HRP-labeled Cells Research Articles

Foxp-Mediated Suppression of N-Cadherin Regulates Neuroepithelial Character and Progenitor Maintenance in the CNS

Neuroepithelial attachments at adherens junctions are essential for the self-renewal of neural stem and progenitor cells and the polarized organization of the developing central nervous system. The balance between stem cell maintenance and differentiation depends on the precise assembly and disassembly of these adhesive contacts, but the gene regulatory mechanisms orchestrating this process are not known. Here, we demonstrate that two Forkhead transcription factors, Foxp2 and Foxp4, are progressively expressed upon neural differentiation in the spinal cord. Elevated expression of either Foxp represses the expression of a key component of adherens junctions, N-cadherin, and promotes the detachment of differentiating neurons from the neuroepithelium. Conversely, inactivation of Foxp2 and Foxp4 function in both chick and mouse results in a spectrum of neural tube defects associated with neuroepithelial disorganization and enhanced progenitor maintenance. Together, these data reveal a Foxp-based transcriptional mechanism that regulates the integrity and cytoarchitecture of neuroepithelial progenitors.

The neuroanatomical and ultrastructural organization of statocyst hair cells in the pond snail,Lymnaea stagnalis

The ultrastructure, neuroanatomy and central projection patterns, including the intercellular connections of the statocyst hair cells of the pond snail, Lymnaea stagnalis, were studied, applying different intra- and extracellular cellular staining techniques combined with correlative light- and electron microscopy. Based on the ultrastructure different hair cells could be distinguished according to their vesicle and granule content, meanwhile the general organization of the sensory neurons was rather uniform, showing clearly separated perinuclear and "vesicular" cytoplasmic regions. Following intra- and extracellular labeling with fluorescence dyes or HRP a typical, local arborization of the hair cells was demonstrated in the cerebral ganglion neuropil, indicating a limited input-output system connected to the process of gravireception. Correlative light- and electron microscopy of HRP-labeled hair cells revealed both axo-somatic and axo-axonic output contacts of hair cell varicosities, and input on sensory axons located far from the terminal arborizations. Our findings suggest (i) a versatile ultrastructural background of hair cells corresponding possibly to processing different gravireceptive information, and (ii) the synaptic (or non-synaptic) influence of gravireception at different anatomical (terminal, axonal and cell body) levels when processed centrally. The results may also serve as a functional morphological background for previously obtained physiological and behavioral observations.

Bilateral innervation of syringeal muscles by the hypoglossal nucleus in the jungle crow (Corvus macrorhynchos)

Bird vocalizations are produced by contractions of syringeal muscles, which are controlled by the hypoglossal nucleus. In oscines, syringeal muscles are controlled by the hypoglossal nucleus ipsilaterally, whereas syringeal innervation is bilateral in non-oscines. We have determined the course of hypoglossal nerves in the jungle crow Corvus macrorhynchos. Our results indicate a cross-over of the hypoglossal nerve from the left side to the right side on the trachea 7 mm rostral to the Musculus sternotrachealis. We also investigated the innervation of the syringeal muscles of jungle crows from the hypoglossal nucleus using the horseradish peroxidase (HRP) method. After HRP was injected into the syringeal muscles on each side, HRP-labeled cells were found bilaterally in the hypoglossal nerve. These results suggest that the syringeal muscles of jungle crows are innervated bilaterally from the hypoglossal nucleus, although these birds are categorized as oscines.

