Complex Neuropil Research Articles

Imp is expressed in INPs and newborn neurons where it regulates neuropil targeting in the central complex

The generation of neuronal diversity remains incompletely understood. In Drosophila, the central brain is populated by neural stem cells derived from progenitors called neuroblasts (NBs). There are two types of NBs, type 1 and 2. T1NBs have a relatively simple lineage, whereas T2NBs expand and diversify the neural population with the generation of intermediate neural progenitors (INPs), contributing many neurons to the adult central complex, a brain region essential for navigation. However, it is not fully understood how neural diversity is created in T2NB and INP lineages. Imp, an RNA-binding protein, is expressed in T2NBs in a high-to-low temporal gradient, while the RNA-binding protein Syncrip forms an opposing gradient. It remains unknown if Imp expression is carried into INPs; whether it forms a gradient similar to NBs; and whether INP expression of Imp is required for generating neuronal identity or morphology. Here, we show that Imp/Syp are both present in INPs, but not always in opposing gradients. We find that newborn INPs adopt their Imp/Syp levels from their parental T2NBs; that Imp and Syp are expressed in stage-specific high-to-low gradients in INPs. In addition, there is a late INP pulse of Imp. We find that neurons born from old INPs (E-PG and PF-R neurons) have altered morphology following both Imp knock-down and Imp overexpression. We conclude that Imp functions in INPs and newborn neurons to determine proper neuronal morphology and central complex neuropil organization.

Neural organization of the third optic neuropil, the lobula, in the highly visual semiterrestrial crab Neohelice granulata.

The visual neuropils (lamina, medulla, and lobula complex) of malacostracan crustaceans and hexapods have many organizational principles, cell types, and functional properties in common. Information about the cellular elements that compose the crustacean lobula is scarce especially when focusing on small columnar cells. Semiterrestrial crabs possess a highly developed visual system and display conspicuous visually guided behaviors. In particular, Neohelice granulata has been previously used to describe the cellular components of the first two optic neuropils using Golgi impregnation technique. Here, we present a comprehensive description of individual elements composing the third optic neuropil, the lobula, of that same species. We characterized a wide variety of elements (140 types) including input terminals and lobula columnar, centrifugal, and input columnar elements. Results reveal a very dense and complex neuropil. We found a frequently impregnated input element (suggesting a supernumerary cartridge representation) that arborizes in the third layer of the lobula and that presents four variants each with ramifications organized following one of the four cardinal axes suggesting a role in directional processing. We also describe input elements with two neurites branching in the third layer, probably connecting with the medulla and lobula plate. These facts suggest that this layer is involved in the directional motion detection pathway in crabs. We analyze and discuss our findings considering the similarities and differences found between the layered organization and components of this crustacean lobula and the lobula of insects.

Neuroarchitecture of the Drosophila central complex: A catalog of nodulus and asymmetrical body neurons and a revision of the protocerebral bridge catalog.

The central complex, a set of neuropils in the center of the insect brain, plays a crucial role in spatial aspects of sensory integration and motor control. Stereotyped neurons interconnect these neuropils with one another and with accessory structures. We screened over 5,000 Drosophila melanogaster GAL4 lines for expression in two neuropils, the noduli (NO) of the central complex and the asymmetrical body (AB), and used multicolor stochastic labeling to analyze the morphology, polarity, and organization of individual cells in a subset of the GAL4 lines that showed expression in these neuropils. We identified nine NO and three AB cell types and describe them here. The morphology of the NO neurons suggests that they receive input primarily in the lateral accessory lobe and send output to each of the six paired noduli. We demonstrate that the AB is a bilateral structure which exhibits asymmetry in size between the left and right bodies. We show that the AB neurons directly connect the AB to the central complex and accessory neuropils, that they target both the left and right ABs, and that one cell type preferentially innervates the right AB. We propose that the AB be considered a central complex neuropil in Drosophila. Finally, we present highly restricted GAL4 lines for most identified protocerebral bridge, NO, and AB cell types. These lines, generated using the split‐GAL4 method, will facilitate anatomical studies, behavioral assays, and physiological experiments.

