Tectoreticular pathways in the turtle, Pseudemys scripta. II. Morphology of tectoreticular cells

The morphology of tectoreticular neurons in turtles was examined with serial section reconstructions of neurons retrogradely filled with HRP. Six classes of tectal neurons project into the three tectobulbar pathways characterized in the preceding paper (Sereno, '85). (1) Large multipolar neurons with somata in the central gray layers, and with moderately branched dendrites sometimes spanning over a millimeter, project into the dorsal tectobulbar pathway, TBd. Their dendrites are covered with fine spicules and tend to arborize in the lower third of the superficial gray layers. (2) Medium-sized neurons with multiple radial dendrites and somata in the central white and upper periventricular layers probably project into the ipsilateral intermediate tectobulbar pathway, TBi. Their dendrites also bear fine spicules and usually reach the tectal surface. (3) Small radial cells in the periventricular layers, and (4) small bitufted radial cells in the superficial gray project into the small caliber component of the ipsilateral ventral tectobulbar pathway, TBv(sm). (5) Medium-sized central gray neurons with stratified dendrites, and (6) medium-sized central gray neurons with horizontal dendrites probably project into the medium caliber component of the ventral tectobulbar pathway, TBv(med). In contrast to TBd and TBi neurons, these last four classes emit a spray of long, filamentous dendritic appendages in the central gray and have dendritic arbors near the top of the superficial gray. The morphology of the neurons described in this and the preceding paper is briefly discussed in light of current ideas about tectally mediated sensorimotor transformations.     

Morphology of retinogeniculate terminals in the turtle, Pseudemys scripta elegans

The dorsal lateral geniculate complex in turtles receives a bilateral, topographic projection from the retina and projects to the telencephalon. This study examined the morphology of individual retinogeniculate terminals that were filled with horseradish peroxidase by injections in the optic tract or optic tectum. A large number of retinogeniculate terminals were successfully filled and detailed drawings were prepared of 87 terminals. Terminals were classified into three types based on the size and number of varicosities in the terminal, and (if a terminal formed a spatially restricted arbor) the volume of the arbor. Type I retinogeniculate terminals form spatially restricted, large-volume arbors with a low density of large varicosities. Type II retinogeniculate terminals form small volume arbors with a high density of small varicosities. Type III retinogeniculate terminals, in contrast to types I and II, do not form spatially restricted arbors. Rather, they consist of sparsely branched axons that parallel the optic tract and contain scattered en passant varicosities. Plots of the distribution of different terminal types throughout the geniculate complex show that all three terminal types occur throughout the rostrocaudal and mediolateral extents of the complex. However, each terminal type has a preferential distribution with type II terminals being concentrated in the outer half of the neuropile, type I terminals in the inner half of the neuropile, and type III terminals in the cell plate. All three types can arise from axons that continue caudally to terminate in the tectum. These findings raise the possibility that various classes of retinal ganglion cells differ in their mode of termination within the geniculate complex, but the precise relation between the three types of retinogeniculate terminals and the classes of ganglion cells remains to be determined.     

Caudal topographic nucleus isthmi and the rostral nontopographic nucleus isthmi in the turtle, Pseudemys scripta

Isthmotectal projections in turtles were examined by making serial section reconstructions of axonal and dendritic arborizations that were anterogradely or retrogradely filled with HRP. Two prominent tectal-recipient isthmic nuclei--the caudal magnocellular nucleus isthmi (Imc) and the rostral magnocellular nucleus isthmi (Imr)--exhibited strikingly different patterns of organization. Imc cells have flattened, bipolar dendritic fields that cover a few percent of the area of the cell plate constituting the nucleus and they project topographically to the ipsilateral tectum without local axon branches. The topography was examined explicitly at the single-cell level by using cases with two injections at widely separated tectal loci. Each Imc axon terminates as a compact swarm of several thousand boutons placed mainly in the upper central gray and superficial gray layers. One Imc terminal spans less that 1% of the tectal surface. Imr cells, by contrast, have large, sparsely branched dendritic fields overlapped by local axon collaterals while distally, their axons nontopographically innervate not only the deeper layers of the ipsilateral tectum but also ipsilateral Imc. Imr receives a nontopographic tectal input that contrasts with the topographic tectal input to Imc. Previous work on nucleus isthmi emphasized the role of the contralateral isthmotectal projection (which originates from a third isthmic nucleus in turtles) in mediating binocular interactions in the tectum. The present results on the two different but overlapping ipsilateral tecto-isthmo-tectal circuits set up by Imc and Imr are discussed in the light of physiological evidence for selective attention effects and local-global interactions in the tectum.     

