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PubMed1989

A reinvestigation of the spinovestibular projection in the cat using axonal transport techniques.

There are numerous discrepancies within the literature concerning the sources of spinovestibular fibers and their distribution in the vestibular complex. Sources of afferents from all spinal levels were sought using the retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase. Following injections of this tracer in all portions of the vestibular complex retrograde labelling was densest at upper cervical levels, especially within the contralateral central cervical nucleus. Labelling was also observed in laminae VI (ipsilaterally), IV, V, VII, and VIII (bilaterally). At progressively more caudal levels, numbers of labelled cells decreased but were similarly distributed in these laminae. The terminal distribution of spinal efferent fibers within the vestibular complex was revealed by injecting wheat germ agglutinin conjugated to horseradish peroxidase or tritiated amino acids into various levels of the spinal cord. These studies showed that all spinal levels project to the descending vestibular nucleus and group x. The f-tail of the descending vestibular complex receives projections from upper cervical and thoracic levels. Terminations within the medial vestibular nucleus arise from both upper cervical and lumbar levels. No conclusive evidence was found supporting the presence of substantial direct spinal projections to the lateral vestibular nucleus, superior vestibular nucleus, or group z. Possible functional roles for the spinovestibular projection in posture and gaze are discussed.

Animals↗
PubMed1998

Autoradiographic visualisation of axonal transport of adenosine A1 receptors along the rat vagus nerve and characterisation of adenosine A1 receptor binding in the dorsal vagal complex of hypertensive and normotensive rats.

The present study had employed in vitro receptor autoradiography with [3H]DPCPX to visualise the presence of adenosine A1 receptors on the rat nodose ganglion, which contains the perikarya of vagal afferent neurons projecting the the nucleus tractus solitarius (NTS). In addition, unilateral vagal ligation resulted in an accumulation of [3H]DPCPX binding adjacent to the ligatures, indication that adenosine A1 receptors are subject to axoplasmic flow along the rat vagus nerve. Radioligand binding assays were utilised to characterise the properties of adenosine A1 receptors in the dorsal vagal complex (NTS, area postrema and dorsal motor nucleus of the vagus) of pup and adult normotensive (Wistar Kyoto, WKY) and hypertensive (spontaneously hypertensive, SHR) rats. Saturation binding indicated that the affinity (KD) of [3H]DPCPX, and the binding site density (Bmax) were not different between the adult WKY and SHR, although the pup SHR had a lower KD value than the pup WKY rat. Competition binding assays revealed complex differences between the two rat strains; however, with respect to hypertension, the affinity of the selective adenosine A1 agonist, cyclohexyladenosine (CHA), was markedly reduced in the membranes from SHR (Ki approximately 93 nM) compared to WKY (approximately 6 nM). Such an observation is consistent with the attenuated responses of SHRs to intra-NTS injections of adenosine.

Adenosine↗
PubMed1976

Transfer RNA may be axonally transported during regeneration of goldfish optic nerves.

If [3H]uridine is injected into the eyes of goldfish during optic nerve regeneration, then the return of fibers to the optic tectum is accompanied by the appearance of [3H]RNA in the tectum. The amount of [3H]RNA arriving in the tectum is consistently greater than in non-regenerating controls and reaches maximum levels (more than 10 times controls) 24 days after optic nerve crush. When [14C]uridine is injected subarachnoidally 1 day prior to sacrificing, the amount of [14C]RNA in the tectum is approximately doubled throughout the regeneration period. In order to characterize the radioactive tectal RNA in these experiments, we have crushed the optic nerves of 15 fish, and 18 days later injected [3H]uridine into both eyes. Five days later [14C]uridine was injected subarachnoidally and all fish were sacrificed a day later. RNA was extracted and fractionated in 2.0% polyacrylamide gels. The amounts of 3H- and 14C-labeled ribosomal as well as small molecular weight RNAs were increased during regeneration. Analysis of the area under the 28S, 18S and 4-7S RNA peaks indicated a small increase in 14C radioactivity in each peak (1.2, 1.5, and 1.5 times control, respectively). On the other hand, 3H radioactivity showed the greatest increase in the 4-7S fraction (8.0 times control) whereas large molecular weight ribosomal fractions were approximately 3 times control. Electrophoresis of the RNA on 10% polyacrylamide gels demonstrated that all of the small molecular weight RNA was confined to the 4S (tRNA) peak. These results suggest that when optic nerves of goldfish regenerate, they may enter the tectum carrying 4S (transfer) RNA.