Neuroarchitecture and neuroanatomy of the Drosophila central complex: A GAL4‐based dissection of protocerebral bridge neurons and circuits

ABSTRACTInsects exhibit an elaborate repertoire of behaviors in response to environmental stimuli. The central complex plays a key role in combining various modalities of sensory information with an insect's internal state and past experience to select appropriate responses. Progress has been made in understanding the broad spectrum of outputs from the central complex neuropils and circuits involved in numerous behaviors. Many resident neurons have also been identified. However, the specific roles of these intricate structures and the functional connections between them remain largely obscure. Significant gains rely on obtaining a comprehensive catalog of the neurons and associated GAL4 lines that arborize within these brain regions, and on mapping neuronal pathways connecting these structures. To this end, small populations of neurons in the Drosophila melanogaster central complex were stochastically labeled using the multicolor flip‐out technique and a catalog was created of the neurons, their morphologies, trajectories, relative arrangements, and corresponding GAL4 lines. This report focuses on one structure of the central complex, the protocerebral bridge, and identifies just 17 morphologically distinct cell types that arborize in this structure. This work also provides new insights into the anatomical structure of the four components of the central complex and its accessory neuropils. Most strikingly, we found that the protocerebral bridge contains 18 glomeruli, not 16, as previously believed. Revised wiring diagrams that take into account this updated architectural design are presented. This updated map of the Drosophila central complex will facilitate a deeper behavioral and physiological dissection of this sophisticated set of structures. J. Comp. Neurol. 523:997–1037, 2015. © 2014 Wiley Periodicals, Inc.

Neuroarchitecture and neuroanatomy of the Drosophila central complex: A GAL4-based dissection of protocerebral bridge neurons and circuits.

Insects exhibit an elaborate repertoire of behaviors in response to environmental stimuli. The central complex plays a key role in combining various modalities of sensory information with an insect's internal state and past experience to select appropriate responses. Progress has been made in understanding the broad spectrum of outputs from the central complex neuropils and circuits involved in numerous behaviors. Many resident neurons have also been identified. However, the specific roles of these intricate structures and the functional connections between them remain largely obscure. Significant gains rely on obtaining a comprehensive catalog of the neurons and associated GAL4 lines that arborize within these brain regions, and on mapping neuronal pathways connecting these structures. To this end, small populations of neurons in the Drosophila melanogaster central complex were stochastically labeled using the multicolor flip-out technique and a catalog was created of the neurons, their morphologies, trajectories, relative arrangements, and corresponding GAL4 lines. This report focuses on one structure of the central complex, the protocerebral bridge, and identifies just 17 morphologically distinct cell types that arborize in this structure. This work also provides new insights into the anatomical structure of the four components of the central complex and its accessory neuropils. Most strikingly, we found that the protocerebral bridge contains 18 glomeruli, not 16, as previously believed. Revised wiring diagrams that take into account this updated architectural design are presented. This updated map of the Drosophila central complex will facilitate a deeper behavioral and physiological dissection of this sophisticated set of structures. J. Comp. Neurol. 523:997–1037, 2015. © 2014 Wiley Periodicals, Inc.

Neural organization of the second optic neuropil, the medulla, in the highly visual semiterrestrial crab Neohelice granulata

Crustaceans are widely distributed and inhabit very different niches. Many of them are highly visual animals. Nevertheless, the neural composition of crustacean optic neuropils deeper than the lamina is mostly unknown. In particular, semiterrestrial crabs possess a highly developed visual system and display conspicuous visually guided behaviors. A previous study shows that the first optic neuropil, the lamina of the crab Neohelice granulata, possesses a surprisingly high number of elements in each cartridge. Here, we present a comprehensive description of individual elements composing the medulla of that same species. Using Golgi impregnation, we characterized a wide variety of cells. Only considering the class of transmedullary neurons, we describe over 50 different morphologies including small- and large-field units. Among others, we describe a type of centrifugal neuron hitherto not identified in other crustaceans or insects that probably feeds back information to every cartridge in the medulla. The possible functional role of such centrifugal elements is discussed in connection with the physiological and behavioral information on visual processing available for this crab. Taken together, the results reveal a very dense and complex neuropil in which several channels of information processing would be acting in parallel. We further examine our results considering the similarities and differences found between the layered organization and components of this crustacean medulla and the medullae of insects.

