Goldfish Optic Tectum Research Articles

Following Activity of Neurons in Optic Tectum of Goldfish with Flashing Stimulation

Following Activity of Neurons in Optic Tectum of Goldfish with Flashing Stimulation

Detecting neuron of on-off response with optic tectum of goldfish

Detecting neuron of on-off response with optic tectum of goldfish

Wireless electrophysiology of the brain of freely swimming goldfish

BackgroundFish are a diverse group of vertebrates with very different brain structures. The study of the neurobiology of fish can thus lead to many important insights on information processing in the brain in a variety of environments. New methodWe developed a novel wireless technique to record extracellular neural signal activity in freely behaving fish. The system is based on a data logger and enables continues recording of up to 2.5h. ResultsFor proof of concept we recorded from the optic tectum of goldfish. We found correlations between the activity of the optic tectum and the light intensity in the room, as expected. Comparison with existing methodsDue to the technical difficulties involved in making electrophysiological recordings of behaving aquatic animals, there is no method for recording electrical neural activity from the brain of freely swimming fish. ConclusionsThis powerful tool should facilitate studies of information processing in behaving fish and other behaving aquatic animals.

Bilateral efferents from nucleus isthmi to the optic tectum in goldfish (Carassius auratus) are spatially restricted

The projection from nucleus isthmi (NI) to the optic tectum (OT) was investigated in the goldfish, Carassius auratus, by retrograde transport of biocytin applied at various sites in a tectal lobe. In previous studies, this projection is described as predominantly from the ipsilateral NI and maps topographically along the approximate rostrocaudal axis of both brain areas. However, the rostromedial tectal lobe, the tectal region representing the binocular visual field, receives afferents from both the ipsilateral and the contralateral NI. The contralateral isthmic neurons are found at the most caudal position in NI and are not topographic with the tectum. The bilateral projection from NI to the tectum may play a role in functions requiring the coordination of both eyes.

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.

Afferent connectivity to different functional zones of the optic tectum in goldfish.

This work studies the afferent connectivity to different functionally identified tectal zones in goldfish. The sources of afferents contributed to different degrees to the functionally defined zones. The dorsocentral area of the telencephalon was connected mainly with the ipsilateral anteromedial tectal zone. At diencephalic levels, neurons were found in three different regions: preoptic, thalamic, and pretectal. Preoptic structures (suprachiasmatic and preoptic nuclei) projected mainly to the anteromedial tectal zone, whereas thalamic (ventral and dorsal) and pretectal (central, superficial, and posterior commissure) nuclei projected to all divisions of the tectum. In the mesencephalon, the mesencephalic reticular formation, torus longitudinalis, torus semicircularis, and nucleus isthmi were, in the anteroposterior axis, topographically connected with the tectum. In addition, neurons in the contralateral tectum projected to the injected zones in a symmetrical point-to-point correspondence. At rhombencephalic levels, the superior reticular formation was connected to all studied tectal zones, whereas medial and inferior reticular formations were connected with medial and posterior tectal zones. The present results support a different quantitative afferent connectivity to each tectal zone, possibly based on the sensorimotor transformations that the optic tectum carries out to generate orienting responses.

Connectivity of the goldfish optic tectum with the mesencephalic and rhombencephalic reticular formation.

The optic tectum of goldfish, as in other vertebrates, plays a major role in the generation of orienting movements, including eye saccades. To perform these movements, the optic tectum sends a motor command through the mesencephalic and rhombencephalic reticular formation, to the extraocular motoneurons. Furthermore, the tectal command is adjusted by a feedback signal arising from the reticular targets. Since the features of the motor command change with respect to the tectal site, the present work was devoted to determining, quantitatively, the particular reciprocal connectivity between the reticular regions and tectal sites having different motor properties. With this aim, the bidirectional tracer, biotin dextran amine, was injected into anteromedial tectal sites, where eye movements with small horizontal and large vertical components were evoked, or into posteromedial tectal sites, where eye movements with large horizontal and small vertical components were evoked. Labeled boutons and somas were then located and counted in the reticular formation. Both were more numerous in the mesencephalon than in the rhombencephalon, and ipsilaterally than contralaterally, with respect to the injection site. Furthermore, the somas showed a tendency to be located in the area containing the most dense labeling of synaptic endings. In addition, labeled boutons were often observed in close association with retrogradely stained neurons, suggesting the presence of a tectoreticular feedback circuit. Following the injection in the anteromedial tectum, most of the boutons and labeled neurons were found in the reticular formation rostral to the oculomotor nucleus. Conversely, following the injection in the posteromedial tectum, most of the boutons and neurons were also located in the caudal mesencephalic reticular formation. Finally, boutons and neurons were found in the rhombencephalic reticular formation surrounding the abducens nucleus. They were more numerous following the injection in the posteromedial tectum. These results demonstrate characteristic patterns of reciprocal connectivity between physiologically different tectal sites and the mesencephalic and rhombencephalic reticular formation. These patterns are discussed in the framework of the neural substratum that underlies the codification of orienting movements in goldfish.

