Transporter Protein-Coupled DPCPX Nanoconjugates Induce Diaphragmatic Recovery after SCI by Blocking Adenosine A1 Receptors.

Respiratory complications in patients with spinal cord injury (SCI) are common and have a negative impact on the quality of patients' lives. Systemic administration of drugs that improve respiratory function often cause deleterious side effects. The present study examines the applicability of a novel nanotechnology-based drug delivery system, which induces recovery of diaphragm function after SCI in the adult rat model. We developed a protein-coupled nanoconjugate to selectively deliver by transsynaptic transport small therapeutic amounts of an A1 adenosine receptor antagonist to the respiratory centers. A single administration of the nanoconjugate restored 75% of the respiratory drive at 0.1% of the systemic therapeutic drug dose. The reduction of the systemic dose may obviate the side effects. The recovery lasted for 4 weeks (the longest period studied). These findings have translational implications for patients with respiratory dysfunction after SCI. SIGNIFICANCE STATEMENT: The leading causes of death in humans following SCI are respiratory complications secondary to paralysis of respiratory muscles. Systemic administration of methylxantines improves respiratory function but also leads to the development of deleterious side effects due to actions of the drug on nonrespiratory sites. The importance of the present study lies in the novel drug delivery approach that uses nanotechnology to selectively deliver recovery-inducing drugs to the respiratory centers exclusively. This strategy allows for a reduction in the therapeutic drug dose, which may reduce harmful side effects and markedly improve the quality of life for SCI patients.

Is there any sense in the Palisade endings of eye muscles?

Palisade endings (PEs), which are unique to the eye muscles, are associated with multiply innervated muscle fibers. They lie at the myotendinous junctions and form a cap around the muscle fiber tip. They are found in all animals investigated so far, but their function is not known. Recently, we demonstrated that cell bodies of PEs and tendon organs lie around the periphery of the oculomotor nucleus in the C- and S-groups. A morphological analysis of these peripheral neurons revealed the existence of different populations within the C-group. We propose that a small group of round or spindle-shaped cells gives rise to PEs, and another group of multipolar neurons provide the multiple motor endings. If PEs have a sensory function, then their cell body location close to motor neurons would be in an ideal location to control tension in extraocular muscles; in the case of the C-group, its proximity to the preganglionic neurons of the Edinger-Westphal nucleus would permit its participation in the near response. Despite their unusual properties, PEs may have a sensory function.

The milk-ejection reflex in the sheep: an anatomical study on the afferent pathway.

The present study is an attempt to reveal the spinal and supraspinal organization of the ascending branch of the milk-ejection reflex in the ewe by means of a tract-tracing technique. For this purpose, injections of horseradish peroxidase (HRP) were performed into the lateral cervical nucleus (LCN) and into the hypothalamic paraventricular nucleus (PVN). Peroxidase injections into the LCN revealed retrogradely labelled neurons in the medial part of laminae I-III of the ipsilateral L3 and L4 spinal segments, while injections of HRP into the PVN revealed retrogradely labelled cells in the contralateral LCN and the medial cuneate nucleus. Taking into account the results obtained, it is concluded that the transmission of the afferent input from the nipples to the PVN is accomplished by at least two pathways: one employing a single relay station located in the medial cuneate nucleus, and another possessing two relay stations located in the medial part of laminae I-III of the dorsal horn of L3 and L4 spinal segments and in the LCN.

Infralimbic cortex projects to all parts of the pontine and medullary lateral tegmental field in cat.

