Total luminous flux: a possible response determinant for the normal monkey.

Thirteen normal monkeys (Macaca mulatta) trained to discriminate between transilluminated figures of equal area and different luminance, and consequently different luminous flux, made similar numbers of errors during training on a new problem with the same luminance values but with targets equated for luminous flux. These findings together with results of "critical trials" suggest that the significant cue in the original problem was luminous flux. This behavior is strikingly similar to what has been reported for the monkey following exclusion of the geniculostriate system.

Early postnatal development of the monkey neostriatum: a Golgi and ultrastructural study.

Paired specimens of the neostriatum were taken from monkeys at zero (newborn), one, two, four, eight, and 16 weeks of age, and prepared for Golgi impregnations and electron microscopy. Light microscopy shows that in the first postnatal week, the structure contains the five neuronal types and four categories of afferent axons described in the adult, as well as some cells too undifferentiated to classify. Most neurons exhibit immature dendritic features, including local enlargements, terminal growth cones with filopodia, and filiform processes. In spiny type I cells, various levels of maturity may coexist in regions of a single dendrite, in different dendrites of the same neuron, and among individual cells. Spine density increases progressively with age, but the relative distribution of spine types remains about the same. Spiny type II neurons show some decline in spine density, and generally mature sooner than spiny type I cells. The long axons of spiny neurons have varicosities which disappear at about eight weeks. In younger animals (newborn and one week), the dendrites of aspiny neurons (types I, II, and III) may have a "spiny" appearance, exhibiting many spine-like and filiform processes. Concurrently, the short axons vary in degree of arborization from very immature to well developed. Electron microscopy corroborates the developmental features recognized in the Golgi material: dendritic and axonal growth cones, filopodia and varicosities, as well as various stages of maturation in somata and dendrites. Degenerating elements, mostly of an axonal nature, are seen up to eight weeks. The synapses which reach maturity at birth are of the asymmetric axospinous type, in which the axonal profile contains small round vesicles, and of the symmetric axodendritic class, with the presynaptic elements having pleomorphic vesicles. Some synapses are slower to mature and appear at one to eight postnatal weeks. These include those made by profiles with pleomorphic vesicles, forming either symmetric contacts with somata and axon initial segments, or asymmetric contacts with spines. The same applies to the asymmetric axodendritic synapses made by elements containing small round vesicles. Finally, profiles containing large round or flat vesicles are the latest to participate in mature synapses formation. Findings indicate that a considerable degree of qualitative and quantitative change takes place in the monkey neostriatal neuropil during early postnatal development, especially in the first eight-week period.

A Golgi and ultrastructural study of the monkey globus pallidus.

Golgi preparations reveal that the most frequent type of pallidal neuron (principal cell), which has been recognized in all previous reports, is large (20-50 microns), fusiform, with dendrites up to 700 microns long. Large neurons of globular shape are less frequently impregnated. The morphology of dendrites varies considerably within the same neuron. Some exhibit numerous spines and protrusions and are seen to terminate in elaborate arborizations. A small interneuron (12 microns), with relatively short dendrites, up to 150 microns, and a short sparsely branching axon is observed less frequently. At least two types of afferent axons are present. A small-diameter fiber from the neostriatum enters the pallidum in bundles and gives rise to numerous thin branching processes with varicosities about 1 micron in size. The axon collaterals are oriented orthogonal to the main axon and parallel to the dendrites of principal cells. A large-caliber fiber with clusters of 2-3 microns swellings can also be seen in close proximity to large pallidal dendrites. Ultrastructurally, principal cell dendrites (trunks, spines, and protrusions) are totally covered by synapsing axon terminals. In contrast, some small dentrites, presumed to belong to interneurons, form very few synapses. At least six categories of profiles containing vesicles are observed. One group has cytologic features of dendrites and participates in serial and triadic synapses with other profiles in the pallidal neuropil. Results suggest that the synaptic organization of the globus pallidus may be viewed as a repetitive, geometric arrangement of striatal and other afferent axons ensheathing and synapsing with the dendrites of principal cells. This pattern is interrupted by the presence of presynaptic dendrites, probably belonging to interneurons, which participate in complex synaptic arrangements.