Changes of Cervical Dorsal Root Ganglia Induced by Compression Injury and Decompression Procedure: A Novel Rat Model of Cervical Radiculoneuropathy

Our study aimed to establish a model of compression injury of cervical dorsal root ganglia (DRG) in the rat and to investigate the pathological changes following compression injury and decompression procedures. Thirty rats were divided into three groups: control group receiving sham surgery, compression group undergoing surgery to place a micro-silica gel on C6 DRG, and decompression group with subsequent decompression procedure. The samples harvested from the different groups were examined with light microscopy, ultrastructural analysis, and horseradish peroxidase (HRP) retrograde tracing techniques. Apoptosis of DRG neurons was demonstrated with TUNEL staining. Changes in PGE2 and PLA2 in DRG neurons were detected with enzyme-linked immunosorbent assay (ELISA). Local expression of vascular endothelial growth factor (VEGF) was monitored with immunohistochemistry. DRG neurons in the compression group became swollen with vacuolar changes in cytoplasm. Decompression procedure partially ameliorated the resultant compression pathology. Ultrastructural examination showed a large number of swollen vacuoles, demyelinated nerve root fibers, absence of Schwann cells, and proliferation in the surrounding connective tissues in the compression group. Compared to the control group, the compression group showed a significant decrease in the number of the HRP-labeled cells and a significant increase in levels of PGE2 and PLA2, in the expression of VEGF protein, and in the number of apoptotic DRG neurons. These findings demonstrate that compression results in local inflammation, followed by increased apoptosis and upregulation of VEGF. We conclude that such a model provides a tool to study the pathogenesis and treatment of cervical radiculoneuropathy.

Cerebellar connections to the rostral reticular nucleus of the thalamus in the rat.

We studied the cerebellar connections to the reticular nucleus thalamus (RNT) by means of retrograde axonal transport of horseradish peroxidase (HRP) in the rat. Specific HRP pressure injections to the rostral RNT (1.6-1.8 mm caudal to bregma) resulted in retrograde labelling of neurones in the cerebellar nuclei. The rostral RNT showed specific topographical organization of its cerebellar connections. Microinjections into the rostral RNT, 1.6 mm caudal to bregma, produced numerous HRP-labelled neurones within the anterior interposed (emboliform nucleus) and scarce HRP-labelled neurones within the lateral (dentate nucleus) cerebellar nuclei, whereas injections into the rostral RNT, 1.8 mm caudal to bregma, produced numerous HRP-labelled neurones within the posterior interposed (globose nucleus) and scarce lightly HRP-labelled neurones within the lateral (dentate nucleus) cerebellar nuclei. Cerebellar connections with the rostral RNT were exclusively ipsilateral to the injection site. No HRP-labelled cells were detected in the medial (fastigial nucleus) cerebellar nucleus. The cerebellar connections reach the RNT via the superior cerebellar peduncle. By contrast, HRP injections into the anterior, posterior interposed and lateral cerebellar nuclei produced no labelled cells within the RNT. This study demonstrates the existence of direct cerebello-RNT but not RNT-cerebellar connections. The presence of the cerebello-RNT connections introduces a new route through which the cerebellum may influence RNT and thus cerebral cortical activity.

The distribution of respiration-related and swallowing-related motoneurons innervating the rat genioglossus muscle

The present study was an attempt to identify the location of genioglossal respiratory and swallowing motoneuron cell bodies within the hypoglossal (XII) nucleus using both electrophysiological and morphological studies. The genioglossus muscle is innervated by the genioglossal branch of the medial XII nerve. At the entrance to the muscle, the genioglossal branch divides in the directions of the mandible and tongue. Five of five rats displayed both respiratory-related and swallowing-related bursts in the medial XII branch towards the mandible. All five rats also displayed swallowing-related bursts in the medial XII branch towards the tongue. In addition, horseradish peroxidase conjugated to wheatgerm agglutinin (HRP:WGA) was injected into the proximal cut ends of each branch. When HRP:WGA was injected into the branch in the direction of the mandible, HRP-labeled cells were detected in the lateral region of the ventromedial subnucleus in the XII nucleus, extending from 0.7 to 1.2 mm rostral to the obex. On the other hand, after injection into the branch in the direction of the mandible, HRP-labeled cells were detected in the ventromedial subnucleus of the XII nucleus, extending from 0.3 to 1.2 mm rostral to the obex. These results provide evidence that genioglossal respiration-related and swallowing-related motoneurons are located in different portions within the ventromedial subnucleus of the XII nucleus.

