Wheat Germ-agglutinated Horseradish Peroxidase Research Articles

Nociceptive Afferents to the Premotor Neurons That Send Axons Simultaneously to the Facial and Hypoglossal Motoneurons by Means of Axon Collaterals

It is well known that the brainstem premotor neurons of the facial nucleus and hypoglossal nucleus coordinate orofacial nociceptive reflex (ONR) responses. However, whether the brainstem PNs receive the nociceptive projection directly from the caudal spinal trigeminal nucleus is still kept unclear. Our present study focuses on the distribution of premotor neurons in the ONR pathways of rats and the collateral projection of the premotor neurons which are involved in the brainstem local pathways of the orofacial nociceptive reflexes of rat. Retrograde tracer Fluoro-gold (FG) or FG/tetramethylrhodamine-dextran amine (TMR-DA) were injected into the VII or/and XII, and anterograde tracer biotinylated dextran amine (BDA) was injected into the caudal spinal trigeminal nucleus (Vc). The tracing studies indicated that FG-labeled neurons receiving BDA-labeled fibers from the Vc were mainly distributed bilaterally in the parvicellular reticular formation (PCRt), dorsal and ventral medullary reticular formation (MdD, MdV), supratrigeminal nucleus (Vsup) and parabrachial nucleus (PBN) with an ipsilateral dominance. Some FG/TMR-DA double-labeled premotor neurons, which were observed bilaterally in the PCRt, MdD, dorsal part of the MdV, peri-motor nucleus regions, contacted with BDA-labeled axonal terminals and expressed c-fos protein-like immunoreactivity which induced by subcutaneous injection of formalin into the lip. After retrograde tracer wheat germ agglutinated horseradish peroxidase (WGA-HRP) was injected into VII or XII and BDA into Vc, electron microscopic study revealed that some BDA-labeled axonal terminals made mainly asymmetric synapses on the dendritic and somatic profiles of WGA-HRP-labeled premotor neurons. These data indicate that some premotor neurons could integrate the orofacial nociceptive input from the Vc and transfer these signals simultaneously to different brainstem motonuclei by axonal collaterals.

Cytoarchitecture of the medial geniculate body and thalamic projections to the auditory cortex in the rufous horseshoe bat (Rhinolophus rouxi)

The auditory cortex in echolocating bats is one of the best studied in mammals, yet the projections of the thalamus to the different auditory cortical fields have not been systematically analyzed in any bat species. The data of the present study were collected as part of a combined investigation of physiological properties, neuroarchitecture, and chemoarchitecture as well as connectivity of cortical fields in Rhinolophus in order to establish a neuroanatomically and functionally coherent view of the auditory cortex in the horseshoe bat. This paper first describes the neuroanatomic parcellation of the medial geniculate body and then concentrates on the afferent thalamic connections with auditory cortical fields of the temporal region. Deposits of horseradish peroxidase and wheatgerm-agglutinated horseradish peroxidase were made into neurophysiologically characterized locations of temporal auditory cortical fields; i.e., the tonotopically organized primary auditory cortex, a ventral field, and a temporal subdivision of a posterior dorsal field. A clear topographic relationship between thalamic subdivisions and specific cortical areas is demonstrated. The primary auditory cortex receives topographically organized input from the central ventral medial geniculate body. The projection patterns to the temporal subdivision of the posterior dorsal field suggest that it is a "core" field, similar to the posterior fields in the cat. Projections to the ventral field arise primarily from border regions of the ventral medial geniculate body. On the whole, the organization of the medial geniculate body projections to the temporal auditory cortex is quite similar to that described in other mammals, including cat and monkey.

