Cluster-and-sheet pattern of enkephalin-like immunoreactivity in the superior colliculus of the cat

The distribution of enkephalin-like immunoreactivity in the superior colliculus has been studied in the cat with the peroxidase-antiperoxidase method. Two striking patterns of immunoreactivity were observed. In the superficial layers there is a thin, dense horizontal band of immunoreactivity in the neuropil of the most dorsal tier of the superficial gray layer (sublamina 1). Because this sublayer corresponds to the zone of densest contralateral retinotectal projection, an intraocular injection of horseradish peroxidase was made in one cat to allow direct comparison of the distributions of opiate-like immunoreactivity and transported tracer in the contralateral superior colliculus. There was a detailed similarity between the two, including the presence of a gap in both at the presumptive site of the optic disc representation. The presence of enkephalin-like immunoreactivity in neural perikarya in and near sublamina 1 of the superficial gray layer, however, raised the possibility that the immunoreactive band is part of an intrinsic opiate system. Deeper in the superficial gray layer there was appreciable but weaker immunoreactivity in the neuropil and fewer immunoreactive neurons. In the intermediate gray layer and, especially medially, even deeper in the superior colliculus, enkephalin-like immunoreactivity was organized into small (100-300 micron wide) patches. In the intermediate gray layer these tended to be arranged periodically, five-seven patches being spaced at 200-600 micron intervals in caudal transverse sections. In some sections adjoining patches appeared to be fused. The patches were absent or difficult to detect in rostral sections. Caudally, they sometimes were adjacent to blood vessels penetrating the intermediate gray layer, but other times were not. Serial section reconstructions suggested that the patches observed in individual sections are part of larger arrays which have the form of anastomotic bands running in longitudinal directions somewhat oblique to the sagittal plane. It is concluded that an opiate mechanism may play a part in controlling the effects of incoming retinal information in the superficial gray layer, directly or indirectly, and that opiate peptides may also act in modulating one or more afferent or efferent systems of the deep collicular layers. Accordingly, from the functional standpoint, enkephalin-like peptides may influence both visual and sensory motor processing in the superior colliculus.     

Calcium-binding protein calretinin immunoreactivity in the dog superior colliculus

We studied calretinin-immunoreactive (IR) fibers and cells in the canine superior colliculus (SC) and studied the distribution and effect of enucleation on the distribution of this protein. Localization of calretinin was immunocytochemically observed. A dense plexus of anti--calretinin-IR fibers was found within the upper part of the superficial gray layer (SGL). Almost all of the labeled fibers were small in diameter with few varicosities. The intermediate and deep layers contained many calretinin-IR neurons. Labeled neurons within the intermediate gray layer (IGL) formed clusters in many sections. By contrast, labeled neurons in the deep gray layer (DGL) did not form clusters. Calretinin-IR neurons in the IGL and DGL varied in morphology and included round/oval, vertical fusiform, stellate, and horizontal neurons. Neurons with varicose dendrites were also labeled in the IGL. Most of the labeled neurons were small to medium in size. Monocular enucleation produced an almost complete reduction of calretinin-IR fibers in the SC contralateral to the enucleation. However, many calretinin-IR cells appeared in the contralateral superficial SC. Enucleation appeared to have no effect on the distribution of calretinin-IR neurons in the contralateral intermediate and deep layers of the SC. The calretinin-IR neurons in the superficial dog SC were heterogeneous small- to medium-sized neurons including round/oval, vertical fusiform, stellate, pyriform, and -horizontal in shape. Two-color immunofluorescence revealed that no cells in the dog SC -expressed both calretinin and GABA. Many horseradish peroxidase (HRP)-labeled retinal ganglion cells were seen after injections into the superficial layers. The vast majority of the double-labeled cells (HRP and calretinin) were small cells. The present results indicate that antibody to calretinin labels subpopulations of neurons in the dog SC, which do not express GABA. The results also suggest that the calretinin-IR afferents in the superficial layers of the dog SC originate from small class retinal ganglion cells. The expression of calretinin might be changed by the cellular activity of selective superficial collicular neurons. These results are valuable in delineating the basic neurochemical architecture of the dog visual system.     