The infralimbic cortex (ILc) in cat is the ventralmost part of the anterior cingulate gyrus. The ILc, together with the amygdala, bed nucleus of the stria terminalis and lateral hypothalamus, is involved in the regulation of fear behavior. The latter three structures are thought to take part in triggering the fear response by means of their projections to the pontine and medullary lateral tegmental field (LTF). The LTF is a large region extending from the parabrachial nuclei rostrally to the spinal cord caudally. It contains almost all the premotor interneurons for the brainstem and for some upper spinal cord motoneurons innervating the muscles of face, head and throat. The question is whether ILc also projects to the LTF. Such a pathway would allow the ILc to influence the fear response by acting directly on these premotor interneurons. Anterograde tracer injections were made in the medial surface of the cortex in four cats. Only when the injection sites involved ILc were anterogradely labeled fibers observed throughout the rostrocaudal extent of the LTF. To verify whether these projections indeed originated from ILc, in two other cases retrograde tracer injections were made in the pontomedullary LTF. The results showed many retrogradely labeled neurons in ILc, but none in adjacent cortical regions. These results show that the ILc projects to the LTF in cat and can possibly modulate the fear response not only via indirect but also via direct routes to the premotor interneurons in the brainstem.

Neurons in the guinea pig (Cavia porcellus) lateral lumbosacral spinal cord project to the central part of the lateral periaqueductal gray matter.

In order to micturate successfully, information from the bladder has to be conveyed to the brainstem. In most experimental animals, this information is relayed, via the lumbosacral spinal cord, to the periaqueductal gray matter (PAG). Although the rat is the most used experimental animal in neurourological research, urodynamic studies show that guinea pig may be a better small experimental animal because its urodynamic profile is, in contrast to that of a rat, similar to that of humans. Therefore, the present study, using anterograde and retrograde tracing, was performed to determine whether the lumbosacral spinal cord projects to the PAG in guinea pig. Results show that neurons in the lateral part of the lumbosacral spinal cord project to the central parts of the PAG. This pathway may convey information about the level of bladder filling to the PAG.

Organization of frontoparietal cortex in the tree shrew (Tupaia belangeri). I. Architecture, microelectrode maps, and corticospinal connections.

Despite extensive investigation of the motor cortex of primates, little is known about the organization of motor cortex in tree shrews, one of their closest living relatives. We investigated the organization of frontoparietal cortex in Belanger's tree shrews (Tupaia belangeri) by using intracortical microstimulation (ICMS), corticospinal tracing, and detailed histological analysis. The results provide evidence for the subdivision of tree shrew frontoparietal cortex into seven distinct areas (five are newly identified), including two motor fields (M1 and M2) and five somatosensory fields (3a, 3b, S2, PV, and SC). The types of movements evoked in M1 and M2 were similar, but M2 required higher currents to elicit movements and had few connections to the cervical spinal cord and distinctive cyto- and immunoarchitecture. The borders between M1 and the anterior somatosensory regions (3a and 3b) were identified primarily from histological analysis, because thresholds were similar between these regions, and differences in corticospinal neuron distribution were subtle. The caudal (SC) and lateral (S2 and PV) somatosensory fields were identified based on differences in architecture and distribution of corticospinal neurons. Myelin-dense modules were identified in lateral cortex, in the expected location of the oral, forelimb, and hindlimb representations of S2, and possibly PV. Evidence for a complex primate-like array of motor fields is lacking in tree shrews, but their motor cortex shares a number of basic features with that of primates, which are not found in more distantly related species, such as rats.

Periparabigeminal and adjoining mesencephalic tegmental field projections to the dorsolateral periaqueductal grey in cat - a possible role for oculomotor input in the defensive system.