Animals↗
PubMed1996

Improved retrograde axonal transport and subsequent visualization of tetramethylrhodamine (TMR) -dextran amine by means of an acidic injection vehicle and antibodies against TMR.

We studied the ability of various dextran amines (DA) to retrogradely label cortical neurons to the full extent of their dendritic configurations. Corticothalamic neurons were labeled by pressure injection of DA into the ventrobasal thalamic nuclei of the rat brain. Of fluorescein-, Texas Red-, Cascade Blue- and tetramethylrhodamine (TMR)-DAs of MW 3000 and TMR-DA of MW 10,000, neurons were most efficiently labeled with TMR-DA of MW 3000. The use of acidic vehicles (pH 1-3) for dissolving TMR-DA enhanced the retrograde labeling, as compared with that of a neutral vehicle. The retrograde labeling with TMR-DA was more clearly demonstrated by using anti-TMR antibodies; the indirect immunofluorescence method with a rhodamine-conjugated secondary antibody and immunoperoxidase method with a peroxidase anti-peroxidase (PAP) complex revealed that the dendrites of many corticothalamic neurons were filled with TMR-DA. The Golgi-like retrograde labeling of TMR-DA visualized by the PAP immunoperoxidase method was comparable with that of biotinylated DA by the avidin-biotinylated peroxidase complex method. Similar Golgi-like dendritic staining was observed among corticospinal neurons after injection of TMR-DA into the corticospinal tract of the spinal cord. Most apical dendrites of corticospinal neurons extended into layer I, whereas those of corticothalamic neurons ended in layer IV or the deep part of layer III. The TMR-DA injection under acidic conditions and immunostaining with the anti-TMR antibodies are considered to be a useful method to visualize the dendrite configuration of cortical projection neurons.

Animals↗
PubMed1978

Mesencephalic and diencephalic afferents to the superior colliculus and periaqueductal gray substance demonstrated by retrograde axonal transport of horseradish peroxidase in the cat.

The mesencephalic and diencephalic afferent connections to the superior colliculus and the central gray substance in the cat were examined by means of the retrograde transport of horseradish peroxidase (HRP). After deep collicular injections numerous labeled cells were consistently found in the parabigeminal nucleus, the mesencephalic reticular formation, substantia nigra pars reticulata, the nucleus of posterior commissure, the pretectal area, zona incerta, and the ventral nucleus of the lateral geniculate body. A smaller number of cells was found in the inferior colluculus, the nucleus of the lateral lemniscus, the central gray substance, nucleus reticularis thalami, the anterior hypothalamic area, and, in some cases, in the contralateral superior colliculus, Forel's field, and the ventromedial hypothalamic nucleus. Only the parabigeminal nucleus and the pretectal area showed labeled cells following injections in the superficial layers of the superior colliculus. In the cats submitted to injections in the central gray substance, labeled cells were consistently found in the contralateral superior colliculus, the mesencephalic reticular formation, substantia nigra parts reticulata, zona incerta and various hypothalamic areas, especially the ventromedial nucleus. In some cases, HRP-positive cells were seen in the nucleus of posterior commissure, the pretectal area, Forel's field, and nucleus reticularis thalami. A large injection in the mediodorsal part of the caudal mesencephalic reticular formation, which included the superior colliculus and the central gray substance, resulted in numerous labeled cells in nucleus reticularis thalami. The findings are discussed with respect to the suggested functional division of the superior colliculus into deep and superficial layers. Furthermore, the possible implications of labeled cells in zona incerta and the reticular thalamic nucleus are briefly discussed.

Afferent Pathways↗
PubMed1977

The olivocerebellar projection in the cat studied with the method of retrograde axonal transport of horseradish peroxidase. VI. The projection onto longitudinal zones of the paramedian lobule.