Evidence for the existence of several different alpha- and beta-subunits of the GABA/benzodiazepine receptor complex from rat brain

gamma-Aminobutyric acid (GABA)/benzodiazepine receptors purified from the brains of young and adult rats were photolabeled by [3H]flunitrazepam or [3H]muscimol. Gel electrophoresis revealed 3 different proteins with apparent molecular weight (Mr) 51,000, 53,000 and 59,000 which were specifically and irreversibly labeled by [3H]flunitrazepam and recognized by the alpha-subunit specific antibody bd-28. Similarly, 3 different proteins with Mrs 51,000, 53,000 and 56,000 were irreversibly labeled by [3H]muscimol and recognized by the beta-subunit-specific antibody bd-17. Comparison of the photolabeling and immunological staining pattern from young and adult rats seems to indicate that proteins labeled by [3H]flunitrazepam and bd-28 are different from those labeled by [3H]muscimol and bd-17.     

Regional and laminar distribution of choline acetyltransferase immunoreactivity in the cat superior colliculus

The pattern of distribution of cholinergic fibers was examined immunohistochemically in the cat superior colliculus by using a monoclonal antibody against choline acetyltransferase (ChAT). In the superficial layers, an obvious immunoreactive zone was found in the rostral two-thirds of the outer portion of the superficial gray layer (SGS), with increasing immunoreactive intensity at the rostral pole of the colliculus. A mesh-like distribution of the immunoreactive fibers was found throughout the deeper portion of this layer with a higher concentration in the caudal levels. In the deeper collicular layers, a number of ChAT-immunoreactive fibers were seen in the outer portion of the intermediate gray layer (SGI) in a patch-like fashion. A few fibers were also immunoreactive in the deeper portion of the SGI and in the medial aspect of the deep gray layer. The density of the immunoreactivity in the deeper layers increased in the caudal levels. After unilateral destruction of the parabigeminal nucleus, the ChAT immunoreactivity was markedly reduced in the rostral aspect of the contralateral SGS, and moderately in the caudal aspect of the ipsilateral SGS.     

Evidence for a neurotransmitter function of acetylcholine in rabbit superior colliculus

Acetylcholinesterase staining and studies on the uptake of [3H]choline into the subsequent efflux of tritium from collicular slices were carried out in order to provide evidence for a neurotransmitter function of acetylcholine in rabbit superior colliculus. Acetylcholinesterase staining was dense and homogeneous in superficial layers whereas the staining was arranged in patches with slightly higher density caudally than rostrally in the intermediate layers. The accumulation of tritium in slices incubated with [3H]choline depended on time, temperature and concentration, and was inhibited by hemicholinium-3. Accumulation was slightly higher in caudal than in rostral slices. Electrical stimulation enhanced tritium outflow from slices preincubated with [3H]choline. Tetrodotoxin and a low calcium medium inhibited the evoked overflow whereas hemicholinium-3 caused an enhancement. Oxotremorine decreased the evoked overflow; atropine prevented this effect. The opioids [D-Ala2, MePhe4, Glycol5]enkephalin, [D-Ala2, D-Leu5]enkephalin and ethylketocyclazocine caused an inhibition. The effects of the latter two agonists were antagonized by naloxone. The GABAB-receptor-agonist (-)-baclofen decreased the evoked overflow at lower concentrations than GABA, whereas the GABAA-receptor-agonist muscimol was ineffective. Serotonin produced an inhibition which was prevented by metitepin, alpha- and beta-adrenoceptor as well as dopamine-receptor ligands caused no change. It is concluded that in the rabbit superior colliculus the pattern of acetylcholinesterase staining is comparable, but not identical to the distribution in other species. The accumulation of [3H]choline, as well as the tetrodotoxin-sensitive and calcium-dependent overflow of tritium upon electrical stimulation (reflecting presumably release of [3H]acetylcholine) indicate that acetylcholine has a neurotransmitter function in this tissue. The release of [3H]acetylcholine was modulated by various transmitter substances and related compounds. The pattern of modulation of release differed from the pattern in other cholinergically innervated tissues.     

