The absence of horseradish peroxidase uptake by cerebellar afferents to the red nucleus of the cat

Horseradish peroxidase (HRP) administered to the red nucleus (RN) failed to be transported retrogradely in the axons of the cerebellorubral projection system. This negative result was independent of the anaesthesia, fixation method, injection time or survival period. The absence of HRP uptake and transport in our work reflects some uncertainty in the general usefulness of the HRP technique.     

Pulvinar-latero posterior afferents to cortical area 7 in monkeys demonstrated by horseradish peroxidase tracing technique

Summary Pulvinar-latero posterior afferents to the parietal cortical area 7 of the monkey have been demonstrated by means of horseradish peroxidase (HRP) tracing technique. Following HRP injection of area 7, labelled neurons have been found in the pulvinar medialis and the nucleus lateralis posterior. The role of these pulvinar projection fibers is discussed with reference to the visual neurons of area 7 recently recorded from.     

Periaqueductal grey projection to the ventrobasal complex in the cat: An horseradish peroxidase study

Horseradish peroxidase (HRP) was injected within the thalamic ventrobasal complex of 14 cats. The aim was to ascertain whether the periaqueductal grey matter (PAG) sends fibres to this complex. Retrogradely labelled cells were found within the PAG following HRP delivery either in the nucleus ventralis posterolateralis (VPL) or ventralis posteromedialis (VPM). PAG-VPL projection is only ipsilateral and arises mainly from lateral PAG. PAG-VPM projection is bilateral and originates from latero-ventral regions of the central grey. The hypothesis that PAG might control the activity of ventrobasal nociceptive neurones is proposed.     

Cortical afferents to the prefrontal cortex of the cat: a study with the horseradish peroxidase technique.

Following horseradish peroxidase (HRP) injections into different areas within the prefrontal cortex (PFC) of the cat, labeled neurons were found in the cingulate and insular cortex. These results demonstrate that the cat's prefrontal cortex is reached directly from these cortical regions, and that the observed cortical projections are similar to those detected in the monkey's prefrontal cortex.     

Cortical neurons projecting to the pontine nuclei in the cat. An experimental study with the horseradish peroxidase technique

Summary Horseradish peroxidase (HRP) injections in various portions of the cat pontine nuclei resulted in retrograde labeling of neurons in layer V of the ipsilateral cerebral cortex.Corticopontine neurons, pyramidal in type, have been found to be labeled in the entire cortex, confirming the previous findings of anterograde degeneration studies. Most (91%) of the labeled cells were 14–26 m in diameter (mean 19.4±4.5 m SD). Small (10–20 m) and medium (20–40 m) cells represent 51.5% and 47.7%, respectively, of the total number of the labeled neurons. The populations of the neurons of various sizes were almost identical in different cortical areas, and were different from the populations of corticoreticular and corticospinal cells.Corticopontine cells were well labeled in experimental cases of 3-days' survival time, confirming the topographical organization established previously by degeneration studies for this projection system. However, in cases of shorter survival time (20–27 h), the number of labeled neurons was very small.The relative paucity of labeled Corticopontine neurons in the sigmoid and lateral gyri is discussed with reference to other cortical descending neurons (e.g., the corticotectal, corticoreticular and corticospinal) which have hitherto been identified morphologically as well as physiologically.Abbreviations AL gyrus lateralis anterior - ASigm gyrus sigmoideus anterior - ASup gyrus suprasylvius anterior - Br.p. brachium pontis - Cor gyrus coronalis - L left - L.m. lemniscus medialis - MEct gyrus ectosylvius medius - MSup gyrus suprasylvius medius - N.dl. nucleus dorsolateralis - N.l. nucleus lateralis - N.m. nucleus medianus - N.p. nucleus peduncularis - N.pm. nucleus paramedianus - N.r.t. nucleus reticularis tegmenti pontis - N.v. nucleus ventralis - Ped corticospinal and corticopontine fibers in cerebral peduncle - PSigm gyrus sigmoideus posterior - R right     