Early postnatal development of the monkey globus pallidus: a Golgi and electron microscopic study.

The globus pallidus of 20 monkeys ranging in age from newborn to 4 months was examined in Golgi-impregnated material and ultrastructurally. There was no discernible difference between the lateral and medial segments of the structure. At the light microscope level, all neuronal types described in the adult are found at birth. The most common, the large fusifom cell, shows initial signs of immaturity such as blunt protrusions and dendritic dilations at bifurcation points, as well as growth cones, filopodia, and filiform processes. These features become more rare with age, and by 4 months, the neurons appear fully mature save for the terminal dendritic arborizations which are still underdeveloped. From the earliest ages examined, the large globular cells and the interneurons are more mature than the previous type. The afferent radial fibers of striatal origin are observed from birth, but they are grouped in bundles only after 8 weeks. The density of their climbing branches increases over time, reaching a mature appearance by 16 weeks. Afferents entering from the ventral surface do not yet show clusters of varicosities at 2 weeks. At the latter age, plexuses of fine beaded fibers are already seen covering large extensions of the nucleus. The fine structure correlates well with the Golgi material. The basic features of the neuropil are present at birth, albeit with immature characteristics such as the incomplete covering of the dendrites with axonal boutons and the low level of myelination of the radial fibers. Growth cones and profiles with signs of degeneration are observed during the first month. In the early ages examined, most dendrites show large varicosities and protrusions, some of which are spinelike and can be postsynaptic to multiple terminals. The other dendritic type, with only an occasional axodendritic synapse, is also seen from birth and increases in size as a function of time. The type I axonal boutons, of probable striatal origin, are quite immature at birth, and their characteristic interdigitations are seen only after the first week. The type II, III, IV, and V boutons appear mature at all ages examined but crest synapses formed by the type III terminals are observed in the later stages of the study. Finally, postsynaptic vesicle-containing profiles are present at 4 weeks, but triadic synaptic arrangements are formed only by 16 weeks.(ABSTRACT TRUNCATED AT 400 WORDS)

GABAergic elements in the neuronal circuits of the monkey neostriatum: a light and electron microscopic immunocytochemical study.

An antibody raised in rabbits against a GABA-BSA conjugate was used together with the PAP technique to label elements in the neostriatum of three Old World monkeys. Light microscopy revealed numerous immunoreactive medium-size neurons of various staining intensities, some of which had indented nuclei, as well as an occasional large cell. The neuropil showed a plexus of fine processes with frequent puncta. Ultrastructurally, the medium-size GABA-positive neurons were of two types: one with smooth nuclei and scanty cytoplasm, similar to spiny I cells, the other with invaginated nuclear envelopes and more abundant perikaryon, resembling the aspiny type. Correspondingly, labeled dendrites were either spiny or varicose. Some stained axons were myelinated, and the boutons had either large and ovoid, or small and pleomorphic vesicles. All of these boutons formed symmetric synapses, the former type with GABA-positive dendritic shafts but also with unlabeled dendrites; the latter type usually with GABA-negative elements, either dendrites, dendritic spines, or somata. Synapses were also observed between unreactive boutons and immunostained dendrites. Terminals with densely packed, small round vesicles that established asymmetric synapses with spines were always GABA-negative. Glial elements were consistently unlabeled, save for some astroglial endfeet. These findings provide positive evidence for the existence of two classes of GABAergic striatal neurons corresponding to a long-axoned spiny I type and an aspiny interneuron. Furthermore, the simultaneous labeling of GABA-immunoreactive presynaptic and postsynaptic profiles offers possible morphologic bases for the various kinds of intrastriatal inhibitory processes, including the feedforward, feedback, and "autaptic" types.

Monoclonal antibodies for ultrastructural visualization of L-baclofen-sensitive GABAB receptor sites.

