Presynaptic modulation of sensory neurons in the segmental ganglia of arthropods

The afferent terminals of arthropod sensory neurones receive abundant input synapses, usually closely intermingled with the sites of synaptic output. The majority of the input synapses use the neurotransmitter GABA, but in some afferents there is a significant glutamatergic or histaminergic component. GABA and histamine shunt afferent action potentials by increasing chloride conductance. Though glutamate can also have this effect in the arthropod central nervous system, its action on afferent terminals appears to be mediated by increases in potassium conductance or by the action of metabotropic receptors. The action of the presynaptic synapses on the afferents are many and varied. Even on the same afferent, they may have several distinct roles that can involve both tonic and phasic patterns of primary afferent depolarisation. Despite the ubiquity and importance of their effects however, the populations of neurones from which the presynaptic synapses are made, remain largely unidentified.     

Grouping and representation of odorant receptors in domains of the olfactory bulb sensory map

Individual glomeruli in the mammalian main olfactory bulb represent a single or at most a few types of odorant receptors. Thus the physical arrangement of glomeruli at the surface of the olfactory bulb can be viewed as a sensory map representing approximately 1,000 types of odorant receptors. This review summarizes the recent advance of the knowledge regarding the spatial organization of the sensory map in the main olfactory bulb. Recent studies show that individual olfactory bulbs contain dual sensory maps, one in the lateral hemisphere and the other in the medial hemisphere of the bulb. The tracings of selective subsets of olfactory axons to their target glomeruli in the olfactory bulb show that glomeruli are parceled into large zones or bands. The spatial arrangement of these zones and bands are stereotypical and conserved across individual mice. Optical imaging studies show that glomeruli in the most rostrodosal zone, zone I, are further parceled into smaller functional domains, and suggest that odorant receptors having a common or similar molecular feature receptive site are grouped together and represented by glomeruli within the functional domain. The possible relation between the functional domain organization and the subjectively perceived odor quality (olfactory submodality) is reviewed.     

Distribution and functional anatomy of amine-containing neurons in decapod crustaceans

One of the lessons learned from studying the nervous systems of phylogenetically distant species is that many features are conserved. Indeed, aminergic neurons in invertebrate and vertebrate systems share a multitude of common characteristics. In this review, the varied roles of serotonin, octopamine, dopamine, and histamine in decapod crustaceans are considered, and the distributions of the amine-containing cells are described. The anatomy of these systems reinforces the idea that amine neurons are involved in widespread modulation and coordination within the nervous system. Many aminergic neurons have long projections, linking multiple regions with a common input, and therefore are anatomically perfected as "gain setters." The developmental patterns of appearance of each amine in the crustacean nervous system are described and compared. The developmental picture suggests that transmitter acquisition is distinctive for each amine, and that the pace of acquisition may be co-regulated with target maturation. The distinctive roles that transmitters play during specific developmental periods may, ultimately, provide important clues to their functional contributions in the mature organism.     

Peripheral synaptic contacts at mechanoreceptors in arachnids and crustaceans: morphological and immunocytochemical characteristics

Two types of sensory organs in crustaceans and arachnids, the various mechanoreceptors of spiders and the crustacean muscle receptor organs (MRO), receive extensive efferent synaptic innervation in the periphery. Although the two sensory systems are quite different-the MRO is a muscle stretch receptor while most spider mechanoreceptors are cuticular sensilla-this innervation exhibits marked similarities. Detailed ultrastructural investigations of the synaptic contacts along the mechanosensitive neurons of a spider slit sense organ reveal four important features, all having remarkable resemblances to the synaptic innervation at the MRO: (1) The mechanosensory neurons are accompanied by several fine fibers of central origin, which are presynaptic upon the mechanoreceptors. Efferent control of sensory function has only recently been confirmed electrophysiologically for the peripheral innervation of spider slit sensilla. (2) Different microcircuit configuration types, identified on the basis of the structural organization of their synapses. (3) Synaptic contacts, not only upon the sensory neurons but also between the efferent fibers themselves. (4) Two identified neurotransmitter candidates, GABA and glutamate. Physiological evidence for GABAergic and glutamatergic transmission is incomplete at spider sensilla. Given that the sensory neurons are quite different in their location and origin, these parallels are most likely convergent. Although their significance is only partially understood, mostly from work on the MRO, the close similarities seem to reflect functional constraints on the organization of efferent pathways in the brain and in the periphery.     