A Quantitative Analysis of Cells in the Ganglion Cell Layer of the Chick Retina

This study investigated the organization of cells in the ganglion cell layer (GCL) using Nissl staining, retrograde cell degeneration with axotomy of the optic nerve, and retrograde cell labeling by injections of horseradish peroxidase (HRP) into the optic nerve of chicks (posthatching day 1 and 8, P-1 and P-8). The total number of cells in the GCL was 6.1 × 10⁶ (P-1) and 4.9 × 10⁶ (P-8), and the cell density was 14,300 cells/mm² (P-1) and 10,400 cells/ mm² (P-8) on average. Two high-density areas, the central area (CA) and the dorsal area (DA), were observed in the central and dorsal retinas in both P-1 (22,000 cells/mm² in CA, 19,000 cells/mm² in DA) and P-8 chicks (19,000 cells/mm² in CA, 12,800 cells/mm² in DA). The cell densities in the temporal periphery (TP) and the nasal (NP) peripheral retinas were 7,800 cells/mm² and 12,500 cells/mm², respectively, in P-1 and 5,000 cells/ mm² and 8,000 cells/mm², respectively, in P-8 chicks. The cell density in the temporal periphery was 35% (P-8) lower than in the nasal periphery in both P-1 and P-8 chicks. Thirty percent (1.9 × 10⁶ cells in P-1) of the total cells in the GCL were resistant to axotomy of the optic nerve. The distribution of the axotomy-resistant cells showed two high-density areas in the central and dorsal retinas, corresponding to the CA (5,800 cells/mm²) and the DA (3,200 cells/mm²). These cells also exhibited a center-peripheral increase (2,200 cells/mm² in the TP) in P-1 chicks, but the high-density area was not found in the dorsal retina of P-8 chicks. From these data and the HRP study, the number of presumptive ganglion cells in P-8 chicks was estimated to be 4 × 10⁶ (8,600 cells/mm² on average), and the density in each area was 13,500 (CA), 10,200 (DA), and 4,300 (TP) cells/mm². The peripheral/ center ratios of the density of ganglion cells were significantly different along the nasotemporal and dorsoventral axes. The density of ganglion cells decreased more rapidly toward the temporal periphery (TP/CA ratio: 0.47 in P-1 and 0.32 in P-8) than toward the nasal periphery (NP/CA ratio: 0.67 in P-1 and 0.52 in P-8). In contrast, there was no significant difference in the peripheral/center ratios between the dorsal retina (DP/CA ratio: 0.6 in P-1 and 0.56 in P-8) and ventral retina (VP/CA ratio: 0.58 in P-1 and 0.51 in P-8). A small peak in the density of the presumptive ganglion cells was detected in the dorsal retina of both P-1 chicks (10,800 cells/mm²) and P-8 chicks (10,200 cells/mm²). The HRP-labeled cells were small in the CA (M ± SD: 35.7 ± 9.1 μm²) and DA (40.0 ± 11.3 μm²), and their sizes increased toward the periphery (63.4 ± 29.7 μm² in the TP) accompanied by a decrease in the cell density. However, the axotomy-resistant cells did not significantly increase in size toward the peripheral retina (12.2 ± 2.2 μm² in the CA, 15.2 ± 3.2 μm² in the DA, 15.1 ± 3.8 μm² in the TP). The characteristic distribution of ganglion cells could be related to visual behavior based upon the specialization of avian visual fields.

Origin of primary sensory neurons innervating the buccal stretch receptor.

