Non-uniform distribution of cellular phenotypes in the optic tectum of the leopard frog

Regional specializations in the retina have been described in a number of species. We have investigated whether such specializations can be found in the optic tectum, an area of the brain responsible for the processing of visual information. Using the tectum of Rana pipiens, we have examined the distribution of three different cell types defined on the basis of their immunoreactivity to somatostatin, substance P, and serotonin antibodies. These three immunoreactive cell populations had differing, nonuniform distributions in the optic tectum. Somatostatin-like irnmunoreactive cells were largely restricted to the caudal one-third of the tectum, whereas both substance P-like immunoreactive (SP-ir) and serotonin-like immunoreactive (5-HT-ir) cells were found unequally represented throughout the tectum. The percentage of SP-ir cells decreased significantly in both the posterior and medial directions from its high in the anterior lateral tectum. Although serotonin-like immunoreactivity was also greatest in the lateral tectum and decreased significantly medially, it was largely constant in the anterior-to-posterior dimension. The populations of SP-ir and 5-HT-ir cells were nonoverlapping. Our results suggest that information may be processed differently in different regions of the optic tectum. © 1995 Wiley-Liss, Inc.     

Connections of the tectum of the rattlesnake Crotalus viridis: an HRP study.

We have studied the connections of the tectum of the rattlesnake by tectal application of horseradish peroxidase. The tectum receives bilateral input from nucleus lentiformis mesencephali, posterolateral tegmental nuclei, anterior tegmental nuclei and periventricular nuclei; ipsilateral input from nucleus geniculatus pretectalis, and lateral geniculate nucleus pars dorsalis; and contralateral input from dorso-lateral posterior tegmental nucleus and the previously undescribed nucleus reticularis caloris (RC). RC is located on the ventro-lateral surface of the medulla and consists of large cells 25--45 micrometer in diameter. Efferent projections from the tectum can be traced to the ipsilateral nucleus lentiformis mesencephali, the ipsilateral lateral geniculate region, anterior tegmental region and a wide bilateral area of the neuropil of the ventral tegmentum and ventral medualla. We have not found any direct tectal projections from the sensory trigeminal nuclei including the nucleus of the lateral descending trigeminal tract (LTTD). We suggest that in the rattlesnake, RC is the intermediate link connecting LTTD to the tectum.     

Afferents of the lamprey optic tectum with special reference to the GABA input: combined tracing and immunohistochemical study

The optic tectum in the lamprey midbrain, homologue of the superior colliculus in mammals, is important for eye movement control and orienting responses. There is, however, only limited information regarding the afferent input to the optic tectum except for that from the eyes. The objective of this study was to define specifically the gamma-aminobutyric acid (GABA)-ergic projections to the optic tectum in the river lamprey (Lampetra fluviatilis) and also to describe the tectal afferent input in general. The origin of afferents to the optic tectum was studied by using the neuronal tracer neurobiotin. Injection of neurobiotin into the optic tectum resulted in retrograde labelling of cell groups in all major subdivisions of the brain. The main areas shown to project to the optic tectum were the following: the caudoventral part of the medial pallium, the area of the ventral thalamus and dorsal thalamus, the nucleus of the posterior commissure, the torus semicircularis, the mesencephalic M5 nucleus of Schober, the mesencephalic reticular area, the ishtmic area, and the octavolateral nuclei. GABAergic projections to the optic tectum were identified by combining neurobiotin tracing and GABA immunohistochemistry. On the basis of these double-labelling experiments, it was shown that the optic tectum receives a GABAergic input from the caudoventral part of the medial pallium, the dorsal and ventral thalamus, the nucleus of M5, and the torus semicircularis. The afferent input to the optic tectum in the lamprey brain is similar to that described for other vertebrate species, which is of particular interest considering its position in phylogeny.     