Monoclonal antibodies were raised against the L-enantiomer of baclofen conjugated by glutaraldehyde to keyhole limpet hemocyanin. Hybridoma clones were selected for their stability and their production of high titers of antibodies directed against the p-chlorophenyl moiety of the L-baclofen molecule. The chosen antibody showed no cross-reactivity with conjugates of GABA and other neurotransmitters to human or bovine serum albumin. Specificity was further confirmed by the ability of L-baclofen-HCl to inhibit the binding of the antibody to L-baclofen-bovine serum albumin conjugate. Immunocytochemical studies were conducted on brain tissue from rats and monkeys injected with baclofen to localize baclofen-sensitive GABAB receptor sites. In these animals, the molecular layer of cerebellar cortex was clearly immunostained and the granular layer showed only some pale immunoreactivity. Ultrastructural observations were conducted in cerebellar cortex, as well as in the substantia nigra and the vestibular nuclei. Discrete labeling of neuronal profiles was observed in these structures, and both immunoperoxidase and colloidal gold methods were employed successfully. Material from saline-injected control animals showed no immunoreactivity at both light and electron microscopic levels. We conclude that the anti-L-baclofen antibody preferentially recognizes the p-chlorophenyl moiety of the baclofen molecule. Antibodies of such specificity are useful tools for the ultrastructural localization of baclofen-sensitive GABAB receptor sites. In general, antibodies directed against accessible moieties of specific neuroactive substances may serve as valuable markers for their sites of action.

A Golgi study of neuronal types in the neostriatum of monkeys.

Examination of the nestriatum of monkeys prepared by the Golgi-Kopsch perfusion method revealed the presence of at least 6 neuronal types. The spiny type I is medium size with a high density of dendritic spines. The axon extends well beyond the dendritic field and gives off many collaterals. The spiny type II is either medium or large size, has long thick dendrites with a relatively low density of spines, and an axon similar to that of the previous type but with fever collaterals. The aspiny type I is medium size with varicose dendrites and a thin axon arborizing in the immediate vicinity of the soma. The aspiny type II is large, with many thick and thin varicose dendrites. The aspiny type III is medium size with smooth dendrites and an axon ramifying profusely within the dendritic field. The neurogliform cell is small with many branching processes. Findings indicate that the neostriatum has 2 distinct types of spiny neurons with long axons (spiny I and II), some of which may contribute to the efferent system. There are also 2 (aspiny I and III) or perhaps as many as 4 categories (aspiny I, II, III and neurogliform) of typical Golgi type II cells. Large neurons belong to 2 separate populations, one with dendritic spines and a long axon (large version of spiny II), and one with varicosities and presumably a short axon (aspiny II). A realistic interpretation of neurophysiologic data on the neostriatum must take into account all cell types instead of the current view of considering it as a pool of interneurons with few output cells.

The magnocellular and parvocellular divisions of the monkey subthalamic nucleus as revealed by cluster analysis of neuronal sizes.

Cluster analysis of neuronal somal sizes in the subthalamic nucleus of rhesus monkeys from newborn to adult age allows the segregation of two territories with predominance of small and large cells, respectively. The topographic distribution of the 'parvocellular' and 'magnocellular' segments is similar when samples are obtained from coronal, horizontal and sagittal series of sections. The parvocellular component occupies the rostral pole, the entire rostrocaudal extent of the medial tip and dorsomedial border, and probably also the caudal cap. The magnocellular segment is in the central core extending to the ventrolateral border except for the medial tip. These findings and their correlation with the results of other morphologic and physiologic studies allow the following conclusions. (1) The monkey subthalamic nucleus contains at least two differentially distributed cell subpopulations. (2) The magnocellular division is more related to the pallido-subthalamic-pallidal loop involving the lateral pallidal segment. (3) The parvocellular division appears strategically located to control the pallidal output to diencephalic and mesencephalic targets. (4) Cluster analysis can reveal the existence of more than one neuronal population in a particular brain structure where an overall unimodal distribution of cell sizes may suggest the presence of a single type.