Structure of the olfactory receptor organs,their GABAergic neural pathways,and modulation of mating behavior,in the giant freshwater prawn,Macrobrachium rosenbergii

In the giant male prawn, Macrobrachium rosenbergii, the olfactory system is thought to be the main pathway for modulating sexual behavior through pheromone perception. In this report, we first used gross anatomical, histological, and SEM methods to describe the structures of the olfactory receptors (sensilla setae), their neural pathways, and possible role in modulating mating behavior. On the surfaces of antennule and antenna filaments there are four types of sensory receptors, viz single spike‐like setae, single flagellum‐like setae, multiple flagella‐like setae, and aesthetascs (ASs). The ASs, which had previously been proposed to be odor receptor setae, are found only on the short filament of lateral antennule (slAn). Each AS on the slAn connects with olfactory receptor neurons (ORNs), whose axons form an outer central antennule nerve (ocAnNv), which then connects with the olfactory neutrophil (ON) of the brain. Thus, the slAn is the major olfactory organ that conveys sensory inputs from each AS to the ON within the deutocerebrum. GABA immunoreactivity was present in ASs, neurons of ORNs, inner central antennular, lateral tegumentary nerve, ocAnNv and the ON, inferring that GABA is the likely neurotransmitter in modulating olfaction. Disruption of the slAn by ablation or covering with Vaseline, resulted in significant reduction of mating behavior, indicating that this organ is crucial for sex pheromone perception. Identification of the active pheromones and further bioassays are now being performed. Microsc. Res. Tech. 76:572–587, 2013. © 2013 Wiley Periodicals, Inc.     

Neuromodulation of arthropod mechanosensory neurons

Arthropod mechanosensory afferents have long been known to receive efferent synaptic connections onto their centrally located axon terminals. These connections cause presynaptic inhibition by attenuating the action potentials arriving at the axon terminals, thus reducing the synaptic potentials in the postsynaptic neurons. This type of inhibition can specifically reduce the excitation of selected postsynaptic neurons while leaving others unaffected. However, recent research has demonstrated that sensory signals detected by arthropod mechanosensory neurons can also be synaptically modulated before they ever arrive at the axon terminals. In arachnids and crustaceans, wide and complex networks of synapses on all parts of the afferent neurons, including the somata and dendrites, provide mechanisms to inhibit or enhance the responses to mechanical stimuli as they are being detected. This modulation will affect the signal transmission to all axonal branches and postsynaptic cells of the affected receptor neuron. In addition to the increased complexity of mechanosensory information transmission produced by these synapses, a variety of circulating neuroactive substances also modulate these neurons by acting on their postsynaptic receptors.     

Impulse conduction of olfactory receptor neuron axons

Compound action potentials were recorded from rat olfactory receptor neuron axons at measured distances from the stimulation electrode along the lateral surface of the main olfactory bulb. Distances were plotted as a function of the latencies measured from stimulus onset to the prominent negative trough of the triphasic compound action potential. A straight line was fitted to these data to calculate impulse conduction velocity, 0.42 +/- 0.01 m/s (n = 25). Two procedures were used to investigate whether those axons that project to caudal regions of the bulb had faster conduction velocities than axons projecting to rostral bulb. First, the stimulating electrode was moved to mid-bulb and the recording electrode was placed on the caudal bulb. Alternatively, axons were stimulated antidromically at the caudal bulb. These two procedures stimulate those axons projecting to caudal bulb and bypass olfactory receptor neuron axons that synapse in the rostral bulb. The mean impulse conduction velocities from these caudal and antidromic recordings were 0.58 +/- 0.19 m/s (n = 8) and 0.57 +/- 0.19 m/s (n = 9), respectively. Though both of these means are higher than the impulse conduction velocity calculated for stimulation at the rostral bulb, the differences were not statistically significant.     

Characterization of chemically defined neurons and their cellular relationships by combined immunocytochemistry and radioautographic localization of transmitter uptake sites

Radioautography and immunocytochemistry may be combined at the light and electron microscopic levels for simultaneously localizing uptake sites for exogenous transmitter molecules [such as (³H)monoamines or (³H)amino acids] and endogenous transmitter-related antigens (classical transmitters and their synthesizing enzymes as well as neuropeptides) in the central nervous system. Silver grain accumulations indicative of transmitter uptake sites are readily distinguishable from immunocytochemical labels of the peroxidase-antiperoxidase (PAP), avitin-biotin, or colloidal gold methods. The combination of uptake radioautography and immunocytochemistry may be applied to the investigation of (1) the chemical identity of (³H) transmitter-accumulating elements, (2) the coexistence of different neurotransmitters within the same neurons, and (3) the cellular basis of interactions between certain neurotransmitters, in particular monoamines, GABA, and neuropeptides. This article describes and evaluates the method and reviews the available experimental data derived from its application.     