Topographic and morphometric effects of bilateral embryonic eye removal on the optic tectum and nucleus isthmus of the leopard frog

Rana pipiens were raised through metamorphosis after extirpation of both eye primordia at Shumway embryonic stage 17 (Shumway '40). The visual connections between the isthmic nuclei and the optic tectum were examined in these animals using horseradish peroxidase (HRP) histochemistry. Isthmo-tectal projections are normally aligned with the primary retinotectal map. We asked whether these connections would develop normal topographic organization in the absence of normal retinal input. HRP was formed into a solid pellet (? 200–500 μm diameter) and inserted into one tectal lobe on the tip of a fine metal probe. The procedure produced relatively restricted retrograde label in somas and dendrites in both isthmi nuclei. In the nucleus isthmus ipsilateral to the tectal lobe receiving the HRP pellet, processes of tecto-isthmi neurons were labeled by anterograde transport. The topography of the isthmo-tectal and tecto-isthmic projections were identical in the developmentally enucleated animals and in normal frogs, even though eye removal severely reduced the volume of the optic tecta and the isthmi nuclei. Thus our analyses indicate that retinal contacts do not play an active role in the development of the positional or polarity cues that are involved in “mapping” projections between central visual nuclei. These results are discussed in the context of peripheral specification of central connections and in terms of models that have recently been proposed to explain the development of the retinotectal system.     

Relative number of cells projecting from contralateral and ipsilateral nucleus isthmi to loci in the optic tectum is dependent on visuotopic location: horseradish peroxidase study in the leopard frog.

The leopard frog optic tectum is the principal target of the contralateral retina. The retinal terminals form a topographic map of the visual field. The tectum also receives bilateral topographic input from a midbrain structure called nucleus isthmi. In this study we determined the relative strength of n. isthmi projections to different loci in the tectum. Horseradish peroxidase (HRP) was applied at single superficial tectal locations in a series of leopard frogs. The application sites were distributed across the tectum. Retrogradely filled cells were counted in ipsilateral and contralateral nucleus isthmi. Although all regions of the tectum receive input from both n. isthmi, the relative number of labeled cells in the two n. isthmi is dependent on visuotopic location. Input to the rostromedial tectum representing the visual field ipsilateral to the labeled tectum comes primarily from the contralateral n. isthmi. Input to the caudolateral tectum representing the visual field contralateral to the labeled tectum originates mostly from the ipsilateral n. isthmi. Tectal application sites representing the visual midline had approximately equal numbers of labeled cells in the two n. isthmi. The results are similar at postapplication survival times ranging from 2 to 14 days. Using application of HRP to rostral tectum and application of nuclear yellow to caudal tectum, we show that the anisotropy in isthmi labeling is not due to take up of these labels by isthmotectal fibers passing through the application sites that terminate elsewhere.     

Afferent connections to the tectum mesencephali in the hagfish, Eptatretus burgeri: an HRP study

The afferent connections to the tectum mesencephali were determined by unilateral injections of HRP in the hagfish, Eptatretus burgeri, whose eyes are degenerate and buried below the skin. Retrogradely labeled cells were observed ipsilaterally in the hypothalamus, lateral part of the pars ventralis thalami of Jansen, and nucl. reticularis mesencephali, and contralaterally in the tectum, nucl. sensorius n. V, area acoustico-lateralis, and "nucl. funiculi dorsalis". Of these non-visual afferents to the tectum, the trigeminal sensory nucleus contributed the greatest number, which ascended along the lemniscus bulbaris after crossing in the commissura ventralis. Retrogradely labeled fibers of the optic nerve could not be recognized in this study, although in a previous study the superficial layer of the tectum mesencephali received a few projections from the contralateral eye. These data are compared with the data from other animals and previous histochemical investigations.     

Field potentials from the frog optic tectum in vitro

A new method for maintaining the isolated frog optic tectum superfused in vitro at a low temperature is described. The preparation remains viable for several hours with electrophysiological responses similar to those of the in vivo tectum. This superfusion technique will allow controlled application of putative neurotransmitters and other substances in known concentrations and therefore makes the in vitro frog optic tectum a useful model system for neuropharmacological investigations into central optic pathways.     