Microinjections (30-50 nl) of a horseradish peroxidase (HRP) suspension of 25% (wt./vol.) were made in different folia of the paramedian lobule of cats, and the sites of occurrence of labeled cells in the inferior olive were precisely determined. In each case only a small number of cells are labeled, aggregated in a minute area. The labeled cells are found within three only of the four olivary areas previously determined (Brodal et al., '75) to project onto the paramedian lobule (fig. 1): one area in the rostral half of the medial accessory olive, another in the dorsal accessory olive (except its caudalmost part), and a third in part of the caudal half of the dorsal lamella of the principal olive. Labeled cells were never found in the fourth area, the ventral lamella. A distinct zonal pattern in the projection is demonstrated (figs. 3, 5B): a middle longitudinal zone of the paramedian lobule receives olivary afferents from the area in the medial accessory olive, a medial zone from part of the projection area in the dorsal accessory olive, a lateral zone from part of the projection area in the dorsal lamella. This zonal projection appears to extend throughout the length of the paramedian lobule (the two caudalmost folia could not be studied). tthe somatotopical pattern in the projections from the accessory olives described previously (Brodal et al., '75) is confirmed. The pattern of a zonal projection obtained with the HRP-method (fig 5B) is simpler than that deduced by Armstrong et al ('74) from recordings of antidromic potentials in the olive (fig 5A). Concerning main points there is satisfactory agreement. The phenomenon that following microinjections of HRP in superficial parts of the folia labeled cells occur within parts only of the regions of the olive which contain labeled cells following large HRP-injections in the paramedian lobule is discussed.

Afferent Pathways↗
PubMed1983

The projection from the motor cortex to the inferior olive in the cat. An experimental study using axonal transport techniques.

The cortico-olivary projection has been investigated in the cat with the methods of retrograde transport of horseradish peroxidase and wheat-germ agglutinin conjugated with horseradish peroxidase as well as autoradiographic techniques using tritium-labelled amino acids. The projection arises from cells in cortical layer V and terminates mainly ipsilaterally and less densely contralaterally. The strongest termination site is in the caudal medial accessory olive adjacent to subnucleus beta. Projections to that area originate in the medial portions of areas 4 and 6 rostral to the cruciate sulcus. Regions of the motor cortex related to axial back and neck, proximal forelimb and face musculature plus the frontal eye field are represented in largely overlapping areas of the caudal medial accessory olive. A second zone of termination is present in the rostral olive at the junction of the ventral lamella of the principal olive and the medial border of the dorsal accessory olive. Projections to that area arise from a central portion of area 4 at the border between the anterior sigmoid gyrus and the presylvian gyrus. This area contains portions of the representation of the muscle groups controlling the neck and proximal forelimb (shoulder and elbow) only. The frontal eye field, which in the cat influences both extraocular and neck musculature, is also an important direct source of input to this portion of the inferior olive. Contralateral terminations are distributed symmetrically. Combining this information with the olivocerebellar distribution, cerebellar cortical areas corresponding to this direct cortical input are defined. Taking into account that the cortico-olivary fibers appear to arise only from those portions of the motor cortex involved in the control of axial and proximal forelimb muscles, it is suggested that the cortico-olivo-cerebellar projections play a preponderant role in the cerebellar control of posture.

Animals↗
PubMed1988

Brainstem afferents to the magnocellular basal forebrain studied by axonal transport, immunohistochemistry, and electrophysiology in the rat.