Slowly developing potentiation in goldfish retino–tectal synapses is masked by prior stimulation; an in vitro study

Field potential responses to local electrical stimulation were analyzed in slices of goldfish optic tectum (OT) in vitro. The post-synaptic component of retino–tectal origin was suppressed by DNQX, suggesting glutamatergic field excitatory post-synaptic potential (fEPSP). Repetitive stimulation at 0.1 Hz for 30 min resulted in a slowly developing potentiation in the fEPSP in a population of slices examined. When stimulation at a lower frequency (0.017 Hz for 30 min) preceded the conditioning, the 0.1 Hz stimulation failed to induce the potentiation even though the 0.017 Hz stimulation by itself caused no lasting changes. Induction of the potentiation was partially blocked by AP5 (100 μM), suggesting an involvement of NMDA receptors, however the effect was not significant. We also examined a possible involvement of adenosine, which is supposed to be co-released with glutamate at synaptic terminals. Exogenously applied adenosine (200 μM) slowly depressed fEPSP, and the depression was blocked by DPCPX (A1 receptor antagonist: 400 nM) but not by DMPX (A2 antagonist: 4 μM). However, DPCPX failed to unmask the potentiation after low-frequency stimulation (0.017 Hz for 30 min). Moreover, the potentiability was not correlated with sensitivity to exogenous adenosine. The A1 depression may not underlie the masking effect.

Tail and eye movements evoked by electrical microstimulation of the optic tectum in goldfish.

This work studies the tail and eye co-ordinated movements evoked by the focal electrical stimulation of the tectum in goldfish. The aim of the study is to understand better those tectal sites and mechanisms that either remain functionally unaltered or are adaptively modified across vertebrates. Stimulation was applied in various tectal zones, and the characteristics of evoked tail and eye movements were examined as a function of the stimulation site over tectal surface and the stimulus parameters. Two types of response were electrically evoked: the former turned the body and the eyes contraversively towards the source of natural stimulus; the second produced initial ipsiversive turning of the body and eyes, followed by several tail beats. Evoking one or other response depended on both the site and parameters of stimulation, and responses were interpreted as orienting- and escape-like, respectively. Depending on the stimulation site, four different zones in the tectum were distinguished: in the medial zone the stimulus elicited eye and tail movements whose size increased with the distance to the rostral pole. The stimulation of the antero-medial zone evoked contraversive or ipsiversive eye saccades but tail movements were similar, irrespective of eye movements. Stimulation within the extreme antero-medial zone evoked convergent eye movements, and tail displacements turning the body either ipsiversively or contraversively. Stimulation of the posterior zone often evoked complex tail movements and pure horizontal eye saccades. Both orienting- and escape-like responses were also dependent on the stimulus parameters. The relationships between stimulus parameters and tail- and eye-orienting movement characteristics suggest that the velocity and duration might be encoded in different aspects of the tectal activity. Current strength also modified the number of tail beats that appeared during escape-like response. In conclusion, the present data suggest the involvement of the optic tectum not only in orienting but also in escape responses and that movements of eye and tail mediating such responses depend on the tectal active locus together with its level of activity.

Spectral sensitivity of the goldfish Torus longitudinalis.

We measured the photopic spectral sensitivity of multiunit activity in the torus longitudinalis and optic tectum of goldfish. Since negative contrast stimuli are most effective for exciting torus longitudinalis, spiking activity was evoked by the shadow of a disc moving through a monochromatic light beam projected upon a screen. The amount of activity evoked in torus longitudinalis generally increased with the monochromatic stimulus radiance at the same rate for all wavelengths, indicating a univariant response. Spiking activity in tectum, however, increased at different rates across the spectrum, indicating color-dependent responses. The action spectra for torus longitudinalis were all similar and relatively flat as expected of a homogeneous, broad-band luminance processing system, and about 1 log unit more sensitive than the tectal action spectra. The latter generally displayed sharp peaks and dips in sensitivity indicative of opponent processing.