A peek into a crab´s optic lobes that support motion detection, visually-guided behaviors and memory

Event Abstract Back to Event A peek into a crab´s optic lobes that support motion detection, visually-guided behaviors and memory Julieta Sztarker1* 1 Universidad de Buenos Aires-CONICET, FBMyC, Argentina Motion detection is crucial to many adaptive behaviors such as prey-capture and predator avoidance. In arthropods, these circuits involve the presence of few, large field lobula giant neurons that are fed by many columnar neurons and process different aspects related to motion detection (e.g. looming detection, detection of small targets, etc). In the highly visual crab, Neohelice granulata, the lobula giant neurons (LGs) have a high capacity for information processing. They are involved in visually-guided escape responses but also support cognitive functions such as learning and memory. By presenting the results of in vivo intracellular recordings I will show that these neurons act individually like a processing unit, collecting a large amount of information. The LGs receive retinotopic information through columnar elements projecting from the medulla. The identity of such elements is largely unknown since most of them are too small for stable intracellular recording. Based on the information available from other arthropods and from our own results describing the neuronal composition of the lamina, the existence of several parallel pathways between the medulla and the lobula is expected (each processing different features of the scene). I analyzed the neuronal composition of the medulla in this species using classical staining techniques. Results reveal a very dense and complex neuropil that shows both differences and commonalities with the insect medulla. Keywords: medulla neurons, Arthropods, Lobula tangential cells, Visual System, learning and memory Conference: International Conference on Invertebrate Vision, Fjälkinge, Sweden, 1 Aug - 8 Aug, 2013. Presentation Type: Oral presentation preferred Topic: Motion vision Citation: Sztarker J (2019). A peek into a crab´s optic lobes that support motion detection, visually-guided behaviors and memory. Front. Physiol. Conference Abstract: International Conference on Invertebrate Vision. doi: 10.3389/conf.fphys.2013.25.00114 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 29 May 2013; Published Online: 09 Dec 2019. * Correspondence: Dr. Julieta Sztarker, Universidad de Buenos Aires-CONICET, FBMyC, Buenos Aires, 1428, Argentina, julieta.sztarker@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Julieta Sztarker Google Julieta Sztarker Google Scholar Julieta Sztarker PubMed Julieta Sztarker Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Astrocyte-like glia associated with the embryonic development of the central complex in the grasshopper Schistocerca gregaria

In this study we employed the expression of the astrocyte-specific enzyme glutamine synthetase, in addition to the glia-specific marker Repo, to characterize glia cell types associated with the embryonic development of the central complex in the grasshopper Schistocerca gregaria. Double labeling experiments reveal that all glutamine synthetase-positive cells associated with the central complex are also Repo-positive and horseradish peroxidase-negative, confirming they are glia. Early in embryogenesis, prior to development of the central complex, glia form a continuous population extending from the pars intercerebralis into the region of the commissural fascicles. Subsequently, these glia redisperse to envelop each of the modules of the central complex. No glial somata are found within the central complex neuropils themselves. Since glutamine synthetase is expressed cortically in glia, it allows their processes as well as their soma locations to be visualized. Single cell reconstructions reveal one population of glia as directing extensive ensheathing processes around central complex neuropils such as the central body, while another population projects columnar-like arborizations within the central body. Such arborizations are only seen in central complex modules after their neuroarchitecture has been established suggesting that the glial arborizations project onto a prior scaffold of neurons or tracheae.

