A pretectofacial projection in the cat: a possible link in the visually-triggered blink reflex pathways

A direct projection from the pretectum to the facial motor nucleus was shown to exist in the cat by the anterograde and retrograde horseradish peroxidase (HRP) methods. Pretectofacial fibers arise from the olivary pretectal nucleus and end mainly in the dorsal division of the facial motor nucleus, bilaterally, with a contralateral predominance. It is known that the olivary pretectal nucleus receives retinal fibers, and that the dorsal division of the facial motor nucleus contains orbicularis oculi motoneurons. Thus, the pretectofacial fibers are assumed to cause protective lid closure with certain visual stimuli.     

Organization of the trigeminal and facial motor nuclei in the hagfish, Eptatretus burgeri: a retrograde HRP study

We studied the trigeminal and facial motor nuclei of the hagfish by the retrograde HRP method. We distinguished 4 components in a single column of the motor nuclei of the trigeminal nerve and the facial nerve, viz., the pars magnocellularis of the trigeminal motor nucleus (mVm), the anterior part of the pars parvocellularis of the trigeminal motor nucleus (mVp1), the posterior part of the pars parvocellularis of the trigeminal motor nucleus (mVp2) and the facial motor nucleus (mVII). Although in Nissl preparations only the mVm could be distinguished from the rest of the nucleus, the boundaries of the other 3 components were clearly demarcated in HRP preparations. Intramuscular injections into two representative antagonistic jaw muscles revealed that there was no apparent topological organization of the neurons pertaining to the opening and closing muscles in the mVm and mVp1, but both antagonistic muscles were innervated bilaterally. Although the hagfish does possess a cartilaginous jaw, the organization pattern of the motor nuclei of the jaw muscles seems to be the most primitive of all living vertebrates.     

Fiber pathways associated with cerebellar self-stimulation in the rat: A retrograde and anterograde tracing study

Intracranial self-stimulation (ICSS) was obtained from an area of cerebellum just rostro-ventral to the fastigial nucleus. The acquisition of cerebellar ICSS was slow, although this depended in part on the type of operant task required. The afferent and efferent fiber connections of the region of cerebellum supporting ICSS were identified using silver-degeneration and horseradish peroxidase tracing techniques. Two major pathways, one ascending to the ventromedial thalamus and one descending to the paramedian reticular formation and the region of the solitary nucleus, are discussed as possible substrates of ICSS at sites in the cerebellum as well as from other pontine and medullary regions.     

Frontal projections to the region of the oculomotor complex in the rat: a retrograde and anterograde HRP study

The retrograde and anterograde capabilities of the horseradish peroxidase (HRP) technique were employed to study frontal projections to the perioculomotor region in the rat. Following HRP microinjections or transcannular HRP gel implants into the oculomotor complex (OMC), the majority of retrogradely labeled pyramidal cells were located in lamina V of the dorsomedial frontal shoulder cortex, i.e., medial precentral and anterior cingulate (PrCm/AC) cortices, the proposed frontal eye field (FEF) in the rat. A smaller number of labeled cells were present in the frontal polar cortex, agranular insular (AI), and lateral precentral (PrCl) cortices. Following HRP gel implants into the PrCM/Ac, anterogradely labeled projections were observed to the dorsal medial subthalamic region (nucleus campi Foreli, NCF) and medial accessory nucleus of Bechterew (MAB), and to other subcortical nuclei known to receive inputs from cortical area 8 in the monkey. These results, taken together with previous anatomical and physiological studies, support the conclusion that the PrCm/AC cortex contains the rat FEF. Its homology with the primate FEF is discussed.     

Identification and somatotopic organization of nuclei projecting via the dorsolateral funiculus in rats: A retrograde tracing study using HRP slow-release gels

Horseradish peroxidase (HRP) in slow-release gel was unilaterally implanted in the transected dorsolateral funiculus (DLF) of either cervical, midthoracic, lumbar or sacral spinal cord levels of adult male rats. Cell mappings were made of all brain areas projecting through the DLF. Following cervical implants, dense labelling was observed within a band along the dorsolateral border of the inferior olive, locus coeruleus, the paralemniscal reticular formation, the mesencephalic central gray, the red nucleus and the paraventricular nucleus of the hypothalamus. The DLF projection from the central gray consisted of a rostrocaudal line of cells extending from the level of the mesen-/diencephalic junction to the rostral red nucleus. The somatotopic organization of labelled nuclei was assessed by comparing the pattern of filled cells following HRP implants at various cord levels. The potential role of these areas in pain modulation was discussed in light of their descending projections through the DLF of the spinal cord.     