The primary sensory neurons innervating mechanoreceptors in oro-facial regions have their cell bodies in either the trigeminal ganglion or the mesencephalic nucleus of the trigeminal nerve. The buccal stretch receptor (BSR), a type of mechanoreceptor in the jaw of rodents, has recently been recognized as signaling the position of the mandible. The location of the primary afferent neurons innervating this receptor is unknown. To investigate the cell bodies of the BSR afferent neurons in rats, we applied wheat germ agglutinin-horseradish peroxidase (WGA-HRP) to the proximal stump of the severed nerve branch of the buccal nerve that supplied the BSR. HRP-labeled cell bodies were observed in the posterolateral portion of the ipsilateral trigeminal ganglion. None was found in the contralateral trigeminal ganglion or in the brainstem. All labeled cell bodies were oval or round and closely resembled pseudo-unipolar neurons. The mean diameter of the labeled somata ranged between 25.5 and 52.5 microm, with small (< or = 30 microm), medium (from 31 to 40 microm), and large somata (> or = 41 microm) accounting for 8.8%, 54.9%, and 36.3%, respectively. Among the myelinated nerve fibers in the branch in which WGA-HRP was applied, 78.5% terminated in the BSR and had larger fiber diameters than the rest, indicating that most of the medium and large HRP-labeled cell bodies were BSR afferents. From these results and the ontogenetic origin of this receptor, it is suggested that the BSR differentiated from the mechanoreceptors in the oral mucosa or the fascia of masticatory muscles.

Origin of Innervation in the Feline Ophthalmic Artery

In this study, we investigate the innervation to the feline ophthalmic artery by the horseradish peroxidase (HRP) tracing method. Five adult cats with body weights ranging between 2.0 and 3.0 kg were used. Under microscopic dissection, the ophthalmic artery was identified and isolated. A gelfoam (Upjohn Co.), 1 × 3 mm in size, containing 0.1 ml of HRP was applied to the prepared artery segment for 2 h. The cat was sacrificed 3 days later. The trigeminal, stellate, superior cervical, middle cervical and nodal ganglia, and oculomotor, trochlear and abducens nuclei were removed, sectioned and stained for HRP-positive cells. HRP-labeled neurons were found in the ipsilateral trigeminal (TRG) and superior cervical ganglia (SCG). The middle cervical, stellate, Edinger-Westphal, trochlear and abducens nuclei were all deemed negative for HRP-labeled cells. In the TRG, HRP-labeled neurons ranged from 21 to 250 (mean ± SE = 93.8 ± 42.5/ganglion). The labeled neurons were distributed primarily in the ophthalmic branch. In the SCG, the HRP-labeled neurons were distributed evenly in the ganglion, ranging from 6 to 180 (mean ± SE = 91.6 ± 31.5/ganglion). Two additional cats having received a sham operation revealed a negative finding. The feline ophthalmic artery is innervated by the ipsilateral TRG and SCG. Such innervation may play a role in regulating blood flow to the optic nerve.

Differential effects of autologous peripheral nerve grafts to the corpus striatum of adult rats on the regeneration of axons of striatal and nigral neurons and on the expression of GAP-43 and the cell adhesion molecules N-CAM and L1

