Biomedical subjects

John G Hildebrand

Publications and source records attributed to John G Hildebrand.

18 recordsLinked to original sources

PubMed2006

Phylogeny of a serotonin-immunoreactive neuron in the primary olfactory center of the insect brain.

Serotonin (5-hydroxytryptamine; 5HT) functions in insects as a neurotransmitter, neuromodulator, and neurohormone. In the sphinx moth Manduca sexta, each of the paired antennal lobes (ALs; the primary olfactory centers in the insect brain) has one 5HT-immunoreactive (5HT-ir) neuron that projects into the protocerebrum, crosses the posterior midline, and innervates the contralateral AL; this is referred to as the contralaterally projecting, serotonin-immunoreactive deutocerebral (CSD) neuron. These neurons are thought to function as centrifugal modulators of olfactory sensitivity. To examine the phylogenetic distribution of 5HT-ir neurons apparently homologous to the CSD neuron, we imaged 5HT-like immunoreactivity in the brains of 40 species of insects belonging to 38 families in nine orders. CSD neurons were found in other Lepidoptera, Trichoptera, Diptera, Coleoptera, and Neuroptera but not in the Hymenoptera. In the paraneopteran and polyneopteran species (insects that undergo incomplete metamorphosis) examined, AL 5HT neurons innervate the ispsilateral AL and project to the protocerebrum. Our findings suggest that the characteristic morphology of the CSD neurons originated in the holometabolous insects (those that undergo complete metamorphosis) and were lost in the Hymenoptera. In a subset of the Diptera, the CSD neurons branch within the contralateral AL and project back to the ipsilateral AL via the antennal commissure. The evolution of AL 5HT neurons is discussed in the context of the physiological actions of 5HT observed in the lepidopteran AL.

Animals↗

PubMed2005

Chemosensory selectivity of output neurons innervating an identified, sexually isomorphic olfactory glomerulus.

The antennal lobe (AL) of insects, like the olfactory bulb of vertebrates, is characterized by discrete modules of synaptic neuropil called glomeruli. In some insects (e.g., moths and cockroaches), a few glomeruli are sexually dimorphic and function in labeled lines for processing of sensory information about sex pheromones. Controversy still exists, however, about whether projection (output) neurons (PNs) of glomeruli in the main AL are also narrowly tuned. We examined this critical issue in the AL of the moth Manduca sexta. We used intracellular recording and staining techniques to investigate the chemosensory tuning of PNs innervating an identifiable, sexually isomorphic glomerulus, G35, in the main AL. We found that the morphological features and chemosensory tuning of G35-PNs were nearly identical in females and males. G35-PNs responded to low concentrations of the plant-derived volatile compound cis-3-hexenyl acetate (c3HA), but the sensitivity threshold of female PNs was lower than that of male PNs. The propionate and butyrate homologs of c3HA could evoke excitatory responses but only at moderate-to-high concentrations. Other plant volatiles did not evoke responses from G35-PNs. Moreover, PNs innervating glomeruli near G35 (in females) showed little or no response to c3HA. Female G35-PNs were hyperpolarized by (+/-)linalool, a compound that excites PNs in an adjacent glomerulus, thus providing evidence for lateral-inhibitory interactions between glomeruli. Our results show that PNs arborizing in an identified glomerulus in the main olfactory pathway are morphologically and physiologically equivalent in both sexes and have characteristic, limited molecular receptive ranges that are highly conserved across individuals.

Acetates↗

PubMed2005

Octopamine-immunoreactive neurons in the brain and subesophageal ganglion of the hawkmoth Manduca sexta.