Brainstem afferents to the magnocellular basal forebrain were studied by using tract tracing, immunohistochemistry and extracellular recordings in the rat. WGA-HRP injections into the horizontal limb of the diagonal band (HDB) and the magnocellular preoptic area (MgPA) retrogradely labelled many neurons in the pedunculopontine and laterodorsal tegmental nuclei, dorsal raphe nucleus, and ventral tegmental area. Areas with moderate numbers of retrogradely labelled neurons included the median raphe nucleus, and area lateral to the medial longitudinal fasciculus in the pons, the locus ceruleus, and the medial parabrachial nucleus. A few labelled neurons were seen in the substantia nigra pars compacta, mesencephalic and pontine reticular formation, a midline area in the pontine central gray, lateral parabrachial nucleus, raphe magnus, prepositus hypoglossal nucleus, nucleus of the solitary tract, and ventrolateral medulla. A similar but not identical distribution of labelled neurons was seen following WGA-HRP injections into the nucleus basalis magnocellularis. The possible neurotransmitter content of some of these afferents to the HDB/MgPA was examined by combining retrograde Fluoro-Gold labelling and immunofluorescence. In the mesopontine tegmentum, many retrogradely labelled neurons were immunoreactive for choline acetyltransferase. In the dorsal raphe nucleus, some retrogradely labelled neurons were positive for serotonin and some for tyrosine hydroxylase (TH); however, the majority of retrogradely labelled neurons in this region were not immunoreactive for either marker. The ventral tegmental area, substantia nigra pars compacta, and locus ceruleus contained retrogradely labelled neurons which were also immunoreactive for TH. Of the retrogradely labelled neurons occasionally observed in the nucleus of the solitary tract, prepositus hypoglossal nucleus, and ventrolateral medulla, some were immunoreactive for either TH or phenylethanolamine-N-methyltransferase. To characterize functionally some of these brainstem afferents, extracellular recordings were made from antidromically identified cortically projecting neurons, mostly located in the HDB and MgPA. In agreement with most previous studies, about half (48%) of these neurons were spontaneously active. Electrical stimulation in the vicinity of the pedunculopontine tegmental and dorsal raphe nuclei elicited either excitatory or inhibitory responses in 21% (13/62) of the cortically projecting neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1987

Anatomical organization of cortico-mesencephalo-olivary pathways in the cat as demonstrated by axonal transport techniques.

Cortical projections arising from areas 4 and 6 and terminating in midbrain cell groups known to project to the inferior olive (IO) have been studied in the cat. Injections of the bidirectional tracers horseradish peroxidase (HRP) and wheat germ agglutinin (WGA) conjugated to HRP were made into the midbrain. All cases of lateralized midbrain injections resulted in virtually ipsilateral labelling of lamina V cortical neurons. Retrogradely labelled neurons in cortical areas 4 and 6 were found after injections located in the interstitial nucleus of Cajal (INC), nucleus of Darkschewitsch (ND), and in the caudal parafascicular (Pf) and subparafascicular (sPf) nuclei (perifascicular region, PF). Injections that were more caudal and within the parvi- and magnocellular red nucleus (RNp and RNm) labelled cells not only in areas 4 and 6 but also in portions of adjacent areas 3a, 3b, 5a, and 7. These midbrain injections also resulted in the anterograde labelling of projections observed to terminate in the ipsilateral IO. The distribution in the midbrain of projections arising from cortical areas 4 and 6, and portions of areas 3a and 3b, was studied with autoradiographic methods. After injections of tritium-labelled amino acids in those cortical areas, a pattern of largely ipsilateral terminations was revealed. Whereas all cortical areas studied labelled the PF, differential grain distributions in central mesencephalic nuclei were apparent after injections in various portions of the motor and adjacent somatosensory cortex. Injections involving the frontal eye fields (FEF) labelled the INC bilaterally, but ipsilateral terminations were densest. These cases also labelled the region of the fields of Forel. When the neck region of the cortex was involved in the injections, the more caudal aspects of the INC (INCc) and the RN were labelled. The cortical areas related to the upper limb gave rise to terminations in the ND and the RNp. Contributions to both ND and RNp inputs from injections in the FEF and neck regions were also occasionally but not consistently noted. A relatively discrete injection in the vibrissae field weakly labelled ND. Additional components of the motor cortical projections to the superior colliculus (SC) and pretectal nuclei were also analysed since those regions also project to the IO. Cortical regions involving the representation of the neck musculature were shown to project principally ipsilaterally to lamina IV of the SC as well as to the anterior pretectal nucleus.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1977

The olivocerebellar projection in the cat studied with the method of retrograde axonal transport of horseradish peroxidase. IV. The projection to the anterior lobe.