The dorsolateral column of the mesencephalic periaqueductal grey (PAGdl) differs from its adjacent columns in terms of afferent and efferent connections and the distribution pattern of different histochemical substances. Functionally, PAGdl is associated with aversive and defensive behaviours, but in an earlier study of this laboratory [E.M. Klop et al. (2005) J. Comp. Neurol., 492, 303-322], it was found that PAGdl specifically receives input from the nucleus prepositus hypoglossi, which plays a role in oculomotor control. In search for other oculomotor-related brainstem structures projecting to PAGdl we studied the projections from the parabigeminal nucleus (PBGN) and its medially adjoining periparabigeminal area (PPBGA). In three cats, injections of wheatgerm agglutinin-horseradish peroxidase involving PAGdl did not, or to only a very limited extent, result in retrogradely labelled neurons in PBGN. When the peripheral parts of PAGdl were involved in the injection site, labelled neurons were located in PPBGA, while after an injection involving only the more central parts of PAGdl they were located in the tegmentum medial to the PPBGA. An anterograde tracing study using [3H]-leucine and biotinylated dextran amine affirmed that neurons in PPBGA project to more peripheral parts of PAGdl, while neurons located in the tegmentum medial to PPBGA project mainly to its central parts. These results provide further evidence for the existence of two different subdivisions of PAGdl. We hypothesize that PAGdl is alerted by sudden changes in the visual field, and that the PAGdl defensive system is inhibited when these changes are caused by eye movements.

Peptidergic and catecholaminergic synaptic contacts onto nucleus preopticus medianus neurons projecting to the subfornical organ in the rat.

The nucleus preopticus medianus (POMe) is known to be a key site in regulation of cardiovascular and body fluid homeostasis. To clarify the regulation mechanism to the POMe, the innervation pattern of synapses made by axon terminals immunoreactive to beta-endorphin, neuropeptide Y and tyrosine hydroxylase onto POMe neurons projecting to the subfornical organ (SFO) was investigated in the rat. After injection of a retrograde tracer, wheat germ agglutinin-conjugated horseradish peroxidase-colloidal gold complex, into the SFO, many neurons were retrogradely labeled in the POMe, more frequently in its dorsal part. Electron microscopy of the POMe revealed that beta-endorphin- and tyrosine hydroxylase-immunoreactive axon terminals formed predominantly axo-somatic synapses, and neuropeptide Y-immunoreactive axon terminals formed more axo-dendritic than axo-somatic synapses with retrogradely labeled neurons. The present localization patterns of POMe neurons retrogradely labeled from the SFO and the type of synapses of axon terminals immunoreactive to three neurochemical markers on these neurons were compared to those of POMe neurons retrogradely labeled from the paraventricular hypothalamic nucleus demonstrated in our previous report.

Striatal projections from the lateral and posterior thalamic complexes. An anterograde tracer study in the cat.

Striatal projections from the lateral intermediate (LI) and posterior (Po) thalamic complexes were studied with the anterograde tracers wheat germ agglutinin-horseradish peroxidase and Phaseolus vulgaris leucoagglutinin. Projections to the lateral part of the head and body of the caudate nucleus (CN) and to the putamen (Pu) were found to arise from the ventral parts of the caudal subdivision of the LI besides the well established sources in the intralaminar and ventral thalamic nuclei. No projections to the CN and only a few to the Pu were found to arise from the medial division of the Po. The presence of terminal and intercalated varicosities in the thalamostriatal fibers suggests that they form both terminal and en passant synapses. Thalamostriatal fibers from these thalamic sectors were unevenly distributed within the CN, with patches of either low-density innervation or with no projections at all interspersed within irregular, more densely innervated areas. The former coincided with the acetylcholinesterase-poor striosomes and the latter areas of dense projection with the extrastriosomal matrix.

Projections of the ventrolateral pontine vocalization area in the squirrel monkey.