Neuronal and transneuronal tracing in the trigeminal system of the rat using the herpes virus suis

The herpes virus suis has been used as a tracer for the pathways in the central nervous system of the rat. The viruses have been inoculated in various peripheral structures innervated by the trigeminal nerve, namely in the cornea by instillation or scarification, in the anterior chamber of the eye by injection and also by subconjunctival injection, nasal instillation and injection in the masseter muscle. The herpes virus suis is easy to detect by immunofluorescence or electron microscopy, the tracing is precise because it does not diffuse, as some other tracers. The virus is replicated at the site of inoculation and at each neuronal relay, thus 'fresh' tracer is continuously brought into the system. The herpes virus suis is transported by retrograde axonal flow. It has been observed in the motor sensory, sympathetic and parasympathetic pathways up to the central nuclei, which demonstrates transneuronal transport. The selectivity of this tracer, applied to the trigeminal pathways, has allowed us to understand the function of the 3 types of neurons present in the trigeminal ganglion, namely to confirm their somatotopy and establish their central projections in the trigeminal system.     

An anterograde HRP study of the retinotectal pathways in albino and pigmented guinea pigs

An uncrossed retinotectal projection was observed in albino guinea pigs using the anterograde HRP method, suggesting that the retinal projection in these animals, similar to that in the pigmented strain, is not strictly crossed as reported previously. It was also noted that a small retinal projection terminated in a restricted region in the most medial portion of the contralateral inferior colliculus, implying that it might be influenced directly by fibers arising from the contralateral eye.     

A morphological evidence of direct connections from the cochlear nuclei to tensor tympani motoneurons in the cat: a possible afferent limb of the acoustic middle ear reflex pathways

The central circuitry of the acoustic middle ear reflex activating the tensor tympani muscle was studied in the cat by tracer methods using horseradish peroxidase (HRP), wheat germ agglutinin (WGA) or HRP conjugated with WGA (WGA-HRP). The results indicate that the dorsal and ventral cochlear nuclei send fibers to motoneurons innervating the tensor tympani muscle, bilaterally with an ipsilateral dominance.     

Neuronal morphology and efferent projections of the dorsal nucleus of the lateral lemniscus in the rat

The dorsal nucleus of the lateral leminsucs (DLL) is the main source of inhibitory influence in the auditory brainstem of mammals. The cytoarchitecture and connectional properties of DLL were established in the cat in contrast to the rat. The goal of the present study was to establish to what extent the anatomical properties of the rat DLL compare to those of the cat, thus providing a basis of interpretation for future functional studies in the rat, an animal model used more and more in the auditory system. DLL of the rat contains four well-differentiated neuronal types, as seen in Nissl-stained material. Type I neurons are large and multipolar with abundant cytoplasm and darkly stained Nissl substance. Type II neurons are large, bipolar and darkly stained in Nissl material. Type III neurons are medium in size and their soma is round or ovoid. Type IV neurons are small and round with scant cytoplasm; they seem to be also the least common neuronal type of the DLL. After Phaseolus vulgaris-leucoagglutinin or biocytin injections in the DLL, fibers and terminals labeled by orthograde transport were observed in the corresponding region of the contralateral DLL and in the inferior colliculus, bilaterally. A few labeled fibers and terminal fields were seen in the deep layers of the superior colliculus bilaterally, as well as in the medial division of the medial geniculate body and, even more rostrally, in the posterior nucleus of the thalamus. Descending projections from DLL terminated in the periolivary regions of the ipsilateral superior olivary complex. Retrograde tracing based on injections of horseradish peroxidase in the various targets of the DLL confirmed the connections established with orthograde labeling. © 1993 Wiley-Liss, Inc.     