Tectal codification of eye movements in goldfish studied by electrical microstimulation

This work compares the tectal codification of eye movements in goldfish with those reported for other vertebrate groups. Focal electrical stimulation was applied in various tectal zones and the characteristics of evoked eye movements were examined as a function of (i) the position of the stimulation over the tectal surface, (ii) the initial position of the eyes and (iii) the parameters (pulse rate, current strength, duration) of the stimulus. In a large medial zone, stimulation within the intermediate and deep layers of the tectum evoked contraversive saccades of both eyes, whose direction and amplitude were roughly congruent with the retinotopic representation of the visual world within overlying layers. These saccades were minimally influenced by the initial position of the eye in the orbit. The topographical arrangement of evoked saccades and body movements suggests that this tectal zone triggers orienting responses in a similar way to those described in other vertebrates. Stimulations applied within the caudal tectum also evoked contraversive saccades, but—in disagreement with the overlying retinotopic map—the vertical component was absent. Taken together with electrically evoked body movements reported in free-swimming fish, these saccades could reveal that this zone is involved in escape responses. When stimulations were applied within the anteromedial zone of the tectum, contraversive movements of both eyes appeared much more dependent on initial eye position. Saccades elicited from this area displayed characteristics of “goal-directed saccades” which were similar to those described in the cat. The generation of goal-directed movements from the anteromedial zone suggests that this portion of the goldfish optic tectum has a different intrinsic organization or is connected with the brainstem saccade generator in a different fashion than the medial zone. Finally, stimulation of the extreme anteromedial zone evoked convergent eye movements. These movements and those reported in free-swimming fish following electrical stimulation of this tectal area suggest that this zone could be involved in feeding responses. The relationships between the parameters of electrical stimulation and the characteristics of elicited saccades suggest that the stimulated location within the tectum determines a constant direction in the evoked saccade, whereas the amount and duration of tectal activity, as mimicked by changes in stimulus parameters, together with the tectal locus, determine the velocity and amplitude of the evoked saccade.

Modulation of Microglial form and Immune Function by Factors Released from Goldfish Optic Nerves

Activation of microglia is associated with neural damage and may aid repair of the CNS. To begin to investigate their role, microglia purified from mouse brain were grown in media conditioned (CM) by goldfish optic nerve (GFON), optic tectum (GFOT), vagal lobe, telencephalon and cerebellum, and medium conditioned by rat optic nerves (RON). Microglia maintained in GFON- or GFOT-CM assumed an ameboid morphology, whereas microglia grown in media conditioned by the other neural tissues produced long, crenellated processes that resembled the ramified microglial form. Microglia maintained in all types of CM functioned as antigen presenting cells in a MHC-restricted manner when tested on conalbumin-specific Thelper (Th) cells, except for microglia maintained in GFON-and GFOT-CM. These studies suggest that GFON, in contrast to RON, produces a substance(s) that affects microglial morphology and immune reactivity, and may promote the vigorous regeneration seen in Gfon after damage.

Cranial meninges of goldfish: age-related changes in morphology of meningeal cells and accumulation of surfactant-like multilamellar bodies.

In the optic tectum of goldfish, the outer, middle and inner layers of the endomeninx were evident in animals ranging in age from 1 month to several years. The outer layer in young animals consisted of closely overlapping cells with intertwined processes, whereas in the older animals it contained large extracellular spaces. The intermediate layer cells were always arranged in a single continuous layer, but in young animals they overlapped extensively with one another toward their edges whereas in the oldest animals they became extremely flat and non-overlapping. The inner layer included an outer tier of cells with their bases adhering to the intermediate layer, and an inner tier of cells detached from both the intermediate layer and the basal lamina overlying the brain parenchyma. Inner layer cells contained many large vacuoles that were in continuity with the extracellular space. With age, the extracellular space and the vacuolar system expanded, and the inner layer evolved into a meshwork of attenuated cytoplasmic processes embedded in the granular extracellular matrix. Another age-related feature was the accumulation adjacent to the basal lamina of uniform disc-shaped membranous structures, resembling multilamellar bodies of lung surfactant. These "disc bodies" were apparently generated by the coalescence of vesicles formed at the surface of the inner layer cells, possibly as a by-product of protein secretion by these cells.