Thalamic projecting neurons in the feline nucleus cuneatus. A combined horseradish peroxidase and high voltage electron microscopic study

Summary The retrograde transport of horseradish peroxidase (HRP) has been employed to identify thalamic projection neurons (TPN) in the feline nucleus cuneatus by means of light microscopy and high voltage electron microscopy. Forty-eight hours after injection of HRP in the contralateral ventrobasal complex of the thalamus, labelled neurons at levels caudal to the obex are concentrated in the cell clusters of the dorsal two-thirds of the nucleus. In plastic sections, labelled TPN are identified by the presence of HRP-positive granules in the perinuclear cytoplasm. TPN are typically about 25 m in diameter, have a round nucleus with a smooth contour and abundant cytoplasm. In contrast, neurons unlabelled after thalamic injection are located at the periphery of clusters of TPN. Unlabelled neurons are characterized by their fusiform shape (hence, round when encountered in cross-section), small diameter (10–15 m), a nucleus with an irregular or highly indented contour, and sparse cytoplasm.At the ultrastructural level, TPN are identified by the presence of HRP-positive, membrane-bound, dense bodies in the perinuclear cytoplasm. Furthermore, the presence of such dense bodies in cross-sections of dendrites allows their identification as processes of TPN. The perikarya of adjacent neurons in a cluster are often closely apposed and separated by an extracellular space of 20 to 25 nm. Adjacent to such sites of apposition, small boutons are often presynaptic to one or both of the neurons. The possible functional implications of such an arrangement are discussed.     

The somatotopic organization of the cat trigeminal ganglion as determined by the horseradish peroxidase technique

The somatotopic organization of the cat trigeminal ganglion has been investigated in the present study by using the horseradish peroxidase (HRP) technique. In separate animals, the corneal, supraorbital, infraorbital, inferior alveolar, or mental branches of the trigeminal nerve have been transected and then soaked in concentrated solutions of HRP. Retrogradely labeled corneal and supraorbital neurons have been found, with extensive overlap between the two cell populations, in the anteromedial region of the trigeminal ganglion. Inferior alveolar and mental neurons have been found to possess similar distributions within the posterolateral part of the trigeminal ganglion. Infraorbital cells have been localized in a central position. The cell bodies of any given nerve are found in at least minimal numbers in all dorsoventral levels of the trigeminal ganglion. However, cell bodies of origin of the supraorbital nerve and the lateral branch of the infraorbital nerve, innervating more posterior or lateral areas of the head and face, are found in greater numbers dorsally. Conversely, cell bodies of origin of the medial branch of the infraorbital nerve, the inferior alveolar nerve, and the mental nerve, supplying more rostral or intraoral areas of the orofacial region, are present in greater numbers ventrally. In contrast, corneal neurons are distributed uniformly in the dorsoventral axis. The ophthalmic and maxillary regions of the trigeminal ganglion appear to be well segregated, whereas the maxillary and mandibular regions exhibit a somewhat greater degree of overlap. Cell bodies of corneal afferent neurons range from 20 to 50 μm in diameter, whereas those of supraorbital, infraorbital, inferior alveolar and mental neurons measure from 20 to 85 μm. It is concluded from the findings of the present work that much of the cat trigeminal ganglion is organized somatotopically in not only the mediolateral axis but also in the dorsoventral axis.     

The topical organization of the afferents to the caudatoputamen of the rat. A horseradish peroxidase study