Patterns of dye coupling involving serotonergic neurons provide insights into the cellular organization of a central complex lineage of the embryonic grasshopper Schistocerca gregaria

All eight neuroblasts from the pars intercerebralis of one protocerebral hemisphere whose progeny contribute fibers to the central complex in the embryonic brain of the grasshopper Schistocerca gregaria generate serotonergic cells at stereotypic locations in their lineages. The pattern of dye coupling involving these neuroblasts and their progeny was investigated during embryogenesis by injecting fluorescent dye intracellularly into the neuroblast and/or its progeny in brain slices. The tissue was then processed for anti-serotonin immunohistochemistry. A representative lineage, that of neuroblast 1-3, was selected for detailed study. Stereotypic patterns of dye coupling were observed between progeny of the lineage throughout embryogenesis. Dye injected into the soma of a serotonergic cell consistently spread to a cluster of between five and eight neighboring non-serotonergic cells, but never to other serotonergic cells. Dye injected into a non-serotonergic cell from such a cluster spread to other non-serotonergic cells of the cluster, and to the immediate serotonergic cell, but never to further serotonergic cells. Serotonergic cells tested from different locations within the lineage repeat this pattern of dye coupling. All dye coupling was blocked on addition of an established gap junctional blocker (n-heptanol) to the bathing medium. The lack of coupling among serotonergic cells in the lineage suggests that each, along with its associated cluster of dye-coupled non-serotonergic cells, represents an independent communicating pathway (labeled line) to the developing central complex neuropil. The serotonergic cell may function as the coordinating element in such a projection system.

Electrolytic lesions within central complex neuropils of the cockroach brain affect negotiation of barriers

Animals must negotiate obstacles in their path in order to successfully function within natural environments. These actions require transitions from walking to other behaviors, many of which are more involved than simple reflexes. For these behaviors to be successful, insects must evaluate objects in their path and then use that information to change posture or re-direct leg movements. Some of this control may occur within a region of the brain known as the central complex (CC). We used discrete electrolytic lesions to examine the role of certain sub-regions of the CC in various obstacle negotiation behaviors. We found that cockroaches with lesions to the protocerebral bridge (PB) and ellipsoid body (EB) exhibit abnormalities in turning and dealing with shelf-like objects; whereas, individuals with lesions to the fan-shaped body (FB) and lateral accessory lobe (LAL), exhibit abnormalities of those behaviors as well as climbing over blocks and up walls to a horizontal plane. Abnormalities in block climbing include decreased success rate, changes in climbing strategy, and delayed response to the block. Increases in these abnormal behaviors were significant in individuals with lesions to the FB and LAL. Although turning abnormalities are present in individuals with lesions to the LAL, EB and the lateral region of the FB, there are some differences in how these deficits present. For instance, the turning deficits seen in individuals with lateral FB lesions only occurred when turning in the direction opposite to the side of the brain on which the lesion occurred. By contrast, individuals with lesions to the EB and LAL exhibited turning abnormalities in both directions. Lesions in the medial region of the FB did not result in directional turning deficits, but in abnormalities in block climbing.

Visual Targeting of Motor Actions in Climbing Drosophila

Drosophila melanogaster flies cross surmountable gaps in their walkway of widths exceeding their body length with an astounding maneuver but avoid attempts at insurmountable gaps by visual width estimation. Different mutant lines affect specific aspects of this maneuver, indicating a high complexity and modularity of the underlying motor control. Here we report on two mutants, ocelliless(1) and tay bridge(1), that, although making a correct decision to climb, fail dramatically in aiming at the right direction. Both mutants show structural defects in the protocerebral bridge, a central complex neuropil formed like a handlebar spanning the brain hemispheres. The bridge has been implicated in step-length control in walking flies and celestial E-vector orientation in locusts. In rescue experiments using tay bridge(1) flies, the integrity of the bridge was reestablished, concomitantly leading to a significant improvement of their orientation at the gap. Although producing directional scatter, their attempts were clearly aimed at the landing site. However, this partial rescue was lost in these flies at a reduced-visibility landing site. We therefore conclude that the protocerebral bridge is an essential part of a visual targeting network that transmits directional clues to the motor output via a known projection system.