Octopamine is a neuroactive monoamine that functions as a neurohormone, a neuromodulator, and a neurotransmitter in many invertebrate nervous systems, but little is known about the distribution of octopamine in the brain. We therefore used a monoclonal antibody to study the distribution of octopamine-like immunoreactivity in the brain of the hawkmoth Manduca sexta. Immunoreactive processes were observed in many regions of the brain, with the distinct exception of the upper division of the central body. We focused our analysis on nine ventral unpaired median (VUM) neurons with cell bodies in the labial neuromere of the subesophageal ganglion. Seven of these neurons projected caudally through the ventral nerve cord. Two neurons projected rostrally into the brain (supraesophageal ganglion), and one of these was a bilateral neuron that sent projections to the gamma-lobe of the mushroom body and the lateral protocerebrum. Octopamine-immunoreactive processes from one or more cells originating in the subesophageal ganglion also form direct connections between the antennal lobes and the calyces of the mushroom bodies.

Animals↗

PubMed2004

Spatial and temporal organization of ensemble representations for different odor classes in the moth antennal lobe.

In the insect antennal lobe, odor discrimination depends on the ability of the brain to read neural activity patterns across arrays of uniquely identifiable olfactory glomeruli. Less is understood about the complex temporal dynamics and interglomerular interactions that underlie these spatial patterns. Using neural-ensemble recording, we show that the evoked firing patterns within and between groups of glomeruli are odor dependent and organized in both space and time. Simultaneous recordings from up to 15 units per ensemble were obtained from four zones of glomerular neuropil in response to four classes of odorants: pheromones, monoterpenoids, aromatics, and aliphatics. Each odor class evoked a different pattern of excitation and inhibition across recording zones. The excitatory response field for each class was spatially defined, but inhibitory activity was spread across the antennal lobe, reflecting a center-surround organization. Some chemically related odorants were not easily distinguished by their spatial patterns, but each odorant evoked transient synchronous firing across a uniquely different subset of ensemble units. Examination of 535 cell pairs revealed a strong relationship between their recording positions, temporal correlations, and similarity of odor response profiles. These findings provide the first definitive support for a nested architecture in the insect olfactory system that uses both spatial and temporal coordination of firing to encode chemosensory signals. The spatial extent of the representation is defined by a stereotyped focus of glomerular activity for each odorant class, whereas the transient temporal correlations embedded within the ensemble provide a second coding dimension that can facilitate discrimination between chemically similar volatiles.

Action Potentials↗

PubMed2004

Glycosylation patterns are sexually dimorphic throughout development of the olfactory system in Manduca sexta.

In the moth Manduca sexta, development of the adult olfactory system depends on complex interactions between olfactory receptor neurons in the antenna, antennal-lobe neurons in the brain, and several classes of glial cells. As one approach to characterizing molecules that may play roles in these interactions, we used lectins to screen antennae and antennal lobes at different stages of adult development. We find that each of the major neural cell types has a distinct pattern of labeling by lectins. Effects of enzymatic and other treatments on lectin labeling lead us to conclude that the predominant lectin ligands are: glycosphingolipids and an O-linked, fucose-containing glycoprotein on axons of olfactory receptor neurons, O-linked glycoproteins on antennal-lobe neurons, and N-linked glycoproteins on all classes of glial cells in the primary olfactory pathway. Wheat germ agglutinin labels all olfactory axons uniformly during much of development, but labeling becomes restricted to the pheromone-responsive olfactory receptor neurons in the adult male. Succinylated WGA reveals differences in these axon classes earlier, as glomerului develop from protoglomeruli. The adult female displays a less pronounced difference in labeling of axons targeting ordinary and sexually dimorphic glomeruli. Differences in labeling of receptor axons targeted to ordinary and sexually dimorphic glomeruli may be correlated with differences in function or connectivity in different regions of the antennal lobe.

Animals↗

PubMed2004

Sensory processing of ambient CO2 information in the brain of the moth Manduca sexta.