Morphology of basal optic tract terminals in the turtle,Pseudemys scripta elegans

The morphologies of axon terminals of retinal ganglion cells projecting to the basal optic nucleus (BON) via the basal optic tract (BOT) were studied in the red-eared turtle. The BOT was visualized on the ventral surface of the brainstem in vitro, and either biotinylated dextran amine was injected extracellularly or neurobiotin was injected into physiologically identified axons during intracellular recordings. Up to 16 hours after tracer injection, the brains were fixed, sectioned parasagittally, and stained for biotin and Nissl substance. The diameters and depths of extracellularly filled axons were measured at three BON sites. Fourteen axons were reconstructed from serial sections with the aid of appropriate computer software. Analysis of extracellularly filled retinal axons revealed that about three times more axons were present just inside the rostral border of the BON compared with its caudal border. Thick (2–4 μm) axons were located within 100 μm from the ventral border, whereas thin (<2 μm) axons were found throughout the nucleus. Only the thinnest axons (<1 μm) extended caudally from the nucleus, indicating that some extracellularly labelled fibers passed through the BON. The intracellularly filled axons were more similar to the thick axons filled extracellularly and arborized entirely within the BON. All of the intracellularly filled axons had thick ventral trunks from which many thin branches extended dorsally and obliquely within the BON. The thin branches bifurcated repeatedly to form bead-like varicosities or boutons that often formed clusters within regions of 150 μm³ or less. These clusters may reflect areas of focused synaptic contact on BON cells with specific direction preferences. J. Comp. Neurol. 393:267–283, 1998. © 1998 Wiley-Liss, Inc.     

Organization of corticogeniculate projections in the turtle, Pseudemys scripta

The efferent pathways from the visual cortex to the dorsal lateral geniculate complex of turtles have been studied by using the orthograde and retrograde transport of horseradish peroxidase (HRP). Injections of HRP in the lateral thalamus retrogradely label neurons throughout the visual cortex. The majority of labeled neurons have somata in layer 2 of the lateral part of dorsal cortex (D2); a minority have somata in layer 3. Labeled neurons in layer 2 tend to have vertically oriented, fusiform somata and dendrites that ascend into layer 1. Labeled neurons in layer 3 have fusiform somata and dendrites, both oriented horizontally. Injections of HRP in visual cortex orthogradely label corticofugal axons. Those projecting to the lateral geniculate complex course laterally from the visual cortex, pass through the striatum (occasionally bearing varicosities), and enter the diencephalon in the ventral peduncle of the lateral forebrain bundle. Individual axons leave the ventral peduncle and run dorsally in the transverse plane, entering the dorsal lateral geniculate complex from its ventral edge. They continue dorsally, principally in the cell plate of the geniculate complex, where they bear varicosities.     

Afferents to the oculomotor nucleus in the goldfish (Carassius auratus) as revealed by retrograde labeling with horseradish peroxidase.

The goal of this work was to compare the distribution and morphology of neurons projecting to the oculomotor nucleus in goldfish with those previously described in other vertebrate groups. Afferent neurons were revealed by retrograde labeling with horseradish peroxidase. The tracer was electrophoretically injected into the oculomotor nucleus. The location of the injection site was determined by the antidromic field potential elicited in the oculomotor nucleus by electrical stimulation of the oculomotor nerve. Labeled axons whose trajectories could be reconstructed were restricted to the medial longitudinal fasciculus. In order of quantitative importance, the afferent areas to the oculomotor nucleus were: (1) the ipsilateral anterior nucleus and the contralateral tangential and descending nuclei of the octaval column. Furthermore, a few labeled cells were found dorsomedially to the caudal pole of the unlabeled anterior octaval nucleus; (2) the contralateral abducens nucleus. The labeled internuclear neurons were arranged in two groups within and 500 microns behind the caudal subdivision of the abducens nucleus; (3) a few labeled cells were observed in the rhombencephalic reticular formation near the abducens nucleus, most of which were contralateral to the injection site. Specifically, stained cells were found in the caudal pole of the superior reticular nucleus, throughout the medial reticular nucleus and in the rostral area of the inferior reticular nucleus; (4) eurydendroid cells of the cerebellum, located close to the contralateral eminentia granularis pars lateralis, were also labeled; and (5) a small and primarily ipsilateral group of labeled cells was located at the mesencephalic nucleus of the medial longitudinal fasciculus. The similarity in the structures projecting to the oculomotor nucleus in goldfish to those in other vertebrates suggests that the neural network involved in the oculomotor system is quite conservative throughout phylogeny. Nevertheless, in goldfish these projections appeared with some specific peculiarities, such as the cerebellar and mesencephalic afferents to the oculomotor nucleus.     

