Visual recovery in goldfish following unilateral optic tectum ablation: Evidence of competition between optic axons for tectal targets

Anatomical studies suggest that regenerating optic axons which invade the ipsilateral lobe of the optic tectum following ablation of the contralateral lobe compete with resident optic axons for synaptic sites on tectal neurons. Invader optic axons are initially uniformly distributed over the entire tectal lobe. With time, the invader and resident optic axons progressively segregate so that the invaders are localized in bands or islands separated by areas that are innervated mainly by the residents. When the resident optic axons are destroyed by ablating the eye opposite to the experimental eye, the invader axons remain continuously distributed and the segregation process apparently does not occur. We investigated the relationship between the segregation process and the recovery of visual function by the invader axons. Visual recovery was measured with a behavioral method in which the index of vision was the occurrence of a branchial suppression response to a moving spot of red light that was classically conditioned to an electric shock stimulus. The minimum time to reappearance of vision following ablation of the contralateral lobe of the tectum in two-eye fish was similar to the reported time of onset of the segregation process. Visual recovery occurred sooner when the opposite eye was removed. The restored vision in both groups disappeared following subsequent ablation of the remaining lobe of the tectum. These results suggest that the goldfish optic tectum normally contains no free synaptic sites for anomalous optic afferents and that the invader axons must compete for targets with the resident optic afferents. The invader axons can apparently remain unconnected or non-functional for several weeks following their arrival in the ipsilateral tectal lobe.     

Topographic projections between the nucleus isthmi and the optic tectum in a teleost,Navodon Modestus

Following horseradish peroxidase injections into the optic tectum of a teleost,Navodon modestus, reciprocal and topographic projections between the nucleus isthmi and the ipsilateral optic tectum were determined. The isthmo-tectal fibers diverge to the optic tectum while maintaining the spatial arrangements of the isthmic cells from which the fibers originate. The tecto-isthmic projections also keep the spatial arrangements in the optic tectum. The tectal fibers converge near the nucleus isthmi and terminate in the non-cellular portion of the nucleus. The reciprocal topography is apparent in the combined results of 9 experiments with one tectal injection in each region. No labeled cells and fibers were found in the contralateral nucleus isthmi.     

Effect of enucleation on choline acetyltransferase activity in layers of goldfish optic tectum

Choline acetyltransferase (ChAT) activity was determined in layers of optic tectum in control goldfish and in goldfish 4-20 days following unilateral enucleation. Significant changes in activity were found in the periventricular (PV) and superficial gray and white (SGW) layers. Within 4 days, ChAT activity in the PV layer on the lesioned side was about 75% of that on the control side. By 20 days, ChAT specific activity in the SGW layer on the lesioned side was about 150-160% of that on the control side. This increase in specific activity in the SGW layer was accounted for by the decrease in volume and in density of the layer after enucleation, so that the total amount of activity in the layer did not change significantly, indicating that the optic terminals contain little to no ChAT activity. ChAT activity in the optic tract was very low and did not decrease after enucleation. These data strongly indicate that the retinotectal pathway in goldfish is not cholinergic and, therefore, that the ChAT activity in the SGW layer is related to sources other than retinal ganglion cells. It is suggested that one such source might be neurons with somata in the PV layer.     

Two distinct visual pathways through the superficial pretectum in a percomorph teleost

The connections of the superficial pretectum and of nucleus isthmi were examined in a percomorph teleost, Lepomis cyanellus. Horseradish peroxidase was injected either with a pin into the parvicellular nucleus of the superficial pretectum or pressure injected into nucleus isthmi; the isthmal injections retrogradely labelled the neurons of the magnocellular nucleus of the superficial pretectum. Two main visual pathways can be recognized: The first projects from the retina to the parvicellular nucleus, and then to the intermediate nucleus of the superficial pretectum, the inferior raphe nucleus, and the trochlear nucleus. The second projects from the retina via the optic tectum to the magnocellular nucleus of the superficial pretectum, and from there to nucleus isthmi and the lateral thalamic nucleus; nucleus isthmi and the lateral thalamic nucleus project back to the optic tectum, and nucleus isthmi also projects back to the magnocellular nucleus. The two pathways are interconnected to some extent because both nucleus isthmi and the optic tectum project to the parvicellular nucleus; nevertheless, we suggest that they may be functionally and evolutionarily distinct. Compared to percomorphs, the first pathway appears reduced in cyprinid teleosts such as goldfish. Furthermore, the magnocellular nucleus of the second pathway is completely different in cyprinids, both in cellular architecture and in efferent connections. A phylogenetic analysis suggests that cyprinid ancestors went through a period of reduced vision and that the magnocellular nucleus of the superficial pretectum in modern cyprinids has been either extensively modified from the primitive condition or lost entirely and replaced by a superficially similar structure.     