Fluorescent compounds as retrograde tracers compared with horseradish peroxidase (HRP). II. A parametric study in the peripheral motor system of the cat

Fluorescent compounds which are currently used as retrograde tracers were tested in the cat peripheral motor system and compared with horseradish peroxidase (HRP). The tracers were either injected into forelimb muscles or applied to the proximal end of transected forelimb nerves. The remaining muscles of the limb has been carefully denervated. Following intramuscular injection all fluorescent compounds labeled spinal cord motoneurons, the DAPI compounds labeled endothelial cells in addition. In the nerve application mode tracer positive motoneurons were only observed when propidium iodide (PI) and the DAPI compounds were used, whereas bisbenzimide (BB), nuclear yellow (NY) and primuline did not label any cells. The fluorescence of BB labeled motoneurons were predominantly located in the cytoplasma. NY positive motoneurons showed a different localization of the fluorescent label between the different neurons of the same motornucleus: in some neurons it was exclusively located in the nucleus, in others predominantly in the cytoplasma, in the majority in both compartments. The intracellular distribution of the BB and the NY label was independent of the pH of the fixation fluid. The fluorescent tracers labeled the motoneuronal cell columns in their complete rostrocaudal extent and in a position identical to the one obtained with HRP. However, some substances (PI, fast blue) labeled less motoneurons of a motornucleus than did HRP, none of the fluorescent tracers labeled more. The results are discussed under several aspects: use of the investigated fluorescent compounds as single tracers; use of several tracers in the same animal to map collateral projections of one and the same neuron; use of several tracers in the same animal to establish the topographical relation between several independent neuronal populations.     

Brain-stem involvement in multiple sclerosis: a comparison between brain-stem auditory evoked potentials and the acoustic stapedius reflex

Summary Brain-stem auditory evoked potentials (BAEPs) and the acoustic stapedius reflex (ASR) were recorded in 68 patients with definite, probable and possible multiple sclerosis (using the definitions of McAlpine). The high incidence of abnormal results, 68% and 60%, respectively, pointed to the diagnostic value of these two measures in detecting brain-stem dysfunction. Combination of the methods increased the diagnostic yield to 85%. Since in part the same brain-stem generator sites underlie BAEPs and the ASR, it was considered that a study of their correlation might serve to increase the reliability and validity of these techniques. There was 71% agreement overall between results from the two measures. Furthermore, 72% of the joint BAEP and ASR abnormalities corresponded in detection of the brain-stem lesion site. It was concluded that the combined approach may supply powerful, complementary information on brain-stem dysfunction, which may aid in establishing the diagnosis of multiple sclerosis.This study was supported by the M. Sackler Foundation for Multiple Sclerosis Research     

Transport of molecules from nose to brain: Transneuronal anterograde and retrograde labeling in the rat olfactory system by wheat germ agglutinin-horseradish peroxidase applied to the nasal epithelium

Transneuronal anterograde labeling with the conjugate wheat germ agglutinin-horseradish peroxidase (WGA-HRP) has been documented in the mammalian and immature avian visual system [6,14]. Transneuronal retrograde labeling was significant only in the chick [6]. The present study was performed to determine whether transneuronal labeling could be shown in the mammalian olfactory system, whether the phenomenon was robust in adults, and whether transneuronal retrograde transport could label several transmitter-specific centrifugal afferent projections to the olfactory bulb. In addition we wished to learn whether molecules that enter the nasal cavity can undergo transport to brain neurons. Gelfoam implants soaked with 1% WGA-HRP, surgically implanted into the nasal cavity, produced transneuronal labeling patterns that affirmed all of these questions. Transneuronal anterograde transport labeled the appropriate zones in the olfactory bulb and in all second order olfactory targets. In addition, there was transneuronal retrograde labeling of neurons in the olfactory bulb, anterior olfactory nucleus and in transmitter-specific projection neurons from the diagonal band (cholinergic), raphe (serotonergic) and locus coeruleus (noradrenergic). Transneuronal labeling was robust and consistent. The patterns of labeling indicated that transneuronal anterograde and retrograde transport occurred along known, specific circuits in the olfactory system. The present results suggest that nasal epithelial application of WGA-HRP may be a useful tool for assessing regeneration of primary olfactory neurons and the status of central circuitry following regeneration. The method should also facilitate the study of central olfactory connections after surgical or genetic lesions of the olfactory bulb. Finally, these experiments suggest the possibility that inhaled molecules including, possibly substances of abuse, may be transported to, and, possibly, influence the function of neurons in the brain, including some (diagonal band, raphe, locus coeruleus) which have extensive projections to wide areas of the CNS.     