A segment of tibial nerve was autografted to the right corpus striatum of deeply anesthetized adult rats; the distal graft was left beneath the scalp. Horseradish peroxidase (HRP) conjugates were injected into the distal graft after 2-30 weeks, and the animals were killed 2-3 days later. Small numbers of neostriatal perikarya were HRP labeled at all survival times; most were large (ca. 20 microm in diameter), and many contained acetycholine esterase (AChE). Many more neurons were labelled in the substantia nigra pars compacta (SNpc) 4 weeks or more after grafting. When the graft encroached on the globus pallidus, numerous pallidal neurons, most of them AChE positive, were also labeled. Nigrostriatal neurons, a population of pallidal cholinergic neurons, and a subclass (or classes) of neostriatal neurons, including cholinergic interneurons, thus can be classified as central nervous system (CNS) neurons with a relatively strong regenerative response. In a second experimental series, animals were killed 1-4 weeks after grafting, and sections were probed for the expression of mRNAs encoding growth-associated protein 43 (GAP-43) and the cell adhesion molecules N-CAM and L1. Subpopulations of mostly large neurons scattered throughout the neostriatum gave moderate signals for GAP-43 and N-CAM mRNAs and a stronger signal for L1 mRNAs. Most SNpc neurons were strongly labeled with all three probes. Neostriatal grafts had no apparent effect on the expression of any of the mRNAs in the SNpc or on L1 and N-CAM mRNAs in the striatum. However, GAP-43 mRNA levels were increased in a few, mainly large neostriatal neurons around the graft tip, resembling the HRP-labeled cells. In contrast, previous work has shown upregulation (from an undetectable level) of GAP-43 and L1 mRNAs in neurons regenerating axons into grafts placed in the thalamus and cerebellum. Thus, GAP-43 and L1 mRNA expression, but not necessarily marked upregulation, may correlate with, and be intrinsic determinants of, the ability of CNS neurons to regenerate their axons.

Identification of Respiratory Neurons Regenerating Axons into Peripheral Nerve Grafts in Adult Rats

The present aimed to identify the origin of medullary and upper cervical respiratory neurons regenerating their axons into the peripheral nerve grafts in adult rats. We employed an antidromic activation technique and a retrograde horseradish peroxidase (HRP) tracing method for determining the origins of the regenerating axons. Autologous segments of the common peroneal nerve were successfully implanted ventrolaterally into the proximal cut end of the C2 spinal cord hemisection (n = 24). Two to 5 months after implantation, spontaneous multi-unit discharges were recorded in all grafts; respiratory-related discharges in 21 (87.5%) grafts; and non-respiratory discharges in 3 (12.5%) grafts. The respiratory discharge patterns were similar to those for normal respiratory efferent neurons in rats. After the recordings were completed, 5 of 182 respiratory units explored in the medullary ventral respiratory cell group (VRG) neurons were antidromically activated by electrical stimulation of the grafts (n = 15). The estimated axonal conduction velocities ranged from 5.6 to 7.4 (mean 6.7) m/s. Retrograde horseradish peroxidase (HRP) labelings applied to the distal cut end of the grafts (n = 13) revealed that HRP-labeled cells were located, predominantly ipsilaterally, in the brainstem up to 5-10 mm distant from the implanted site. Some HRP-labeled cells were observed in the region of the nucleus ambiguus where many respiratory neurons exist. These results demonstrate that peripheral nerve grafts implanted in the C2 segment can induce axonal regeneration of medullary VRG neurons conveying funcional efferent signals.

Relationship between Behavioral Recovery from Unilateral 6-OHDA Lesion of the Substania Nigra and Changes in the Tuberomammillary-Striatal Projection as Measured by HRP-Labelling

The tuberomammillary nucleus (TM) is the only known neuronal cell group in the brain which contains histamine. There is evidence for a reciprocal relationship between the effects of histamine and dopamine on behavior. A unilateral lesion of dopamine cells in the Substantia Nigra (SN) results in asymmetrical behavior, and recovery from this asymmetry was previously found to be associated with asymmetrical changes in the nigro-striatal projection as determined by anatomical tracing technique. In view of the apparent opposing interaction between the TM and the SN, we examined whether the tuberomammillary-striatal projection also shows changes after a unilateral lesion of the SN. Rats had 6-OHDA injected into one substantia nigra and were tested for behavioral asymmetry over a 15 day period. Horseradish peroxidase (HRP) was then deposited in the caudate-putamen (CPU) ipsilateral to the lesion. Controls devoid of a lesion had the tracer deposited in either the right of left CPU. More HRP-labeled cells were found in the anterior part of TM ipsilateral to the lesion in the animals which failed to recover from lesion-induced asymmetry in turning behavior, as compared to normal controls or animals which showed behavioral recovery. The labeled cells in the anterior part of the ipsilateral TM were smaller in animals examined 15 days after the lesions than in the control animals. The results indicate an increase in HRP uptake or transport in the anterior part of the tuberomammillary-striatal projection predominantly ipsilateral to the lesion in animals which failed to recover from lesion induced asymmetry. Previously, we had found an opposite effect in the nigro-striatal projection, namely an increase in HRP uptake or transport only in animals which recovered from their lesion induced asymmetry. The results are discussed in terms of the evidence for a reciprocal relationship between the TM- and SN-striatal systems.