In four squirrel monkeys (Saimiri sciureus), the tracer biotin dextranamine (BDA) was injected into the ventrolateral pons at a site at which injection of the glutamate antagonist kynurenic acid blocked vocalization electrically elicited from the periaqueductal gray (PAG). Anterograde projections could be traced into all cranial motor and sensory nuclei involved in phonation, that is, the nucleus ambiguus, facial, hypoglossal and trigeminal motor nuclei, the motorneuron column in the ventral gray substance innervating the extrinsic laryngeal muscles, the nucleus retroambiguus, solitary tract and spinal trigeminal nuclei. Projections were also found into a number of auditory nuclei, namely the nucleus cochlearis-complex, superior olive, ventral and dorsal nuclei of the lateral lemniscus and inferior colliculus. Furthermore, there were projections into the reticular formation of the lateral and dorsocaudal medulla and lateral pons, into nucleus gracilis, inferior and medial vestibular nuclei, lateral reticular nucleus, ventral raphe, pontine gray, superior colliculus, PAG and mediodorsal thalamic nucleus. Injection of the tracer wheat germ agglutinin-conjugated horseradish peroxidase into the ventrolateral pontine vocalization-blocking area in one animal yielded retrograde labeling throughout the PAG. Injection of BDA into a vocalization-eliciting site of the PAG in another animal yielded projections into the ventrolateral pontine vocalization-blocking area. It is concluded that the ventral paralemniscal area in the ventrolateral pons represents a relay station of the descending periaqueductal vocalization-controlling pathway.

Vibration-induced disruption of retrograde axoplasmic transport in peripheral nerve.

Hand-arm vibration syndrome (HAVS) results from excessive exposure to hand-transmitted vibration. Whether the peripheral nerve damage characteristic of HAVS is a direct result of vibration or is secondary to vascular insufficiency remains unclear. The purpose of this study was to explore the effect of vibration exposure on axoplasmic transport in peripheral nerves and soleus motor neurons. Sciatic nerves and motor neurons from rats following two 5-h periods of vibration exposure demonstrated disruption in retrograde transport compared to normal. After 10 days of vibration (5 h/day), axoplasmic transport failed to recover within 24-48 h in most rats. This study demonstrates that disrupted axoplasmic transport is an early consequence of short-term vibration exposure. The effects of vibration on axoplasmic transport also appear to be cumulative. This study provides a new biological way to evaluate measures to prevent early vibration injury.

Ultrastructure of the rostral ventral respiratory group neurons in the ventrolateral medulla of the rat.

The neurons in the ventrolateral medulla that project to the spinal cord are called the rostral ventral respiratory group (rVRG) because they activate spinal respiratory motor neurons. We retrogradely labeled rVRG neurons with Fluoro-Gold (FG) injections into the fourth cervical spinal cord segment to determine their distribution. The rostral half of the rVRG was located in the area ventral to the semicompact formation of the nucleus ambiguus (AmS). A cluster of the neurons moved dorsally and intermingled with the palatopharyngeal motor neurons at the caudal end of the AmS. The caudal half of the rVRG was located in the area including the loose formation of the nucleus ambiguus caudal to the AmS. We also labeled the rVRG neurons retrogradely with wheat germ agglutinin-horseradish peroxidase (WGA-HRP) to determine their ultrastructural characteristics. The neurons of the rVRG were medium to large (38.1 x 22.1 microm), oval or ellipsoid in shape, and had a dark cytoplasm containing numerous free ribosomes, rough endoplasmic reticulum (rER), mitochondria, Golgi apparatuses, lipofuscin granules and a round nucleus with an invaginated nuclear membrane. The average number of axosomatic terminals in a profile was 33.2. The number of axosomatic terminals containing round vesicles and making asymmetric synaptic contacts (Gray's type I) was almost equal to those containing pleomorphic vesicles and making symmetric synaptic contacts (Gray's type II). The axodendritic terminals were large (1.55 microm), and about 60% of them were Gray's type I. The rVRG neurons have ultrastructural characteristics, which are different from the palatopharyngeal motor neurons or the prorpiobulbar neurons.

Vestibular projection to the periarcuate cortex in the monkey.