A direct hypothalamic projection to the superior salivatory nucleus neurons in the rat. A study using anterograde autoradiographic and retrograde HRP methods

The location of the superior salivatory nucleus and terminal labelings of the hypothalamic descending fibers were demonstrated in the nucleus reticularis parvocellularis using HRP and the autoradiographic techniques, respectively. When both techniques were used in the same animals, some HRP-labeled neurons were seen among the accumulations of silver grains, suggesting pericellular terminations. The present study demonstrates that the hypothalamic efferents project directly to the superior salivatory nucleus innervating salivary and lacrimal glands.     

Projection neurons of the basolateral amygdala: A correlative Golgi and retrograde tract tracing study

This study analyzed the projection neurons of the anterior subdivision of the rat basolateral amygdaloid nucleus (BLa) by correlating the morphology of Golgi-stained neurons with the morphology of neurons that were retrogradely labeled by injections into the main terminal fields of BLa. In each animal multiple injections of horseradish peroxidase (HRP) and wheat germ agglutinin-conjugated HRP were made into the prefrontal cortex and rostral striatum. These injections labeled approximately 85% of BLa neurons. The great majority of labeled neurons were the same shape and relative size as the pyramidal (class I) neurons described in previous Golgi studies. The unlabeled neurons appeared to correspond to the nonpyramidal (class II and class III) neurons described in Golgi studies. Thus this investigation provides experimental evidence that the pyramidal neurons are the main projection neurons of BL, whereas most of the nonpyramidal cells are local circuit neurons.     

The cutaneous contribution to the hamstring flexor reflex in the rat: an electrophysiological and anatomical study

The location and properties of the cutaneous receptive fields responsible for detecting the flexor withdrawal reflex in the posterior head of biceps femoris (pBF) and semitendinosus (ST) components of the hamstring muscle have been examined in unanaesthetized decerebrate rats, spinalized at T10-T11. Single alpha-motoneurone efferents were recorded from the nerve to pBF and the principal head of ST and their responses to ipsi- and contralateral hindlimb skin stimulation investigated. The efferents to both muscles characteristically had a low or absent background discharge and they all had mechanoreceptive fields on the ipsilateral foot. The mechanical threshold of these fields was high with no response to light touch or brush. Fifty-four percent of these units also had a smaller and weaker contralateral mechanoreceptive field. The only apparent difference between ST and pBF efferents was that more ST efferents had contralateral fields than pBF units. Noxious, hot and cold thermal stimuli applied to the ipsilateral foot activated 56% of the efferents. Mustard oil, a chemical irritant, produced a long-lasting flexor response when applied to the ipsilateral foot. The responses of these efferents to stimulation of A beta, A delta and C cutaneous afferents in the sural nerve were also studied. Short latency reflexes were elicited in all efferents by A beta inputs, longer latency reflexes were elicited in 64% by A delta inputs and very long latency responses with long afterdischarges were found in 73% of the units to C inputs. Retrograde labelling of the hamstring motoneurones with WGA-HRP indicated that they lay in ventrolateral lamina IX extending from the caudal portion of the third lumbar segment to the junction of the 5th and 6th lumbar segments. Transganglionic labelling of small diameter primary afferent terminals in the dorsal horn of cutaneous nerves innervating the foot revealed that the longitudinal distribution corresponded closely with that of the hamstring motor nucleus. The flex-or reflex in the spinal rat provides a useful model therefore, for studying how the input in nociceptive afferents is processed and transformed within the spinal cord, to produce appropriate outputs.     

The trajectory of the sympathetic nerve fibers to the rat cochlea as revealed by anterograde and retrograde WGA-HRP tracing

Wheat germ agglutinin-horseradish peroxidase conjugate was injected in the unilateral superior cervical ganglion (SCG), and the projection pathways of postganglionic sympathetic nerve fibers innervating the cochlea were traced in the rat. The labeled axons advanced along the internal carotid artery (ICA), and a few advanced caudally in the major petrosal nerve (MPN) and entered the facial nerve, while the majority ran rostral to the pterygopalatine ganglion at the point where they crossed the MPN in the carotid canal. The rest of the labeled fibers remained on the surface of the ICA and advanced to the cranial cavity. Most of the labeled fibers along the facial nerve joined the cochlear nerve and finally reached the osseous spiral lamina through the spiral ganglion. Some of the labeled fibers ran along the anterior inferior cerebellar artery from the basilar artery which was previously thought to have been the only pathway. We could not find any labeled fiber on the modiolar artery from anterior inferior cerebellar artery in the cochlea. These observations are consistent with our hypothesis that the sympathetic fibers innervating the neural tissues or related structures follow nerve fibers and meninges as matrices of projection pathways rather than arteries.     