Insects use information about CO2 to perform vital tasks such as locating food sources. In certain moths, CO2 is involved in oviposition behavior. The labial palps of adult moths that feed as adults have a pit organ containing sensory receptor cells that project into the antennal lobes, the sites of primary processing of olfactory information in the brain. In the moth Manduca sexta and certain other species of Lepidoptera, these receptor cells in the labial-palp pit organ have been shown to be tuned to CO2, and their axons project to a single, identified glomerulus in the antennal lobe, the labial-palp pit organ glomerulus. At present, however, nothing is known about the function of this glomerulus or how CO2 information is processed centrally. We used intracellular recording and staining to reveal projection (output) neurons in the antennal lobes that respond to CO2 and innervate the labial-palp pit organ glomerulus. Our results demonstrate that this glomerulus is the site of first-order processing of sensory information about ambient CO2. We found three functional types of CO2-responsive neurons (with their cell bodies in the antennal lobe or the protocerebrum) that provide output from the antennal lobe to higher centers in the brain. Some physiological characteristics of those neurons are described.

Action Potentials↗

PubMed2004

Learning modulates the ensemble representations for odors in primary olfactory networks.

Recent evidence suggests that odor-driven responses in the insect antennal lobe (AL) can be modified by associative and nonassociative processes, as has been shown in the vertebrate olfactory bulb. However, the specific network changes that occur in response to olfactory learning remain unknown. To characterize changes in AL network activity during learning, we developed an in vivo protocol in Manduca sexta that allows continuous monitoring of neural ensembles and feeding behavior over the course of olfactory conditioning. Here, we show that Pavlovian conditioning produced a net recruitment of responsive neural units across the AL that persisted after conditioning. Recruitment only occurred when odor reliably predicted food. Conversely, when odor did not predict food, a net loss of responsive units occurred. Simultaneous measures of feeding responses indicated that the treatment-specific patterns of neural recruitment were positively correlated with changes in the insect's behavioral response to odor. In addition to recruitment, conditioning also produced consistent and profound shifts in the temporal responses of 16% of recorded units. These results show that odor representations in the AL are dynamic and related to olfactory memory consolidation. We furthermore provide evidence that the basis of the learning-dependent changes in the AL is not simply an increase in activity in the neural network representing an odorant. Rather, learning produces a restructuring of spatial and temporal components of network responses to odor in the AL.

Animals↗

PubMed2004

Floral CO(2) emission may indicate food abundance to nectar-feeding moths.

As part of a study of the roles of the sensory subsystem devoted to CO(2) in the nectar-feeding moth Manduca sexta, we investigated CO(2) release and nectar secretion by flowers of Datura wrightii, a preferred hostplant of Manduca. Datura flowers open at dusk and wilt by the following noon. During the first hours after dusk, when Manduca feeds, the flowers produce considerable amounts of nectar and emit levels of CO(2) that should be detectable by moths nearby. By midnight, however, both nectar secretion and CO(2) release decrease significantly. Because nectar production requires high metabolic activity, high floral CO(2) emission may indicate food abundance to the moths. We suggest that hovering moths could use the florally emitted CO(2) to help them assess the nectar content before attempting to feed in order to improve their foraging efficiency.

Animal Feed↗

PubMed2004

Enantioselectivity of projection neurons innervating identified olfactory glomeruli.

Projection neurons (PNs) with arborizations in the sexually dimorphic "lateral large female glomerulus" (latLFG) in the antennal lobe (AL) of the moth Manduca sexta previously were shown to respond preferentially to antennal stimulation with (+/-)linalool, a volatile compound commonly emitted by plants. In the present study, using intracellular recording and staining techniques, we examined the responsiveness of latLFG-PNs to the enantiomers, (+)linalool and (-)linalool and found that (1) latLFG-PNs are more responsive to antennal stimulation with (+)linalool than with (-)linalool, (2) PNs with arborizations in a glomerulus adjacent to the latLFG are preferentially responsive to (-)linalool, and (3) PNs with arborizations confined to other glomeruli near the latLFG are equally responsive to both enantiomers of linalool. Structure-activity studies showed that the hydroxyl group in this tertiary terpene alcohol is the key feature of the molecule determining the response of enantioselective PNs to linalool. In contrast, the responses of non-enantioselective PNs are less dependent on the alcoholic functionality of linalool. Our findings show that PNs innervating a uniquely identifiable glomerulus respond preferentially to a particular enantiomer of an odor substance. Moreover, PNs with arborizations in a glomerulus adjacent to the latLFG, although less sensitive than latLFG-PNs to linalool, respond preferentially to the opposite enantiomer, demonstrating that information about stimulus-absolute configuration can be encoded in different olfactory glomeruli.

