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Crustaceans and insects share many similarities of brain organization suggesting that their common ancestor possessed some components of those shared features. Stomatopods (mantis shrimps) are basal eumalacostracan crustaceans famous for their elaborate visual system, the most complex of which possesses 12 types of color photoreceptors and the ability to detect both linearly and circularly polarized light. Here, using a palette of histological methods we describe neurons and their neuropils most immediately associated with the stomatopod retina. We first provide a general overview of the major neuropil structures in the eyestalks lateral protocerebrum, with respect to the optical pathways originating from the six rows of specialized ommatidia in the stomatopod's eye, termed the midband. We then focus on the structure and neuronal types of the lamina, the first optic neuropil in the stomatopod visual system. Using Golgi impregnations to resolve single neurons we identify cells in different parts of the lamina corresponding to the three different regions of the stomatopod eye (midband and the upper and lower eye halves). While the optic cartridges relating to the spectral and polarization sensitive midband ommatidia show some specializations not found in the lamina serving the upper and lower eye halves, the general morphology of the midband lamina reflects cell types elsewhere in the lamina and cell types described for other species of Eumalacostraca.

To further our understanding of the corticospinal projection (CSP) from the hand/arm representation of the primary motor cortex (M1), high-resolution anterograde tracing methodology and stereology were used to investigate the terminal distribution of this connection at spinal levels C5 to T1. The highest number of labeled terminal boutons occurred contralaterally (98%) with few ipsilaterally (2%). Contralaterally, labeled boutons were located within laminae I–X, with the densest distribution found in lamina VII and, to a lesser extent, laminae IX and VI. Fewer terminals were found in other contralateral laminae. Within lamina VII, terminal boutons were most prominent in the dorsomedial, dorsolateral, and ventrolateral subsectors. Within lamina IX, the heaviest terminal labeling was distributed dorsally. Ipsilaterally, boutons were found in laminae V–X. The most pronounced distribution occurred in the dorsomedial and ventromedial sectors of lamina VII and fewer labeled boutons were located in other ipsilateral laminae. Segmentally, contralateral lamina VII labeling was highest at levels C5–C7. In contrast, lamina IX labeling was highest at C7–T1 and more widely dispersed among the quadrants at C8–T1. Our findings suggest dominant contralateral influence of the M1 hand/arm CSP, a contralateral innervation pattern in lamina VII supporting Kuypers (1982) conceptual framework of a “lateral motor system,” and a projection to lamina IX indicating significant influence on motoneurons innervating flexors acting on the shoulder and elbow rostrally (C5–C7), along with flexors, extensors, abductors and adductors acting on the digits, hand and wrist caudally (C8–T1). J. Comp. Neurol. 521:4205–4235, 2013. © 2013 Wiley Periodicals, Inc.

The spinothalamic tract (STT), well known for its role in the relay of information about noxe, temperature, and crude touch, is usually associated with projections from lamina 1, but spinothalamic neurons in other laminae have also been reported. In cat, no complete overview exists of the precise location and number of spinal cells that project to the thalamus In the present study the laminar distribution of retrogradely labeled cells in all spinal segments (C1-Coc2) was investigated after large WGA-HRP injections in the thalamus. The results show that this distribution of STT cells differed greatly between the different spinal segments. Quantitative analysis showed that there exist at least five separate clusters of spinothalamic neurons. Lamina I neurons in cluster A and lamina V neurons in cluster B are mainly found contralaterally throughout the length of the spinal cord. Cluster C neurons are located bilaterally in the ventrolateral part of laminae VI-VII and lamina VIII of the C1-C3 spinal cord. Cluster D neurons were found contralaterally in lamina VI in the C1-C2 segments, and cluster E neurons were located mainly contralaterally in the medial part of laminae VI-VII and lamina VIII of the lumbosacral cord. Most spinothalamic neurons are not located in the enlargements and most spinothalamic neurons are not located in lamina 1, as suggested by several other authors. The location of the spinothalamic neurons shows remarkable similarities, but also differences, with the location of spino-periaqueductal gray neurons.