GABAB receptor intracellular trafficking after internalization in Paramecium

The number of neurotransmitter receptors on the plasma membrane is regulated by the traffic of intracellular vesicles. Golgi-derived vesicles provide newly synthesized receptors to the cell surface, whereas clathrin-coated vesicles are the initial vehicles for sequestration of surface receptors, which are ultimately degraded or recycled. We have previously shown that GABAB receptors display a punctuate vesicular pattern dispersed on the cell surface and throughout the cytoplasm and are internalized via clathrin-dependent and -independent endocytosis. Here we have studied constitutive GABAB receptor trafficking after internalization in Paramecium primaurelia by confocal laser scanning microscopy and multiple immunofluorescence analysis. After internalization, receptors are targeted to the early endosomes characterized by the molecular markers EEA1 and rab5. Some of these receptors, destined for recycling back to the plasma membrane, traffic from the early endosomes to the endosomal recycling compartment that is characterized by the presence of rab4-immunoreactivity (IR). Receptors that are destined for degradation exit the endosomal pathway at the early endosomes and traffic to the late endosome-lysosome pathway. In fact, some of the GABAB-positive compartments were identified as lysosomal structures by double staining with the lysosomal marker LAMP-1. GABAB vesicle structures also colocalize with TGN38-IR and rab11-IR. TGN38 and rab11 are proteins found in association with post-Golgi and recycling endosomes, respectively.     

Confocal laser scanning microscopy: using cuticular autofluorescence for high resolution morphological imaging in small crustaceans

The utility of cuticular autofluorescence for the visualization of copepod morphology by means of confocal laser scanning microscopy (CLSM) was examined. Resulting maximum intensity projections give very accurate information on morphology and show even diminutive structures such as small setae in detail. Furthermore, CLSM enables recognition of internal structures and differences in material composition. Optical sections in all layers and along all axes of the specimens can be obtained by CLSM. The facile and rapid preparation method bears no risk of artefacts or damage occurring to the preparations and the visualized specimens can be used for later analyses allowing for the investigation of irreplaceable type specimens or parts of them. These features make CLSM a very effective tool for both taxonomical and ecological studies in small crustaceans; however, the maximum thickness of the specimens is limited to a few hundred micrometers. Three‐dimensional models based on CLSM image stacks allow observation of the preparations from all angles and can permit, improve and speed up studies on functional morphology. The visualization method described has a strong potential to become a future standard technique in aquatic biology due to its advantages over conventional light microscopy and scanning electron microscopy.     

Development of the vomeronasal receptor epithelium and the accessory olfactory bulb in sheep

The morphological development of the vomeronasal organ (VNO) and accessory olfactory bulb (AOB) of the sheep from anlage to birth were studied by classical and histochemical methods using embryos and fetuses obtained from an abattoir with ages estimated from crown-to-rump length. Both VNO and AOB developed in a biologically logical sequence and completed their morphological development around day 98, at entry into the last third of the gestation period. A lectin with specificity for oligomeric N-acetylglucosamine labeled the sensory epithelium of the VNO, the vomeronasal nerves, and the nervous and glomerular layers of the AOB before birth. These results suggest that the vomeronasal system, which is well developed and functional in adult sheep, may be able to function at or even before birth in these animals (whereas in rodents, for example, this is precluded by the AOB not completing its development until after birth).     

Ciliated and microvillar receptor cells degenerate and then differentiate in the olfactory epithelium of rainbow trout following olfactory nerve section.