Early postnatal development of the monkey visual system. I. Growth of the lateral geniculate nucleus and striate cortex.

The postnatal growth of the dorsal lateral geniculate nucleus (LGNd) and the striate cortex (SCx) was compared in the same monkeys, by estimating LGNd volume, and the volume, surface area, and thickness of the SCx at birth, 1, 2, 4, 8 and 17 weeks. Shrinkage during histologic manipulations was determined in individual animals, and the above measurements were adjusted accordingly so that final volumes reflected a common, and, therefore, comparable state before processing. The volume of the LGNd increases approximately 17% between 2 and 4 weeks, and this growth primarily reflects that of the parvocellular laminae, the magnocellular components contributing a stable amount in absolute terms. Lamina 1 is larger than lamina 2 at all ages sampled. In contrast, the SCx expands about 75% in volume from birth to the oldest age examined without reaching an asymptote during the period of study. During the first 2 postnatal months, the growth results from increases in the thickness of the SCx whereas in the second 2 months it is caused by expansion in surface area. A comparison of exposed vs buried SCx does not reveal differences in the developmental patterns of regions subserving central vs peripheral visual fields, respectively, the exposed cortex being consistently greater in volume and area but thinner than the buried segment. No significant right/left asymmetries are found across the subjects in either of the structures studied. The findings indicate that the early postnatal development of the monkey visual system proceeds in a sequential fashion with the LGNd preceding that of the SCx.

Early postnatal development of monkey subthalamic nucleus: a light and electron microscopic study.

The subthalamic nuclei of 9 rhesus monkeys, ranging in age from newborn to 17 weeks, were examined at the light and/or electron microscopic levels, using computer assisted quantitative methods. The volume of the structure does not change significantly over the period of study. The mean cross-sectional area of neuronal somata, however, decreases by 33%, and most markedly during the first month. This is paralleled by a similar change in the mean area of cell nuclei but the perikaryon/nucleus ratio increases steadily after the first week. There is an overall decline in total cell numbers from newborn to 17 weeks. Ultrastructural features include dendritic growth cones in the neonatal monkey, and signs of axonal degeneration during the entire period. In addition to conventional axosomatic and axodendritic synapses, there are also synaptic junctions between vesicle-containing profiles which are seen only after the first month. The distributions of plaque diameters were reconstructed by the Coupland stereologic method from linear measurements of synaptic profiles and used to calculate synaptic densities and estimate the total number of synapses. This number is stable during the first postnatal month, declines markedly in the second month, and to a lesser degree thereafter, reaching a value of 55% of that at birth by 16 weeks. Findings indicate the occurrence of substantial changes within the subthalamic nucleus during the first 4 postnatal months, the most prominent of which is a marked synapse elimination.

Early postnatal development of the monkey visual system. II. Elimination of retinogeniculate synapses.

Profiles of retinal terminals, and of their synaptic and non-synaptic contacts, were measured in electron micrographs from magnocellular and parvocellular laminae of the dorsal lateral geniculate nucleus (LGNd) in newborn, 1-,4-,8- and 17-week-old rhesus monkeys. Morphologic criteria, i.e., the presence of pale mitochondria and large round vesicles, were used to identify the profile of retinal origin. Size-frequency histograms were stereologically reconstructed and used to calculate the density of retinal boutons and synaptic and non-synaptic plaques. The density values were adjusted for laminar growth to yield estimates of total numbers of these elements. Numerical estimates indicate bouton proliferation during the first week, followed by substantial reductions in bouton number accompanied by profound decreases in synapse number and cumulative synaptic area. In magnocellular layers, the reduction in synapse number is more pronounced after the eighth week, whereas the decrements in both features in the parvocellular laminae occur before this time. This synapse elimination process may be due entirely to retinal bouton retraction in parvocellular layers, but involves additional retinal synapse loss in the magnocellular segment. The parvocellular division shows a further size contraction of the remaining synapses. Immature synapses predominate in the LGNd throughout the 4-month period, and no quantitative evidence for direct transformation of immature to mature contacts is obtained. Non-synaptic junctions are stable in number but of increasing size in magnocellular layers, whereas substantial increases in number and area are found in parvocellular laminae. The preceding modifications in synaptic organization of the monkey LGNd occurring during the initial postnatal period may provide morphologic bases for the physiological and behavioral changes observed in this species during the same interval. Our data underscore the conclusion that synaptic reorganization occurs over a prolonged period, probably extending beyond 4 months, and involving the process of synapse elimination.