Afferent connections of the optic tectum in lampreys: an experimental study

Tectal afferents were studied in adult lampreys of three species (Ichthyomyzon unicuspis, Lampetra fluviatilis, and Petromyzon marinus) following unilateral BDA injections into the optic tectum (OT). In the secondary prosencephalon, neurons projecting to the OT were observed in the pallium, the subhipoccampal lobe, the striatum, the preoptic area and the hypothalamus. Following tectal injections, backfilled diencephalic cells were found bilaterally in: prethalamic eminence, ventral geniculate nucleus, periventricular prethalamic nucleus, periventricular pretectal nucleus, precommissural nucleus, magnocellular and parvocellular nuclei of the posterior commissure and pretectal nucleus; and ipsilaterally in: nucleus of Bellonci, periventricular thalamic nucleus, nucleus of the tuberculum posterior, and the subpretectal tegmentum, as well as in the pineal organ. At midbrain levels, retrogradely labeled cells were seen in the ipsilateral torus semicircularis, the contralateral OT, and bilaterally in the mesencephalic reticular formation and inside the limits of the retinopetal nuclei. In the hindbrain, tectal projecting cells were also bilaterally labeled in the dorsal and lateral isthmic nuclei, the octavolateral area, the sensory nucleus of the descending trigeminal tract, the dorsal column nucleus and the reticular formation. The rostral spinal cord also exhibited a few labeled cells. These results demonstrate a complex pattern of connections in the lamprey OT, most of which have been reported in other vertebrates. Hence, the lamprey OT receives a large number of nonvisual afferents from all major brain areas, and so is involved in information processing from different somatic sensory modalities.     

Cholinergic innervation of the optic tectum in the frog Rana pipiens

An immunohistochemical method for choline acetyltransferase (ChAT) identifies presumably cholinergic axons in two retino-receptive laminae in the optic tectum of the frog Rana pipiens. Following eye enucleation there is no loss of immunoreactive axons in the optic tectum. Following unilateral ablation of the nucleus isthmi there is a near-total loss of ChAT-positive axons in the superficial cholinergic lamina contralaterally and in the deeper cholinergic lamina ipsilaterally. Thus, the cholinergic innervation of the tectum appears to derive from the nucleus isthmi. However, ChAT-positive staining of the basal optic nucleus does depend upon an intact retinal input and could derive from either retinal axons or some system trophically dependent on them.     

Cytoarchitecture of the tectum mesencephali in salamanders: a Golgi and HRP study.

The tectum mesencephali of salamanders shows a morphology that has long been considered primitive when compared with that of frogs. The alternative hypothesis is that the salamander brain is secondarily simplified. In order to test these two hypotheses, the cytoarchitecture of the tectum and the projections of tectal neurons were studied in 11 species of salamanders. Application of the Golgi method reveals three major morphological types. Type 1 has a very wide dendritic arborization mostly confined to the deep fiber layers, and somata are always located within the most superficial part of the periventricular gray matter. Type 2 possesses a wide to medium-size dendritic arborization. In subtype 2a the somata are located in the uppermost part of the gray, and dendrites always reach the uppermost layer of retinal afferents; in subtype 2b the somata are found in deeper parts of the gray, and dendrites arborize in the deeper layers of retinal afferents; and in subtype 2c the somata are also located in deeper parts, but the wide dendritic arborization is confined to deep fiber layers. Type 3 shows the narrowest dendritic arbors that always reach the upper two tectal fiber layers. The somata are found at any depth of the gray matter. HRP experiments reveal a correlation between morphological differences and the projections of tectal neurons. Type 1- and type 2c-like cells constitute the uncrossed tecto-bulbo-spinal tract, whereas type 1- and type 2a-like cells and migrated large spindle-shaped cells (Salamandra) constitute the crossed tecto-bulbo-spinal tract. Type 3-like neurons project to thalamic, pretectal, and isthmic termination sites. The HRP experiments also demonstrate the existence of two classes of mesencephalic trigeminal cells. A comparison shows that salamanders and frogs possess very similar functional and morphological types of tectal cells. However, tectal cells of salamanders show a "juvenile" morphology, and the number of migrated cells is about 10 times higher in frogs compared to salamanders. Both phenomena are seen as the result of secondary simplification of brain structures in the context of paedomorphosis.     