We used scanning (SEM) and transmission (TEM) electron microscopy to examine ultrastructural changes in the olfactory epithelium (OE) of rainbow trout following unilateral olfactory nerve section. Both ciliated receptor cells (CRC) and microvillar receptor cells (MRC) degenerated and subsequently differentiated from unidentified precursor cells. The following changes took place in fish that were held at 10 degrees C at the stated period following olfactory nerve section: on day 7, MRC and CRC contained intracellular vacuoles; on day 12, the olfactory knobs appeared disrupted; by day 26, olfactory receptor cells were absent from the OE; on day 42, there were receptor cell bodies and a few CRC with short cilia at the apical surface; and on day 55, a small number of both CRC and MRC had differentiated. By day 76, both CRC and MRC repopulated the OE. Degenerative changes in the cytoplasm of the sustentacular cells (SC) and ciliated nonsensory cells (CNC) were observed in the first 26 days following olfactory nerve section, but these cells remained intact throughout the experiment. The degeneration and subsequent differentiation of CRC and MRC supports and extends previous observations that both cell types are olfactory receptor neurons with axons that extend along the olfactory nerve to the olfactory bulb.     

Active dendritic properties constrain input-output relationships in neurons of the central olfactory pathway in the crayfish forebrain

Parasol cells are multimodal sensory interneurons of the hemi-ellipsoid body in the decapod forebrain. In reptant crustaceans, the hemi-ellipsoid body resides in the base of the eyecup, as an appendage to the terminal medulla. Parasol cells exhibit periodic depolarizations at a frequency of 0.5-1.0 Hz. I have investigated the role of these periodic depolarizations and their superimposed impulse bursts in affecting the input/output properties of these neurons. Parasol cells receive input from photic, olfactory, and mechanosensory pathways. Strong stimulation over any one of these pathways can lead to the generation of one or more impulse bursts in a subset of parasol cells, timed to occur at the peak of successive periodic depolarizations. A role for the periodic depolarizations in the function of the parasol cells has yet to be established. I suggest the possibility that they may act as a nonlinear amplifier that boosts spatially-summated excitatory synaptic potentials from strong or appropriate stimuli above threshold for burst generation. Another possibility includes modification of voltage-sensitive ion channels in the dendritic membrane, permitting a more effective spread of excitatory synaptic currents to impulse or burst initiating zones. Impulse bursts may be a highly effective mode of output for these neurons, especially so as they occur synchronously in a subset of cells in response to strong sensory input. Furthermore, backfiring of bursts into the dendritic tree has a brief (2-3 second) but effective suppressive action upon weak sensory input, which can thereby be masked by stronger, burst-generating input. This masking phenomenon is seen in other arthropod sensory interneurons, where its physiological basis appears to be a transient accumulation of intracellular Ca(++) ions that open calcium-sensitive potassium channels.     

Five-colour in vivo imaging of neurons in Caenorhabditis elegans

In the last few years variants of the ‘green fluorescent protein’ (GFP) with different spectral properties have been generated. This has greatly increased the number of possible applications for these fluorochromes in cell biology. The significant overlap of the excitation and emission spectra of the different GFP variants imposes constraints on the number of variants that can be used simultaneously in a single sample. In particular, the two brightest variants, GFP and YFP, are difficult to separate spectrally. This study shows that GFP and YFP can be readily separated with little spectral overlap (cross‐talk) with the use of a confocal microscope equipped with an acusto‐optical beam splitter and freely adjustable emission windows. Under optimal recording conditions cross‐talk is less than 10%. Together with two other fluorescent proteins and the lipophilic dye DiD a total of five different colours can now be used simultaneously to label in vivo distinct anatomical structures such as neurons and their processes. Spatial resolution of the confocal microscope is sufficient to resolve the relative position of labelled axons within a single axon bundle. The use of five distinct marker dyes allows the in vivo analysis of the Caenorhabditis elegans nervous system at unprecedented resolution and richness in detail at the light microscopic level.     

The aesthetasc concept: structural variations of putative olfactory receptor cell complexes in Crustacea.

The structure of the aesthetascs has been investigated in the prawn Macrobrachium rosenbergii (larvae and juveniles), the opossum shrimp Neomysis integer, the euphausid Meganyctiphanes, and in the water-fleas Daphnia magna and D. longispina. The aesthetascs, that are thought to represent olfactory receptors, exhibit a considerable structural variation, ranging from the well known aesthetascs of higher crustaceans (lobster, crab, crayfish) to the corresponding sensilla found in the water-fleas and the males of opossum shrimps. The two following morphological characteristics of the aesthetascs are thought to indicate an olfactory function: the shape of the cuticular hair that is long and essentially hose-shaped, and the thin, loosely arranged cuticle of at least the outer part of the cuticular hair. The presence of other structural elements such as sensory cells, cilia, and enveloping cells are vital for the olfactory function, but the development is variable, which makes their use in the morphological definition of aesthetascs problematic.     