Trigeminal Inputs to Eyeblink Motoneurons in the Rabbit

The rabbit nictitating membrane and eyeblink response is widely used in studies of classical conditioning. Eyeblinks involve coordinated activation of the orbicularis oculi motoneurons (OOcVII) and accessory abducens motoneurons (AccVI) which close the external eyelids and nictitating membrane, respectively, and inhibition of levator palpebrae motoneurons (LPIII) whose activity raises the upper eyelid. The identification of blink interneurons that may coordinate these responses is an important step in the analysis of mechanisms supporting eyeblink conditioning as they are likely to receive convergent inputs from circuitry associated with learned as well as unlearned responses. We first investigated the distribution of OOcVII motoneurons in the facial nucleus and LPIII motoneurons in the oculomotor nucleus by retrograde tracing of wheat germ-agglutinated horseradish peroxidase (WGA–HRP) injected into the appropriate muscles. We then used an anterograde tracing method to locate trigeminal and paratrigeminal inputs to OOcVII, to AccVI nucleus, and to LPIII. Injections of WGA–HRP were placed into the principal trigeminal nucleus (Vp) and into all divisions of the spinal trigeminal nucleus. We found an area in Vp and the adjacent rostral parts of pars oralis of the spinal trigeminal nucleus that gave clear projections to OOcVII and AccVI motoneurons and adjacent to LPIII motoneurons in the contralateral oculomotor nucleus. We suggest that neurons in this premotor blink area in rabbits can coordinate learned and reflex blink responses involving the external eyelidsandthe nictitating membrane. In addition, there are direct projections from the pars interpolaris and pars caudalis of the spinal trigeminal nucleus to the facial nucleus that may mediate short latency responses of the external eyelid orbicularis oculi muscle alone.

Effects of bilateral lesions of the dorsolateral funiculi and dorsal columns at the level of the low thoracic spinal cord on the control of locomotion in the adult cat. I. Treadmill walking

1. A quantitative and longitudinal analysis of locomotion was made after bilateral lesions of the dorsolateral funiculi (DLF) and/or the dorsal columns (DC) in the lower thoracic cord (T12 or T13) in five adult cats. All cats were chronically implanted several weeks before the spinal cord lesion to permit the recording of electromyographic (EMG) activity from selected flexor and extensor muscles of the fore- and hindlimbs of each side. This allowed each cat to act as its own control when comparing the pattern and amplitude of EMG activity before and after the lesion. All experiments were also videotaped to allow an analysis of the kinematic changes before and after the lesions. Kinematic data were only analyzed for the side of the cat facing the camera; for all cats this was the left side. 2. After recovery periods of 2-5 mo, wheat-germ-agglutinated horseradish peroxidase (WGA-HRP) was injected caudal to the lesion site (normally at L2). The extent of the lesion was verified both histologically and by evaluating the number of HRP-labeled neurons in different supraspinal structures. These analyses showed that the cortico- and rubrospinal tracts (CST and RST, respectively) were completely interrupted, bilaterally, in two of five of the cats; in one of these cats the DCs were also interrupted. In the other three cats there was more variable damage, and the CST and RST were only completely interrupted on the right side of one of these animals. The DCs were completely sectioned in two of these cats. 3. During the 1st wk subsequent to the lesion, most cats had difficulty in supporting their weight and in walking. However, within 10 days all were able to walk, unaided, for extended periods on the treadmill at speeds of at least 0.35 m/s. In the two cats with the complete, bilateral DLF lesions, the animals dragged both their left and right hindpaws along the treadmill belt during the swing phase of the step cycle (paw drag) throughout the testing period of 3-5 mo. In the other three cats, paw drag in either hindlimb was only seen in the 1st 2-3 wk after the lesions, with the exception of the cat with the complete lesion of the DLF on the right side, which showed sustained paw drag in the right hindlimb throughout the testing period. 4. Significant increases in step cycle and swing duration following the lesion were observed only in the two cats with the largest lesions. In all five cats, statistical comparisons of the slopes from a linear regression analysis showed that the relationship between swing and step cycle duration was unchanged by the lesions. 5. Joint angles in the left hindlimb of the two cats with the largest lesions were generally smaller (more flexed) than in the prelesion controls. This was particularly true for the knee and ankle joints. The other three cats showed changed joint angle values for the hip, knee, and ankle only in the 1st 2-3 wk after the lesions. All of the cats, except the one with the least damage to the left DLF, exhibited increased joint excursions at the metatarsophalangeal (MTP) joint of the left limb throughout the recovery period. 6. There were sustained changes in the coupling between the hip and the knee of the left limb, together with smaller changes in the coupling of the knee and ankle in the two cats with the complete lesions of the DLF. In contrast to the prelesion controls, flexion at the ankle occurred before swing onset in these same two cats. The changes in the coordination between the hip and the knee were associated with changes in the temporal coupling between the hip flexor, sartorius (Srt), and the knee flexor, semitendinosus (St). Despite some improvement, the coupling between these two muscles never fully recovered to the prelesion values. Similar, but smaller, changes in the delay between Srt and St were also seen in two of the other three cats. (ABSTRACT TRUNCATED)

The overrepresentation of the fovea and adjacent retina in the striate cortex and dorsal lateral geniculate nucleus of the macaque monkey.