青蛙前视盖与视顶盖间神经传导的细胞内电生理研究

目的已有许多研究报告了青蛙的前视盖对视顶盖起抑制作用,但关于此神经活动的特性尚不清楚。本研究探讨了这种复杂的神经活动的机理。方法用细胞内记录方法,通过电刺激前视盖的神经细胞核来记录视顶盖细胞的神经活动。结果前视盖的电刺激在同侧视顶盖主要唤起了两种神经反应：一种是兴奋性（excitator ypostsynaptic potential,EPSP）和抑制性突触后电位（an inhibitory postsynaptic potential,IPSP）同时出现,另一种是单纯的IPSP,后者在本记录中占主导地位。另外我们也记录到了某些投射到前视盖的视盖投射细胞的神经电位。它揭示了视顶盖和前视盖之间存在着交叉性的相互作用。短潜时的EPSP可能是通过单突触进行传导的,而大多数的IPSP是通过多突触方式进行神经信息传递的。几乎98％被记录的视盖细胞对前视盖的刺激显示出了抑制性反应。结论前视盖的神经细胞对视顶盖的神经活动发挥了强烈的抑制性作用。     

青蛙前视盖与视顶盖间神经传导的细胞内电生理研究(英文)

目的已有许多研究报告了青蛙的前视盖对视顶盖起抑制作用,但关于此神经活动的特性尚不清楚。本研究探讨了这种复杂的神经活动的机理。方法用细胞内记录方法,通过电刺激前视盖的神经细胞核来记录视顶盖细胞的神经活动。结果前视盖的电刺激在同侧视顶盖主要唤起了两种神经反应:一种是兴奋性(excitatory postsynaptic potential,EPSP)和抑制性突触后电位(an inhibitory postsynaptic potential,IPSP)同时出现,另一种是单纯的IPSP,后者在本记录中占主导地位。另外我们也记录到了某些投射到前视盖的视盖投射细胞的神经电位。它揭示了视顶盖和前视盖之间存在着交叉性的相互作用。短潜时的EPSP可能是通过单突触进行传导的,而大多数的IPSP是通过多突触方式进行神经信息传递的。几乎98%被记录的视盖细胞对前视盖的刺激显示出了抑制性反应。结论前视盖的神经细胞对视顶盖的神经活动发挥了强烈的抑制性作用。     

Laminar analysis of the synaptic organization of the frog optic tectum]

Distribution of axo-axonic and axo-dendritic synapses, nervous terminals and neuron somata in the depth of the optic tectum was studied in frog Rana temporaria L. in norm and 6-9, 60 and 134 days after contralateral eye enucleation. In outer plexiform layer 9 density of synapses was found to be maximal near the surface of the tectum; it decreased in deeper levels. In the outer zone of layer 9 (about 0.30 micrometer from the surface) many axo-axonic synapses were found. Terminals of myelinated optic fibres of large diameter ("dark" terminal degeneration) were widely distributed in layer 9. Density of axo-dendritic synapses in deep plexiform layer 5 was similar to that in layer 9. Numerous nervous terminals containing not only light synaptic vesicles, but also granular vesicles were seen in layer 5 and in adjacent zones.     

Peripheral nerve grafts to the frog optic tectum: a morphological study of foreign axon regeneration in the central nervous system