Afferent innervation of the rat pancreas: retrograde tracing and immunohistochemistry in the dorsal root ganglia.

This study was undertaken to determine the segmental organization of the dorsal root ganglion (DRG) cells that give rise to pancreatic afferents containing a certain neuropeptide in the rat. These cells were examined using retrograde tracing combined with immunohistochemistry. Injection of horseradish peroxidase (HRP) into the pancreas resulted in the labeling of cells in bilateral T5-L2 DRGs, with most labeled cells lying at T10-T11. Injection into the duodenal (right), splenic (left), and entire lobes consistently produced more labeled cells significantly in the right, left, and right DRGs, respectively. Calcitonin gene-related peptide (CGRP)-, substance P (SP)-, somatostatin (SOM)-, and galanin (GAL)-immunoreactive (IR) cells in the DRGs (T9-T12) were found in -52, 17, 8, and 6%, respectively, but neuropeptide Y- and vasoactive intestinal polypeptide-IR cells were not found. About 88% of HRP-labeled cells in DRGs (T9-T12) contained CGRP, and approximately 16% of them contained SP. Although SOM- and GAL-IR cells were localized in the DRGs, these cells innervating the pancreas could not be found. In brief, these results show that bilateral (not similar in cell number on each side) DRG cells innervate the duodenal or splenic pancreas, and the majority of these cells that project to the pancreas contain CGRP and SP.

Cortico-cortical connections from somatosensory areas to the motor area of the cortex following peripheral nerve lesion in the cat.

Cortico-cortical connections of the forelimb digital area of the motor cortex (MCx) following peripheral nerve lesion were examined using horseradish peroxidase (HRP) in adult cats. A small amount of HRP injection was made into the digital area of the MCx in the control animals with intact nerves. HRP labelled cells were found in the first somatosensory area (SI; areas 1-2, 3a and 3b), secondary somatosensory area (SII) and area 5 of the ipsilateral cortex. In addition, a small number of HRP labelled cells were found in the contralateral cortex located in area 4 (MCx) and area 3a of SI. In the experimental animals, peripheral nerves were cut and after 2 or 3 months of survival, HRP was injected to the corresponding area of the MCx. The HRP labelled cells were found in SI (areas 2, 3a, 3b), SII, SIII, SIV and SV and in areas Id and 5 of the ipsilateral cortex. Furthermore, HRP-labelled cells were found in area 4 of the MCx, in SI (3a), SII, SIV and Id on the contralateral side of the cortex. HRP-labelled cells were located in layers II, III and V. Most of these cells were found in layer III.

The ipsilateral and contralateral connections of the fifth somatosensory area (SV) in the cat cerebral cortex.

THE ipsilateral and contralateral corticocortical connections to the fifth somatosensory area (SV) in the feline cortex were determined from the location of retrogradely labelled cells following a single injection of HRP into SV. The injection was made into physiologically defined components of the body representation in SV. After injection of HRP into the face regions of SV, HRP-labelled cells were located ipsilaterally in areas 6 beta, 3b and 1-2 of the primary somatosensory (SI), in the second somatosensory (SII), third somatosensory (SIII), and fourth somatosensory (SIV) areas, along the ansate sulcus, and in areas 5a and 6a beta of the ipsilateral cortex, as well as in area 1-2 of SI and in SV of the contralateral cortex. On the other hand, after HRP had been injected into the trunk/hindlimb area, HRP-labelled cells were located in areas 3a, 1-2 of SI, in area 5, in SII, in SIII and in SIV of the ipsilateral cortex, as well as in area 1-2 of SI, and in SV of the contralateral cortex. The extent of these interconnections suggests that SV receives multiple sensory inputs and may function to integrate this information.

