendritic field was 1,492 microns, the mean mediolateral extent was 1,784 microns, and the mean dorsoventral extent was 1,562 microns. There were no type I neurons with axon collaterals. In contrast, type II neurons (reticuloreticular neurons) had antidromic spike potentials produced by stimulation of the bulbar reticular formation (BRF) and sent axons directly to the BRF. In comparison with type I neurons, most type II neurons had smaller ellipsoidpolygonal somata (mean, 40.2 microns), thinner axons (average diameter, 2.32 microns), and smaller, slightly oblate dendritic fields. The mean anteroposterior extent of the dendritic field was 1,264 microns; the mean mediolateral extent was 1,511 microns; and the mean dorsoventral extent was 1,226 microns. Also in contrast to type I neurons, 36% of type II neurons had axon collaterals.     

Morphological and functional properties of trigeminal nucleus oralis neurons projecting to the trigeminal motor nucleus of the cat

Horseradish peroxidase (HRP) was injected into the somata located in the rostrodorsomedial part (Vo.r) of the trigeminal nucleus oralis; an axonal projection to the trigeminal motor nucleus (Vmo) was demonstrated in two Vo.r neurons. The two neurons differed in their morphological and functional properties. The first Vo.r neuron responded to stimulation of low-threshold mechanoreceptors and its stem axon gave off massive axon collaterals that issued terminal branches to the dorsolateral subdivision of Vmo, Vo.r, and the medial and lateral parts of the lower brainstem reticular formation. The second Vo.r neuron was activated by stimulation of the tooth pulp or lingual nerve at twice longer latency than that of the first neuron. This stem axon was divided into two main ascending and one descending branches, and one of the main ascending branches was further bifurcated into two branches. The main non-bifurcated ascending branch gave off 4 collaterals, two of which sent terminal branches into the dorsolateral subdivision of Vmo and others into the Vo.r and juxta-trigeminal regions. The somato-dendroarchitectonic differences were also described in the two Vo.r neurons stained.     

Morphologic features and electrical membrane properties of projection neurons in the marginal layer of the medullary dorsal horn of the rat

Possible correspondence between morphologic features and electrical membrane properties of projection neurons in lamina I [the marginal zone (MZ)] of the caudal subnucleus of the spinal trigeminal nucleus [the medullary dorsal horn (MDH)] was examined by using intracellular recordings and biocytin-injections combined with histochemical and immunohistochemical staining techniques. The experiments were done in horizontal slice preparations of the rat brain. Thirteen MZ neurons were recorded stably and stained successfully. These neurons were confirmed to send their axons to the brain regions outside the MDH by camera lucida reconstruction. They were divided into two types on the basis of branching patterns of their axons within the MDH: Type I projection (P-I) neurons (n = 7 neurons) had main axons that rarely emitted axon collaterals within the MDH, whereas type II projection (P-II) neurons (n = 6 neurons) had main axons that emitted many axon collaterals within laminae I, II (substantia gelatinosa), and III (magnocellular part) of the MDH and also to the spinal tract of the trigeminal nerve; these axon collaterals usually constituted a dense mesh of axonal processes within laminae I and II of the MDH, especially in lamina II. About half of the neurons of each type showed immunoreactivity for the neurokinin-1 receptor. Resting membrane potentials were significantly more positive in P-I neurons than in P-II neurons. The P-II neurons had higher input resistance, a longer membrane time constant, and a higher threshold for spike than P-I neurons. In response to weak, long depolarizing current pulses, P-II neurons often showed slow ramp depolarization; the same neurons exhibited delayed repolarization to the resting potential (slow after depolarization) after the offset of the long depolarizing current pulses. Neither the slow-ramp depolarization nor the slow after depolarization was observed in P-I neurons. Slow return to resting membrane potential after offset of hyperpolarizing current pulses also was observed frequently in P-II neurons but not in P-I neurons. The results indicate that P-II neurons differ in their membrane properties compared with P-I neurons, and P-II neurons may be involved in the local circuit mechanism within the MDH more deeply than P-I neurons.     

Projection subtypes of rat neostriatal matrix cells revealed by intracellular injection of biocytin.