The central part of the retina, which includes the fovea, is substantially overrepresented in the topographic map of the retina in the striate cortex. We tested whether this simply reflects the uneven distribution of ganglion cells in the retina in accordance with the "principle" of peripheral scaling, or whether there is additional expansion of the fovea and adjacent retina in the retinocortical projection. Wheatgerm agglutinated horseradish peroxidase was injected into the striate cortex of three rhesus macaque monkeys so as to surround the representation of the fovea at a mean eccentricity of 8.6 degrees, and the retinae were processed histochemically to stain the retrogradely and transneuronally labelled ganglion cells which projected topographically to the injection sites. This enabled regions of the striate cortex to be related precisely to corresponding regions of the dorsal lateral geniculate nucleus and retina. Mathematical models of the distribution of ganglion cells in the retina, clipped, three-dimensional computer reconstructions of the striate cortex and lateral geniculate nucleus, and counts of neurons in the latter, were used to calculate the proportion of neurons allocated to the marked perifoveal region at each stage of projection. This was used to calculate the relative allocation of neurons to the representation of the fovea and surrounding retina among the different stages of the visual pathway. The values obtained showed that the cortical representation of the perifovea was expanded two to three times more than could be accounted for on the basis of ganglion cell topography in the retina, and that the expansion occurred both between the retina and the thalamus, and between the thalamus and the cortex. These results are inconsistent with the idea that peripheral scaling is a general principle of sensory representation in the cortex. They could also explain why many visual thresholds, including hyperacuities, cannot be accounted for by peripheral factors such as ganglion cell density.

Monosynaptic input from Leu5-enkephalin-immunoreactive terminals to vagal motor neurons in the nucleus ambiguus: comparison with the dorsal motor nucleus of the vagus.

Vagal motor neurons in the rat dorsal motor nucleus of the vagus (DMN) are known to receive direct synaptic input from enkephalin-containing terminals. We examined 1) whether the vagal motor neurons within the nucleus ambiguus (NA) also received monosynaptic input from enkephalin-immunoreactive terminals and 2), if so, whether their ultrastructural relations differed from those in the DMN. In both regions, terminals containing Leu5-enkephalin-like immunoreactivity (LE-LI) were examined in relation to motor neurons identified by retrograde transport of wheat germ-agglutinated horseradish peroxidase (WGA-HRP) applied to the cut end of the cervical vagus nerve in single sections of the medulla oblongata of adult rats. By light microscopy, the most significant overlap between varicose processes with LE-LI and WGA-HRP-containing neurons was seen in the rostral compact portion of the NA and the DMN at the level of the obex. Thus, only these regions were examined by electron microscopy. The most distinguishing ultrastructural feature of WGA-HRP-labeled neurons in the NA compared to the DMN was their higher incidence of nonsynaptic appositions with other neurons. In both the NA and the DMN, terminals with LE-LI formed primarily symmetric synapses on smaller (presumably distal) dendrites; many of these dendrites, as well as most target perikarya, contained WGA-HRP. Additionally, in the compact portion of the NA compared to the DMN 1) multiple LE-labeled terminals more frequently contacted single perikarya or dendrites and 2) single terminals with LE-LI more commonly showed two contacts or active zones and contained more abundant LE-immunoreactive large (80-100 nm) dense-core vesicles (dcvs). In contrast to small (40-50 nm), clear vesicles, which were usually aggregated near active zones, the immunoreactive dcvs were usually located near glial processes distal to these zones. These results indicate that enkephalin immunoreactivity is intensely localized to dcvs within terminals that may have direct inhibitory (symmetric synapses) actions on vagal motor neurons in both the compact portion of the NA and the DMN. Moreover, because numbers of dcvs and active zones have been equated with synaptic strength, our findings suggest enhanced potencies of enkephalin-immunoreactive terminals in the compact portion of the NA. Our findings support a prominent role for enkephalin in the coordinated activity of esophageal motor neurons located in the compact portion of the NA.

Inhibitory synaptic inputs to the oculomotor nucleus from vestibulo-ocular-reflex-related nuclei in the rabbit.

