Distribution and structure of identified tonic and phasic synapses between L-neurones in the locust ocellar tract.

The ultrastructures and distributions of the discrete anatomical synapses which constitute two distinct types of output connections made by individual ocellar L-neurons, L1-3, are described. Outputs to neurones L4-5 are excitatory and transmit tonically, whereas reciprocal connections among the three L1-3 neurones are inhibitory and incapable of transmission for longer than a few milliseconds. The tonically transmitting synapses are located in the lateral ocellar tract and are made between the axons of L1-3, which do not receive inputs, and short branches of L4-5, which make no outputs. Each excitatory connection is composed of a few hundred discrete anatomical synapses, each characterised by a bar-shaped presynaptic density which is 0.15-1.5 microns in length and associated with a large number of round synaptic vesicles. Two postsynaptic profiles are apposed to each presynaptic density. Associated with tonic synapses are abundant invaginations of the presynaptic membrane. Synapses of the reciprocal, inhibitory, phasic connections occur in the protocerebral arbors of L1-3, among numerous output synapses of these neurones. Each phasic connection is composed of a few tens of discrete anatomical synapses. Each bar-shaped presynaptic density is associated with two postsynaptic profiles, and is 0.1-1.0 microns long. Compared with the tonic, excitatory connection, there are fewer vesicles and fewer invaginations of the presynaptic membrane associated with each synapse.     

Nontopographic projection of olfactory sensory neurons in the crayfish brain

We have used tritiated leucine to trace the input projection pattern of olfactory sensory neurons in crayfishes. The olfactory neurons are associated with cuticular sensilla on the external antennular filaments. Each sensillum, or aesthetasc, harbors the distal dendritic segments of about 175 bipolar sensory neurons, the cell bodies for which are grouped in a subcuticular ensemble or ganglion. About 150-175 individual ganglia may be found on each antennule in an adult crayfish. When an aesthetasc is exposed to tritated leucine, the tracer is taken up by the associated olfactory sensory neurons and is transported along the axons to their central terminations within the glomeruli of the ipsilateral olfactory lobe. We tested the possibility that the sensory neurons from specific aesthetascs project to specific glomeruli. By restricting access of the leucine to small groups of aesthetascs, we exposed less than 2% of the olfactory sensory neurons to the tracer. Nonetheless, all glomeruli were labeled following such treatment. We conclude that the sensory neurons are generally distributed to the olfactory glomeruli. If each neuron terminates in a single glomerulus, these data support a divergent pattern of sensory projection from individual ganglia to all regions of the olfactory lobe.     

Organization of a sensory neuropile in the auditory pathway of two groups of Orthoptera

The anterior intermediate sensory neuropile (aISN) is a prominent neuropile in the ventral nerve cord of locusts and bushcrickets. Previous studies have shown that it receives its main sensory input from auditory receptors. In this paper we examine the structural and physiological relationship between tympanal receptor terminations and the dendrites of sound-sensitive interneurones in the homologous neuropile of locusts and bushcrickets. Each individual receptor fibre of the bushcricket terminates in a somewhat different target area of the neuropile. The ordering is with respect to the characteristic frequency of the fibres (tonotopic) in the anterior-posterior and dorsoventral axis. In the locust, representatives of the four tympanal receptor groups branch in different areas of the aISN. Most of the dorsal neuropilar region, and the anterior ventral region, do not receive input from tympanal receptors. The dendrites of identified sound-sensitive interneurones were examined in the context of this afferent projection. Local interneurones as well as intersegmental interneurones in bushcrickets have dendritic branches in the whole aISN or part of it and thus overlap with at least some receptors. By recording intracellularly from their main neurites, short-latency synaptic potentials were found in response to receptor spikes indicating monosynaptic input. The tuning of these neurones could be predicted by their dendritic morphology. In contrast, in the locust only local and bisegmental neurones are monosynaptically connected with tympanal receptors, but not the studied intersegmental neurones. This is consistent with the finding that most or all branches of intersegmental neurones lie in the dorsal area of neuropile where no receptors terminate. Anatomical and physiological evidence is presented for identified local neurones providing the excitatory and inhibitory synaptic input for such intersegmental neurones. The difference in the basic wiring diagram in the homologous neuropile of the two orthopteran groups is discussed with respect to the possible different roles that sound plays in their behaviour.     