Individual neostriatal-matrix spiny neurons were stained intracellularly with biocytin after intracellular recording in vivo, and their axons were traced into the globus pallidus (GP), entopeduncular nucleus (EP), and/or substantia nigra (SN). The locations of the neurons within the matrix compartment of the neostriatum (NS) were established by immunocytochemical counterstaining of sections containing the cell bodies using antibodies for calbindin-D28K. This allowed nearly complete visualization of the axonal projections of single NS neurons. On the basis of their intrastriatal axonal arborizations, matrix spiny neurons could be divided into 2 types. One type, which was the more common, had local axonal arborizations restricted to the region of the dendritic field, often with axon collaterals arborizing within the dendritic field of the cells of origin. A second, less common, cell type in the matrix had local axon collaterals distributed widely in the NS. Among matrix neurons with restricted local collateral fields, 3 subtypes could be distinguished on the basis of their efferent axonal projections. Type I cells projected only to the GP. Type IIa cells projected to the GP, EP, and SN pars reticulata. Type IIb cells projected to the GP and SN but not to the EP. The shapes and densities of the GP arborizations varied in the 3 cell types, with the cells projecting only to the GP (type I) projecting more heavily and filling a larger volume there than type II cells. The dendrites and intrastriatal axon collaterals of 3 subtypes were similar in morphology. The class of matrix spiny neurons with intrastriatal axon collaterals distributed widely in the NS were observed to project to the GP. Projections beyond the GP were not identified for this cell type, but could not be ruled out. Somatodendritic morphologies of neurons did not differ according to the projection site. These results demonstrate that NS matrix spiny cells are more heterogeneous in their efferent projection patterns than previously suspected on the basis of retrograde axonal tracing and immunocytochemical studies. As predicted by those previous studies, there is a class of matrix neurons that projects only to the GP. Presumably, these cells contain enkephalin. Cells projecting to the SN and EP, and so presumably containing substance P, give off a small projection to the GP, as well, and differ in their collateralization patterns within the 3 major target nuclei.     

Postnatal changes in the termination pattern of recurrent axon collaterals of triceps surae alpha-motoneurons in the cat

alpha-Motoneurons innervating the triceps surae and short plantar muscles were stained intracellularly with horseradish peroxidase (HRP) in 0-44-day-old kittens and adult cats. The terminal arborizations of the recurrent axon collaterals in the spinal cord were studied in the light microscope (LM). The short plantar motoneurons lacked axon collaterals in all age groups. With a few exceptions in the youngest kittens (0-1 days of age), the projection field of the axon collaterals of triceps surae motoneurons did not change during development. The exceptional motoneurons had axon collaterals projecting ventromedial to the adult termination areas in Rexed's laminae VII and IX. Within all parts of the projection field, there was a substantial postnatal reduction in the number of axon collateral swellings, interpreted as synaptic terminals, and a total elimination of short and thin axonal processes without swellings. The findings are discussed in relation to earlier demonstrated loss of synaptic terminals on the motoneurons and elimination of polyneuronal innervation of muscle fibers postnatally.     

Principal neurons of the basilar pons as the source of a recurrent collateral system

The basilar pontine nuclei in the opossum are composed of two general categories of neurons, intrinsic cells and the principal or projection neurons. Observations from Golgi material indicate that principal neurons whose primary axons project to the cerebellar cortex may also give rise to recurrent branches distributing within the pontine gray. Such collaterals were observed to arise near the soma and at some distance from the cell body of the parent axon. The electron microscopic correlate of such a system was identified in the basilar pontine neuropil in animals subjected to lesions of the cerebellar cortex. These lesions destroyed mossy terminals and their parent axons and thus initiated a retrograde reaction in basilar pontine projection neurons which manifested itself in the form of morphologic alterations observed in somata, dendrites, and a class of axonal boutons. Similar altered axon terminals were not observed in control material and did not correspond to the terminals of cerebello-pontine axons described in previous work. It was therefore suggested that such boutons represented the terminals of the recurrent collateral system observed in Golgi material.     

Local circuitry of identified projection neurons in cat visual cortex brain slices

The relationship between pyramidal cell morphology and efferent target was investigated in layer 6 of cat primary visual cortex (area 17). Layer 6 has 2 projections, one to the lateral geniculate nucleus (LGN) and another to the visual claustrum. The cells of origin of each projection were identified by retrograde transport of fluorescent latex microspheres. The labeled cells were visualized in brain slices prepared from area 17, using an epifluorescence compound microscope modified for intracellular recording. Individual retrogradely labeled cells were penetrated and intracellularly stained with Lucifer yellow to visualize the patterns of axons and dendrites associated with each projection. The neurons that give rise to the 2 projections had very different patterns of dendrites and local axonal collaterals, but the patterns within each group were highly stereotyped. The differences between their axonal collaterals were particularly dramatic. Claustrum projecting cells had fine, horizontally directed collaterals that arborized exclusively in layer 6 and lower layer 5. Most LGN projecting cells had virtually no horizontal arborization in layer 6. Instead, they sent widespread collaterals vertically, which arborized extensively in layer 4. The apical dendrites of the 2 groups also differed markedly. Claustrum projecting cells had apical dendrites reaching to layer 1, with branches in layer 5 only, while LGN projecting cells never had an apical dendrite reaching higher than layer 3, with side branches in layers 5 and 4. Therefore, each efferent target must receive inputs from neurons whose synaptic connections within area 17 are significantly different from those of neurons projecting to other targets. This further suggests that distinct visual response properties should be associated with each projection. In addition to the claustrum and LGN projecting cells, about 20% of layer 6 pyramidal neurons lacked an efferent axon. Morphologically, most resembled LGN projecting neurons, but a few had characteristics of claustrum projecting cells. These neurons may represent cells that either failed to make an efferent connection or cells that lost an efferent axon during development. Their frequency suggests that such intrinsic, presumably excitatory, neurons may play a significant role in cortical processing.     