Following injections of horseradish peroxidase (HRP) in the cerebellar cortex of the anterior lobe of the cat, the distribution of labeled cells in the inferior olive was mapped. The findings largely confirm those made previously in studies of olivary retrograde cell loss following cerebellar ablations (Brodal, '40b). In addition, they reveal further olivary areas projecting onto the anterior lobe, and permit a more detailed analysis of the pattern in this projection. Concerning major points the results are in agreement with physiological studies by Armstrong et al. ('74). They bring supporting evidence for a longitudinal zonal pattern in the anterior lobe (fig. 6C). The middle zone of the vermis receives its fibers from a large central area in the caudal half of the medial accessory olive, a lateral zone of the vermis from the lateral half of the dorsal accessory olive. Both olivary areas project to the corresponding cerebellar zone throughout lobules V-I. The lateralmost part of the anterior lobe (lobules IV-V) receives afferents from an area in the dorsal lamella of the principal olive. The intermediate part of lobules IV-V receives afferents from the medial half of the dorsal accessory olive and from an area in the rostral half of the medial accessory olive. There is suggestive evidence that the latter projects to a middle zone, the former to a medial and a lateral zone within the intermediate part as found physiologically. Conclusions concerning projections to the intermediate part of lobules III-II could not be made. The findings in this and preceding studies with the HRP-method show that the concept of a longitudinal pattern in the cerebellum is scarcely generally valid of the entire olivocerebellar projection. Within the projections of the lateral half of the dorsal accessory olive and the area in the rostral part of the medial accessory olive there appears to be a topical relation with the folial pattern in the anterior lobe. An analysis of the findings with reference to the afferents traced anatomically to the various olivary areas permits some conclusions as to the functional role of the olivary areas. Comparison with Oscarsson's ('73) diagram of the sites of termination of two of the spinal-olivary pathways (his DF-SOCP) and VF-SOCP) permits an anatomical explanation as concerns the projections to the vermis, while correlations as concerns the intermediate part are less satisfactory.

Animals↗
PubMed1976

The olivocerebellar projection in the cat studied with the method of retrograde axonal transport of horseradish peroxidase. III. The projection to the vermal visual area.

Horseradish peroxidase (HRP) was injected separately in one of the cerebellar lobules VI, VIIA, VIIB, VIIIA or VIIIB (together corresponding to the vermal visual area) in 17 cats. After 1-3 days the distribution of labeled cells in the inferior olive was mapped. In spite of some overlapping it is clear that the various lobules of the vermal visual area receive fibers from separate parts of a horseshoeshaped region in the caudal half of the contralateral medial accessory olive (fig. 5C). The projection area of lobule VIIA is found caudomedially and overlapping with the area supplying lobule VIIB. This in addition receives a few fibers from the nucleus beta. Fibers terminating in lobule VIIIA arise caudolaterally as do fibers destined for lobule VIIIB. A central part of the total projection area projects to lobule VI. Following injections leading to a similar extent of cortical staining in lobules VI, VII or VIII the projection of labeled cells in the corresponding projection areas differ markedly. In the area of lobule VII apparently all cells are labeled, in the area of lobule VI the density of labeled cells is considerably less, and in that of lobule VIII there are rather few labeled cells. In a few cases with widespread staining of the cerebellar visual area there was spreading of HRP to the nucleus fastigii. The projection to this form the olive was therefore investigated to avoid erroneous conclusions. In the discussion it is pointed out that on most points our findings agree fairly well with the results of studies of the olivocerebellar projection undertaken with other methods (studies of retrograde cellular changes, electrophysiological methods). No support for a longitudinal subdivision of lobules VI-VIII was found. Studies of the available literature indicate that the areas in the medial accessory olive projecting onto lobules VI-VIII probably do not receive direct afferents from regions which are known to be concerned in the transmission of visually evoked impulses. Fibers to the olive from the superior colliculus appear to pass to the nucleus beta only. This projects mainly to the uvula, to a little extent only to lobule VII. However, it may be imagined that visual impulses may reach the vermal visual area via the inferior olive by way of intercalated neurons, for example in the mesencephalic RF. Major contingents of afferents to the olivary regions projecting onto the vermal area come from the spinal cord, the motor cortex and the periaqueductal gray.