Studies of the pathways involved in the vestibulo-ocular reflex have suggested that the projection from the superior vestibular nucleus to the ipsilateral oculomotor nucleus is inhibitory, whereas the medial vestibular nucleus, the abducens nucleus and the contralateral superior vestibular nucleus most likely exert excitatory effects on oculomotor neurons. In order to determine directly the termination pattern and the neurotransmitter of these afferents, we studied their input to the oculomotor nucleus in the rabbit at the light microscopic level with the use of anterograde tracing of Phaseolus vulgaris-leucoagglutinin combined with retrograde tracing of horseradish peroxidase from the extraocular muscles, and at the ultrastructural level with the use of anterograde tracing of wheatgerm-agglutinated horseradish peroxidase combined with GABA and glycine postembedding immunocytochemistry. The general ultrastructural characteristics of the neuropil and the types of boutons observed in the rabbit oculomotor nuclei are in general agreement with the descriptions for the oculomotor complex of other mammals. The superior vestibular nucleus projected bilaterally to the superior rectus and inferior oblique subdivisions, and ipsilaterally to the inferior rectus and medial rectus subdivision; the medial vestibular nucleus projected bilaterally to the medial rectus, inferior oblique, inferior rectus and superior rectus subdivisions with a strong contralateral predominance. The abducens nucleus projected contralaterally to the medial rectus subdivision. More than 90% of all the anterogradely labeled terminals from the ipsilateral superior vestibular nucleus were GABAergic. These terminals were characterized by flattened vesicles and symmetric synapses, and they contacted somata, as well as proximal and distal dendrites of motoneurons. All terminals derived from the medial vestibular nucleus the abducens nucleus and the contralateral superior vestibular nucleus were non-GABAergic. These non-GABAergic terminals showed spherical vesicles and asymmetric synapses, and they contacted predominantly distal dendrites. None of the anterogradely labeled terminals from the studied vestibular nuclei or abducens nucleus were glycinergic. The present study provides the first direct anatomical evidence that most, if not all, of the synaptic input from the superior vestibular nucleus to the ipsilateral oculomotor nucleus is GABAergic, and that the medial rectus subdivision is included in the termination area. Furthermore, it confirms that the projections from the medial vestibular nucleus, the abducens nucleus and the contralateral superior vestibular nucleus are exclusively non-GABAergic.

Organization of the colliculo-suprageniculate pathway in the cat: a wheat germ agglutinin-horseradish peroxidase study.

A wheat germ-agglutinated horseradish peroxidase (WGA-HRP) tracing technique was used to label the cell bodies of neurons in the superior colliculus that send projections into the visually sensitive region of the suprageniculate nucleus (Sg) in the feline thalamus. After determination of the position of the Sg by detecting characteristic single-unit responses to moving visual stimuli, WGA-HRP was injected into the Sg in five pentobarbital-anesthetized cats. The animals were than sacrificed, and serial frozen sections of the midbrain were processed for the demonstration of peroxidase activity. A total of 2,736 WGA-HRP-stained neurons were located within the ipsilateral superior colliculus (SC), and a few labeled cells were consistently found bilaterally in the external nuclei of the inferior colliculus. In each cat, a small but significant fraction of the labeled cells were encountered contralateral to the injection. Medial SC neurons tended to project to the posterior Sg, and lateral SC neurons tended to project to more rostral Sg. However, labeled cells were distributed homogeneously along the rostrocaudal extent of the SC, indicating the absence of a well defined topographic relationship. Nor was the Sg injection site location related to the laminar distribution of SC projection neurons. In all cases, the majority of the labeled cells were found in layer IV (49.0%), with fewer cells in layers III (17.5%) layer V (20.0%), and layer VI (11.8%). No labeled cells were located in layer I, although a few were located in the deep part of layer II. Five types of SC projection cells were distinguished morphologically. Of the 258 labeled cells that could be characterized, 25% were stellate cells, 25% vertical cells, 20% granular cells, 17% triangular cells, and 12% horizontal cells. The average diameter of 226 cells ranged between 8 and 47 microns. We conclude that a mixed population of SC cells projects to the Sg; the morphological heterogeneity and the distribution of these cells suggests that several functionally different pathways may be involved in the colliculothalamic pathway and in the processing of visual input in the SC.

Connections to cerebellar cortex (Larsell's HVI) in the rabbit: a WGA-HRP study with implications for classical eyeblink conditioning.