Anatomic evidence suggestive of dendrodendritic synapses in the opossum basilar pons

The basilar pontine nuclei in the opossum are composed of two general categories of neurons, intrinsic cells and the principal or projection neurons. Observations from Golgi material indicate that principal neurons whose primary axons project to the cerebellar cortex may also give rise to recurrent branches distributing within the pontine gray. Such collaterals were observed to arise near the soma and at some distance from the cell body of the parent axon. The electron microscopic correlate of such a system was identified in the basilar pontine neuropil in animals subjected to lesions of the cerebellar cortex. These lesions destroyed mossy terminals and their parent axons and thus initiated a retrograde reaction in basilar pontine projection neurons which manifested itself in the form of morphologic alterations observed in somata, dendrites, and a class of axonal boutons. Similar altered axon terminals were not observed in control material and did not correspond to the terminals of cerebello-pontine axons described in previous work. It was therefore suggested that such boutons represented the terminals of the recurrent collateral system observed in Golgi material.     

Morphology of masticatory motoneurons stained intracellularly with horseradish peroxidase

Masticatory motoneurons were identified electrophysiologically and stained with horseradish peroxidase (HRP). The masseter motoneurons could be divided into 3 groups on the basis of their dendritic morphology. In contrast, the digastric or mylohyoid motoneurons showed a similar dendritic configuration. These neurons had much developed dendritic trees in the dorsomedial than ventrolateral direction. The first group of the masseter motoneurons had their dendritic trees which extended radially in all directions with a slight preference to project rostrally. These somata were located in the center of the subdivision containing the masseter motoneurons. In the second group, their dendritic arbores had a polarity extending hemispherically. These neuronal somata were located in the medial, ventral, and lateral regions of the subdivision. For the masseter motoneurons in the two groups and jaw-opening motoneurons, the dendritic swellings were frequently observed in the distal branches. The third group had their dendritic trees which were much simpler in configurations with less tapering or branching than those of other neurons examined. Furthermore, a wide variety of dendritic spines and appendages, and no dendritic swellings, observed in the third group were distinct from other neurons stained. The dendritic trees of the jaw-closing and -opening motoneurons were confined to the individual subdivisions. There were no instances in which axon collaterals were observed for well-stained 16 axons.     

The intercalated cells of the amygdala

The intercalated cell groups, or massa intercalata, of the amygdala have been studied in rodent brains with Golgi methods. They also have been examined in gallocyanin-chromalum-, AChE-, and Timm-stained rat brains. The Golgi data indicate that the intercalated cells are not confined to a series of isolated cell clumps but form a neuronal net that covers the rostral half of the lateral-basolateral nuclear complex, stretches across a major portion of rostral amygdala, and continues rostrally beneath the anterior commissure. There are two general types of intercalated neuron--medium and large neurons. The medium intercalated neurons are more common. They have round to elongate somata, 9-18 microns in diameter, and round to bipolar dendritic trees, depending on their location. Most of the dendrites are spine-bearing, as are 20% of the somata. Their axons often have locally ramifying collaterals. The parent axons apparently terminate in either the lateral-basolateral or central nuclei and some of them appear to enter the external capsule. There is a unique medium intercalated neuron that has nearly spine-free, varicose dendrites and an axon that is typical of short axon (Golgi II) cells. There are two varieties of large intercalated neuron-spiny and aspiny. Most of them are aspiny, although they usually have a few spines scattered along their dendrites. Both varieties have elongate, sometimes round, somata that can be as much as 60 microns long. Their dendrites are long, thick, and have few branch points. Only the initial part of the large aspiny cell axon has been impregnated. The large spiny cell axons have several local collaterals; the destination of the parent axons is unknown. The intercalated cells occur along fiber bundles, which are probably afferent to them. The axons that travel among the intercalated cells give off short collaterals and boutons en passant. The sources of these fibers are not known. From the published experimental data, it is likely that they originate in the piriform and entorhinal cortices, the lateral preoptic area, lateral hypothalamus, and ventral pallidum. Axon collaterals of basolateral nucleus pyramidal cells appear to terminate among the intercalated cells. It is suggested that the intercalated cells serve as sites for integration of the output of these various areas and, in turn, communicate it to the lateral-basolateral and central amygdaloid nuclei. The intercalated cells closely resemble neurons in the corpus striatum. Thus the question is raised and discussed of whether the intercalated cells are a ventral extension of the corpus striatum.