Distribution of visual callosal neurons in normal and strabismic cats.

It has been suggested that synchronous activation of cortical loci in the two cerebral hemispheres during development leads to the stabilization of juvenile callosal connections in some areas of the visual cortex. One way in which loci in opposite hemispheres can be synchronously activated is if they receive signals generated by the same stimulus viewed through different eyes. These ideas lead to the prediction that shifts in the cortical representation of the visual field caused by misalignment of the visual axes (strabismus) should change the width of the callosal zone in the striate cortex. We tested this prediction by using quantitative techniques to compare the tangential distribution of callosal neurons in the striate cortex of strabismic cats to that in normally reared cats. Animals were rendered strabismic surgically at 8-10 days of age and were allowed to survive a minimum of 18 weeks, at which time multiple intracortical injections of the tracer horseradish peroxidase (HRP) were used to reveal the distribution of callosally projecting cells in the contralateral striate cortex. HRP-labeled cells were counted in coronal sections, and data from four animals with divergent strabismus (exotropia) and four with convergent strabismus (esotropia) were compared to those from four normally reared animals. Although our data from strabismic cats do not differ markedly from those reported previously, we find that the distribution of callosal cells in the striate cortex of these cats does not differ significantly from that in our normally reared control cats. These results do not bear out the prediction that surgically shifting the visual axes leads to stabilization of juvenile callosal axons in anomalous places within the striate cortex.

Nimodipine accelerates axonal sprouting after surgical repair of rat facial nerve

Facial-facial anastomosis (FFA), i.e., suture of transected facial nerve, was performed in adult Wistar rats. For 10-112 d post-operation (DPO), half of the animals received standard food (placebo) and half received food pellets containing 1000 ppm nimodipine, a Ca2+ channel blocker. The time course of mimetic reinnervation between these two groups was compared by counting all retrogradely labeled motoneurons after injection of horseradish peroxidase (HRP) into the whiskerpad. In unoperated animals, injection of HRP labeled 1280 +/- 113 motoneurons. After FFA, this number dropped to zero, and the first HRP-labeled facial motoneurons reappeared in both placebo- and nimodipine-treated animals at 14 DPO. The treatment with nimodipine yielded two beneficial effects. (1) It accelerated axonal sprouting until 28 DPO. Whereas the number of HRP-labeled cells in the placebo group was 171 +/- 9 (mean +/- SD) at 16 DPO, 372 +/- 43 at 21 DPO, and 636 +/- 187 at 28 DPO, the number of sprouted motoneurons in nimodipine-treated rats was twice as high: 386 +/- 34 at 16 DPO, 620 +/- 28 at 21 DPO, and 756 +/- 257 at 28 DPO. (2) Nimodipine reduced the polyneuronal innervation of the target muscles. Whereas the number of HRP-labeled cells in the placebo group increased to 1430 +/- 36 at 56 DPO and 1600 +/- 31 at 112 DPO, the number of labeled motoneurons in nimodipine-treated rats remained almost within the normal range: 1315 +/- 31 at 56 DPO and 1354 +/- 33 at 112 DPO.

Cholinergic projections to the anterior thalamic nuclei in the rat: a combined retrograde tracing and choline acetyl transferase immunohistochemical study.