Synapses between GABA-immunoreactive axonal and dendritic elements in monkey substantia nigra.

gamma-Aminobutyric acid (GABA)-positive elements in the monkey substantia nigra pars reticulata (SNr) were identified using the peroxidase-antiperoxidase immunocytochemical technique with a direct GABA antibody. Light-microscopic examination revealed fine-caliber immunostained fibers piercing the cerebral peduncle and entering the SNr, where they formed a dense plexus. Ultrastructural observations included boutons with densely packed ovoid synaptic vesicles forming synapses with labeled and unlabeled dendritic elements. Most terminals contained small dark mitochondria, although some endings held larger, paler and fewer of these organelles. The synapses observed between GABA-immunoreactive profiles may represent the morphologic basis for disinhibition of pars compacta neurons reported to follow neostriatal or pallidal stimulation.

The internal organization of the pallidum in mammals.

A survey of morphologic data on the pallidum indicate that this structure contains at least two types of efferent neurons probably related to its GABA mediated inhibitory and Substance P mediated excitatory action on target cells. In addition, there are short-axoned neurons with presynaptic dendrites forming triadic synapses of unknown significance. Characterization of afferent axon terminals suggest that the majority originate in the striatum, are gabaergic and inhibitory. Enkephalins may coexist in these afferents. Other striatal fibers are excitatory and may contain Substance P. Possible afferents from the subthalamic nucleus form inhibitory synapses and their action may be mediated by glycine. Brain stem afferents from the substantia nigra, pars compacta (dopaminergic), raphe nuclei (serotoninergic) and n. tegmentalis pedunculopontinus (transmitter unknown) are excitatory on pallidal cells on morphologic and/or physiologic grounds.

Postnatal differentiation of "presynaptic dendrites" in the lateral geniculate nucleus of the rhesus monkey.

The most characteristic synaptic arrangement in the LGN is the triadic unit, in which a retinal terminal is presynaptic to a principal cell (P cell) and to a Golgi interneuron (I cell) dendrite, which contains synaptic vesicles and is in turn presynaptic to the same P-cell element. The ontogenetic differentiation of these "presynaptic dendrites" was studied in monkey LGN by standard and quantitative electron microscopy. The dendrites and axonal arborization of I cells are well developed in the newborn monkey. Scattered synaptic vesicles are present in the dendrites, but these profiles exhibit only postsynaptic sites. The dendrodendritic synapse of the triadic arrangement is missing, although contacts between P-cell and I-cell dendrites can be observed. Conversely, the I-cell axons in the newborn establish numerous synapses with dendrites and perikarya of P cells. At about 2 weeks of age, presynaptic sites appear in the I-cell dendrites, resulting in the formation of synaptic triads. Parallel to the development of "axonal" properties in the dendrites, the number of true I-cell axonal profiles decreases sharply. These transformations become progressively more frequent with age, and beyond 8 weeks the LGN ultraarchitectonics approaches that of the mature animal. The percentage of the surface of interneuron processes occupied by presynaptic sites is similar at all developmental stages at about the 2.6% level. The relative contribution of presynaptic dendrites and of axons changes, however, so that the actual length of contacts in the mature monkey is only one-half that of the newborn for the axonal sites, and over 50 times longer for the dendritic sites. The correlation of these findings with electrophysiologic and behavioral references suggests that some but not all axonal functions may be taken over by the dendrites, that the possible inhibitory phasing of P-cell discharge would appear only after the second postnatal week, and that the triadic arrangement may not be indispensable for brightness or total luminous flux discrimination but could be required for more complex forms of visually guided behavior.