Conditioned eyeblink responses are presumably learned in the cerebellum and relayed to motoneurons by way of the red nucleus. Projections from the red nucleus to cerebellar cortex (Larsell's lobule HVI) could be important for shaping temporally adaptive features of the conditioned response. Rabbits that had pipettes containing wheat germ agglutinated horseradish peroxidase (WGA-HRP) implanted unilaterally into HVI showed retrograde labeling of neurons within subregions of the contralateral red nucleus implicated in eyeblink conditioning by lesioning and recording studies. Retrogradely labeled neurons were also observed in the pontine nuclei, inferior olive, and spinal trigeminal nucleus pars oralis. Projections to HVI provide a possible neural substrate for implementing time-derivative computational models of learning in the cerebellum. Time-derivative models are capable of describing the timing and topography of conditioned responses.

Olivary projecting neurons in the nucleus of Darkschewitsch in the cat receive excitatory monosynaptic input from the cerebellar nuclei

The direct projection from the cerebellar nuclei to the inferior olive is GABAergic. In the present study, we investigated the projection from the cerebellar nuclei to the mesodiencephalic junction which is known to provide an excitatory projection to the inferior olive. The mesodiencephalic junction was studied in cat following anterograde transport of wheatgerm agglutinated horseradish peroxidase from the cerebellar nuclei in combination with: (1) retrograde transport of gold-lectin conjugate from the inferior olive; and (2) postembedding GABA-immunocytochemistry. Light microscopic analysis revealed that overlap of the anterograde and retrograde labeling was especially prominent in the nucleus of Darkschewitsch. Electron microscopic examination of this area showed: (1) that many cerebellar terminals made synaptic contacts with neurons that project to the inferior olive; (2) that virtually all cerebellar terminals were non-GABAergic and displayed an excitatory morphology; and (3) olivary projecting neurons were non-GABAergic. It is concluded that the indirect cerebellar projection to the inferior olive via the nucleus of Darkschewitsch is disynaptic and excitatory.

Reinnervation by axon collaterals from single facial motoneurons to multiple muscle targets following axotomy in the adult guinea pig.

To study the process of recovery from facial palsy experimentally, the location of cranial motoneurons supplying the posterior belly of the digastric muscle (PDG) and the extratemporal portion of the facial nerve trunk was examined in a double-labeling paradigm using two retrograde tracers in the adult guinea pig of which the facial nerve had been surgically injured. In different stages after the induced facial palsy had recovered functionally (4-13 weeks after the surgical operation), wheat germ-agglutinated horseradish peroxidase (WGA-HRP) was injected into the PDG and Fluoro-Ruby (FR) was applied to the proximal cut end of the extratemporal portion of the facial nerve trunk. Distribution of neurons retrogradely labeled with WGA-HRP and/or FR was plotted in the brainstem and compared with that of the controls. In the intact cases, HRP-labeled neurons were restrictedly seen in the accessory facial nucleus (Acs7), while FR-labeled neurons were found within the main facial nucleus (FMN). In the axotomized cases: (1) HRP-labeled neurons were seen diffusely in the Acs7 as well as in the FMN, where normal myotopical representation no longer seemed to be maintained. (2) FR-labeled neurons were also observed diffusely in the FMN and the Acs7. (3) A considerable number of neurons were doubly labeled with WGA-HRP and FR in both the Acs and the FMN in cases with shorter survival periods (4-7 weeks), but not in cases with longer survival periods (12-13 weeks). Thus, new findings show that connections are temporarily maintained by single, facial motoneurons with axon collaterals to multiple muscle targets in adult mammals.(ABSTRACT TRUNCATED AT 250 WORDS)

Ultrastructure and synaptic associations of auditory thalamo-amygdala projections in the rat

Projections from the acoustic thalamus to the lateral nucleus of the amygdala (AL) have been implicated in the formation of emotional memories. In order to begin elucidating the cellular basis of emotional learning in this pathway, the ultrastructure and synaptic associations of acoustic thalamus efferents terminating in AL were studied using wheat-germ agglutinated horseradish peroxidase (WGA-HRP) and Phaseolus vulgaris Leucoagglutinin (Pha-L) as ultrastructural anterograde axonal markers. The tracers were injected into those areas of the thalamus (medial division of the medial geniculate body and posterior intralaminar nucleus, MGM/PIN) known both to project to AL and to receive afferents from the inferior colliculus. Terminals labeled with WGA-HRP or Pha-L in AL contained mitochondria and many small, round clear vesicles and 0-3 large, dense-core vesicles. Most labeled terminals formed asymmetric synapses on unlabeled dendrites; of these the majority were on dendritic spines. These data demonstrate that projections from the acoustic thalamus form synapses in AL and provide the first characterization of the ultrastructure and synaptic associations of sensory afferent projections to the amygdala.