Quantitative analyses were performed on the dendrites and somata of 25 electrophysiologically identified preganglionic neurons (PGN) obtained from the sacral spinal cord of the cat by intracellular injection of Neurobiotin or horseradish peroxidase. Total dendritic length and surface area were measured for each dendrite. The sizes of the stem dendrites measured at their base were positively correlated with the sizes of the entire tree and numbers of end branches. Total surface area of somata and dendrites averaged 39,138 μm2; 90.7% of that was from the dendrites. To obtain measurements of the relative contributions of PGN dendrites to specific regions of the spinal cord, the percentage of each dendrite occupying eight spinal cord regions was recorded. Sixty-three percent of the dendrites projected dorsal to their somata, whereas an average of 33.3% of dendrites were located in the white matter, most of them in the lateral and dorsolateral funiculi. The neurons within this sample formed a continuum with some neurons having a large percentage of dendrites in lamina I but little in the white matter, whereas at the other end of the continuum were cells with the reverse configuration. The intermediate neurons had dendrites in both locations. Taken together, these data indicate a heterogeneous population of PGN in the lateral band of the sacral parasympathetic nucleus. J. Comp. Neurol. 437:56–69, 2001. © 2001 Wiley-Liss, Inc.

The first optic neuropil of the compound eye, the lamina, of the skipper butterfly Parnara guttata, was examined by light microscopy after Golgi-impregnation and by electron microscopy (EM) to clarify the cellular and synaptic organization. In the lamina, five different types of lamina neurons (L neurons) were characterized by using Golgi-impregnation. By EM, each cartridge was found to contain all nine receptor axons from an ommatidium, five L neurons, and a few putative centrifugal elements. Axons from photoreceptors (retinula cells) R2, R3, R4, R6, R7, and R8 terminate as short visual fibers (svfs) in the lamina cartridge. Those from R1, R5, and R9 penetrate the lamina and terminate in the medulla as long visual fibers (lvfs). In the cartridges, the synaptic contacts were formed from svfs onto L neurons, from the lvfs of R1 and/or R5 to the lvf of R9 and L neurons, and from the lvf of R9 to L neurons. The putative centrifugal fibers also make synapses to svfs and L neurons. At the most distal level of the cartridge, one of the centrifugal fibers containing dense-core vesicles makes presynaptic contacts to the putative long collaterals of the L neuron. A novel characteristic feature of this lamina is that svfs of R3 and R7 and the lvfs of R1 or R5 have long collaterals extending into neighboring cartridges. Presynaptic contacts were confirmed in such long collaterals from the svf. These results imply that receptor axons provide direct intercartridge connections as well as providing indirect connections to neighboring cartridges by way of their input upon L neurons. J. Comp. Neurol. 408:107–124, 1999. © 1999 Wiley-Liss, Inc.

The organization of multiple motor areas in the cerebral cortex has been investigated frequently in primates but rarely in nonprimate species. To compare sensorimotor areas in cats and primates, the cytoarchitecture of frontal and parietal areas of the cat cerebral cortex was described and mapped from coronal sections stained with cresyl violet or thionine. Multiple subdivisions of areas 4 and 6 were recognized; of these, the cytoarchitecture of area 4γ is similar to that of area 4 described in other carnivores and in primates and is characterized by giant pyramidal cells in multiple rows or clusters in lamina V. In other subdivisions of area 4 (4δ, 4sfu, and 4fu), giant pyramidal cells are few or absent in lamina V, and these subdivisions resemble area 6 of primates. Area 6 of the cat cortex is heterogeneous, and differences in laminar appearance and size of pyramidal cells in lamina V distinguish its four subdivisions (6aα, 6aβ, 6aγ, and 6iffu). The adjoining prefrontal areas are distinguishable from area 6 by the presence of a thin internal granular lamina (lamina IV) and the reduced size of pyramidal cells in lamina V. Laminae are poorly differentiated in the cingulate areas, where a rostral and caudal subdivision can be distinguished on the basis of the absence or presence of lamina IV. Area 3a is characterized by a thin lamina IV and is located between frontal agranular and parietal granular (well-defined lamina IV) fields (3b, 1, 2, 2pri, 5, and 7). Insular cortex can be subdivided into granular and agranular fields.