Uptake, release, and modulation of release of noradrenaline in rabbit superior colliculus

The noradrenaline content, the uptake of [3H]noradrenaline, and the release of previously incorporated [3H]noradrenaline were studied in slices of rabbit superior colliculus. The concentration of endogenous noradrenaline was higher in superficial than in deep layers of the superior colliculus. Upon incubation with [3H]noradrenaline, tritium was accumulated by a mechanism that was strongly inhibited by oxaprotiline but little inhibited by 6-nitroquipazine. Electrical stimulation at 0.2 or 3 Hz increased the outflow of tritium from slices preincubated with [3H]noradrenaline; the increase was almost abolished by tetrodotoxin or a low calcium medium. Clonidine reduced the evoked overflow of tritium, whereas yohimbine increased it and antagonized clonidine. The evoked overflow was also reduced by the dopamine D2-receptor-selective agonists apomorphine and quinpirole, an effect antagonized by sulpiride. The preferential opioid kappa-receptor agonist ethylketocyclazocine produced an inhibition that was counteracted by naloxone. Nicotine accelerated the basal outflow of tritium; part of the acceleration was blocked by hexamethonium. The muscarinic agonist oxotremorine slightly diminished the electrically evoked overflow, and its effect was abolished by atropine. The oxaprotiline-sensitive uptake of [3H]noradrenaline as well as the tetrodotoxin-sensitive and calcium-dependent overflow of tritium upon electrical stimulation (presumably reflecting the release of [3H]noradrenaline) indicate that noradrenaline is a neurotransmitter in the superior colliculus. The release of [3H]noradrenaline is modulated through alpha 2-adrenoceptors as well as dopamine D2-receptors, opioid kappa-receptors and nicotine and muscarine receptors. No clear evidence was found for modulation through beta-adrenoceptors, D1-receptors, serotonin receptors, opioid mu- or delta-receptors or receptors for GABA or glutamate. Only the alpha 2-adrenoceptors receive an endogenous agonist input, at least under the conditions of these experiments. The pattern of presynaptic modulation resembles that found for noradrenaline release in other rabbit brain regions, suggesting that all noradrenergic axons arising in the locus coeruleus possess similar presynaptic receptor systems.     

Distribution of corticotectal cells in macaque

We compared the cortical inputs to the superficial and deep compartments of the superior colliculus, asking if the corticotectal system, like the colliculus itself, consists of two functional divisions: visual and visuomotor. We made injections of retrograde tracer extending into both superficial and deep layers in three colliculi: the injection site involved mainly the upper quadrant representation in one case, the lower quadrant representation in a second case, and both quadrants in a third. In a fourth colliculus, the tracer injection was restricted to the lower quadrant representation of the superficial layers. After injections involving both superficial and deep layers, labeled cells were seen over V1, many prestriate visual areas, and in prefrontal and posterior parietal cortex. Both the density of labeled cells and the degree of visuotopic order as inferred from the distribution of labeled cells in cortex varied among areas. In visual areas comprising the lower levels of the cortical hierarchy, visuotopy was preserved, whereas in "higher" areas the distribution of labeled cells did not strongly reflect the visuotopic location of the injection. Despite the widespread distribution of labeled cells, there were several areas with few or no labeled cells: MSTd, 7a, VIP, MIP, and TE. In the case with an injection restricted to superficial layers, labeled cells were seen only in V1 and in striate-recipient areas V2, V3, and MT. The results are consistent with the idea that the corticotectal system consists of two largely nonoverlapping components: a visual component consisting of striate cortex and striate-recipient areas, which projects only to the superficial layers, and a visuomotor component consisting of many other prestriate visual areas as well as frontal and parietal visuomotor areas, which projects to the deep compartment of the colliculus.     

Subcellular localization of GABAA/benzodiazepine receptor-like immunoreactivity in the superficial gray layer of the rat superior colliculus

The monoclonal antibody bd-17, which recognizes the β₂ and β₃-subunits of GABA_A/benzodiazepine receptors, was used to determine the cellular and subcellular localization of receptor-like immunoreactivity in the superficial gray layer of the rat superior colliculus. In numerous dendrites, very strong immunostaining was present in the cytoplasm and on the postsynaptic dendritic membrane of synaptic junctions. The extrasynaptic portion of the dendritic membrane also very often showed [β₂ + β₃]-like immunoreactivity. However, due to methodological limitations, it could not be stated with certainty whether presynaptic β₂- and β₃-subunits of GABA_A/benzodiazepine receptors actually occur in this mesencephalic visual structure. In conclusion, these results strongly suggest that synaptic and non-synaptic GABA_A/benzodiazepine receptors are present in the superficial gray layer of the rat superior colliculus. These receptors may modulate neuronal cell activity in different ways, depending on their location.     