Projection from the external pallidum to the reticular thalamic nucleus in the squirrel monkey

Small injections of biocytin in the external segment of the pallidum (GPe) of the squirrel monkey ( Saimiri sciureus) led to anterograde labeling of fibers in the thalamic reticular nucleus (NRT). These fibers reached NRT by coursing along the ventral tipp of the internal capsule or by directly piercing the internal capsule more dorsally. They arborized profusely within the entire rostrocaudal extent of the nucleus. Within NRT, biocytin-labeled fibers were long, slightly varicosed, and emitted numerous short collaterals whose terminal portions consisted of clusters of large varicosities. Some of these varicosities were closely apposed to cell bodies and proximal dendrites of NRT neurons. Small injections of wheat germ-agglutinated horseradish peroxidase in the rostral pole of NRT led to retrograde cell labeling within the entire rostrocaudal extent of GPe. These retrogradely-labeled cells did not display immunoreactivity for choline acetyltransferase. Hence, beside the well-established projection from the internal pallidum to the thalamus, our findings support the existence of another pallidothalamic projection whereby GPe neurons could exert a powerful influence upon the thalamocortical neurons via a relay in NRT.

Differential organization of crossed and uncrossed projections from the superior olive to the inferior colliculus in the mole

Projections from the medial (MSO) and lateral (LSO) superior olivary nuclei to the inferior colliculus (IC) were examined in the mole. In each mole, Fluoro-Gold was injected into one IC and wheat germ-agglutinated horseradish peroxidase (WGA-HRP) into the other IC; most MSO neurons were double-labeled bilaterally, while most LSO neurons were single-labeled bilaterally. The results indicate that the bilateral projections from the MSO and LSO to the IC are mainly established by single MSO neurons with axon collaterals to the IC on both sides, and by two separate populations of LSO neurons with axons to the contralateral or ipsilateral IC.

The topographical projection of the nucleus isthmi pars parvocellularis (Ipc) onto the tectum opticum in the pigeon

The projection of the nucleus isthmi pars parvocellularis (Ipc) onto the tectum opticum was studied with tectal injections of three different fluorescent tracers and wheat germ-agglutinated horseradish peroxidase (WGA-HRP) in 21 pigeons. In frontal sections obtained, the relationship betwee position and size of the tectal injection and the location of the retrogradely labelled Ipc neurons were analyzed. The results reveal a topographically organized system with reciprocal projections in which the dorsoventral axis of the tectum receives afferents from the lateromedial axis of the Ipc. The caudorostral tectal axis is innervated by neurons situated along the caudorostral axis of the Ipc. Specific points on the tectal surface do not correspond to single points within the Ipc, but to column-like strings of Ipc neurons extending through the dorsoventral aspect of the nucleus.

Commissural connections between the auditory cortices of the rat

The pattern of commissural connections of the rat auditory cortex (AC) was investigated with injections of wheat germ agglutinated horseradish peroxidase into the AC. Homotopic and heterotopic patches of neurons were retrogradely labeled in the contralateral hemisphere. Each injection labeled neurons at the corresponding contralateral site, i.e. the homotopic site. In addition, retrogradely labeled neurons were found at non-corresponding locations in contralateral AC, i.e. at heterotopic locations. The pattern of heterotopic labeling changed systematically with the injections. Mapping rules were established that led to the parcellation of areas 41 and 36 into 6 fields. Four fields were defined in Krieg's area 41 (primary AC) and two fields in Krieg's area 36 (secondary AC). In area 41 the heterotopic connection is not reciprocal; in area 36, however, heterotopic projections are organized reciprocally. Contrary to the visual cortex, homotopic and heterotopic projection neurons were equally distributed across the cortical laminae. With double-label experiments it could be shown that a considerable number of the neurons in area 41 bifurcate and project to homotopic as well as to heterotopic sites in the contralateral hemisphere. We conclude that in the AC there are several subtypes of neurons projecting to the contralateral hemisphere; it would be of interest whether these anatomical differences are manifested by physiological differences.