Eye movement related neurons in the cat pontine reticular formation: projection to the flocculus

This study examines projection to the cerebellar flocculus of eye movement-related neurons in the median and paramedian part of the cat pontine tegmentum between the trochlear and the abducens nucleus. They were identified by rhythmic activity related to horizontal vestibular nystagmus induced by sinusoidal rotation. These neurons were classified into several groups by their discharge patterns during nystagmus, using criteria of earlier studies on saccadic eye movements and vestibular nystagmus in the monkey. Electrical stimulation of the ipsilateral flocculus elicited antidromic spike responses in a number of burst-tonic neurons and long-lead and medium-lead burst neurons. These neurons were located in and around the medial longitudinal fasciculus, the nucleus raphe pontis and the nucleus reticularis tegmenti pontis. A few neurons tested were also activated antidromically by stimulation of the contralateral flocculus. In contrast, no pauser neurons were activated from the ipsi-lateral flocculus. It is concluded that eye movement-related neurons in the medial pontine tegmentum, except for pauser neurons, directly project to the flocculus and may convey information about eye movements of visual and vestibular origins to the flocculus.     

Neuropeptide Y-immunoreactive amacrine cells in the retina of the turtle Pseudemys scripta elegans

Antiserum directed against neuropeptide Y selectively labeled certain amacrine cells in the turtle retina. The cell types, sizes, dendritic stratification, regional distribution, and degrees of immunolabeling were examined. The results indicated that three morphologically distinct cell types were labeled: types A, B, and C. Computer rotation of digitized data from camera lucida drawings was used to study dendritic stratification. The type A somata were large (11.5 micron in diameter), well-stained, and located in the third tier of the inner nuclear layer. Type A somata gave rise to well-stained processes which arborized within the inner plexiform layer in strata 1 and 3 and at the border between strata 4 and 5. Processes in stratum 1 were sparse and delicate with small boutons. Processes in stratum 3 were numerous and often coarse, with many large and small boutons. At the border between strata 4 and 5 the processes were frequently numerous but slender, with many small boutons. Occasional immunolabeled processes were found in the ganglion cell layer. The somata of the type B cells were smaller (9.0 micron in diameter) and gave rise to single labeled processes which descended into the inner plexiform layer and divided quickly into two secondary processes. These secondary processes gave rise to lightly labeled dendritic fields which arborized primarily in strata 2 and 4. The type C cells were usually observed at the periphery of the retina and had large somata (12.0 micron in diameter) with simple, but very elongated, dendritic arborizations in strata 1, 4, and 5. Observations also showed that type A and B cells were often found in close proximity to each other and suggested that dendrites of these cells made contact with each other. The labeled neurons were distributed relatively evenly throughout the retina except for the visual streak where they were sparse. This study indicates that neuropeptide Y-like immunoreactivity is found in more than one anatomically distinct class of amacrine cells in the turtle retina.     

Morphology of lumbar motoneurons innervating hindlimb muscles in the turtle Pseudemys scripta elegans: an intracellular horseradish peroxidase study