We examined lamina I trigemino- and spinothalamic tract (TSTT) terminals labeled with Phaseolus vulgaris leucoagglutinin in the nucleus submedius (Sm), a nociceptive relay in the cat's thalamus. Volume-rendered (three-dimensional) reconstructions of ten lamina I TSTT terminals identified with light and electron microscopy were built from serial ultrathin sections by computer, which enabled the overall structures of the terminal complexes to be characterized in detail. Two fundamentally different terminations were observed: compact clusters of numerous boutons, which predominate in the dense focus of a lamina I terminal field in the Sm, and boutons-of-passage, which are present throughout the terminal field and predominate in its periphery. Reconstructions of cluster terminations reveal that all boutons of each cluster make synaptic contact with protrusions and branch points on a single dendrite and involve presynaptic dendrites (PSDs) in triadic arrangements, providing a basis for the secure relay of sensory information. In contrast, reconstructions show that boutons-of-passage are generally characterized by simple contacts with PSDs, indicating an ascending inhibitory lamina I influence. These different synaptic arrangements are consistent with physiological evidence indicating that the morphologically distinct nociceptive-specific and thermoreceptive-(cold)-specific lamina I TSTT neurons terminate differently within the Sm. Thus, a suitable structural substrate exists in the cat's Sm for the inhibitory effect of cold on nociception, a behavioral and physiological phenomenon of fundamental significance. We conclude that the Sm is more than a simple relay for nociception, and that it may be an integrative comparator of ascending modality-selective information that arrives from neurons in lamina I.

The present study is the first comprehensive mapping of glial fibrillary acidic protein (GFAP)-immunopositive structures in the avian brain. Two main types of GFAP-immunopositive elements were observed: (1) nonbranching fibers, occasionally twisted or varicose, and (2) star-shaped cells. Long immunostained fibers originate from the lateral ventricle to form three bundles. Fibers of the dorsal group, emanating from the dorsal/lateral corner of the ventricle, course in lateral, anterior, and ventral directions forming a semidome, which separates the outer pallial (lateral cortical) regions from the underlying striatal mass. The middle group of fibers is directed anteriorly and laterally corresponding to the laminae frontales superior and suprema. The ventral fiber bundle is conical and traverses the lobus parolfactorius, crossing also the lamina medullaris dorsalis (the latter consisting mainly of star-shaped cells). The hippocampus, septum, and hypothalamus also contain straight radial fibers. In some areas, given their variable orientation, the fibers cannot be regarded as merely persisting radial glia. In the telencephalon, the nuclei basalis, accumbens, ectostriatum, paleostriatum primitivum, and the ventral paleostriatum are particularly rich in GFAP-positive cells, whereas the neostriatum, hyperstriatum, and paleostriatum augmentatum are almost devoid of GFAP labelling. Certain nuclei of the thalamus and the lower brainstem are conspicuous by their low levels of GFAP immunoreactivity. The Bergmann glia were GFAP-immunonegative.

The ventral lateral geniculate nucleus (vLGN) of the tree shrew (Tupaia belangeri) was differentiated into multiple subdivisions (dorsal cap, intergeniculate leaflet, parvicellular segment, and internal and external magnocellular laminae, the latter being further divisible into a lateral and medial division) on the basis of retinal projections, immunochemistry, and histochemistry. Retinal projections traced with intravitreal injections of wheat germ agglutinin conjugated horseradish peroxidase revealed direct bilateral input to all subregions of the vLGN, except for the internal magnocellular lamina (which received only contralateral input) and the parvicellular segment (which was not retinorecipient). Furthermore, retinal inputs clearly distinguished the relatively heavily retinorecipient intergeniculate leaflet from the less prominently labeled dorsal cap. Immunohistochemical localization of Neuropeptide Y (NPY) perikarya revealed their prominence in the intergeniculate leaflet and the external magnocellular laminae with a concentration along the optic tract. NPY immunoreactive fibers were seen in all but the parvicellular subregion. Gamma amino butyric acid immunoreactivity was seen throughout the vLGN, but was most concentrated in the dorsal cap and the magnocellular laminae, followed by the intergeniculate leaflet. Histochemical studies of cytochrome oxidase and nicotinamide adenosine dinucleotide phosphate (NADPH)-diaphorase localization revealed similar patterns of dense reactivity within the external magnocellular lamina, intergeniculate leaflet and dorsal cap, and somewhat less dense, but substantial reactivity in the internal magnocellular lamina. Within the external magnocellular lamina, cells reactive for cytochrome oxidase were noted in the lateral portion bordering the optic tract, whereas those specific for NADPH-diaphorase were dispersed throughout the lamina. Poor reactivity for both histochemical markers was evident in the parvicellular segment. Overall, the markedly different patterns of retinal input and neurochemical organization between the subdivisions of the tree shrew vLGN suggest their involvement in diverse functions. Furthermore, the basic similarity of the organization of the tree shrew vLGN to that of the taxonomically unrelated ground squirrel may indicate a common mammalian scheme.