Acyclic Monoterpenes↗

PubMed2004

Floral CO2 reveals flower profitability to moths.

The hawkmoth Manduca sexta (Lepidoptera: Sphingidae), an experimentally favorable Lepidopteran that is highly sensitive to carbon dioxide (CO2), feeds on the nectar of a range of flowering plants, such as Datura wrightii (Solanaceae). Newly opened Datura flowers give off dramatically elevated levels of CO2 and offer ample nectar. Thus, floral CO2 emission could indicate food-source profitability. This study documents that foraging Manduca moths prefer surrogate flowers that emit high levels of CO2, characteristic of newly opened Datura flowers. We show for the first time that CO2 may play an important role in the foraging behavior of nectar-feeding insects.

Animals↗

PubMed2003

Coordination of central odor representations through transient, non-oscillatory synchronization of glomerular output neurons.

At the first stage of processing in the olfactory pathway, the patterns of glomerular activity evoked by different scents are both temporally and spatially dynamic. In the antennal lobe (AL) of some insects, coherent firing of AL projection neurons (PNs) can be phase-locked to network oscillations, and it has been proposed that oscillatory synchronization of PN activity may encode the chemical identity of the olfactory stimulus. It remains unclear, however, how the brain uses this time-constrained mechanism to encode chemical identity when the stimulus itself is unpredictably dynamic. In the olfactory pathway of the moth Manduca sexta,we find that different odorants evoke gamma-band oscillations in the AL and the mushroom body (a higher-order network that receives input from the AL), but oscillations within or between these two processing stages are not temporally coherent. Moreover, the timing of action potential firing in PNs is not phase-locked to oscillations in either the AL or mushroom body, and the correlation between PN synchrony and field oscillations remains low before, during, and after olfactory stimulation. These results demonstrate that olfactory circuits in the moth are specialized to preserve time-varying signals in the insect's olfactory space, and that stimulus dynamics rather than intrinsic oscillations modulate the uniquely coordinated pattern of PN synchronization evoked by each olfactory stimulus. We propose that non-oscillatory synchronization provides an adaptive mechanism by which PN ensembles can encode stimulus identity while concurrently monitoring the unpredictable dynamics in the olfactory signal that typically occur under natural stimulus conditions.

Animals↗

PubMed2003

Role of neurosecretory cells in the photoperiodic induction of pupal diapause of the tobacco hornworm Manduca sexta.

In the tobacco hornworm, Manduca sexta, pupal diapause can be induced by exposure of fifth-instar larvae to a short-day photoperiod. We studied the effect of surgical ablation of tissues containing the neurosecretory cells of the brain of fifth-instar larvae on the photoperiodic induction of pupal diapause. At the end of the experiments, we immunostained the neurosecretory cells to determine the success of the ablations. Under long-day conditions (LD 16:8 at 22 degrees C), all intact larvae, most of the sham-operated larvae, and control-operated larvae developed into nondiapausing pupae. Under short-day conditions (LD 10:14 at 22 degrees C), most intact, sham-operated, and control-operated larvae developed into diapausing pupae. Removal of type-II cells did not interfere with the photoperiodic response. Under long-day conditions, elimination of type-Ia(1) cells did not affect the incidence of nondiapausing pupae. When type-Ia(1) cells were removed under short-day conditions, however, the incidence of nondiapausing pupae was higher (51%, n = 41) than that of the intact (16%, n = 75), sham-operated (24%, n = 88), control-operated larvae (5%, n = 40), and larvae with type-II cells removed (11%, n = 27). Thus, removal of type-Ia(1) cells can impede induction of diapause. These results indicate that the type-Ia(1) neurosecretory cells have an important role in the induction of pupal diapause.

Animals↗

PubMed2003

Electroantennographic and behavioral responses of the sphinx moth Manduca sexta to host plant headspace volatiles.