Origin of sympathetic and sensory innervation of the elbow joint in the rat: A retrograde axonal tracing study with wheat germ agglutinin conjugated horseradish peroxidase

After injection of wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) into the elbow joint of adult rats, labeled neurons were found in the stellate and the T2-T4 ganglia of the ipsilateral sympathetic trunk, and also in dorsal root ganglia at the C4–T4 levels. Most labeled sympathetic cells, 90% or more, were located in the stellate ganglion. The sensory innervation to the joint originated mainly from the dorsal root ganglia at the levels of C7–T1.     

Afferents to the zona incerta in the rat: a combined retrograde and anterograde study

In a first set of experiments, the retrograde transport of horseradish peroxidase (HRP) was utilized to investigate the afferent projections to the zona incerta (ZI) in the hooded rat. HRP was introduced in its crystalline form into various sectors of the ZI of seven subjects. The largest contingent of afferents arises from the following centers: the cingulate and somatosensory cortices, central amygdaloid nucleus, ventromedial hypothalamic nucleus, posterior thalamic nucleus, anterior pretectal nucleus, peripeduncular area, deep and intermediate layers of the superior colliculus, dorsal and ventral parabrachial nuclei, principal and interpolar trigeminal subnuclei, and cuneate nucleus. Other centers less systematically or more sparsely labeled were the lateral hypothalamic area, ventrobasal complex, lateral geniculate nucleus pars ventralis, medial geniculate nucleus, interstitial nucleus of Cajal, Darkschewitsch nucleus, perirubral fields, cuneiform, tegmental pedunculopontine, and deep mesencephalic reticular nuclei, pontine reticular nucleus pars oralis, lateral and interpositus cerebellar nuclei, and gracile nucleus. In a second set of experiments, an anterograde tracer (WGA-HRP) was injected in several centers projecting to the ZI in order to localize their terminal fields within this structure. It has been thus possible to distinguish a ventral zone (ventral sector of pars caudalis and pars ventralis) in which the somesthetic (somatosensory cortex, trigeminal complex, and dorsal column nuclei (DCN), collicular, and cerebellar projections terminate, from a dorsal zone (pars dorsalis) to which a limbic input (cingulate cortex and ventromedial hypothalamic nucleus) is directed. In most cases, the labeled terminal fields consisted of well-delimited, narrow bands disposed obliquely, parallel to the cerebral peduncle or the internal capsule. The contingent of somatosensory afferents is relatively large and there is a high degree of overlapping between the different somatosensory terminal fields within the ventral ZI. This suggests a participation of this structure in the treatment of somesthetic information and/or in the transmission of noxious stimuli.     

Striatonigral and pallidonigral pathways studied by a combination of retrograde horseradish peroxidase tracing and a pharmacohistochemical method for γ-aminobutyric acid transaminase

The pharmacohistochemical neuronal staining method for gamma-aminobutyric transaminase (GABA-T) combined with retrograde horseradish peroxidase (HRP) staining was used to define more precisely the descending striatonigral and pallidonigral pathways. Previous studies have established that GABA-T intensive cells in the basal ganglia and other structures correspond with reported glutamic acid decarboxylase (GAD)-containing cells and are therefore presumed to use GABA as their neurotransmitter. Following injection of HRP into the substantia nigra, many HRP-labeled cells were detected in the caudate-putamen and globus pallidus. Two separate groups of cells were doubly labeled for GABA-T and HRP and seemed to represent two distinct GABA-T-rich descending pathways to the substantia nigra. One component came from medium-sized cells in the lateral aspect of the globus pallidus. It represented a majority of all descending cells from that nucleus. The other came from the lateral aspect of the caudate-putamen and represented only a minority of descending cells from that structure. These data suggest that the majority of striatonigral fibers are non-GABA containing while the majority of pallidonigral fibers are GABA-containing. The precise location of the GABA-T intensive cells making up these two pathways helps to explain much confusing data in the literature on the source of descending GABA fibers to the substantia nigra.