Assessment of Congo red as a fluorescence marker for the exoskeleton of small crustaceans and the cuticle of polychaetes

In this study, the potential of the common dye Congo red as a fluorescence marker for chitin in the exoskeleton of small crustaceans and collagen in the polychaete cuticle was tested. The Congo red staining turned out to be rather efficient and yielded intensively fluorescing structures, which made a very detailed visualization by confocal laser scanning microscopy possible. The excellent results are comparable to those described for the utilization of other efficient fluorescence dyes and intense autofluorescence. The application of Congo red is easy, the fluorescence of this dye is very stable, and the excitation maximum of the structures stained with Congo red is in a range, which is covered by the lasers of most of the confocal laser scanning microscopes. These advantageous properties make the fluorescence staining by Congo red a method of choice for the detailed visualization of the external morphology of small crustaceans and polychaetes.     

Drosophila as a focus in olfactory research: mapping of olfactory sensilla by fine structure, odor specificity, odorant receptor expression, and central connectivity.

This review intends to integrate recent data from the Drosophila olfactory system into an up-to-date account of the neuronal basis of olfaction. It focuses on (1) an electron microscopic study that mapped a large proportion of fruitfly olfactory sensilla, (2) large-scale electrophysiological recordings that allowed the classification of the odor response spectra of a complete set of sensilla, (3) the identification and expression patterns of candidate odorant receptors in the olfactory tissues, (4) central projections of neurons expressing a given odorant receptor, (5) an improved glomerular map of the olfactory center, and (6) attempts to exploit the larval olfactory system as a model of reduced cellular complexity. These studies find surprising parallels between the olfactory systems of flies and mammals, and thus underline the usefulness of the fruitfly as an olfactory model system. Both in Drosophila and in mammals, odorant receptor neurons appear to express only one type of receptor. Neurons expressing a given receptor are scattered in the olfactory tissues but their afferents converge onto a few target glomeruli only. This suggests that in both phyla, the periphery is represented in the brain as a chemotopic map. The major difference between mammals and fruitflies refers to the numbers of receptors, neurons, and glomeruli, which are largely reduced in the latter, and particularly in larvae. Yet, if activated in a combinatorial fashion, even this small set of elements could allow discrimination between a vast array of odorants.     

Phylogeny of the vomeronasal system and of receptor cell types in the olfactory and vomeronasal epithelia of vertebrates.

In this paper, the evolutionary origin of the vomeronasal system as a discrete sensory system separate from olfaction is examined. The presence of a discrete vomeronasal system appears to be a derived character in tetrapods, and its presence in larval amphibians indicates that the system did not arise as a terrestrial adaptation. The vomeronasal system has been lost independently in several taxa, including crocodilians, some bats, cetaceans, and some primates. The presence of microvillar receptor cells in the vomeronasal epithelium appears to be the ancestral condition for tetrapods, and alternative hypotheses concerning the ancestral condition for receptor cell types in the vertebrate olfactory epithelium are discussed. Finally, the possibility that the vomeronasal system is present in some fishes in a form that has not been recognized is discussed in relation to the phylogenetic distribution of receptor cell types in vertebrates.     

The olfactory organ of the trout Salmo trutta fario: A novel localization for a progestin receptor

A progestin receptor (PR) has been detected in the olfactory organ of the trout Salmo trutta fario. The specificity of this receptor was high for 17α,20β‐dihydroxy‐4‐pregnen‐3‐one (17α,20β‐DP), but it also bound 17α‐hydroxy‐progesterone (17α‐OHP) and 21‐hydroxyprogesterone (21‐OHP), even when present at low concentrations (10‐fold in relative binding affinity assay). Progesterone (P) competed effectively at much higher concentrations (1,000‐fold in relative binding affinity assay). Immunohistochemical studies carried out with three different monoclonal antibodies against human progesterone receptor (hPR), chicken progesterone receptor hinge region (cPR), and chicken progesterone receptor A/B domain (PR22), revealed that immunoreactivity was present in the epithelium of the olfactory organ of females and males of the trout Salmo trutta fario only against hPR. Western blotting showed two hPR immunoreactive bands of about 62 and 66 kDa. Finally, a portion of the cDNA of about 300 nucleotides extending over the DNA binding domain and the ligand binding domain was cloned and sequenced, revealing a high degree of sequence homology of the PR in Salmo trutta fario with the PR in other teleosts. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.