Visual afferents from an eye in the terrestrial slug Limax valentianus

Terrestrial gastropods have a lens-bearing eye on the tip of their tentacles. There are two morphologically distinct photoreceptors, called Type-I and Type-II photoreceptors, in the retina. Type-I photoreceptors are equipped with highly developed photoreceptive microvilli in their outer rhabdomeric segment, whereas Type-II photoreceptors have short and fewer microvilli. Although both types of photoreceptors send afferent projections directly to the brain, their destinations in the brain, called optic neuropiles, have not been sufficiently investigated. Our recent studies revealed that there are commissural fibers in the cerebral ganglia that transmit photic information acquired by bilateral eyes. Moreover, some of the retinal photoreceptors are connected by gap junctions to the photosensitive brain neurons, suggesting the functional interaction of the photic information between the eye and brain photoreceptors, as well as between bilateral eyes. However, it has not been clarified which type of retinal photoreceptors send commissural projections to the contralateral hemiganglion nor interact with the brain photoreceptors. In the present study, we demonstrated by molecular histological analyses and tracer injections that (1) Type-I and Type-II photoreceptors send glutamatergic afferent projections to the medial and lateral lobes of the ipsilateral optic neuropile, respectively, (2) direct synaptic interaction between bilateral optic nerves occurs in the medial lobe of the optic neuropile, and (3) brain photosensory neurons form gap junctions with the medial lobe of the contralateral optic neuropile. These results reveal an ordered pattern of afferent projections from the retina and provide insight into the different functional roles of retinal photoreceptors.     

Neurocytology of the cerebral ganglion of Fasciola hepatica (Platyhelminthes)

An ultrastructural study of the organization and fine structure of the nervous system of the parasitic flatworm Fasciola hepatica was undertaken. The brain consists of paired cerebral ganglia, located just posterior to the oral sucker, that are connected by a transverse commissure which crosses over the dorsal surface of the pharynx. The cell bodies of the cerebral ganglia are not organized into a clearly defined rind around the neuropile but are loosely scattered around and within the neuropile area. The neuropile consists of two morphologically distinct types of unmyelinated nerve processes. The small nerve processes, with smooth cell membranes, are less than 2 micron in diameter, whereas the "giant" processes are greater than 12 micron in diameter and have extensively invaginated cell membranes. Giant processes make up the bulk of the nerve fibers in the transverse commissure and longitudinal nerve cords. Four morphological types of vesicles are present in the small processes; small clear vesicles (which were found associated with synapses), spheroidal and ellipsoidal dense vesicles, and dense-core vesicles. Two types of synapses are found in the neuropile: simple synapses characterized by pre- and postsynaptic membrane densities, clusters of small clear vesicles, and a clear synaptic cleft; and wedge-shaped synapses or divergent diads each having one presynaptic process synapsing onto two postsynaptic processes. Synaptic contacts were observed only between small processes; no synapses were observed on the cell bodies or on giant processes.     