Fine structure of the optic fiber termination layer in the tectum of the teleost Rutilus: a stereological and morphometric study

The stratum fibrosum et griseum superficiale (SFGS) of the Rutilus optic tectum, which receives a massive fiber projection from the contralateral retina, was studied by electron microscopy. The qualitative and quantitative analysis of the laterodorsal (LD) portion of the stratum involved both a stereological examination of the different elements and a morphometric study of the various profiles containing synaptic vesicles (PCSVs). The relative volume of each element in the LD SFGS was as follows: myelinated and unmyelinated axons, 6.6%; PCSVs, 38%; dendrites without vesicles, spines, and cell bodies, 41.7%; glia, 10.5%. With the fixation employed, 35% of PCSVs showed spheroidal synaptic vesicles. These profiles could be subdivided into three types: (1) S1 (23.5%) represented optic terminals, since they degenerated after retinal ablation or were labeled after intraocular injection of HRP or [3H] proline. Three subgroups of S1 were identified: S1m--profiles containing clear mitochondria;S1c--profiles that were contiguous with S1m and lacked mitochondria;S1i--isolated profiles without mitochondria. (2) S2 (9.3%) were characterized mainly by their dark mitochondria. (3) S3 (2.2%) corresponded to small nonvisual terminals that were isolated and lacked mitochondria. The PCSVs with pleiomorphic synaptic vesicles (65%) were subdivided into three groups: P1 (38%), P2 (19%), and P3 (8%). P1 and P2 were axonal in nature; P2 could be distinguished from P1 by a greater density of synaptic vesicles. P3 was of dendritic origin. Analysis of synaptic patterns revealed a small number of serial synapses. The presynaptic elements were optic boutons, whereas the intermediate profiles were dendrites with synaptic vesicles (P3). Results are compared with ultrastructural data obtained in the superficial tectal layers of other teleosts and other vertebrate groups.     

Identification of retinotectal projection pathway in the deep tectal laminae in the chick

The optic tectum is a visual center of nonmammalian vertebrates that receives retinal fibers in a retinotopic manner. It has been accepted that retinal fibers project to some superficial laminae of the tectum, but do not go through lamina g of stratum griseum et fibrosum superficiale (SGFS). By a novel fiber-tracing method, we found a novel pathway of retinal fibers that run through deep laminae of the tectum. The retinal fibers that would run through the newly identified pathway first run caudally along the medial edge after invading the tectum, turn laterally, and extend toward the lateral side through the deep pathway. The deep pathway runs through stratum album centrale and stratum fibrosum periventriculare. The fibers that run through the deep pathway do not enter the stratum opticum, where the conventional retinal fibers run. As development proceeds, these fibers decrease and disappear by the adult stage. By the new method, we found that some of the conventional retinal fibers transiently run through lamina g of SGFS and invade laminae h/i. In conclusion, we found distinct but transient retinal fiber pathway in the deep tectal laminae, which have not been thought to be retinorecipient.     

Organization of retinal axons within the optic nerve,optic chiasm,and the innervation of multiple central nervous system targets Rana pipiens

Light microscopic analysis of the optic nerve, chiasm, and optic tracts of Rana pipiens after the anterograde and retrograde transport of horseradish peroxidase has shown that retinal ganglion-cell axons reach the optic nerve head in chronotopically organized fascicles that form bands across the intraocular optic nerve. These bands of fascicles are divided along the midline in a “zone of reorganization” to create two full maps of the retinal surface; however, this map is discontinuous in that nasal and temporal quadrants are adjacent to one another. In the intracranial portion of the optic nerve, axons undergo another reorganization such that peripheral retinal axons shift position and become localized laterally and ventrally, whereas centrally placed axons become localized dorsally. Within this reorganization, the nerve is reconfigured into laminae of axons, and each lamina consists of age-related axons organized into two retinal maps. In the ipsilateral chiasm, axons diverge to form three central, optic tracts: the medial optic tract, the projection to the corpus geniculatum, and the basal optic root. Ipsilateral axons leave the chiasm at the same level of the chiasm as do their contralateral counterparts. The remaining axons converge in the lateral diencephalon to form a fourth fascicle, the marginal optic tract. Thus, within the optic chiasm, a sequence of positional transformations occur that result in the formation of multiple optic pathways. The various changes in axonal trajectory always coincide with changes in the orientation of cell groups that lie within the nerve and optic chiasm. J. Comp. Neurol. 402:222–237, 1998. © 1998 Wiley-Liss, Inc.     