Coupled gas chromatography with electroantennographic detection (GC-EAD) using antennae of adult female Manduca sexta was employed to screen for olfactory stimulants present in headspace collections from four species of larval host plants belonging to two families: Solanaceae--Lycopersicon esculentum (tomato), Capiscum annuum (bell pepper), and Datura wrightii; and Martyniaceae--Pronboscideaparviflora. Headspace volatiles were collected from undamaged foliage of potted, living plants. GC-EAD revealed 23 EAD-active compounds, of which 15 were identified by GC-mass spectrometry. Identified compounds included aliphatic, aromatic, and terpenoid compounds bearing a range of functional groups. Nine EAD-active compounds were common to all four host plant species: (Z)-3-hexenyl acetate, nonanal, decanal, phenylacetaldehyde, methyl salicylate, benzyl alcohol, geranyl acetone, (E)-nerolidol, and one unidentified compound. Behavioral responses of female moths to an eight-component synthetic blend of selected tomato headspace volatiles were tested in a laboratory wind tunnel. Females were attracted to the blend. A comparison of responses from antennae of males and females to bell pepper headspace volatiles revealed that males responded to the same suite of volatiles as females, except for (Z)-3-hexenyl benzoate. EAD responses of males also were lower for (Z)-and (E)-nerolidol and one unidentified compound. Electroantennogram EAG dose-response curves for the 15 identified EAD-active volatiles were recorded. At the higher test doses (10-100 microg), female antennae yielded larger EAG responses to terpenoids and to aliphatic and aromatic esters. Male antennae did respond to the higher doses of (Z)-3-hexenyl benzoate, indicating that they can detect this compound. On the basis of ubiquity of the EAD-active volatiles identified to date in host plant headspace collections, we suggest that M. sexta uses a suite of volatiles to locate and identify appropriate host plants.

Animals↗

PubMed2003

Localization of myoinhibitory peptide immunoreactivity in Manduca sexta and Bombyx mori, with indications that the peptide has a role in molting and ecdysis.

For normal development of Manduca sexta larvae, the ecdysteroid titer must drop following its sudden rise at the start of the molting cycle; this sudden decline in titer may be due to myoinhibitory peptide I (MIP I), which has an inhibitory effect on the release of ecdysone by the prothoracic glands of Bombyx mori in vitro. Using an antiserum to MIP, we have demonstrated secretion of an MIP-like peptide by the epiproctodeal glands of Manduca sexta, which are located on each proctodeal nerve, just anterior to the rectum. These MIP-immunoreactive glands are also present in B. mori. In fourth-instar larvae of M. sexta, the epiproctodeal glands show a distinct cycle of synthesis and sudden release of MIP that coincides with the time of the rapid decline in ecdysteroid titer. The function of the epiproctodeal glands appears to be the timely release of MIP during the molting cycle, so as to inhibit the prothoracic glands and thus to facilitate the sudden decline in ecdysteroid titer. In addition, we have found that MIP immunoreactivity is co-localized with that of crustacean cardioactive peptide (CCAP) in the 704 interneurons; these peptides appear to be co-released at the time of ecdysis. It is known that CCAP can initiate the ecdysis motor program; our results suggest that MIP may also be involved in activating ecdysis behavior.

Animals↗

PubMed2002

Pheromonal and host-odor processing in the insect antennal lobe: how different?

In the olfactory bulb of vertebrates and the antennal lobe of insects, precise connections between sensory receptor cells and olfactory glomeruli form the basis of a highly organized chemotopic map at the first stage of central processing in the brain. Beyond this basic level of organization, the olfactory system is typically separated into two subsystems: a 'main' olfactory pathway that detects and processes information about most environmental odorants, and an 'accessory' olfactory pathway that is devoted to information about social signals such as sex pheromones. A growing number of studies show, however, that it is not always possible to draw clear functional distinctions between the two subsystems. These findings have led some to speculate that the organizational principles by which olfactory stimuli are represented across glomeruli may be more similar in these two olfactory subsystems than previously thought.