Evidence for intrinsic expression of enkephalin-like immunoreactivity and opioid binding sites in cat superior colliculus

We have investigated the cellular localization of opioid peptides and binding sites in the cat's superior colliculus by testing the effects of retinal deafferentation and intracollicular excitotoxin lesions on patterns of enkephalin-like immunostaining and opiate receptor ligand binding. In normal cats, enkephalin-like immunoreactivity marks a thin tier in the most dorsal stratum griseum superficiale, small neurons of the stratum griseum superficiale, and patches of fibers in the intermediate and deeper gray layers. Eliminating crossed retinotectal afferents by contralateral eye enucleation had little immediate effect on this pattern, although chronic eye enucleation from birth did reduce immunoreactivity in the superficial layers. By contrast, fiber-sparing destruction of collicular neurons by the excitotoxins N-methyl-D-aspartate and ibotenic acid virtually eliminated enkephalin-like immunoreactivity in the neuropil of the upper stratum griseum superficiale, presumably by killing enkephalinergic cells of the superficial layers. Such lesions did not eliminate the patches of enkephalin-like immunoreactivity in the deeper layers. In normal cats, opiate receptor ligand binding is dense in the stratum griseum superficiale, particularly in its upper tier, and moderately dense in the intermediate gray layer. Contralateral eye removal had no detectable effect on the binding pattern, but excitotoxin lesions of the colliculus dramatically reduced binding in both superficial and deep layers. Some ligand binding, including part of that in the upper stratum griseum superficiale, apparently survived such lesions. Similar effects were observed in the lateral geniculate nucleus: enucleation produced no change in binding, whereas excitotoxin lesions greatly reduced specific opiate binding. We conclude that in the superficial collicular layers, both enkephalin-like opioid peptides and their membrane receptors are largely expressed by neurons of intrinsic collicular origin. The close correspondence between the location of these intrinsic opioid elements and the tier of retinal afferents terminating in the upper stratum griseum superficiale further suggests that opiatergic interneurons may modulate retinotectal transmission postsynaptically.     

The cerebellar projections to the superior colliculus and pretectum in the cat: An autoradiographic and horseradish peroxidase study

Efferent projections from the cerebellar nuclei to the superior colliculus and the pretectum have been studied using both retrograde and orthograde labeling techniques in the cat. In order to identify what parts of the cerebellar nuclei project to the superior colliculus and the pretectum, the retrograde horseradish labeling technique was employed. In another set of experiments, tritiated amino acids were injected into each of the cerebellar regions from which the cerebello-tectal and cerebellopretectal projections arise, and the laminar and spatial distributions of orthograde labeling in the superior colliculus and the pretectum were compared.The results showed that the cerebello-tectal projections arise from two different regions of the cerebellar nuclei: the caudal half of the medial nucleus and the ventrolateral part of the posterior interposed nucleus. Fibers arising from the medial nucleus distribute bilaterally in the superficial zone of the intermediate gray layer in the superior colliculus, while those originating from the posterior interposed nucleus terminate contralaterally in the deeper aspect of the intermediate gray layer and in the deep gray and white layers. Although the lateral nucleus does not contribute to the cerebello-tectal projection, it projects profusely to the pretectum contralaterally. The origin of the cerebello-pretectal projection lies in the parvicellular part of the lateral nucleus. Among several pretectal nuclei, the posterior pretectal, the medial pretectal nucleus and the reticular part of the anterior pretectal nucleus receive the cerebellar afferents.The findings of the differential projections from the cerebellum to the superior colliculus and the pretectum suggest that the cerebellum exerts a regulatory influence on visuo-motor and somato-motor transfer in these midbrain structures by differential circuits.     