Embryonic origin of the Drosophila brain neuropile

Neurons of the Drosophila larval brain are formed by a stereotyped set of neuroblasts. As differentiation sets in, neuroblast lineages produce axon bundles that initially form a scaffold of unbranched fibers in the center of the brain primordium. Subsequently, axons elaborate interlaced axonal and dendritic arbors, which, together with sheath-like processes formed by glial cells, establish the neuropile compartments of the larval brain. By using markers that visualize differentiating axons and glial cells, we have analyzed the formation of neuropile compartments and their relationship to neuroblast lineages. Neurons of each lineage extend their axons as a cohesive tract ("primary axon bundle"). We generated a map of the primary axon bundles that visualizes the location of the primary lineages in the brain cortex where the axon bundles originate, the trajectory of the axon bundles into the neuropile, and the relationship of these bundles to the early-formed scaffold of neuropile pioneer tracts (Nassif et al. [1998] J. Comp. Neurol. 402:10-31). The map further shows the growth of neuropile compartments at specific locations around the pioneer tracts. Following the time course of glial development reveals that glial processes, which form prominent septa around compartments in the larval brain, appear very late in the embryonic neuropile, clearly after the compartments themselves have crystallized. This suggests that spatial information residing within neurons, rather than glial cells, specifies the location and initial shape of neuropile compartments.     

Some scanning electron microscopic studies of the surface of coronal sections cut from the rat brain

The cut surfaces of coronal sections of the rat cerebral hemispheres have been examined with the scanning electron microscope. Under low magnification the structures of the brain are easily identified. Using the focusing power of the microscope and the ability to tilt the specimen in its column, under higher magnification the neurones, glial cells, cell processes, bundles of myelinated axons and the cerebral blood vessels can be examined in great detail. This method of study should prove of value in the future for the investigation of brains which have been subjected to experimental procedures and those affected by disease.     

Antennal neuropile in the brain of the crayfish: morphology of neurons

The cellular composition of the antennal neuropile of the crayfish is described. As a context for this work the distribution of neuronal cell bodies throughout the supraoesophageal ganglion (brain) is also described. The neuronal cell bodies in the brain are concentrated in 19 distinct clusters. Three paired clusters are located on the dorsal side of the brain, four paired and one midline cluster bend around the brain laterally and frontally respectively. Fewer than ten somata lie outside of these clusters. The antennal neuropile is composed of primary afferent terminals, efferents, and projecting and local interneurons. The structures of individual neurons of all four types were determined by filling them with Lucifer yellow, and an overview of the neuropile structure was obtained with cobalt backfills of selected nerves. The antennal afferents are concentrated in four main tracts that run medially in the outer layer of the antennal neuropile. Up to 11 orthogonal side branches occur at equal distances (25-35 microns) along the main branches and penetrate the neuropile. The efferents contribute very thin dendrites to the antennal neuropile. The majority of the neuronal mass of the antennal lobe consists of projecting and local interneurons. The branching pattern of the interneurons within the antennal neuropile also shows an orthogonal arrangement of main branches and higher-order branches. Thus the antennal neuropile displays a strong geometrical regularity: Main processes of all four types of neurons run in bundles the length of the long axis of the neuropile (lateral to medial inside the brain) giving rise to orthogonal side branches at regular intervals. This branching pattern leads to a striped appearance of the antennal lobe.     

External ions and membrane potential of leech neuropile glial cells

The ionic mechanism of a membrane effect of 5-hydroxytryptamine (5-HT) on neuropile glial (NG) cells in ganglia of the medicinal leech was investigated with conventional single-barrelled microelectrodes. Control experiments were made with double-barrelled ion-selective microelectrodes. 5-Hydroxytryptamine hyperpolarized the NG-cell membrane and increased the conductance considerably. Methysergide, a potent 5-HT antagonist, blocked the 5-HT-induced hyperpolarization completely. When leech ganglia were superfused with physiological bathing media free of 5-HT, the NG-cell membrane conductance returned to the original value, but the membrane potential recovered only partially from the hyperpolarization in most experiments. In glial membranes artificially depolarized by means of constant-current injection, the amplitude of the 5-HT response increased. The amplitude decreased with membrane hyperpolarization and reversed at —73 mV, close to the potassium equilibrium potential. The reversal potential changed by 52 mV when the extracellular potassium concentration was altered by a factor of 10. We conclude that 5-HT increases the potassium conductance of NG-cell membranes.