Contrast sensitivity of neurons in pigeon optic tectum

An analysis of contrast sensitivity of neurons in the optic tectum of intact and wulst-ablated pigeons was carried out using moving light bars.

(1) Changes of contrast from 0.1% to 89%, obtained with different bar luminances, induced increases of response amplitude linearly related to log contrast.

(2) The units differed in their contrast threshold but 90.7% had a contrast threshold below 2.5%.

(3) Both contrast threshold and threshold-saturation range values were independent of the location of the units in the tectal layers.

(4) Contrast threshold remarkably increased on passing from low photopic to scotopic adaptation levels.

(5) The relationship between unit responses and contrast values, as well as the contrast threshold value, did not differ between intact and wulst-ablated animals.

Retinal removal up-regulates cannabinoid CB(1) receptors in the chick optic tectum

The endocannabinoid system has been implicated in several neurobiological processes, including neurodegeneration and neuroprotection. The aim of this study was to evaluate the effects of unilateral retinal ablation on the expression of the cannabinoid receptor subtype 1 (CB(1)) at both protein and mRNA levels in the optic tectum of the adult chick brain. After different survival times postlesion (2-30 days), the chick brains were subjected to immunohistochemical, immunoblotting, and real-time PCR procedures to evaluate CB(1) expression. TUNEL and Fluoro-Jade B were used to verify the possible occurrence of cell death, and immunostaining for the microtubule-associated protein MAP-2 was performed to verify possible dendritic remodeling after lesions. No cell death could be observed in the deafferented tectum, at least up to 30 days postlesion, although Fluoro-Jade B could reveal degenerating axons and terminals. Retinal ablation seems to generate an increase of CB(1) protein in the optic tectum and other retinorecipient visual areas, which paralleled an increase in MAP-2 staining. On the other hand, CB(1) mRNA levels were not changed after retinal ablation. Our results reveal that CB(1) expression in visual structures of the adult chick brain may be negatively regulated by the retinal innervation. The increase of CB(1) receptor expression observed after retinal removal indicates that these receptors are not presynaptic in retinal axons projecting to the tectum and suggests a role of the cannabinoid system in plasticity processes ensuing after lesions.     

Central retinal area is not the site where ganglion cells are generated first

The development of retinal ganglion cells (RGC) was studied in the chick from stage 18 to adulthood. Our main objectives were to identify the retinal site where the first RGCs differentiate, to locate this site relative to the optically defined central retinal area, and to map the spatial arrangement of the RGC field at different stages in development. The eyes of the experimental animals were fixed and serially sectioned. The borders of RGC fields were determined from the presence of either ganglion cell perikarya or ganglion cell axons. In seven cases between stages 21 and 26, the borders of the RGC fields were confirmed electron microscopically. The serial sections together with the RGC fields were then reconstructed in three dimensions. The reconstructed retinae were projected onto a plane by using the radially equidistant polar azimuthal projection. First, RGCs appear dorsal to the apex of the optic fissure. Ganglion cell development then initially spreads out symmetrically with respect to the optic fissure. However, from stage 29 on, the nasal half of the retina expands much more than the temporal half. This asymmetrical growth entails that the optic fissure is eventually located in the temporal half of the retina in the mature animal. The RGC fields of the embryonic stages were superimposed on the retina of a visually active animal according to their real size and position. It turned out that the central retinal area was at least 2 mm away from the site where the first RGCs were generated. It is not before stage 28 that the prospective central retinal area is included into the expanding ganglion cell field. The fact that RGCs at the central retinal area are generated 2.5 days later than first RGCs near the apex of the optic fissure has important implications for the formation of the retinotectal projection. © 1993 Wiley-Liss, Inc.     