Time of origin of neurons of the rat superior colliculus in relation to other components of the visual and visuomotor pathways

Summary Groups of pregnant rats were injected with two successive daily doses of ³H-thymidine from gestational day 12 and 13 (E12+E13) until the day before parturition (E21+22) in order to label all the multiplying precursors of neurons. At 60 days of age the proportion of neurons generated (or no longer labelled) on specific days was determined in the separate layers of the superior colliculus. Neurogenesis begins with the production of a few large multipolar neurons in layers V and IV on day E12; the bulk (87%) of these cells are generated on day E13. This early-produced band of large neurons, the intermediate magnocellular zone, divides the superior colliculus into two cytogenetically distinct regions. In both the deep and the superficial superior colliculus neuron production is relatively protracted. In the deep superior colliculus neuron production peaks on day E15 in layer VII, on day E15 and E16 in layer VI, and on day E16 (the large neurons excluded) in layer V, indicating an inside-out sequence. In the superficial superior colliculus peak production time of layer III cells is on day E15 and of layer IV cells on day E16; peak production time of both layer I and II is on day E16 but in the latter region neuron production is more prolonged and ends on day El8. One interpretation of these results is that the two pairs of superficial layers are produced in an outside-in sequence. These three cytogenetic subdivisions of the superior colliculus may be correlated with its structural-functional parcellation into an efferent spinotectal, a deep somatomotor and a superficial visual component.A comparison of neurogenesis in different components of the visuomotor and visual pathways of the rat indicates that the motor neurons of the extraocular muscles, the abducens, trochlear and oculomotor nuclei, and neurons of the nucleus of Darkschewitsch are produced first. Next in line are source neurons of efferents to the bulb and the spinal cord: those of the Edinger-Westphal nucleus and the intermediate magnocellular zone of the superior colliculus. These are followed by the relay neurons of the dorsal nucleus of the lateral geniculate body. The neurons of the superficial superior colliculus and of the visual cortex implicated in visual sensori-motor integrations are produced last.Abbreviations A aqueduct - ap stratum album profundum (layer VII) - bi brachium of the inferior colliculus - c caudal - CGd central gray, pars dorsalis - CGl central gray, pars lateralis - CGv central gray, pars ventralis - dm deep magnocellular zone - EW Edinger-Westphal nucleus - gi stratum griseum intermediale (layer IV) - gp stratum griseum profundum (layer VI) - gs stratum griseum superficiale (layer II) - IC inferior colliculus - im intermediate magnocellular zone - LGd lateral geniculate nucleus, pars dorsalis - ll lateral lemniscus - lm stratum lemnisci (layer V) - MG medial geniculate nucleus - ND nucleus of Darkschewitsch - NO nucleus of the optic tract - op stratum opticum (layer III) - ot optic tract - r rostral - SC superior colliculus - vIII third ventricle - ZO stratum zonale (layer I) - III oculomotor nucleus - IV trochlear nucleus - Vm mesencephalic nucleus of the trigeminal - VI abducens nucleus     

Muscimol in the deep layers of the superior colliculus/mesencephalic reticular formation blocks expression but not acquisition of fear-potentiated startle in rats

Deposits of the retrograde tracer Fluoro-Gold into the ventrolateral nucleus reticularis pontis caudalis labeled neurons in the deep layers of the superior colliculus/mesencephalic reticular formation (deep SC/Me). To test the involvement of this area in the fear-potentiated startle effect, rats were implanted with cannulas into the deep SC/Me and trained for fear-potentiated startle after infusion of the GABA(A) agonist muscimol (0.1 microg/0.5 microl). Two days later, they were tested for fear-potentiated startle. Rats then received a 2nd training session without any infusions, and 2 days later they were reinfused with muscimol (0.1 microg/0.5 microl) and tested for fear-potentiated startle. Local infusion of muscimol into the deep SC/Me completely blocked the expression but not the acquisition of fear-potentiated startle. These results indicate that a synapse in the midbrain is critical for the expression of fear-potentiated startle.     