6,7-Dinitroquinoxaline-2,3-dione but not MK-801 exerts a protective effect against kainic acid neurotoxicity in the goldfish retina

Recent findings indicated that the excitotoxicity of glutamate analogues was prevented in the mammalian nervous system by N-methyl- d-aspartate (NMDA) antagonists. The neurodegenerative effects of kainic acid, and the putative protection of MK-801 and 6,7-dinitroquinoxaline-2,3-dione (DNQX), were investigated by morphological studies showing the toxicity of kainic acid to the neurons of the inner nuclear layer, and measuring choline acetyltransferase and glutamate decarboxylase activities in the retina. In addition, the proliferation of Miller retinal cells was assumed as an index of neuronal degeneration and was quantified by counting glial fibrillary acidic protein immunopositive cells. Our observations suggest that the non-NMDA receptor antagonist DNQX exerted a protective effect on goldfish retinal neurons, while MK-801 did not prevent the neurotoxicity induced by kainic acid in the goldfish retina. This finding is in agreement with previous work on kainic acid toxicity in the goldfish optic tectum.

Further characterization of axonally transported proteoglycans

We report further analysis of axonally transported proteoglycans in soluble and membranous subfractions of goldfish optic tectum. Distribution of transported 35SO4 radioactivity was 35.2% soluble, 63.4% Triton-NaCl extractable and 1.4% unextracted. Proteoglycans isolated on DEAE cellulose were treated with chondroitinase AC or nitrous acid and remaining heparan sulfate proteoglycans (HSPGs) and chondroitin sulfate proteoglycans (CSPGs) were sized on Sepharose CL-6B. Kav values and estimated molecular weights were: Soluble CSPG-0.36 (160 kDa), Triton-NaCl extracted CSPG-.031 (200 kDa), Soluble HSPG-0.37 (150 kDa), Triton-NaCl extracted HSPG-0.37 (150 kDa). For constituent CS and HS chains the Kav values and estimated molecular weights on CL-6B were: Soluble CS-0.55 (15 kDa), Triton-NaCl extracted CS-0.55 (15 kDa), Soluble HS-0.59 (13 kDa) and Triton-NaCl extracted HS-0.65 (9 kDa). CS was shown to be sulfated exclusively at carbon 4 for both soluble and Triton NaCl extracted fractions.

Axonal Transport of Proteoglycans in Regenerating Goldfish Optic Nerve

Labeling of goldfish optic nerve and tectum proteoglycans (PGs) was quantified following intraocular injection of 35SO4 and [3H]proline or [3H]glucosamine. Both intact animals and animals which had survived for periods of 10 to 119 days after an optic nerve crush lesion were examined. Regenerating retinal ganglion cell (RGC) axons reached the rostral pole of the tectum by 10 days postcrush and by 21 days had densely innervated the optic synaptic laminae. If the contralateral tectum had been removed, the regenerating RGC axons innervated the remaining ipsilateral tectum with a delay of approximately 14 days. There was a biphasic increase in the synthesis and transport of PGs during optic fiber regeneration which was not affected by the removal of the tectum. More highly sulfated PGs were preferentially off-loaded from the orthograde transport pool proximally in the optic nerve, both in the unoperated animals and during regeneration. These PGs also had longer and/or more glycosaminoglycan (GAG) chains than those off-loaded distally, in the tectum. During early regeneration, the synthesis and transport of chondroitin sulfate PGs (CSPGs) increased more than those of heparan sulfate PGs, and during the period of optic fiber invasion of the synaptic laminae, the PGs retained in the nerve had a higher content of CSPGs than those transported into the tectum. Removal of the contralateral tectum at the time of nerve crush resulted in a decrease in the size and/or numbers of GAGs and overall sulfation of PGs in the nerve by 21 days postoperatively. In addition, in the absence of the rectum, the HS/CS ratio in the nerve remained at preoperative levels until 21 days postoperatively at which time it began to fall. Newly synthesized PGs transported in the regenerating retinotectal projection undergo changes reflecting regulation of transcription/translation and of post-translational modification at the level of glycosyltransferase and sulfotransferase activities and by mechanisms intrinsic to the orthograde and retrograde transport system of RGC axons. These changes are regulated by both target-dependent and target-independent signals.