Optomotor-blind expression in glial cells is required for correct axonal projection across the Drosophila inner optic chiasm

In the Drosophila adult visual system, photoreceptor axons and their connecting interneurons are tied into a retinotopic pattern throughout the consecutive neuropil regions: lamina, medulla and lobula complex. Lamina and medulla are joined by the first or outer optic chiasm (OOC). Medulla, lobula and lobula plate are connected by the second or inner optic chiasm (IOC). In the regulatory mutant In(1)omb H31 of the T-box gene optomotor-blind ( omb), fibers were found to cross aberrantly through the IOC into the neuropil of the lobula complex. Here, we show that In(1)omb H31 causes selective loss of OMB expression from glial cells within the IOC previously identified as IOC giant glia (ICg-glia). In the absence of OMB, ICg-glia retain their glial cell identity and survive until the adult stage but tend to be displaced into the lobula complex neuropil leading to a misprojection of axons through the IOC. In addition, adult mutant glia show an aberrant increase in length and frequency of glial cell processes. We narrowed down the onset of the IOC defect to the interval between 48 h and 72 h of pupal development. Within the 40 kb of regulatory DNA lacking in In(1)omb H31 , we identified an enhancer element (ombC) with activity in the ICg-glia. ombC-driven expression of omb in ICg-glia restored proper axonal projection through the IOC in In(1)omb H31 mutant flies, as well as proper glial cell positioning and morphology. These results indicate that expression of the transcription factor OMB in ICg-glial cells is autonomously required for glial cell migration and morphology and non-autonomously influences axonal pathfinding.

Afferent and efferent connections of the rat retrotrapezoid nucleus

The rat retrotrapezoid nucleus (RTN) contains candidate central chemoreceptors that have extensive dendrites within the marginal layer (ML). This study describes the axonal projections of RTN neurons and their probable synaptic inputs. The ML showed a dense plexus of nerve terminals immunoreactive (ir) for markers of glutamatergic (vesicular glutamate transporters VGLUT1-3), gamma-aminobutyric acid (GABA)-ergic, adrenergic, serotonergic, cholinergic, and peptidergic transmission. The density of VGLUT3-ir terminals tracked the location of RTN chemoreceptors. The efferent and afferent projections of RTN were studied by placing small iontophoretic injections of anterograde (biotinylated dextran amine; BDA) and retrograde (cholera toxin B) tracers where RTN chemoreceptors have been previously recorded. BDA did not label the nearby C1 cells. BDA-ir varicosities were found in the solitary tract nucleus (NTS), all ventral respiratory column (VRC) subdivisions, A5 noradrenergic area, parabrachial complex, and spinal cord. In each target region, a large percentage of the BDA-ir varicosities was VGLUT2-ir (41-83%). Putative afferent input to RTN originated from spinal cord, caudal NTS, area postrema, VRC, dorsolateral pons, raphe nuclei, lateral hypothalamus, central amygdala, and insular cortex. The results suggest that 1) whether or not the ML is specialized for CO(2) sensing, its complex neuropil likely regulates the activity of RTN chemosensitive neurons; 2) the catecholaminergic, cholinergic, and serotonergic innervation of RTN represents a possible substrate for the known state-dependent control of RTN chemoreceptors; 3) VGLUT3-ir terminals are a probable marker of RTN; and 4) the chemosensitive neurons of RTN may provide a chemical drive to multiple respiratory outflows, insofar as RTN innervates the entire VRC.

Class-specific features of neuronal wiring.

Brain function relies on specificity of synaptic connectivity patterns among different classes of neurons. Yet, the substrates of specificity in complex neuropil remain largely unknown. We search for imprints of specificity in the layout of axonal and dendritic arbors from the rat neocortex. An analysis of 3D reconstructions of pairs consisting of pyramidal cells (PCs) and GABAergic interneurons (GIs) revealed that the layout of GI axons is specific. This specificity is manifested in a relatively high tortuosity, small branch length of these axons, and correlations of their trajectories with the positions of postsynaptic neuron dendrites. Axons of PCs show no such specificity, usually taking a relatively straight course through neuropil. However, wiring patterns among PCs hold a large potential for circuit remodeling and specificity through growth and retraction of dendritic spines. Our results define distinct class-specific rules in establishing synaptic connectivity, which could be crucial in formulating a canonical cortical circuit.