金黄地鼠上丘和外侧膝状体之间的联系

本实验采用了顺行和逆行追踪技术,对金黄地鼠上丘与丘脑视核团的纤维联系进行了实验观察。一、用尼氏和Loyez染色法,对4只正常金黄地鼠上丘和外侧膝状体背核和腹核的正常结构做了观察。二、3~H-亮氨酸和3~H-脯氨酸注入动物上丘不同部位(4只,存活期一天)后,可见神经末梢标记于同侧的外侧膝状体背核和腹核的外侧部位。若注射部位在上丘外侧,其投射部位在外侧膝状体背核和腹核的尾外侧;注射部位移向内侧,投射部位移向吻外侧。三、将HRP注入外侧膝状体背核(4只,存活期一天)或腹核(2只,存活期一天)或后外侧核(2只,存活期一天)后,在同侧上丘浅层见有标记神经元。在上丘深层则未见。本实验说明了上丘浅层的神经元对同侧的外侧膝状体背核和腹核有局部定位的投射,并按视野和视网膜将该投射作定位排列。     

Ionotropic glutamate receptor GluR2/3-immunoreactive neurons in the cat, rabbit, and hamster superficial superior colliculus

Ionotropic glutamate receptor (GluR) subtypes occur in various types of cells in the central nervous system. We studied the distribution of AMPA glutamate receptor subtype GluR2/3 in the superficial layers of cat, rabbit, and hamster superior colliculus (SC) with antibody immunocytochemistry and the effect of enucleation on this distribution. Furthermore, we compared this labeling to that of calbindin D28K and parvalbumin. Anti-GluR2/3-immunoreactive (IR) cells formed a dense band of labeled cells within the lower superficial gray layer (SGL) and upper optic layer (OL) in the cat SC. By contrast, GluR2/3-IR cells formed a dense band within the upper OL in the rabbit and within the OL in the hamster SC. Calbindin D28K-IR cells are located in three layers in the SC: one within the zonal layer (ZL) and the upper SGL in all three animals, a second within the lower OL and upper IGL in the cat, within the IGL in the rabbit and within the OL in the hamster, and a third within the deep gray layer (DGL) in all three animals. Many parvalbumin-IR neurons were found within the lower SGL and upper OL. Thus, the GluR2/3-IR band was sandwiched between the first and second layers of calbindin D28K-IR cells in the cat and rabbit SC while the distribution of GluR2/3-IR cells in the hamster matches the second layer of calbindin D28K-IR cells. The patterned distribution of GluR2/3-IR cells overlapped the tier of parvalbumin-IR neurons in cat, but only partially overlapped in hamster and rabbit. Two-color immunofluorescence revealed that more than half (55.1%) of the GluR2/3-IR cells in the hamster SC expressed calbindin D28K. By contrast, only 9.9% of GluR2/3-IR cells expressed calbindin D28K in the cat. Double-labeled cells were not found in the rabbit SC. Some (4.8%) GluR2/3-IR cells in the cat SC also expressed parvalbumin, while no GluR2/3-IR cells in rabbit and hamster SC expressed parvalbumin. In this dense band of GluR2/3, the majority of labeled cells were small to medium-sized round/oval or stellate cells. Immunoreactivity for the GluR2/3 was clearly reduced in the contralateral SC following unilateral enucleation in the hamster. By contrast, enucleation appeared to have had no effect on the GluR2/3 immunoreactivity in the cat and rabbit SC. The results indicate that neurons in the mammalian SC express GluR2/3 in specific layers, which does not correlate with the expression of calbindin D28K and parvalbumin among the animals.     

Tectorecipient zone of cat lateral posterior nucleus: evidence that collicular afferents contain acetylcholinesterase

Summary The superficial layers of the cat's superior colliculus innervate the medial subdivision of the thalamic lateral posterior nucleus (LPm). LPm is set off from adjoining thalamic zones by its denser staining for acetylcholinesterase (AChE). We sought to learn whether the tectal afferents to LPm might themselves be the source of the enzyme staining by examining the effects of collicular lesions on the thalamic staining pattern. Large excitotoxin lesions of the colliculus largely eliminated AChE staining in the ipsilateral LPm. By contrast, fibersparing lesions of LPm itself left AChE staining nearly unchanged. Destruction of collicular neurons by excitotoxins dramatically reduced AChE staining in fibers of the brachium and superficial gray layer of the superior colliculus. The reduction was especially pronounced in the lower part of the superficial gray layer, in which LP-projecting collicular neurons are located. These results are consistent with the view that LP-projecting collicular neurons synthesize AChE and account for much of the histochemically detectable enzyme present both in the lower superficial gray layer and in LPm. In the colliculus, the excitotoxin lesions spared AChE staining in a thin sheet at the upper border of the superficial gray layer and in the enzyme-positive patches in the intermediate layers. This surviving tectal AChE thus is probably presynaptic and could be contained at least partly in cholinergic afferents from the parabigeminal nucleus and pontomesencephalic tegmentum. The collicular lesions had no obvious effect on AChE staining in the parabigeminal nucleus or in the C-laminae or ventral division of the lateral geniculate nucleus.     