The aim of the present study was to assess (1) whether the various brain areas known to send projections to the neostriatum of the rat (neocortex, thalamus, substantia nigra, ventral tegmental area and dorsal raphe nucleus) project to all parts of this structure, and (2) whether the subcortical projections show a topical organization. For these purposes, small deposits of horseradish peroxidase were delivered by iontophoretic application, so that the whole extent of the caudatoputamen could be covered in a total of 40 rats.Labeled cortical cells were present mainly in lamina V, and showed a roughly topographical organization. Small numbers of labelled cells were observed in the basal nucleus of the amygdala after injections into the dorsal and central parts of the caudatoputamen. The cells of origin of thalamic afferents to the neostriatum were found not only in the intralaminar nuclei, but also in various other anterior, ‘midline’, and posterior nuclei (e.g. the medial part of the medial geniculate body). In the thalamostriatal projection a topical organization was demonstrated, consisting of oblique thalamic zones, which cross the borders of several thalamic nuclei and project to different parts of the neostriatum. In the substantia nigra and ventral tegmental area many retrogradely labelled cells were present. This nigrostriatal projection appears to be organized along an oblique longitudinal neostriatal axis. The nucleus raphes dorsalis was labelled most abundantly after caudal and ventrolateral injections into the caudatoputamen.It is concluded that, despite the homogeneous cytoarchitectonic structure of the caudatoputamen in the rat, this brain area is rather heterogeneous as regards its afferent connections. In fact each part of the neostriatum receives a specific and unique combination of afferents. The main changes in the input of the neostriatum appear to occur along an oblique longitudinal axis, from the most rostromedial and dorsal part to the caudolateral and ventral part. Such a topographical organization suggests that the neostriatum is likely to be involved in very complex integrative functions involving several brain areas.     

Convergence of afferent fibers from the entopeduncular nucleus and the substantia nigra pars reticulata onto single neurons in the ventromedial thalamic nucleus: an electron microscope study in the cat

Convergence of afferent fibers from the entopeduncular nucleus (EP) and the substantia nigra pars reticulata (SNr) onto single neurons in the ventromedial thalamic nucleus (VM) was shown electron microscopically in the cat. Axon terminals from the EP were anterogradely labeled with wheat germ agglutinin-horseradish peroxidase (WGA-HRP) injected into the EP. Axon terminals from the SNr were degenerated by injecting ibotenic acid into the SNr. Both WGA-HRP-labeled axon terminals and degenerating ones were found on single neuronal profiles in the VM.     

The striatonigral projection and nigrotectal neurons in the rat. A correlated light and electron microscopic study demonstrating a monosynaptic striatal input to identified nigrotectal neurons using a combined degeneration and horseradish peroxidase procedure

In a light and electron microscopic study of the substantia nigra of the rat, the distribution and morphology of nigrotectal neurons and the pattern of termination of striatonigral fibres have been examined following the placement of horseradish peroxidase injections in the superior colliculus and kainic acid lesions in the dorsal striatum. In confirmation of previous findings, nigrotectal neurons which had been identified by the retrograde transport of horseradish peroxidase from the superior colliculus had mainly medium sized somata, varied from fusiform to stellate in shape and were found in mainly ventral regions of the rostral two-thirds of the substantia nigra pars reticulata. On electron microscopic examination, single and multiple (from two to six) degenerating striatonigral boutons were found in synaptic contact with the soma, proximal mainstem dendrites and small dendrites (but mainly on small dendrites) of labelled nigrotectal and unlabelled nigral neurons in the ventral region of the pars reticulata. In addition, a small number of degenerating striatonigral boutons formed axoaxonic synapses with degenerating or normal boutons which were presynaptic to nigral dendrites. Almost all of the identified striatonigral synapses were of the symmetrical type, although a few degenerating boutons established asymmetrical synaptic contacts on unlabelled dendrites. These findings provide evidence of a monosynaptic input from the dorsal striatum to nigrotectal projection neurons in the substantia nigra and thus demonstrate the existence of a bineuronal pathway from the striatum through the substantia nigra to the superior colliculus. The possible significance of the pattern of termination of striatonigral fibres in the substantia nigra is discussed with reference to the known dendritic arborization of nigral neurons.     

Trigeminal primary afferent neurons projecting directly to the solitary nucleus in the cat: A transganglionic and retrograde horseradish peroxidase study

After applying horseradish peroxidase to peripheral branches of the trigeminal nerve in the cat, the lingual and pterygopalatine nerves were found to contain fibers which ended ipsilaterally in the rostral portions of the solitary nucleus (SN); massively in the medial and ventrolateral SN, moderately in the intermediate and interstitial SN and sparsely in the ventral SN. The rostralmost part of the SN was free from labeled terminals. After injecting the enzyme into the SN portions rostral to the area postrema, small neurons were scattered in the maxillary and mandibular divisions of the trigeminal ganglion.     