Animals↗

PubMed2002

Local inhibition modulates odor-evoked synchronization of glomerulus-specific output neurons.

At the first stage of olfactory processing in the brain, synchronous firing across glomeruli may help to temporally bind multiple and spatially distributed input streams activated by a given odor. This hypothesis, however, has never been tested in an organism in which the odor-tuning properties of several spatially identifiable glomeruli are known. Using the sphinx moth, an insect that meets these specific criteria, we recorded odor-evoked responses simultaneously from pairs of projection neurons (PNs) innervating the same or different glomeruli in the macroglomerular complex (MGC), which is involved in processing pheromonal information. PNs that branched in the same glomerulus and were activated by the same pheromone component also showed the strongest coincident responses to each odor pulse. Glomerulus-specific PN pairs were also inhibited by the pheromone component that selectively activated PNs in the neighboring glomerulus, and about 70% of all intraglomerular pairs showed increased synchronization when stimulated with a mixture of the two odorants. Thus, when two adjacent glomeruli receive their inputs simultaneously, the temporal tuning of output from each glomerulus is enhanced by reciprocal and inhibitory interglomerular interactions.

Animals↗

PubMed2002

Developmental changes in the electrophysiological properties and response characteristics of Manduca antennal-lobe neurons.

Using whole cell patch-clamp recordings, we have examined changes in the electrophysiological properties and response characteristics of antennal lobe (AL) neurons associated with the metamorphic adult development of the sphinx moth, Manduca sexta. Whole cell current profiles and electrical excitability were examined in dispersed AL neurons in vitro, and in medial-group AL neurons in situ in semi-intact brain preparations. Around stages 2-4 of the 18 stages of metamorphic adult development, whole cell current profiles were dominated by large outward (K+) currents. Calcium-dependent action potentials could be elicited at this stage, but only a small percentage of cells exhibited sodium spikes. From stages 3 to 10, there was a rapid increase in the proportion of AL neurons exhibiting rapidly activating, transient sodium currents, and many cells in vitro exhibited spontaneous bursts of spike activity at this time. As development progressed, action-potential waveforms became shorter in duration and larger in amplitude. Cell-type-specific differences in the prevalence of spontaneous activity, and in the electrophysiological properties and response characteristics of AL neurons, were most apparent late in metamorphosis. While removal of antennal sensory input to the ALs early (stage 1-2) in metamorphosis had no detectable effect on the development of cell excitability, a significantly higher percentage of neurons in vitro from stage 4 pupae exhibited sodium-based action potentials following the addition of serotonin to the culture medium. Characteristic forms of electrical excitability in developing Manduca AL neurons, and their modulation by serotonin, seem likely to play a central role in the functional development of the ALs.

Animals↗

PubMed2002

Developmental changes in the density of ionic currents in antennal-lobe neurons of the sphinx moth, Manduca sexta.

Early in metamorphic adult development, action potentials elicited from Manduca sexta antennal lobe neurons are small in amplitude, long in duration, and calcium dependent. As development proceeds, the action potential waveform becomes larger in amplitude, shorter in duration, and increasingly sodium dependent. Whole cell voltage-clamp analysis of Manduca antennal-lobe neurons in vitro has been used to identify voltage-activated currents that contribute to developmental changes in the electrical excitability of these cells. Proximal Branching neurons [putative projection (output) neurons] and Rick Rack neurons (putative local antennal-lobe interneurons) are examined in detail early (pupal stage 5) and late (pupal stage 14) in adult metamorphosis. In both cell types, four voltage-gated and two calcium-dependent ionic currents have been identified. Cell-type-specific changes in the density of sodium, calcium, and potassium currents correlate temporally with changes in cell excitability and spike waveform. Developmental changes in ionic current profiles are accompanied also by the emergence of cell-type-specific response characteristics in the cells. Together with the accompanying paper, this study provides an important foundation for examining the impact of developmental changes in electrical excitability on the growth, electrical properties and connectivity of neurons in central olfactory pathways of the moth.

Action Potentials↗