Retrograde transport of horseradish peroxidase (HRP) was combined with choline acetyltransferase (ChAT) immunohistochemistry to study cholinergic projections to the anterior thalamic nuclei in the rat. Small iontophoretic injections of HRP placed into different subdivisions of the anterior thalamic nuclear complex resulted in distinct patterns of retrograde labelling in two major cholinergic cell groups of the mesopontine tegmentum, the laterodorsal tegmental nucleus (LDTg), in which a majority of the labelled cells was located, and the pedunculopontine tegmental nucleus (PPT). After injections into the posterior subdivision of the anteroventral thalamic nucleus (AVp), double-labelled neurons were present predominantly in the ipsilateral LDTg while a smaller number was found in the PPT. In the ipsilateral LDTg, 60-70% of ChAT-positive neurons were HRP-labelled, and 90-95% of the HRP-labelled neurons were ChAT-positive. In the contralateral LDTg, 30-40% of ChAT-positive neurons were HRP-labelled. After injections in the medial subdivision of the anteroventral thalamic nucleus (AVm), the pattern of labelling in LDTg was similar to that detected after injections in the AVp. The number of double-labelled neurons in the LDTg and PPT was much lower after injections into AVm than after injections into AVp. When injections were confined to the anterodorsal thalamic nucleus (AD), no HRP-labelled cells were present in the LDTg or PPT. These results show that the LDTg and PPT are the sources of the cholinergic input to the rat anterior thalamus. The major projection from LDTg and PPT is to the AVp, whereas there is a lighter cholinergic projection to the AVm. The AD does not receive a projection from cholinergic cells in the mesopontine tegmentum.

Sources of subcortical afferents to the macaque's dorsal lateral geniculate nucleus.

The dorsal lateral geniculate nucleus (dLGN) is the thalamic region responsible for transmitting retina signals to cortex. Brainstem pathways to this nucleus have been described in several species and are believed to control the retinocortical pathway depending on the state of the animal (awake, asleep, drowsy, etc.). The purpose of this study was to determine all of the subcortical sources of afferents to the dLGN in a higher primate, the macaque monkey, whose visual system is similar to that of humans. Injections of horseradish peroxidase (HRP), with or without conjugation to wheat germ agglutinin, were made into the dLGNs of seven macaque monkeys, followed by perfusion, brain sectioning, and analyses of neurons in the brainstem, thalamus, and hypothalamus that contained the retrogradely transported marker. The reticular nucleus of the thalamus, pedunculopontine nucleus, parabigeminal nucleus, pretectal nucleus of the optic tract, superior colliculus, dorsal raphe nucleus, and tuberomammillary region of the hypothalamus contained many retrogradely labeled neurons ipsilateral to the injections. In the contralateral brainstem, HRP-labeled cells were found only in the pedunculopontine nucleus, nucleus of the optic tract, and dorsal raphe nucleus. The number of labeled neurons on the contralateral side was about one-half of that in corresponding ipsilateral nuclei. The locus coeruleus contained no labeled neurons in four of the macaques that had injections limited to the dLGN. There are seven subcortical regions that send afferents to the dLGNs of macaque monkeys. Except for the locus coeruleus, these are the same as observed for other species, such as the cat and rat, and indicate the possible sources of subcortical control over the dLGNs of humans.

Localization of trigeminal sensory innervation to the tongue and pharynx of the cat using horseradish peroxidase as tracer.

Trigeminal sensory innervation to the tongue and pharynx was investigated in the cat, using horseradish peroxidase (HRP) as a retrograde tracer. Following HRP application to the tongue, cells were labeled primarily in the posterolateral portion of the trigeminal ganglion. The pharynx was divided according to classical methodology into three regions: the epipharynx (nasopharynx), mesopharynx (oropharynx), and hypopharynx. When HRP was applied to the epipharynx and mesopharynx, HRP-labeled cells were found in the anteromedial and posterolateral portions of the trigeminal nerve, respectively. When HRP was applied to the hypopharynx, a single labeled cell was found in the trigeminal ganglion. These results indicate that trigeminal sensory nerve fibers in the cat branch to the tongue, epipharynx, and mesopharynx, but rarely to the hypopharynx.