Animals↗
PubMed1988

Anterograde and retrograde axonal transport of Phaseolus vulgaris leucoagglutinin (PHA-L) from the globus pallidus to the striatum of the rat.

Iontophoretic administration of PHA-L into the globus pallidus of rats resulted in the labeling of neuronal perikarya in the striatum as well as axons and terminals in the striatum, entopeduncular nucleus, subthalamus and substantia nigra. The labeled striatal perikarya were densely stained in Golgi fashion with virtually complete filling of the dendrites and spines. It is concluded that the striatal cells were filled by the retrograde transport of PHA-L and represent either striatopallidal cells, or striatonigral cells whose axons were interrupted as they passed through the injection site. The anterogradely labeled axon terminals in the striatum were observed in close apposition to the dendrites of the retrogradely labeled neurons suggesting the existence of synaptic contacts between the two groups of cells. This study demonstrates that PHA-L can be transported retrogradely as well as anterogradely following iontophoretic injections.

Animals↗
PubMed1986

The long-term effects of a single injection of beta,beta'-iminodipropionitrile on slow axonal transport in the rat.

Following a single intraperitoneal injection of beta,beta'-iminodipropionitrile (IDPN) simultaneously with or 5 weeks after injection of [35S]methionine into the ventral horn area of rat spinal cord, the changes of slowly migrating axonal proteins were analyzed electrophoretically up to 10 weeks after labeling, and the following results were obtained. After a single injection of IDPN, only the transport of neurofilament proteins is inhibited, leaving that of tubulin and actin almost unaffected, though a small portion of the former was retarded through the interaction with neurofilaments. The inhibitory effect of IDPN on neurofilament transport is not a complete blockage, but a slowing of the rate of transport to about a half of the control with a possible short halting period just after IDPN treatment. The dose-response data indicate a threshold between 0.5 and 1.0 g/kg of body weight, increasing the dose above which does not further affect the neurofilament transport. The transport of neurofilaments is uniformly impaired by IDPN along the whole axon.

Animals↗
PubMed1972

Axonal transport of tritium-labeled putrescinein the embryonic visual system of zebrafish.

The transport of [(3)H]putrescine is demonstrated by autoradiography in the retino-tectal tract of Brachydanio rerio embryos. Transport of [(3)H]putrescine appears to be more rapid than that of tritium-labeled protein and is not inhibited by a colchicine effect on axonal neurotubules as is protein transport. The radioactivity transported to the brain is found, on electrophoresis, in the putrescine fraction.

Animals↗
PubMed1976

Axonal transport of [3H]serotonin in an identified neuron of Aplysia californica.

A population of characteristic ellipsoidal dense-core vesicles was identified in axons of the giant cerebral neuron of the mollusc Aplysia. We injected [3H]serotonin into the cell body of this identified serotonergic neuron in the isolated central nervous system in order to study the subcellular components associated with the neurotransmitter. Subcellular fractionation by differential centrifugation indicated that injected serotonin was rapidly taken up into particulate form. [3H]Serotonin appeared in the axons within 2 h after injection, and export continued at a constant rate of 6% of the total in the neuron/h thereafter. The dependence of the total amounts of [3H]serotonin which appeared in the axons in 6 h (export from the cell body) on the amounts injected was consistent with the idea that export is a saturable process, possibly depending on the capacity of somatic vesicles or of some unidentified carrier for serotonin. [3H]Serotonin moved into both major branches of the axon, where it was translocated rapidly. The transmitter, which was shown by autoradiography to be restricted to the axons of the injected cell, was distributed along axons in accumulations of radioactivity; in contrast, its precursor, [5-3H]hydroxytryptophan, moved slowly along axons in a smooth, declining curve, its kinetics consistent with diffusion. Quantitative electron microscope autoradiography revealed that the dense-core vesicles and the cytosol of axons fixed with glutaraldehyde were labeled with [3H]serotonin.

5-Hydroxytryptophan↗