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。     

Origins of serotonin-like immunoreactivity in the optic tectum of rana pipiens

We have previously identified a population of serotonin-like immunoreactive (5-HT-ir) retinal ganglion cells in Rana pipiens. In this study, we examined serotonin-like immunoreactivity (5-HTLI) in a probable target of those cells, the optic tectum. We observed both 5-HT-ir fibers and cell bodies in this structure. 5-HT-ir cells were located in the cellular layers of the tectum, layers 2, 4, and 6, and scattered in its superficial layers. 5-HT-ir fibers in the tectum displayed a laminated organization and were located in tectal layers 3, 5, 6, 7, and 9. Retrograde labelling experiments showed that 5-HT-ir retinal ganglion cells projected to the optic tectum. However, these experiments also demonstrated that serotonergic neurons in the midbrain tegmentum, the nucleus isthmi, and the medulla did so as well. 5-HT-ir fibers seen in lamina A of layer 9 were very much reduced in density in animals in which the optic nerve had been lesioned for 3–6 months. Immunoreactive fibers in lamina B of layer 9 were not affected by the lesion. Our results suggest that 5-HT-ir fibers in lamina A of layer 9 are mainly of retinal origin, whereas those in lamina B originate from other brain areas. The 5-HT-ir tectal cells located in the cellular layers probably contribute the 5-HT-ir fibers seen in layers 3, 5, 6, and 7. © 1995 Wiley-Liss, Inc.     

Connections of eye-saccade-related areas within mesencephalic reticular formation with the optic tectum in goldfish

Physiological studies demonstrate that separate sites within the mesencephalic reticular formation (MRF) can evoke eye saccades with different preferred directions. Furthermore, anatomical research suggests that a tectoreticulotectal circuit organized in accordance with the tectal eye movement map is present. However, whether the reticulotectal projection shifts with the gaze map present in the MRF is unknown. We explored this question in goldfish, by injecting biotin dextran amine within MRF sites that evoked upward, downward, oblique, and horizontal eye saccades. Then, we analyzed the labeling in the optic tectum. The main findings can be summarized as follows. 1) The MRF and the optic tectum were connected by separate axons of the tectobulbar tract. 2) The MRF was reciprocally connected mainly with the ipsilateral tectal lobe, but also with the contralateral one. 3) The MRF received projections chiefly from neurons located within intermediate and deep tectal layers. In addition, the MRF projections terminated primarily within the intermediate tectal layer. 4) The distribution of labeled neurons in the tectum shifted with the different MRF sites in a manner consistent with the tectal motor map. The area containing these cells was targeted by a high-density reticulotectal projection. In addition to this high-density topographic projection, there was a low-density one spread throughout the tectum. 5) Occasionally, boutons were observed adjacent to tectal labeled neurons. We conclude that the organization of the reticulotectal circuit is consistent with the functional topography of the MRF and that the MRF participates in a tectoreticulotectal feedback circuit.     

Telencephalic terminals in the major retinal synaptic lamina of the goldfish optic tectum

Light and electron microscopic degeneration studies were used to examine the telencephalotectal pathway in goldfish. Both techniques showed that each telencephalic lobe sent bilateral projections to several tectal laminae. Degenerating synaptic terminals and fibers were observed in the major retinal projection lamina as well as in other tectal laminae. The terminals contained round to oval synaptic vesicles, asymmetric synapses and contacted relatively small postsynaptic profiles.     

Positional specificities of retinal growth cones in the mouse superior colliculus.

In the developing retinotectal system, repulsive topographic tectal cues have been demonstrated to contribute to the final mapping. Here, we describe a novel response of nasal axons to growth-promoting cues expressed by anterior tectal cells. In in vitro experiments, contact of fibres from the nasal (but not temporal) pole of the mouse retina with anterior (but not posterior) tectal membranes leads to their adopting very elongated and filopodial morphologies, and to increase their growth rates. As previously demonstrated, fibres from the temporal pole of the retina are collapsed by posterior tectal membranes in vitro. In addition, a study of retinal growth cone morphologies in vivo, at early stages of target invasion, shows that growth cones of nasal fibres have streamlined morphologies, usually indicative of active elongation growth modes, in the anterior part of the embryonic mouse tectum, and more elaborate morphologies posteriorly. Vice versa, temporal fibres have mainly elaborate growth cones anteriorly, and collapsed growth cones posteriorly. These experiments demonstrate that nasal retinal fibres respond preferentially to permissive or growth-promoting cues in the embryonic mouse tectal environment, both in vitro and in vivo. This phenomenon might contribute to ingrowth of retinal fibres in their target area, and to promote the homing of nasal fibres towards the posterior aspect of the tectum, which is their normal target region.     