The fine structure and synaptic architecture of the afferent terminations in dorsal horn lamina II are studied using a combined light and electron microscopic procedure after anterograde labelling with horseradish peroxidase. Vibratome parasagittal sections, stained with heavy metal intensified diaminobenzidine after tracer application to the dorsal roots, were flat-embedded in Epon. The five types of labelled terminal arbors occurring in lamina IIi (Cruz et al., '87: J. Comp. Neurol. 261:221-236) were drawn and relocated in 5-microns sections cut serially from the thick sections. Ultrathin sections were then cut from the 5-microns sections so that the terminal fibers and swellings observed in the light microscope could be traced in the electron microscope. The flame-shaped arbors arose from fine myelinated stem fibers. Terminal strands generated large oval central terminals of type II synaptic glomeruli (CII), which established frequent axoaxonal contacts. Similar terminals have been labelled in the cat after tracer injections into hair-follicle fibers (Rethelyi et al., '82: J. Comp. Neurol. 207:381-393). The other four plexuses arose from unmyelinated stem fibers. The swarms of ultrafine boutons consisted of extremely thin terminal fibers generating very small, round, or polygonal glomerular terminals containing tightly packed agranular synaptic vesicles of variable size and one mitochondrion at best. The terminal strands of the bouquet plexus bore long and scalloped central varicosities of type I synaptic glomeruli (CI) with pleomorphic agranular vesicles and a relative abundance of dendroaxonal contacts. These features, together with the location in dorsal lamina IIi, suggest their belonging to the fluoride resistant acid phosphatase (FRAP)-reactive population. The boutons of the undulating fibers and those of the lateral plexus were, like those of the bouquets, scalloped and elongated rostrocaudally (CI), but contained a few large granular vesicles. The occurrence of the swarm, undulating, and lateral plexuses in ventral lamina IIi, which seems to lack FRAP or peptidergic terminals, suggests an origin from other, still unidentified neurochemical populations of fine primary afferents.

Spinal cord projections from lumbar dorsal root ganglia (DRGs) were investigated in adult rats following injections of choleragenoid horseradish peroxidase (B-HRP) into each of the six lumbar DRGs. This method is known to label primarily thick fibers. Labeling was found in all laminae except in the outer part of lamina II. Labeled fibers and terminal-like structures were found 8–13 segments rostral and 1–5 segments caudal to the injected DRGs. A somatotopic organization was revealed in laminae III, where the labeling seemed to be organized in mediolateral zones. Some of these protruded as fingerlike processes through segments rostral and caudal to the root entry level. An interdigitating pattern for these fingerlike processes was seen between some DRGs, while an extensive overlap was found between other DRGs. Many zones were found to correspond to the known central projections of peripheral sensory nerves supplied by the injected ganglion. This suggests that the central projection of a DRG is highly related to the projections of the peripheral nerves included in the DRG. The projections to lamina IV were organized in a similar manner as in lamina III, even though the projections showed a higher degree of overlap than in lamina III. No clear somatotopic organization was found in laminae V-IX. Provided that the topographical relationship between central projections of single peripheral nerves and of DRGs correspond to their peripheral projections, the results of this study, together with results of earlier studies suggest that the outlines of dermatomes are highly related to the territories of peripheral nerves included in the dermatomes.

