Afferent and efferent connections of the mesencephalic reticular formation in goldfish.

The physiology of the mesencephalic reticular formation (MRF) in goldfish suggests its contribution to eye and body movements, but the afferent and efferent connections underlying such movements have not been determined. Therefore, we injected the bidirectional tracer biotinylated dextran amine into functionally identified MRF sites. We found retrogradely labelled neurons and anterogradely labelled boutons within nuclei of the following brain regions: (1) the telencephalon: a weak and reciprocal connectivity was confined to the central zone of area dorsalis and ventral nucleus of area ventralis; (2) the diencephalon: reciprocal connections were abundant in the ventral and dorsal thalamic nuclei; the central pretectal nucleus was also reciprocally wired with the MRF, but only boutons were present in the superficial pretectal nucleus; the preoptic and suprachiasmatic nuclei showed abundant neurons and boutons; the MRF was reciprocally connected with the preglomerular complex and the anterior tuberal nucleus; (3) the mesencephalon: neurons and boutons were abundant within deep tectal layers; reciprocal connections were also present within the torus semicircularis and the contralateral MRF; neurons were abundant within the nucleus isthmi; and (4) the rhombencephalon: the superior and middle parts of the reticular formation received strong projections from the MRF, while the projection to the inferior area was weaker; sparse neurons were present throughout the reticular formation; a reciprocal connectivity was observed with the sensory trigeminal nucleus; the medial and magnocellular nuclei of the octaval column projected to the MRF. These results support the participation of the MRF in the orienting response. The MRF could also be involved in other motor tasks triggered by visual, auditory, vestibular, or somatosensory signals.

Eye movements evoked by electrical microstimulation of the mesencephalic reticular formation in goldfish.

Anatomical studies in goldfish show that the tectofugal axons provide a large number of boutons within the mesencephalic reticular formation. Electrical stimulation, reversible inactivation and cell recording in the primate central mesencephalic reticular formation have suggested that it participates in the control of rapid eye movements (saccades). Moreover, the role of this tecto-recipient area in the generation of saccadic eye movements in fish is unknown. In this study we show that the electrical microstimulation of the mesencephalic reticular formation of goldfish evoked short latency saccadic eye movements in any direction (contraversive or ipsiversive, upward or downward). Movements of the eyes were usually disjunctive. Based on the location of the sites from which eye movements were evoked and the preferred saccade direction, eye movements were divided into different groups: pure vertical saccades were mainly elicited from the rostral mesencephalic reticular formation, while oblique and pure horizontal were largely evoked from middle and caudal mesencephalic reticular formation zones. The direction and amplitude of pure vertical and horizontal saccades were unaffected by initial eye position. However the amplitude, but not the direction of most oblique saccades was systematically modified by initial eye position. At the same time, the amplitude of elicited saccades did not vary in any consistent manner along either the anteroposterior, dorsoventral or mediolateral axes (i.e. there was no topographic organization of the mesencephalic reticular formation with respect to amplitude). In addition to these groups of movements, we found convergent and goal-directed saccades evoked primarily from the anterior and posterior mesencephalic reticular formation, respectively. Finally, the metric and kinetic characteristics of saccades could be manipulated by changes in the stimulation parameters. We conclude that the mesencephalic reticular formation in goldfish shares physiological functions that correspond closely with those found in mammals.

Visual orienting response in goldfish: a multidisciplinary study.

The neural basis underlying the orienting response has been thoroughly studied in frontal-eyed mammals. However, in non-mammalian species, including fish, it remains almost unknown. Therefore, we studied the contribution of the optic tectum and the mesencephalic reticular formation to the performance of the orienting response in goldfish, using behavioural, physiological, and anatomical tracer techniques. The appearance of a visual stimulus (a pellet of food) in the environment of a goldfish evoked a turn of the body to reorient the line of sight. Left-tectal lobe ablation abolished the orienting turn response towards the contralateral hemifield. Electrical microstimulation of the optic tectum suggested the presence of a motor map, which is in correspondence with the overlying visual representation, as previously reported in other vertebrates. The tracer biotin-dextran amine was injected into different functionally identified tectal zones. The results showed that rostral and caudal poles of the mesencephalic reticular formation receive outflow mainly from the rostral and caudal tectal poles, respectively. This suggests that the tectal wiring with downstream structures is site-dependent. Furthermore, the electrical activation of rostral and caudal mesencephalic reticular formation revealed a different contribution to vertical and horizontal orienting eye movements. We conclude that the basic neural system coding the orienting response appears early in phylogenesis, although some specific characteristics are selected by adaptive pressure.

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.

Neural substrata underlying tectal eye movement codification in goldfish.

The optic tectum encodes orienting eye saccades in a spatially ordered map. To investigate whether the functional properties of each tectal site are related to a particular pattern of connectivity with downward structures in the brainstem, two sets of experiments were carried out. First, biotinylated dextran amine (BDA) was injected at different tectal sites along the anteroposterior axis. Electrical stimulation at these sites evoked saccades whose horizontal component amplitudes increased with the distance to the rostral pole. In the second experiment, BDA and fluoro-ruby (FR) were injected at different tectal sites along the mediolateral axis. Electrical stimulation here evoked saccades with different upward and downward directions, but similar horizontal component amplitudes. A major finding of the first experiment was that a topographic link of the tectum exists with the mesencephalic reticular formation, but that such a connection was absent or very attenuated for the rhombencephalic reticular formation. In the second set of experiments, the clusters of BDA and FR boutons left by the mediolateral tectal sites were separated in the rostral mesencephalon, at the level of the nucleus of the medial longitudinal fasciculus, but overlapped in the caudal mesencephalon and rhombencephalon. These data provide evidence that decodification of tectal motor commands is based, at least in part, on the connectivity of each tectal locus on downward structures with the brainstem.

Connectivity of the tectal zones coding for upward and downward oblique eye movements in goldfish.

Deep layers of the goldfish tectum code movements in a topographically ordered motor map. This work studies the relationship between tectal sites (coding eye movements with different vertical directions) and the distributions of boutons (left by their projections), within rostral mesencephalic structures and rhombencephalic reticular formations. These regions have been involved in the generation of the vertical and horizontal components of eye movement, respectively, as suggested by the Cartesian hypothesis of de-codification of tectal signal. With this aim, discrete injections of biotinylated dextran amine (BDA) and Fluoro-Ruby (FR) were made into functionally identified tectal sites, coding oblique eye movements with similar amplitude of the horizontal component but opposite upward and downward vertical directions, and the distribution of synaptic endings was determined. The main findings of the present work were as follows: 1) within the tectal descending tract, axons were organized according to the location of injected sites within the tectum; 2) BDA and FR boutons were distributed in separate clusters within the medial longitudinal fasciculus and oculomotor nuclei, as well as in the nearby mesencephalic reticular formation; and 3) the regions containing both types of bouton overlapped moderately within the mesencephalic reticular formation at the isthmus level. Overlapping was more extended at the different levels of the rhombencephalic reticular formation, although a shift in the distribution of both types of bouton was always observed. These results suggest that, within the vertical generator, the endings were separated to contact the different neuronal population that codes the upward and downward components of movements. In contrast, in the horizontal generator, tectal endings more likely converge on the same neuronal population to code the horizontal component of movements, irrespective of whether the oblique movements were directed upward or downward.