An ascending visual pathway to the dorsal telencephalon through the optic tectum and nucleus prethalamicus in the yellowfin goby Acanthogobius flavimanus (Temminck & Schlegel, 1845)

Dual visual pathways reaching the telencephalon appear to be an ancient vertebrate trait, but some teleost fish seem to possess only one pathway via the optic tectum. We undertook the present study to determine if and when this loss occurred during evolution. Tracer injection experiments to the optic nerve, the optic tectum, and the dorsal telencephalon were performed in the present study, to investigate ascending visual pathways to the dorsal telencephalon in an acanthopterygian teleost, the yellowfin goby Acanthogobius flavimanus (Temminck & Schlegel, 1845). We confirmed the presence of a nucleus prethalamicus (PTh) in the goby, which has been convincingly identified only in holocentrids, suggesting that this nucleus is present in other acanthopterygians. We found that the optic tectum projects to the PTh bilaterally. The PTh projects in turn to the dorsal telencephalon, ipsilaterally. These results suggest that the yellowfin goby possesses only an extrageniculate‐like pathway, while a geniculate‐like pathway could not be identified. This situation is common with that of holocentrids and may be a character common in acanthopterygians. It is possible that a geniculate‐like system was lost in the common ancestor of acanthopterygians, although the scenario for the evolution of ascending visual systems in actinopterygians remains uncertain due to the lack of precise knowledge in a number of actinopterygian taxons.     

Kainate-induced lesion in the optic tectum: dependency upon optic nerve afferents or glutamate

Kainic acid is known to induce characteristic lesions in neurons receiving an intact input with presumed glutamate-mediated neurotransmission. There are indications for glutamate as a transmitter of retinal afferent terminals in the pigeon optic tectum. After tectal injection of kainic acid (0.5–2.0 μg in 0.5 μl) the optic tectum was studied by light and electron microscopy and the following changes were observed: (a) within 1–48 h important neuropil vacuolization predominantly in lower part of layer 5. Such vacuoles were sometimes postsynaptic to identified retinal afferent terminals: (b) within 1 h to 21 days progressive neuronal cell loss throughout the tectal layers. These toxic effects were not observed 2–12 weeks after contralateral retinal ablation but could partially be restored by combined glutamate (0.2 mg) and kainate injection. Thus in the pigeon tectum, kainic acid neurotoxicity is dependent upon an intact retinal input, a finding consistent with a special role for glutamate — possibly as a transmitter — in retinal terminals.     

Spatiotemporal profile of synaptic activation produced by the electrical and visual stimulation of retinal inputs to the optic tectum: a current source density analysis in the pigeon (Columba livia)

The optic tectum of the pigeon is a highly organized, multilayered structure that receives a massive polystratified afference of at least five different populations of retinal ganglion cells and gives rise to various anatomically segregated efferent systems. The synaptic organization of retino-tectal circuitry is, at present, mostly unknown. To investigate the spatiotemporal profile of synaptic activation produced by differential (electrical and visual) stimulation of the retinal inputs, we performed a high-spatial-resolution current source density analysis in the optic tectum of the anaesthetized pigeon. Electrical stimuli consisted of brief pulses of different durations applied to the optic nerve head, while visual stimuli consisted of light flashes of different intensities. Electrical stimulation generated sinks confined to retinorecipient layers. The temporal structure, spatial location and thresholds of these sinks indicated that they are all due to primary tectal synapses of retinal fibers with different conduction velocities. Sinks evoked by the fastest retinal axons were more superficially located than sinks produced by slower retinal fibers. Visual stimulation, on the other hand, resulted in a more complex pattern of current sinks, with various sinks located in the retinorecipient layers and also well below. Visual stimulation induced action potentials at superficial as well as deep tectal levels. We conclude that electrical stimulation activates most of the populations of ganglion cells as well as their primary tectal synapses, but is unable to elicit a significant activation of secondary tectal synapses. Visual stimulation, on the contrary, activates just some of the incoming retinal populations, but in a way that produces noticeable secondary activation of intratectal circuits. Laminar segregation of retinally evoked tectal activity, as reported here, has also been found in other vertebrates. Similarities and differences with previous studies are discussed.