Evidence for the participation of nigrotectal γ-aminobutyrate-containing neurones in striatal and nigral-derived circling in the rat

The γ-aminobutyrate-containing nature of nigrotectal neurones and the possible involvement of the tectum in circling behaviour were investigated in the rat. Electrolytic or kainic acid lesions of the substantia nigra reduced γ-aminobutyrate levels on average by 19–29% in intermediate and deep, but not superficial superior colliculus. Placement of lesions or injection of muscimol (40 ng) into these γ-aminobutyrate-innervated layers of superior colliculus gave only weak ipsilateral posturing or circling that was intensified by apomorphine, but which strongly antagonized contraversive apomorphine-induced circling in 6-hydroxydopamine pretreated rats (lateral > medial sites). Contraversive circling to unilateral intranigral muscimol (40 ng) was significantly attenuated by lesions or muscimol injections placed in the ipsi- or contralateral superior colliculus. Picrotoxin (40 ng) and tetanus toxin (30 mouse LD₅₀ doses) evoked explosive motor behaviour from medial colliculus and vigorous contraversive circling when injected into the lateral colliculus. The latter offset ipsiversive asymmetries to kainate (0.8 μg) in the corresponding substantia nigra. Bilateral intratectal picrotoxin produced hyperactivity that reversed haloperidol catalepsy. Similar bilateral administration of muscimol did not produce catalepsy but a state of frozen immobility. Kainic acid introduced into the superior colliculus gave mixed excitatory-inhibitory responses initially followed by ipsiversive circling only and loss of tectal perikarya. None of these drug effects occurred from the overlying cerebral cortex or subjacent tegmentum.We propose that separate medial ‘non-postural’ and lateral ‘postural’ tectal locomotor regions may exist in the superior colliculus that are situated within a striato-nigrotectal outflow system capable of influencing the animal's motor activity and posture.     

Cerebellar benzodiazepine receptor distribution: an autoradiographic study of the normal C57BL/6J and Purkinje cell degeneration mutant mouse

The distribution of [3H]flunitrazepam binding sites in the cerebella of normal mice and Purkinje cell degeneration mutant mice was studied by light microscopic autoradiography. In the cerebellar cortex of normal mice, a high density of [3H]flunitrazepam binding was observed over the molecular layer, an intermediate density over the Purkinje cell layer and a low density over the granule cell layer; the white matter was devoid of labeling. The deep cerebellar nuclei were labeled to an intermediate density. In the 54-day-old Purkinje cell degeneration mutant cerebellum, which is depleted of Purkinje cells, a 36% reduction in labeling density of the cerebellar cortex was observed. The density was reduced by approximately equal amounts in both the molecular and granule cell layers; labeling in the deep cerebellar nuclei was, however, substantially increased.     

Inhibition of the substantia nigra suppresses absences and clonic seizures in audiogenic rats, but not tonic seizures: evidence for seizure specificity of the nigral control

GABAergic inhibition of the substantia nigra pars reticulata has been shown to suppress seizures in most models of epilepsy involving forebrain networks, such as absences or clonic seizures. No such antiepileptic effects were observed, however, in genetically audiogenic rats exhibiting tonic seizures generated in the brainstem. This suggests a constitutive dysfunction of the nigral GABAergic neurotransmission in this strain of rat or a selective action of the nigral control on specific networks. In the present study, we first confirmed that bilateral injection of muscimol (700 pmol/side) in the substantia nigra had no effect in Wistar rats with audiogenic seizures (Wistar AS). [3H]Muscimol autoradiography suggested a 40% reduced density of GABA(A) receptors in the substantia nigra of Wistar AS, whereas no change was observed in the cortex and the superior colliculus (superficial and intermediate layers), as compared to control animals. In Wistar AS where 40 repetitions of audiogenic stimulations progressively induced generalised convulsive seizures with both tonic and clonic components, bilateral injection of muscimol (350 pmol/side) in the substantia nigra suppressed the clonic component but had no effect on tonic seizures. In hybrid rats issued from cross-breeding between Wistar AS and rats with spontaneous absence seizures, bilateral injection of muscimol (18 pmol/side) in the substantia nigra abolished cortical spike-and-wave discharges, but had no effect on tonic audiogenic seizures at doses up to 700 pmol/side. These results show that despite a decreased number of GABA(A) receptors in the substantia nigra, inhibition of this structure in Wistar AS still leads to inhibition of seizures involving forebrain structures. These results confirm that GABAergic inhibition of the substantia nigra has antiepileptic effects through the control of forebrain circuits. They suggest that this control mechanism has no inhibitory effect on circuits underlying audiogenic tonic seizures.     