Restriction of peroxidase-mediated antibody reactivity to single neurons by local hydrogen peroxide production

Current electrophysiological and imaging methods have begun to target the subcellular structure and function of single CNS neurons. Although physiological imaging methods now permit the resolution of activity of single presynaptic and postsynaptic elements, it has not been possible to unequivocally examine the array of proteins expressed at these same structures. This problem arises from the inability of current immunocytochemical techniques to differentiate between a process of the neuron of interest and the surrounding neuropil belonging to other cells. Thus, we have sought to develop an antibody staining method which would restrict reactivity to only a single neuron. Our assay involves preloading a single neuron with the coupling reagent biocytin. Following fixation, the injected biocytin is then complexed with avidin-linked glucose oxidase, providing a means of locally generating hydrogen peroxide within a cell of interest which catalyses the peroxidase-mediated (coupled to primary antibody) staining reaction. We have used this method successfully with antibodies to a number of neuronal markers (neuron-specific enolase, neurofilament, microtubule-associated protein and the glutamate receptor GluR2/3). Our staining method enables subcellular resolution of immunocytochemical markers within a single neuron without confounding staining of neighboring cells. We anticipate that this approach will facilitate the study of neuronal phenotype in fine dendritic processes in electrophysiologically characterized neurons in specimens with a complex neuropil, such as brain slices or high-density cultures.

Ultrastructure of the stomatogastric ganglion neuropil of the crab, Cancer borealis.

The stomatogastric ganglion (STG) of the crab, Cancer borealis, contains the neural networks responsible for rhythmic pattern generation of the foregut. Neuron counts indicate that the STG of C. borealis has 25-26 neurons, 4-5 fewer than that found in lobsters. We describe the ultrastructural features of the ganglion by focusing on those that may be involved in storage, release, or range of action of peptide modulators, including a lacunar system and multiple types of intercellular junctions. In the neuropil, we identify five synaptic profile classes that contain the invertebrate presynaptic apparatus (dense bars, small clear vesicles), two of which also contain dense core (modulator-containing) vesicles. These latter two are comprised of multiple immunocytochemical classes that are not easily distinguished by structural criteria. In addition, we find neurohemal-like profiles that contain primarily dense core vesicles. Our finding that multiple profile types in the STG possess modulator-containing vesicles coincides with immunocytochemical results better than do previous ultrastructural studies that report only one such profile type. We show that a single modulatory input, stomatogastric nerve axon 1, makes only classical synapses and not neurohemal-like profiles, although some modulators are found in both these profile types. These data provide the groundwork for understanding the architecture of modulatory input-target interactions and suggest ways that the specificity of modulatory effects within a complex neuropil may be attained.

Functional organization of cotransmission systems: lessons from small nervous systems.

Small invertebrate nervous systems allow one to ask a series of questions concerning the functional roles of cotransmitters. This review outlines some of the implications of cotransmission for target selectivity in complex neuropils. We suggest the possibility that a unique constellation of cotransmitters in individual identified modulatory neurons allows a specificity of action even when peptides may act over an extended distance, and when individual modulatory substances may be released from several modulatory neurons.

Organization of the commissural fibers in the adult brain of the locust

The brain (supraoesophageal ganglion) is the most complex of the segmental ganglia composing the nerve cord of the locusts Schistocerca gregaria and Locusta migratoria. In this paper, we describe the ground plan of the commissures crossing the midline of the brain and propose a nomenclature with the aim of making a complex neuropil more understandable at the level of individual neurons. For developmental and comparative reasons the neuroarchitecture of the brain is related to the neural axis, not to the body axis. We have identified 73 commissural fiber bundles belonging to the adult brain, and these are named according to their location (ventral, dorsal, anterior, posterior, medial) with respect to the central complex as reference point. Reconstructions of identified neurons from intracellular stainings, cobalt backfills, or immunohistochemical studies demonstrate the various configurations in which fibers cross the brain.