Drosophila mutants black and ebony show pigmentation defects in the adult cuticle, which disclose their cooperative activity in β-alanyl-dopamine formation. In visual signal transduction, Ebony conjugates β-alanine to histamine, forming β-alanyl-histamine or carcinine. Mutation of ebony disrupts signal transduction and reveals an electroretinogram (ERG) phenotype. In contrast to the corresponding cuticle phenotype of black and ebony, there is no ERG phenotype observed when black expression is disrupted. This discrepancy calls into question the longstanding assumption of Black and Ebony interaction. The purpose of this study was to investigate the role of Black and Ebony in fly optic lobes. We excluded a presynaptic histamine uptake pathway and confirmed histamine recycling via carcinine formation in glia. β-Alanine supply for this pathway is independent of enzymatic synthesis by Black and β-alanine synthase Pyd3. Two versions of Black are expressed in vivo. Black is a specific aspartate decarboxylase with no activity on glutamate. RNA in situ hybridization and anti-Black antisera localized Black expression in the head. Immunolabeling revealed expression in lamina glia, in large medulla glia, in glia of the ocellar ganglion, and in astrocyte-like glia below the ocellar ganglion. In these glia types, Black expression is strictly accompanied by Ebony expression. Activity, localization, and strict coexpression with Ebony strongly indicate a specific mode of functional interaction that, however, evades ERG detection. J. Comp. Neurol., 521:1207–1224, 2013. © 2012 Wiley Periodicals, Inc.

Five intracellularly HRP-stained Renshaw cells were subjected to light microscopic analysis of the trajectories, branching patterns, and projections of the axonal systems. The cell bodies were located ventrally in lamina VII. In three neurons the axon originated from the cell body and in the remaining two cells from a dendrite. After a 600-870-microns distance the axons entered the ventral funiculus, where all of them continued rostrally. Two axons also gave off a caudal branch in the funiculus. The diameters of the main axons varied between 2.1 and 10.0 microns. The main axons gave off one to four first-order collaterals before entering the ventral funiculus and up to three collaterals could be seen to originate from the same node of Ranvier. In the ventral funiculus up to five first-order collaterals could be traced from the same main axon. The axon collateral trees were often very extensive and daughter branches up to the 22nd order were observed. The distance between two successive branching points varied between 4 and 410 microns. A large number of boutonlike swellings were found along (59%) or at the ends of the collateral branches. At the most, 1,278 swellings originated from a single axon collateral tree. Most of the swellings were located in lamina IX, but they also appeared ventrally and dorsolaterally in lamina VII.

Tectal projection neurons were labeled by retrograde transport of horseradish peroxidase (HRP) or cobaltic-lysine. The tracer substances were delivered iontophoretically or by pressure injection or diffusion into various regions of the brain or spinal cord. Histochemical procedures allowed identification of labeled cells projecting to the injected regions. Many neurons were filled with cobaltic-lysine, resulting in a Golgi-like staining. After cobalt injections in the diencephalon most of the labeled cells, identified as small piriform neurons, were located in layer 8 of the tectum. Two types of small piriform neurons were distinguished. Type 1 neurons have flat dendritic arborizations confined to lamina D, while the dendrites of type 2 cells may span all of the superficial tectal strata. In smaller numbers large piriform, pyramidal, and ganglionic cells of the periventricular tectal layers were labeled after diencephalic injections. Rhombencephalic cobalt and HRP injections labeled cells whose axons form the tectobulbospinal tract. The neurons most frequently labeled were large ganglionic cells. Ipsilaterally, the majority of their somata were located in layer 7, and their dendrites arborized mainly in lamina F. Contralaterally, labeled ganglionic cell somata occupied the top of layer 6, and most of their dendritic end-branches entered lamina B. The possible functional significance of this anatomical arrangement is discussed. After tectal cobalt injections the topography of the tectoisthmic projection and the terminals of tectal efferent fibers in the diencephalon and brainstem were observed. It is concluded that the organization of frog tectofugal pathways is very similar to that of mammals.