The neurons of the substantia nigra and zona incerta which project to the cat superior colliculus are GABA immunoreactive: a double-label study using GABA immunocytochemistry and lectin retrograde transport

The neurotransmitter cytochemistry of neurons in the substantia nigra and zona incerta which project to the cat superior colliculus was examined. Neurons in both structures were double-labeled with an antibody to the transmitter GABA and a retrograde tracer, [3H]n-acetylated wheat germ agglutinin, injected into the superior colliculus. All cells in the zona incerta and substantia nigra which projected to the superior colliculus were labeled by the GABA antiserum. Most other neurons within the zona incerta and virtually all within the substantia nigra pars reticulata and pars lateralis were also labeled by the GABA antibody. By contrast, neurons in the substantia nigra pars compacta were not labeled by either the GABA antibody or wheat germ agglutinin. Nigrotectal cells in the substantia nigra were of medium to large size and most had stellate-shaped cell bodies. Zona incerta cells projecting to the superior colliculus were also of medium to large size, but most had horizontal fusiform cell bodies. This study demonstrates two new findings: (1) that all nigrotectal neurons in cat are immunoreactive to a GABA antibody and probably contain the neurotransmitter GABA; and (2) that these GABA immunoreactive neurons in cat are found not only in the substantia nigra pars reticulata but also within the pars lateralis. Zona incerta cells projecting to the superior colliculus have a different morphology but are also apparently GABAergic. These data provide an anatomical substrate for the known inhibitory action of the nigrotectal pathway on superior colliculus neurons.     

The cholinergic innervation of normal and transplanted superior colliculus in the rat: an immunohistochemical study

The distribution of choline acetyltransferase was determined in normal and transplanted rat superior colliculus with choline acetyltransferase immunohistochemistry. This distribution was compared to the pattern of histochemically detected acetylcholinesterase activity. To determine cholinergic input to the superficial superior colliculus, double labelling experiments combining retrograde tracing methods and choline acetyltransferase immunohistochemistry were carried out. No choline acetyltransferase-containing neurons were observed in the rat superior colliculus. A dense network of choline acetyltransferase-immunoreactive fibres and terminals was seen in the intermediate layers of the normal superior colliculus. The distribution was patchy and very similar to the pattern of acetylcholinesterase activity. Occasional fibres and terminals were seen in the deep tectal laminae. The superficial layers contained a low number of choline acetyltransferase-stained fibres and terminals but a comparatively high level of acetylcholinesterase activity. Following a unilateral injection of a tracer into the superficial superior colliculus, retrogradely labelled choline acetyltransferase-immunoreactive neurons were found in the dorsal and ventral subnuclei of the ipsilateral parabigeminal nucleus. As in the normal superior colliculus, choline acetyltransferase-positive neurons were not found in tectal transplants. However, choline acetyltransferase-immunoreactive fibres and terminals were seen in grafts but only in those which had extensive connections with the host midbrain. The pattern of staining most closely resembled that seen in the intermediate layers of the normal superior colliculus. The similar arrangement of choline acetyltransferase and acetylcholinesterase activity in the intermediate layers of normal rat superior colliculus provides further evidence for cholinergic innervation to these layers, probably originating in the dorsal and pedunculopontine tegmental nuclei. The data from the double labelling experiments indicate that the choline acetyltransferase-immunoreactive terminals observed in the superficial layers represent the terminal field of an ipsilateral cholinergic projection from the parabigeminal nucleus. Tectal grafts receive cholinergic innervation from the host. The evidence suggests that much of this input derives from the cholinergic nuclei in the brainstem tegmentum which normally project to the intermediate tectal layers.