Retinal projection to the nucleus of the optic tract in the cat as revealed by retrograde transport of horseradish peroxidase

Retrograde transport of horseradish peroxidase injected iontophoretically into the nucleus of the optic tract of cats revealed that the direction-selective cells in this pretectal nucleus receive direct retinal projections from small retinal ganglion cells, the so-called gamma-cells. These cells from a horizontal band on the contralateral retina. Few labeled cells are found in the ipsilateral temporal retina. The input from the contralateral retina is 10 times more numerous than from the ipsilateral one. In both retinae the highest concentration of labeled cells is near the area centralis.     

The thalamic projection to cat visual cortex: ultrastructure of neurons identified by Golgi impregnation or retrograde horseradish peroxidase transport

Specific afferents to the primary visual cortex of the cat were identified by electron microscopy after electrolytic lesions of the lateral geniculate nucleus. Postsynaptic target neurons were marked by either Golgi impregnation or horseradish peroxidase retrogradely transported from the opposite visual cortex. The type and the location of identified neurons were determined by light microscopy before further investigation by electron microscopy. Different pyramidal neurons have relatively homogeneous ultrastructural characteristics, but non-pyramidal neurons, divisible into large and small, spiny, sparsely-spiny and non-spiny, vary in their synaptology. Thalamo-cortical afferents have been found to all neuronal types examined in layer IV and lower layer III, including horseradish peroxidase-filled callosal cells. These include pyramidal cells of layer III which receive thalamo-cortical afferents mainly on spines of apical and basal dendrites, but also on basal dendritic shafts. Layer V pyramids receive thalamo-cortical input to apical dendritic spines. All types of layer IV non-pyramidal neurons studied are contacted by geniculate terminals on dendritic spines and shafts and directly on the cell body.We conclude that input to the visual cortex is organized on both hierarchical and parallel bases.     

Topographical organization of the projections from the cerebral cortex to the head of the caudate nucleus. A horseradish peroxidase study in the cat

The topographical organization of the projections from the cerebral cortex to the head of the caudate nucleus was studied in the cat using the horseradish peroxidase method. Various amounts of horseradish peroxidase were injected into several sites of the head portion of the caudate nucleus at about the frontal level where its cross section was widest. Injections of small amounts of horseradish peroxidase retrogradely labeled neurons in rather limited cortical areas bilaterally, showing the localized organization of the projections. Neurons in the lateral portions of the ventral bank of the cruciate sulcus and in the dorsal bank (areas 4 gamma and 4 delta) were labeled after horseradish peroxidase injections into the dorsolateral part of the head of the caudate nucleus. Neurons in the intermediate portions of the ventral bank (areas 6 a delta and 6 infra fundum) were strongly labeled after dorsolateral or ventrointermediate injections, and neurons in the medial portion (area 6a beta), after dorsomedial, dorsointermediate, ventrointermediate or central injections. These findings indicate that areas 4 gamma and 4 delta project to the dorsolateral part of the caudate nucleus, areas 6a delta and 6 infra fundum to the lateral half, and area 6a beta to a more medial portion. Other findings revealed that the gyrus proreus projects to the medial part of the caudate nucleus and the anterior cingulate gyrus to the dorsal region.     

Ultrastructural and electrophysiological properties of accessory abducens nucleus motoneurones: An intracellular horseradish peroxidase study in the cat

The physiological and morphological (light and electron microscopy) properties of six retractor bulbi motoneurones were analysed using the technique of intracellular recording and intracellular labelling with horseradish peroxidase. The retractor bulbi motoneurones were identified by antidromic invasion and orthodromic responses following stimulation of trigeminal afferents were studied. Two of these motoneurones were examined ultrastructurally. Terminal boutons forming synapses with labelled soma, labelled proximal and distal dendrites were characterized. Serial sections allowed the axon hillock to be analyzed and the initial segment of a presumed motoneurone to be observed in the section where the injected motoneurone was described. The ultrastructure of unidentified elements observed in the accessory abducens nucleus is stressed.     