The proximal stump of a transected mandibular nerve was grafted onto the rostrodorsal surface of the optic tectum in adult Rana pipiens to investigate the morphologic characteristics of nonspecific axonal regeneration in a highly organized region of central nervous system (CNS). Within the first 3 weeks postgraft surgery (WPS), the nerve-tectum interface became firmly established. Concomitant with this was an invasion of the host tectum by a small number of fine "pioneerlike" axons from the nerve. By 6 WPS there developed a concerted instreaming of a large number of peripheral fibers. Once within the CNS, the foreign axons distributed themselves throughout the rostrocaudal extent of the tectum, but primarily its dorsal aspect within superficial layers 8 and 9. Presence of intact optic fibers at the time of mandibular fiber invasion served somewhat to restrict the regenerating aberrant axons in their course through layer 9. This restriction could be avoided by removal of the optic input either before or during peripheral ingrowth. However, once peripheral fibers had entered and established themselves in the host environment, no subsequent manipulation of the retinotectal projection had any effect. The aberrant growth pattern, which appeared remarkably stable after 6 WPS, consisted of a plexus of medium- and fine-caliber peripheral axons. Many of these fibers had numerous branches and "en passant" varicosities, the latter encompassing a variety of shapes and sizes. Terminal swellings and arborizations were also found. When comparing the regeneration of optic and mandibular nerve fibers in the tectum, two distinctions were made. Whereas optic axons revealed a fascicular and layered organization, mandibular axons showed a highly segregated and disordered growth pattern. These characteristic differences were maintained even when the two fiber systems were allowed to coregenerate into the same target tectum. Thus, each of the two groups of axons interacts with the tectal substrate in a distinct manner, apparently independent of the other.     

Peripheral nerve grafts to the frog optic tectum: A morphological study of the axon reaction in trigeminal motor and sensory neurons

The mandibular branch of the trigeminal nerve was severed and the proximal stump was grafted onto the optic tectum in adult Rana pipiens. The resultant changes occurring in the cell bodies of origin in the ipsilateral trigeminal motor and mesencephalic nuclei were studied qualitatively and quantitatively. Nucleolar, nuclear, and somal cross-sectional areas increased in size significantly approximately 3 days after surgery and peaked at 6 weeks postsurgery. This swelling, in which the nucleolus was most severely affected, gradually reversed itself and disappeared by 24 weeks after surgery. Despite the cell enlargement, cytoplasmic basophilia was maintained or even slightly increased. These morphologic changes suggest a strong anabolic reaction. Two differences were found between the motoneurons and the sensory neurons. First, the morphometric cell changes occurred at a faster rate in neurons of the trigeminal motor nucleus than in those of the mesencephalic nucleus. The time course of the motoneuron response correlated well with that of axonal regeneration from the nerve graft. Second, there was a delayed loss of mesencephalic nucleus cells between 12 and 24 weeks after surgery, whereas cells of the trigeminal motor nucleus were maintained at all survival times studied. Taken together with sensory cell loss in the trigeminal ganglion, this suggests a greater viability of regenerating motoneurons.     

Intracellular and current source density analysis of pretectal input to the optic tectum of the frog

Objective

Electrophysiological examination of the ipsilateral pretectotectal projection has proved that pretectal cells elicit strong suppressive responses to the ipsilateral tectum. However, the neural mechanisms underlying the contralateral pretectotectal prejection are still obscure. The present study aimed to examine the synaptic nature of pretectal nuclei and contralateral tectal cells, and to demonstrate the spatiotemporal pattern of neuronal activity in the 2 main brain structures.

Methods

Intracellular recording and current source density (CSD) analysis were used to test the complexity of neuronal mechanism of pretectotectal information transfer.

Results

The pretectal stimulation elicited only one type of response on the contralateral tectum, the inhibitory postsynaptic potential (IPSP). The majority of contra-induced IPSPs were assumed to be polysynaptically driven. In the CSD analysis, only one sink with short latency was observed in each profile. The ipsilateral projection produced a prominent monosynaptic sink in layer 8 of tectum. Recipient neurons were located in layers 6 and 7 of tectum. The result confirmed former findings from ipsilateral intracellular recordings.

Conclusion

These results suggest the following neuronal circuit: afferents from the pretectal nuclei broadly inhibit both tectal neuron, and since no second sink occurs in tectal layers, the pretectotectal excitatory afferents probably do not extend over the whole tectum, but are within limited state. The results of intracellular recording and CSD analysis further provide evidence of how pretectal afferent activity flows within the tectal laminae.