Vestibular inputs to the cerebral cortex are important for spatial orientation, body equilibrium, and head and eye movements. We examined vestibular input to the periarcuate cortex in the Japanese monkey by analyzing laminar field potentials evoked by electrical stimulation of the vestibular nerve. Laminar field potential analysis in the depths of the cerebral cortex showed that vestibular-evoked potentials consisted of early-positive and late-negative potentials and early-negative and late-positive potentials in the superficial and deep layers of the periarcuate cortex, respectively, with latencies of 4.8-6.3 ms, suggesting that these potentials were directly conveyed to the cortex through the thalamus. These potentials were distributed continuously in the fundus, dorsal and ventral banks of the spur and the bottom of the junctional part of the arcuate sulcus and spur. This vestibular-projecting area overlapped the cortical distribution of corticovestibular neurons that were retrogradely labeled by tracer injection into the vestibular nuclei (previously reported area 6 pa), and also the distribution of smooth pursuit-related neurons recorded in the periarcuate cortex including area 8 in a trained monkey. These results are discussed in relation to the function of vestibular information in control of smooth pursuit and efferents of the smooth pursuit-related frontal eye field.

Anatomic organization of the ascending branch of the milk-ejection reflex in sheep: primary afferent neurons.

The present study examines the anatomic characteristics of the primary afferent neurons that innervate the nipples and pseudonipples of ewes and the nipples of lambs. For this purpose, horseradish-peroxidase coupled to wheat germ agglutinin (WGA-HRP) was injected intradermally into the whole extent, the tip, or the base of the nipples and pseudonipples, as well as into a region of the posterior surface of the udder. After survival periods of 72-96 hours, dorsal root ganglia (DRG), segments of the spinal cord and medulla oblongata were sectioned and reacted histochemically with tetramethylbenzidine to reveal the transganglionically transported tracer. Injections of WGA-HRP in the nipples and pseudonipples of the ewe resulted in labeled cells in the second to fifth ipsilateral lumbar spinal ganglia (L(2)-L(5)) and third and fourth (L(3) and L(4)) lumbar spinal ganglia, respectively. Labeled cells after WGA-HRP injections in the nipples of the lamb were found in the ipsilateral L(3)-L(5) spinal ganglia. Central projections of the DRG-labeled cells were found in the medial part of laminae I-III of the ipsilateral L(3) and L(4) spinal segments (ewe and lamb) and in the ipsilateral dorsal column nuclei (ewe). Central projections of the DRG-labeled cells after injections in the pseudonipples of the ewe were located in the medial part of laminae I-III of the ipsilateral L(3) spinal segment. The results of this study demonstrate that, whereas the innervation of the nipples of the ewe originates from four successive lumbar spinal ganglia (L(2)-L(5)), the innervation of the nipples of the lamb and the pseudonipples of the ewe originates from three (L(3)-L(5)) and two (L(3) and L(4)) successive ganglia, respectively.

Nigrothalamostriatal and nigrothalamocortical pathways via the ventrolateral parafascicular nucleus.

The present tract-tracing study in the rat indicated that neurons in the ventrolateral part of the parafascicular thalamic nucleus (PF), where nigral fibers from the dorsolateral part of the substantia nigra pars reticulata (SNr) terminated, sent their axons to the ventrolateral part of the striatum as well as to the rostrolateral part of the lateral agranular cortex (AGl). We further demonstrated that symmetrical synaptic contacts were made between these nigral axons and striatum- or AGl-projecting PF neurons. Since the dorsolateral part of the SNr, ventrolateral part of the striatum and rostrolateral part of the AGl are responsible regions for orofacial behaviors, the nigrothalamostriatal and nigrothalamo-cortical pathways via the ventrolateral part of the PF may play a role in the control of orofacial motor function.

Brainstem origin of duodenal vagal preganglionic parasympathetic neurons. A WGA-HRP study in the ferret (Mustela Putorius Furo), a human model.