Identification of axon terminals of the cerebello-olivary fibers in the cat: an electron microscope study using the anterograde horseradish peroxidase method

After injection of horseradish peroxidase (HRP) into the lateral cerebellar nucleus of the cat, axon terminals containing electron-dense HRP-granules were identified contralaterally within the principal olive. These terminals contained spherical synaptic vesicles, and made asymmetrical synaptic contacts with dendritic profiles of small or medium size.     

Synaptic terminals in the dorsal lateral geniculate nucleus from neurons of the thalamic reticular nucleus: A light and electron microscope autoradiographic study

(³H)-proline was injected in the caudodorsal part (visual segment) of the thalamic reticular nucleus to study its projection to the dorsal lateral geniculate nucleus. This was done by autoradiographic tracing of anterograde axonal transport of tritium at the light- and electron microscopic level. The results of the light-microscope autoradiography show that connections of the thalamic reticular nucleus are distributed along lines of projections in the dorsal lateral geniculate nucleus, indicating a retinotopic arrangement of this projection. The results of the electron microscope autoradiography provide direct evidence that axons of cells in the thalamic reticular nucleus terminate in the dorsal lateral geniculate nucleus as synaptic boutons that contain flattened synaptic vesicles, dark mitochondria and establish symmetrical synapses with perikarya and with proximal, intermediate and distal dendrites. They do not take part in intraglomerular synapses (as boutons with pleomorphic synaptic vesicles do) and are not postsynaptic to other vesicle-containing boutons in the dorsal lateral geniculate nucleus.The present results, taken in conjunction with physiological studies that have shown postsynaptic inhibitory effects of the thalamic reticular nucleus on dorsal lateral geniculate nucleus relay cells in the rat, establish a correlation of an inhibitory synapse with the presence of flattened synaptic vesicles in the corresponding synaptic boutons. Also, the observation that thalamic reticular nucleus terminals in the dorsal lateral geniculate nucleus avoid forming synapses with boutons containing pleomorphic vesicles, believed to be synaptic processes of interneurons, is indicative that the inhibitory effects are exerted monosynaptically on geniculate relay cells.     

Identification and distribution of the spinal and hypophyseal projection neurons in the paraventricular nucleus of the rat. A light and electron microscopic study with the horseradish peroxidase method

Summary The distribution of labeled neurons in the paraventricular nucleus of the hypothalamus (PVN) was studied following injections of horseradish peroxidase (HRP) into the spinal cord (C8 to T1) or the hypophysis in the rat. Injections were also made in the spinal cord in another group of animals, which were subjected to water deprivation for a period of 3 days, and the PVN of these animals was examined with the electron microscope.Spinal projection neurons (paraventriculospinal tract, PVST, neurons) formed two groups; the dorsal and the ventral groups. They were located within the parvocellular part of the PVN and fused into one at the caudal level. The neurons of the dorsal group were well assembled whereas those of the ventral group were intermingled with paraventriculohypophyseal tract (PVHT) neurons, which were concentrated in the magnocellular part. Electron microscopic observations revealed that HRP-labeled neurons after spinal injections did not contain neurosecretory granules and that they were not affected by water deprivation. On the other hand, neurons containing a number of neurosecretory granules displayed a significant degree of dilatation of the endoplasmic reticulum as the result of water deprivation. These neurons contained no HRP granules.The present findings suggest that the PVST neurons are distinct from the PVHT neurons and that the neuronal groups of both systems form different cell columns within the nucleus.Abbreviations C caudal - D dorsal - Mgc magnocellular part - NH neurohypophysis - PVHT paraventriculohypophyseal tract - PVN paraventricular nucleus - PVST paraventriculospinal tract - R rostral - SC spinal cord - V ventral - III third ventricle