Motoneurons in the turtle lumbar spinal cord, electrophysiologically identified as innervating a muscle belonging to a functional group, were injected with horseradish peroxidase by electrophoresis. A total of 45 motoneurons were reconstructed from transverse sections. Eleven motoneurons were identified as innervating knee extensor muscles, eight as innervating hip retractor and knee flexor muscles, 14 as supplying ankle and/or toe extensors, and 12 as innervating ankle and/or toe flexor muscles. The cell bodies were elongated and spindle-shaped in the transverse plane. The mean equivalent soma diameter was calculated to be 33.4 micrometers. The mean axon conduction velocity was 15.7 m/second. Significant, though rather weak, positive correlations were found between soma diameter, axon diameter, and axon conduction velocity. The axons of the reconstructed motoneurons did not reveal a recurrent axon collateral. However, a few unidentified motoneurons did possess such collaterals. The dendritic trees were restricted to the ipsilateral side of the cord, but reached out in lateral, ventral, and ventromedial directions to the subpial surface. Easily recognizable and characteristic dendrites were found both in the dorsal dendritic tree and in the dorsomedial dendritic tree. Correlations were calculated between the soma diameter and (1) the number of first-order dendrites, (2) the mean diameter of the first-order dendrites, and (3) the combined diameter of the first-order dendrites. In each case no correlations or only weak correlations were found. Fair correlations were observed between the diameter of a first-order dendrite and the number of terminal dendritic branches (r = .61) and the combined dendritic length (r = .78). However, correlations between the combined diameter of all first-order dendrites per neuron and the total number of terminal dendritic branches and the total combined dendritic length of a neuron were extremely weak. The overall appearance of turtle spinal motoneurons is comparable to that observed in other "lower" vertebrates such as frog and lizard. However, similarities are also observed between certain morphometric parameters in turtle and cat lumbar motoneurons.     

Quantitative autoradiography of muscarinic and benzodiazepine receptors in the forebrain of the turtle, Pseudemys scripta

The distribution of muscarinic and benzodiazepine receptors was investigated in the turtle forebrain by the technique of in vitro receptor autoradiography. Muscarinic binding sites were labeled with 1 nM 3H-quinuclidinyl benzilate (3H-QNB), and benzodiazepine sites were demonstrated with the aid of 1 nM 3H-flunitrazepam (3H-FLU). Autoradiograms generated on 3H-Ultrofilm apposed to tissue slices revealed regionally specific distributions of muscarinic and benzodiazepine binding sites that are comparable with those for mammalian brain. Dense benzodiazepine binding was found in the anterior olfactory nucleus, the lateral and dorsal cortices, and the dorsal ventricular ridge (DVR), a structure with no clear mammalian homologue. Muscarinic binding sites were most dense in the striatum, accumbens, DVR, lateral geniculate, and the anterior olfactory nucleus. Cortical binding sites were studied in greater detail by quantitative analysis of autoradiograms generated by using emulsion-coated coverslips. Laminar gradients of binding were observed that were specific for each radioligand; 3H-QNB sites were most dense in the inner molecular layer in all cortical regions, whereas 3H-FLU binding was generally most concentrated in the outer molecular layer and was least dense through all layers in the dorsomedial cortex. Because pyramidal cells are arranged in register in turtle cortex, the laminar patterns of receptor binding may reflect different receptor density gradients along pyramidal cell dendrites.     

Localization of aspartate-like immunoreactivity in the retina of the turtle (Pseudemys scripta).

Aspartate has been reported to be a putative excitatory neurotransmitter in the retina, but little detailed information is available concerning its anatomical distribution. We used an antiserum directed against an aspartate-albumin conjugate to analyze the anatomy, dendritic stratification, and regional distribution of cell types with aspartate-like immunoreactivity in the turtle retina. The results showed dramatic differences in immunoreactivity in the peripheral versus the central retina. Strong aspartate-like immunoreactivity was shown in the peripheral retina, with many well-labeled processes in the inner plexiform layer. Many bipolar, horizontal, amacrine, and ganglion cells, some photoreceptors, and some unidentified cells were strongly immunoreactive in the peripheral retina. In contrast, although the central retina showed well-labeled horizontal cells, there was only light labeling in the inner plexiform layer with weakly immunoreactive amacrine and ganglion cells and no labeled bipolar cells. There were several strongly immunoreactive efferent nerve fibers which left the optic nerve head and arborized extensively in the retina. At the electron microscopic level, electron-dense reaction product was associated with synaptic vesicles at bipolar and amacrine cell synapses in the inner plexiform layer. These results suggest that aspartate may be involved in many diverse synaptic interactions in both the outer plexiform layer and the inner plexiform layer of the turtle retina.     