Laminar organization of the substantia nigra pars reticulata in the cat

Using a semihorizontal section plane tangential to the ventral surface of the cerebral peduncle, the authors re-examined cyto-, myelo- and dendroarchitecture, acetylcholinesterase activity, afferent fibers, and efferent projection neurons of the substantia nigra pars reticulata. In the semihorizontal section plane, the substantia nigra pars reticulata was a disc-shaped nucleus and contained two to three myelinated fiber bundles running from anteromedial to posterolateral. Bands of high acetylcholinesterase activity existed parallel to the anteromedial-posterolateral direction. The Golgi silver impregnation study revealed that many nigral neurons extended their varicose dendrites anteromedially and posterolaterally. In cases with injections of wheat germ agglutinated horseradish peroxidase into the neostriatum or injections of tritiated leucine into the subthalamic nucleus, anterogradely labeled afferent fibers and axon terminals in the substantia nigra pars reticulata were organized into bands in the same anteromedial-posterolateral direction. In cases with injections of wheat germ agglutinated horseradish peroxidase into either the superior colliculus, the pedunculopontine tegmental nucleus or the ventromedial nucleus of the thalamus, retrogradely labeled neurons were also clustered along the anteromedial-posterolateral direction with their dendrites extending anteromedially and posterolaterally. The present findings strongly suggest that the substantia nigra pars reticulata has a laminar organization.

Secondary vestibulocerebellar mossy fiber projection to the caudal vermis in the rabbit

The vestibulocerebellar projection in the rabbit has been investigated by the anterograde axonal transport of tritiated leucine, wheat germ-agglutinated horseradish peroxidase, and Phaseolus vulgaris leucoagglutinin. Mossy fiber terminals originating from all vestibular nuclei, with the exception of the lateral vestibular nucleus of Deiters, were found bilaterally in the lobules X, IX, and VIII of the caudal vermis, without a clear difference in laterality. Most of the vestibular mossy fiber terminals in the caudal vermis originated in the superior and caudal medial vestibular nuclei. Application of the different tracers led to similar results. The labeled terminals were always most numerous in the lobules X and IXd. Small to moderate numbers of mossy fiber terminals were found in the lobules IXa, b, c, and lobule VIII. The greatest change in the density of terminals occurred in most cases around the apex of lobule IXd and not in the depth of the posterolateral fissure between lobules X and IX. The mossy fiber terminals were not distributed equally over the cortex but showed a preference for the proximal parts of the individual lobules. In all experiments, the terminals exhibited a certain degree of clustering in the mediolateral direction, but the clusters were not arranged in longitudinal zones continuous over successive folia.

Development of the spinal projections of the nucleus paraventricularis hypothalami of the rat: an intra-uterine WGA-HRP study

The development of the spinal projections of the nucleus paraventricularis hypothalami (PVH) was studied by injecting wheatgerm-agglutinated horseradish peroxidase into the upper thoracic spinal cord of fetal rats at developmental ages ranging from embryonic day 16 (E16) to E21, and into the thoracic spinal cord of rat pups and adult rats. At E18 the first retrogradely labeled cells are seen in the PVH. These cells are located dorsally in the PVH, adjacent to the ventricular matrix. At E20 a more ventrally located group of retrogradely labeled cells appears. This ventral group joins caudally with the dorsal group of labeled cells. The dorsal group of labeled cells is closely packed, while the ventral group of labeled cells is intermingled with unlabeled cells. After E20 this spatial configuration remains essentially constant. Therefore descending projections from all spinal projecting cell groups in the PVH have reached lower cervical spinal cord levels at E20.

Striped pattern of labeling in ground squirrel superior colliculus following intraocular HRP injections

In the 13-lined ground squirrel ( Citellus tridecemlineatus), intravitreal injections of horseradish peroxidase (HRP) or wheat germ-agglutinated HRP (WGA-HRP) tracers produced a striped pattern of labeling in the contralateral superficial gray (SGS) of the superior colliculus in most, but not all, cases. The stripes extended dorsoventrally throughout the SGS in a rostral-medial to caudal-lateral orientation. In general, the striped labeling was more distinct and was observed more frequently with the HRP, rather than the WGA-HRP, tracer.