The projections of vagal brainstem neurons to the duodenal segment of the gastrointestinal tract were studied in the ferret using the WGA-HRP neurohistochemical technique. Fourteen adult ferrets with weights ranging from 800 gm to 1500 gm were used for the study. The muscular wall of the duodenum of six ferrets was injected with 0.1 ml of 5% WGA-HRP in 0.5 M sodium chloride. The eight remaining ferrets were used as controls. Two of these had injections of 0.1 ml normal saline into the muscular wall of the duodenum. The second set of two ferrets was injected with 0.1 ml of 5% WGA-HRP in buffer after bilateral truncal vagotomy. The third set of two ferrets received intraperitoneal injection of 0.1 ml of 5% WGA-HRP while, in the last set, the tracer was injected into the hepatic portal vein. Following the injections, the ferrets were allowed to survive for 48-72 hours after which each ferret was perfused transcardially first with normal saline followed by a fixative containing 1% paraformaldehyde and 1.25% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4 at room temperature and finally with 10% buffered sucrose at 4 degrees C. Transverse serial frozen sections of the brainstem were then taken and processed for WGA-HRP neurohistochemistry and were analyzed under light and dark-field illuminations. The analyses of the sections taken from the six ferrets injected with WGA-HRP revealed neurons labelled with the tracer in the dorsal motor nucleus of the vagus nerve (DMNV). Sections taken from the control ferrets did not reveal any WGA-HRP labelled neurons in the brainstem.

Descending projections to the inferior colliculus from the posterior thalamus and the auditory cortex in rat, cat, and monkey.

Projections from the posterior thalamus and medial geniculate body were labeled retrogradely with wheat germ agglutinin conjugated to horseradish peroxidase injected into the rat, cat, and squirrel monkey inferior colliculus. Neurons were found ipsilaterally in the (1) medial division of the medial geniculate body, (2) central gray, (3) posterior limitans nucleus, and the (4) reticular part of the substantia nigra. Bilateral projections involved the (5) peripeduncular/suprapeduncular nucleus, (6) subparafascicular and posterior intralaminar nuclei, (7) nucleus of the brachium of the inferior colliculus, (8) lateral tegmental/lateral mesencephalic areas, and (9) deep layers of the superior colliculus. The medial geniculate projection was concentrated in the caudal one-third of the thalamus; in contrast, the labeling in the subparafascicular nucleus, substantia nigra, and central gray continued much further rostrally. Robust anterograde labeling corresponded to known patterns of tectothalamic projection. Biotinylated dextran amine deposits in the rat inferior colliculus revealed that (1) many thalamotectal cells were elongated multipolar neurons with long, sparsely branched dendrites, resembling neurons in the posterior intralaminar system, and that other labeled cells were more typical of thalamic relay neurons; (2) some cells have reciprocal projections. Similar results were seen in the cat and squirrel monkey. The widespread origins of descending thalamic influences on the inferior colliculus may represent a phylogenetically ancient feedback system onto the acoustic tectum, one that predates the corticocollicular system and modulates nonauditory centers and brainstem autonomic nuclei. Besides their role in normal hearing such pathways may influence behaviors ranging from the startle reflex to the genesis of sound-induced seizures.

Organization of reciprocal connections between the perigeniculate nucleus and dorsal lateral geniculate nucleus in the cat: a transneuronal transport study.

Cells of the cat's perigeniculate nucleus (PGN), part of the visual sector of the thalamic reticular nucleus (TRN), provide GABAergic inhibition to the A and A1 layers of the dorsal lateral geniculate nucleus (LGNd) and, therefore, may control information flow from the retina to the cortex. Previous electrophysiological experiments suggested that the PGN may be subdivided on the basis of ocular dominance thus reflecting the afferent and efferent projections with lamina A and A1 of the LGNd. The present study utilized the ability of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) to be transported transneuronally following intraocular injections in four cats to examine whether there is any anatomical evidence for eye specific layers within the PGN. Sections were processed with tetramethylbenzidine. Light WGA-HRP transneuronal labeling of LGNd collaterals and somata were seen in the PGN and very light labeling (but not somata) was seen in the TRN. Neither the cells of the PGN projecting to the LGNd nor the LGNd relay collaterals within the PGN were clearly organized into nonoverlapping laminae related to the eye specific layers of the LGNd. However, parts of the PGN immediately adjacent to the LGNd appear devoid of connections with lamina A1 thus creating a thin monocular segment for the contralateral eye.