Kainic acid neurotoxicity in the goldfish optic tectum No protection by the NMDA receptor antagonist MK801 and partial protection by abolition of a tectal excitatory input

Some features of the mechanism of kainic acid neurotoxicity were tested after the injection of this substance in the optic tectum of goldfish. A systemic treatment with the N-methyl- d-aspartate (NMDA) receptor antagonist MK801 did not influence the degree of toxicity, assessed by a decrement of enzymatic neuronal markers; instead, a significant neuronal rescue was obtained after the abolition of the excitatory input from the torus longitudinals to the tectum.

Characterization of a novel set of membrane antigens associated with axonal growth. III: Expression in the regenerating goldfish optic nerve and tectum

In the preceding two papers in this series, a polyclonal antiserum (3070) and several monoclonal antibodies (MAbs 4 and 199) were used to define a novel set of laminin-binding membrane antigens in chick brain. In chick brain, 3070 and MAb 4 antigens stained growing axons and migrating nascent neurons transiently; though projection neurons were intensely labelled at all stages, normal glial cells were stained faintly or not at all. To learn if these antigens are associated more generally with active states of nerve growth, the present paper examined the expression of 3070 and MAb 4 antigens during regeneration of the goldfish optic nerve. Only a few optic fibers or glial cells were stained by 3070 antiserum in the resting (uninjured) optic nerve, but 10 days following a unilateral nerve crush, when the growth response is maximal in these fish, distal regenerating neuritic sprouts and glial cell bodies and processes were profusely labelled. The intensity of labelling was diminished by 135 days post-crush, when most active growth was completed. Since 3070/MAb 4 antigens exhibit properties expected for functional laminin receptors, these findings suggest several possible roles that they could play to aid nerve regeneration.

Extracellular proteins of goldfish optic tectum labeled by intraocular injection of 3H-proline

A promiment group of soluble glycoproteins with a molecular weight of 30K–40K and p I 5.0–5.6 was detected in various parts of the goldfish brain as well as in the optic nerves. Since these proteins are readily liberated from the tissue, we have designated them exoglycoproteins (X-GPs). The X-GPs in the optic tectum were found to be labeled after intraocular injection of radioactive tracers, in a manner consistent with the labeling of proteins transported in the optic axons. However, the labeling of X-GPs was blocked by intracranial injection of a protein synthesis inhibitor, whereas the labeling of axonally transported proteins was unaffected. This shows that the X-GPs can be synthesized locally within the brain. Nevertheless, when protein synthesis in the retina was blocked, the labeling of the X-GPs in the tectum was prevented, like the labeling of axonally transported proteins. Thus precursors for the synthesis of X-GPs can be derived from materials transported in the optic axons. This synthesis can occur in nonneuronal cells, as indicated by the incorporation of labeled amino acid into X-GPs in optic nerves directly exposed to the label. The synthesis of X-GPs was increased in regenerating nerves, suggesting that these proteins may play a role in regeneration. Partial amino acid sequencing of the proteins showed that they are identical to the proteins previously identified as “ependymins,” which have been implicated in neuronal plasticity. There are minor differences in amino acid sequence among some individual spots.

Pharmacologic evidence for NMDA, APB and kainate/quisqualate retinotectal transmission in the isolated whole tectum of goldfish

The optic tectum of goldfish with intact optic and toral marginal fiber tracts was isolated in a perfusion chamber where the effectiveness of antagonists was tested on synaptic field potential responses to stimulation of each afferent system. There were 3 main conclusions about excitatory synapses. First, monosynaptic activation of retinotectal synapses was not detectably antagonized by d-tubocurarine, implying there is no nicotinic cholinergic component to optic transmission nor strong cholinergic gating of optic terminals. Second, a significant component of retinotectical transmission was shown to be mediated by kainate and quisqualate receptors since 6,7-dinitroquinoxaline-2,3-dione and kynurenate strongly suppressed the optic field potential. In addition, activation of these synapses involves two previously undescribed N-methyl- d-aspartate (NMDA) and APB receptor subtypes since optic field potentials were partially suppressed by 2-amino-5-phosphonovalerate (APV), 2-amino-4-phosphonobutyrate (APB) and MK-801. This is the first evidence that APB receptors exist in the visual system central to the retina. Together, these results are consistent with the possibility that retinal ganglion cells use multiple glutamate receptor subtypes. Third, the optic tectum contains a population of intrinsic glutaminergic synapses activated by a non-optic input, the marginal fibers, which can be suppressed by both APV and kynurenate. The existence of tectal NMDA receptors whicha are not at primary optic synapses implies that APV used to interfere with rearrangement of optic fibers during development may act not only at afferent synapses but also at a more central component of the circuit.