To test the differentiation of visual interneurons that had their targets removed before axogenesis, we ablated neuronal precursors in brains of first instar fly larvae by using a laser microsurgery unit. We describe ablations that resulted in the elimination of the third neuropil region (the lobula complex) of the optic lobes. Neural differentiation in the more peripheral second and first neuropil regions (the medulla and the lamina) was thus studied in the absence of the lobula complex. It was found that the medulla neuropil differentiated with normal columnar and layered organization. The neuropil, however, folded along its central surface. The only connection between the medulla and more central neuropil (the midbrain) was via a bundle of axons (the Cuccati bundle) present also in the normal optic lobes. Some types of neurons that normally connect the medulla and the lobula complex could be identified. These appeared to end in a disorganized neuropil mass in the center of the folded medulla. The differentiation of the lamina neuropil also appeared normal in flies with the lobula complex eliminated and the medulla folded. Also, in optic lobes where the medulla was severely disorganized and/or reduced due to laser ablations, the lamina neuropil appeared more or less normal. The results suggest that lamina and medulla nerve cells can differentiate and develop normal neuropil patterns in absence of their appropriate targets.

A projection from the subiculum to the retrosplenial cortex (area 29a and b) was studied by autoradiographic tracing of anterogradely transported proteins following injections of radioactive amino acids into ventral and dorsal parts of the subiculum. Only the dorsal parts of the subiculum projected to the retrosplenial cortex. The projection was exclusively ipsilateral, and topographically arranged along both the longitudinal septotemporal axis and the transverse plane of the subiculum. The termination in the retrosplenial cortex was confined to the relatively cell-free superficial layer (lamina 1) and the underlying cell-rich layers (laminae 3-4). Between these two terminations a narrow zone (lamina 2) was only weakly labeled. The projection constitutes an output route for hippocampal activity in addition to the fimbria-fornix system and the subicular projections to the parahippocampal areas.

The cell bodies and dendritic trees of five lumbosacral Renshaw cells of adult cats were studied in the light microscope (LM) after intracellular injection with horseradish peroxidase (HRP). The cell bodies were all located in the ventral part of lamina VII. The dendrites extended up to 0.7 mm from the cell body into the neighbouring parts of laminae VIII and IX as well as into more dorsal parts of lamina VII. The dendritic branching was sparse and about half the dendrites were unbranched. The mean diameter of the cell body was positively correlated to both the combined and mean diameters of the first-order dendrites. Between four and eight dendrites originated from the cell bodies. The number of dendritic end-branches, the combined dendritic length, the mean dendritic length from the cell body to the termination of the end branches, the distance from the cell body to the termination of the most remote end-branch, the dendritic surface area, and the dendritic volume all correlated positively with the diameter of the parent first-order dendrite. The dendritic tapering was somewhat more pronounced in the Renshaw cells than previously observed in alpha- and gamma-motoneurons. The present data are discussed in relation to previous morphological observations on Renshaw cells and alpha- and gamma-motoneurons.

Extensive terminal branchings of fine fibers in the substantia gelatinosa of Golgi-Kopsch preparations of the adult cat spinal cord were subjected to a semi-quantitative analysis. transverse sections suggest that these fibers are probably unmyelinated primary afferent elements of dorsal root origin. In transverse sections these elements pass medially and ventrally and shortly disappear due to a change in orientation. Similar thin fibers in sagittal sections can be followed for several hundred microns as they give rise to side branches that also run mainly in a longitudinal direction. The side branches divide in turn to produce preterminal axon arborizations. The arborizations were distributed in 150 mum wide zones in the dorsal horn region corresponding to Rexed's lamina II. The end terminals are large bulbs, usually preceded by two to three equally large en passant enlargements. Seven to eight terminals stem from each side branch. The terminals and enlargements are arranged in narrow (16-26 mum thick) sagittal sheets. The terminals of several side branches often converge upon a common region so that clusters of terminals occur within the sagittal sheet. It is proposed that these observations are consistent with the substantia gelatinosa (lamina II) as the termination of unmyelinated (C) primary afferent fibers and that the latter are the only type of primary fibers ending in this portion of the spinal cord.