Brainstem afferents to the oculomotor omnipause neurons in monkey

To determine how saccade-related areas in the brainstem address the saccade generator, we examined the afferents to the nucleus raphe interpositus. This region contains the omnipause neurons, which are pivotal in the generation of saccades. Horseradish peroxidase injected iontophoretically into the nucleus raphe interpositus retrogradely labeled a variety of brainstem nuclei. The greatest numbers of labeled neurons were in the paramedian pontomedullary reticular formation, in the nuclei reticularis gigantocellularis, and paragigantocellularis lateralis. Labeling was more modest but consistent in the interstitial nucleus of Cajal and the adjacent mesencephalic reticular formation, the middle gray of the superior colliculi, the region dorsolateral to the nucleus reticularis tegmenti pontis, and the medial vestibular nucleus. A few neurons were labeled around the habenulopeduncular tract and in the medial portion of the nucleus of the fields of Forel, in the nucleus reticularis medullaris ventralis, and in the spinal nucleus of the trigeminal nerve, the cochlear nucleus, and the superior olivary complex. The distribution and density of labeling suggest that omnipause neurons in the monkey are more intimately connected with other oculomotor structures than those in the cat. In addition, the rhombencephalic reticular afferents to the monkey omnipause neurons are more concentrated in their immediate vicinity than in the cat. The label consistently found dorsolateral to the nucleus reticularis tegmenti pontis may be a newly discovered link in saccade generation.     

Anatomy and physiology of saccadic burst neurons in the alert squirrel monkey. I. Excitatory burst neurons

Saccadic burst neurons in the pontine reticular formation have been implicated in the generation of saccades in the horizontal plane on the basis of lesion and extracellular recording studies in the cat and monkey. In the present study, saccadic burst neurons were anatomically and physiologically characterized with intraaxonal recording and injection of horseradish peroxidase in the alert squirrel monkey. A population of burst neurons were found that appear analogous to the excitatory burst neurons (EBNs) described previously in the cat. All neurons are located in the caudal pontine reticular formation and have a major axonal projection to the ipsilateral abducens nucleus. Additional projections were found to the medial vestibular nucleus, the nucleus prepositus, and regions of the pontine and medullary reticular formation rostral, ventral, and caudal to the abducens. All neurons fire exclusively during saccades and have a discharge pattern similar to that of medium-lead burst neurons described previously in the cat and monkey. In most neurons the saccadic burst begins 5-15 msec before saccade onset. Linear relationships exist between burst duration and saccade duration, number of spikes in the burst and saccade amplitude, and firing frequency and instantaneous velocity. Physiological activity of each neuron shows the closest relationship with the amplitude of the saccade component in a particular direction. For all neurons, this on-direction is in the ipsilateral hemifield and is predominantly horizontal, but may have either an upward or downward vertical component. These results support a major role for the EBNs in the monkey in generating the saccadic burst in abducens motoneurons, as well as in contributing to the oculomotor activity in other classes of premotor neurons.     

Electrophysiological and neuropharmacological study of tectoreticular pathways in lampreys

Tectoreticular (TR) cells along the diencephalic–mesencephalic border are the origin of prominent crossed and uncrossed pathways that project to the middle (MRRN) and posterior (PRRN) rhombencephalic reticular nuclei in juvenile and adult lampreys [I.C. Zompa, R. Dubuc, Diencephalic and mesencephalic projections to rhombencephalic reticular nuclei in lampreys, Brain Res. (1998) in press.]. This study investigated the synaptic contacts between TR axons and the reticular cells. Intracellular recordings were carried out in reticular neurones (n=124) while microstimulating the TR regions. Tectoreticular inputs were recorded in all reticular cells studied (248 PSPs); although stronger responses were evoked in the MRRN neurones. The majority of responses were excitatory, but increasingly mixed and inhibitory when recorded in the middle and caudal part of the reticular nuclei. The excitation had the shortest onset latencies and sharpest slopes measured in both reticular nuclei, while the inhibition was longer and smoother. The characteristics of TR inputs to different reticular cell types is also presented. The transmission of evoked responses was isolated to the crossed and uncrossed TR pathways by studying the effects of 1% Xylocaine ejections and surgical lesions. The TR inputs were transmitted to reticular cells through monosynaptic and polysynaptic contacts. The synaptic transmission involved excitatory amino acids, acting through AMPA and NMDA receptors, while the inhibition was glycinergic. Comparisons with other sensory systems in lampreys are discussed.     