Motoneurons of twitch and nontwitch extraocular muscle fibers in the abducens, trochlear, and oculomotor nuclei of monkeys.

Eye muscle fibers can be divided into two categories: nontwitch, multiply innervated muscle fibers (MIFs), and twitch, singly innervated muscle fibers (SIFs). We investigated the location of motoneurons supplying SIFs and MIFs in the six extraocular muscles of monkeys. Injections of retrograde tracers into eye muscles were placed either centrally, within the central SIF endplate zone; in an intermediate zone, outside the SIF endplate zone, targeting MIF endplates along the length of muscle fiber; or distally, into the myotendinous junction containing palisade endings. Central injections labeled large motoneurons within the abducens, trochlear or oculomotor nucleus, and smaller motoneurons lying mainly around the periphery of the motor nuclei. Intermediate injections labeled some large motoneurons within the motor nuclei but also labeled many peripheral motoneurons. Distal injections labeled small and medium-large peripheral neurons strongly and almost exclusively. The peripheral neurons labeled from the lateral rectus muscle surround the medial half of the abducens nucleus: from superior oblique, they form a cap over the dorsal trochlear nucleus; from inferior oblique and superior rectus, they are scattered bilaterally around the midline, between the oculomotor nucleus; from both medial and inferior rectus, they lie mainly in the C-group, on the dorsomedial border of oculomotor nucleus. In the medial rectus distal injections, a "C-group extension" extended up to the Edinger-Westphal nucleus and labeled dendrites within the supraoculomotor area. We conclude that large motoneurons within the motor nuclei innervate twitch fibers, whereas smaller motoneurons around the periphery innervate nontwitch, MIF fibers. The peripheral subgroups also contain medium-large neurons which may be associated with the palisade endings of global MIFs. The role of MIFs in eye movements is unclear, but the concept of a final common pathway must now be reconsidered.

The fine organization of nigro-collicular channels with additional observations of their relationships with acetylcholinesterase in the rat.

The nigro-collicular pathway that links the basal ganglia to the sensorimotor layers of superior colliculus plays a crucial role in promoting orienting behaviors. This connection originating in the pars reticulata and lateralis of the substantia nigra has been shown in rat and cat to be topographically organized. In rat, a functional compartmentalization of the substantia nigra has also been shown reflecting that of the striatum. In light of this, we reinvestigated the topographical arrangement of the nigro-collicular pathway by examining the innervation of each nigral functional zone. We performed small injections of either biocytin or wheatgerm agglutinin conjugated with horseradish peroxidase restricted to identified somatic, visual and auditory nigral zones. Frontally cut sections showed that innervations provided by the three main nigral zones form a mosaic of complementary domains stratified from the stratum opticum to the ventral part of the intermediate collicular layers, with the somatic afferents sandwiched between the visual and the auditory ones. When reconstructed from semi-horizontal sections, nigral innervations organized in the form of a honeycomb-like array composed of 100 cylindrical modules covering three-quarters of the collicular surface. Such a modular architecture is reminiscent of the acetylcholinesterase lattice we previously described in rat intermediate collicular layers. In the enzyme lattice, the surroundings of the cylindrical modules are composed of a mosaic of dense and diffuse enzyme subdomains. Thus, we compared the distribution of the overall nigral projection and of its constituent channels with the acetylcholinesterase lattice. The procedure combined axonal labelling with histochemistry on single sections for acetylcholinesterase activity. The results demonstrate that the overall nigral projection overlaps the acetylcholinesterase lattice and its constituent channels converge with either the dense or the diffuse enzyme subdomains. The stereometric arrangement of the nigro-collicular pathway is suggestive of an architecture promoting the selection of collicular motor programs for different classes of orienting behavior.