Cerebellar Purkinje cell markers are expressed in retinal bipolar neurons

Previous studies have been directed at the elucidation of neuron-specific gene expression in the mammalian central nervous system. In particular, we have identified a series of marker molecules that are expressed in cerebellar Purkinje cells with varying degrees of specificity. Here, we show by light microscopic immunocytochemistry and Northern transfer and hybridization that two of these markers, namely, L7 and PEP19, are expressed in the retina of mouse and rabbit, while a third marker, cerebellin, is absent. Light and electron microscopic immunocytochemistry proves that L7-like immunoreactivity is restricted to rod bipolar cells, while PEP 19-like immunoreactivity is distributed in both rod and cone bipolars. PEP19 is also expressed by subsets of amacrine and ganglion cells. The density of PEP19-positive bipolar cells is greater than that of L7-positive bipolar cells, although the density of each is approximately equal in central and peripheral portions of the retina. An antiserum to a fourth Purkinje cell marker, vitamin D-dependent calcium-binding protein-28 kD (CaBP), reveals primarily axonless horizontal cells, but also subsets of rod bipolar, amacrine, and, in the mouse but not in the rabbit, ganglion cells. The processes of immunoreactive cell bodies form discrete bands in the internal plexiform layer, and mixtures of the antisera help distinguish their identity. Thus, these Purkinje cell markers can be used at the electron microscopic level to unravel the extremely complex neuropil of this retinal layer. Furthermore, knowledge of the retinal distribution of this panel of molecules is of general value for future studies of retinal neuronal typology and can serve to map the densities of subsets of bipolar cells throughout the retina. The expression of L7 and PEP19 in bipolar cells and in Purkinje cells suggests a biochemical relationship between these two spatially distant neuronal populations.

A light- and electron-microscopic investigation of the optic tectum of the frog, Rana pipiens, II: The neurons that give rise to the crossed tecto-bulbar pathway

The superficial layers of the frog's optic tectum, Potter's (1969) layers A-G, comprise a complex neuropil made up of many afferent axons, the somata of a few neurons, and many dendrites from the neurons located in the deeper layers. Different types of retinal axons are believed to terminate in different layers (Maturana et al., 1960; Kuljis & Karten, 1988; Sargent et al., 1989), but little is known about the relationships between each type of input and the dendrites of the deep tectal neurons that extend into these superficial layers. The present study used the method of retrograde transport of horseradish peroxidase to study the synaptic contacts on the dendrites of the neurons that give rise to the crossed tecto-bulbar pathway. These cells have apical dendrites that ascend through the superficial retino-recipient layers. The somata of the cells that give rise to the crossed tecto-bulbar pathway are located in the superficial half of layer 6, preferentially clustered along the caudal, lateral, and rostral margins of the tectum. The somata of these cells range from 8-30 microns in diameter. Their axons are large (2-4 microns in diameter) myelinated fibers that arise from either their somata or proximal dendrites. Their axons travel within the deep medullary layer to leave the tectum at the lateral margin. Their dendritic arbors extend obliquely through the superficial layers to reach layer B where they turn and extend within the layer for up to 0.5 mm. The somata of these cells receive only a scant synaptic input. In contrast, their dendrites receive input in every layer, but the nature of this input varies from layer to layer. Synaptic terminals that resemble retinal ganglion cell boutons contact the labeled dendrites in layers B, F, and G. This indicates that the dendrites may receive monosynaptic input from several types of retinal ganglion cells. Terminals with small, flattened vesicles also contact the dendrites of these cells in each layer. In layer F and below, the terminals with flattened vesicles constitute 15% of the contacts; above layer F they constitute only 5-8% of the contacts. Terminals with medium-sized, flattened vesicles also contact the dendrites of these cells in every layer and constitute a large proportion of their input (33-95%). The latter terminals resemble those that are often postsynaptic to retinal terminals.