Frontal eye field projection to the paramedian pontine reticular formation traced with wheat germ agglutinin in the monkey

Injections of the retrograde tracer [125I]wheat germ agglutinin have been placed in different areas of the paramedian pontine reticular formation (PPRF), a well known premotor center for gaze control. Experiments in 5 monkeys revealed 3 major sources of input: (1) bilateral projections from the so-called frontal eye field (FEF), which is situated in the frontal cortex around the arcuate sulcus; (2) the intermediate and deep layers of mainly the contralateral superior colliculus; and (3) ipsilateral projections from brainstem structures such as the accessory oculomotor nuclei (nucleus interstitialis of Cajal, nucleus of Darkschewitsch, and nucleus of the posterior commissure), the mesencephalic reticular formation, the vestibular nuclei, the nucleus prepositus hypoglossi, and the cerebellar fastigial nucleus. The results are compared with previous anatomical investigations and confirm the electrophysiologically demonstrated FEF-PPRF-abducens disynaptic pathway.     

Dendrite distribution of identified motoneurons in the lumbar spinal cord of the turtle Pseudemys scripta elegans

Motoneurons in the turtle lumbar spinal cord were injected with HRP by electrophoresis after being electrophysiologically identified as innervating a muscle belonging to a functional group. The distribution of dendrites was studied in transverse reconstructions of 45 motoneurons, including 11 motoneurons identified as innervating knee extensor muscles, eight motoneurons innervating hip retractor and knee flexor muscles, 14 motoneurons innervating ankle and/or toe extensors and 12 motoneurons innervating ankle and/or toe flexor muscles. The dorsal dendritic tree of motoneurons innervating distally positioned musculature (ankle and/or toe extensors and flexors) was observed to contain significantly less terminal dendritic branches compared to the dorsal dendritic trees of motoneurons innervating proximally situated (hip and knee) muscles. The distribution of dendrites within the white matter was studied by measuring the total projected length of the dendritic branches within empirically defined sectors in the transverse plane. This kind of analysis also revealed differences between the dorsal dendrites of motoneurons innervating distally and proximally positioned muscles conforming to the counts of terminal dendritic branches. It is suggested that these apparent differences in the size of the dorsal dendrite may be related to the number of synapses made by primary afferents. In the white matter, the highest dendritic density for all four groups of mononeurons was found within the central part of the lateral funiculus. However, only in the ventral funiculus could slight indications be found that the dendritic density of functionally different motoneuron groups may bear some relation to the locations of the terminations of the descending pathways known to establish monosynaptic contacts with lumbar mononeurons.     

Eye movements evoked by electrical stimulation in the superior colliculus of rats and hamsters

Although the organization of the sensory representations in the rodent superior colliculus is known to be similar to that of more advanced mammals, little is known about collicular motor organization in these animals. Since it has been suggested that rodents make head, rather than eye, movements to facilitate visual orientation, fundamental organizational dissimilarities could exist in the superior colliculi of different species to underlie these different orientation preferences. However, we observed that electrical stimulation of the rodent colliculus evoked the same pattern of movements observed in other mammals. Conjugate, contraversive saccadic eye movements were evoked and were found to be topographically organized. In addition, vibrissa and pinna movements could also be evoked. Although some species differences in evoked movements were evident, the same pattern of motor representations appears to exist in widely disparate mammalian species.     

Distribution of vesicular glutamate transporters in the brain of the turtle (Pseudemys scripta elegans)

The distribution of glutamatergic neurons has been extensively studied in mammalian and avian brains, but its distribution in a reptilian brain remains unknown. In the present study, the distribution of subpopulations of glutamatergic neurons in the turtle brain was examined by in situ hybridization using probes for vesicular glutamate transporter (VGLUT) 1–3. Strong VGLUT1 expression was observed in the telencephalic pallium; the mitral cells of the olfactory bulb, the medial, dorsomedial, dorsal, and lateral parts of the cerebral cortex, pallial thickening, and dorsal ventricular ridge; and also, in granule cells of the cerebellar cortex. Moderate to weak expression was found in the lateral and medial amygdaloid nuclei, the periventricular cellular layer of the optic tectum, and in some brainstem nuclei. VGLUT2 was weakly expressed in the telencephalon but was intensely expressed in the dorsal thalamic nuclei, magnocellular part of the isthmic nucleus, brainstem nuclei, and the rostral cervical segment of the spinal cord. The cerebellar cortex was devoid of VGLUT2 expression. The central amygdaloid nucleus did not express VGLUT1 or VGLUT2. VGLUT3 was localized in the parvocellular part of the isthmic nucleus, superior and inferior raphe nuclei, and cochlear nucleus. Our results indicate that the distribution of VGLUTs in the turtle brain is similar to that in the mammalian brain rather than that in the avian brain.