Neural regulation of a complex behavior: body patterning in cephalopod molluscs

Unshelled cephalopods have a remarkable ability to alter theirappearance, using textural, postural, and chromatic elementsto generate a myriad of body patterns. Of the unshelled cephalopods,it is generally acknowledged that cuttlefish express the mostdetailed and widest range of body patterns, including staticand dynamic patterns. In this paper we present data on the neuronalmechanisms underlying this amazing behavior, focusing on theneuroregulation of the chromatic elements, the chromatophoreorgans, in the European cuttlefish Sepia officinalis. Cephalopodchromatophore organs, including those in Sepia, are unlike thosein any other animal taxa; each consists of a pigment-containingchromatophore cell that expands in response to the coordinatedactivation of a set of radial muscles which are directly attachedto the chromatophore cell. We show that the chromatophore musclesare regulated by 2 different excitatory transmitters, glutamateand the family of FMRFamide-related peptides (FaRPs). Glutamatemediates rapid and transient chromatophore cell expansion whereasthe FaRPs are responsible for slower, more sustained responses.Using retrograde dye filling, immunocytochemical and in situhybridization techniques, we demonstrate that the cell bodiesof the glutamatergic and FaRPs-containing motoneurons innervatingthe fin chromatophore muscles are primarily localized to theposterior chromatophore and fin lobes in the posterior subesophagealmass of the Sepia brain. Data are also presented showing thatsome fin chromatophore motoneurons have multiple axons in differentnerve branches, which accounts for overlapping chromatophoremotor fields by adjacent peripheral nerves.     

Aminergic neurons in the brain of blowflies and Drosophila: dopamine- and tyrosine hydroxylase-immunoreactive neurons and their relationship with putative histaminergic neurons

Summary The distribution and morphology of neurons reacting with antisera against dopamine (DA), tyrosine hydroxylase (TH) and histamine (HA) were analyzed in the blowflies Calliphora erythrocephala and Phormia terraenovae. TH-immunoreactive (THIR) and HA-immunoreactive (HAIR) neurons were also mapped in the fruitfly Drosophila melanogaster. The antisera against DA and TH specifically labeled the same neurons in the blowflies. About 300 neurons displayed DA immunoreactivity (DAIR) and THIR in the brain and subesophageal ganglion of the blowflies. Most of these neurons were located in bilateral clusters; some were distributed as bilateral pairs, and two ventral unpaired median (VUM) neurons were seen in the subesophageal ganglion. Immunoreactive processes were found in all compartments of the mushroom bodies except the calyces, in all divisions of the central body complex, in the medulla, lobula and lobula plate of the optic lobe, and in non-glomerular neuropil of protocerebrum, tritocerebrum and the subesophageal ganglion. No DA or TH immunoreactivity was seen in the antennal lobes. In Drosophila, neurons homologous to the blowfly neurons were detected with the TH antiserum. In Phormia and Drosophila, 18 HA-immunoreactive neurons were located in the protocerebrum and 2 in the subesophageal ganglion. The HAIR neurons arborized extensively, but except for processes in the lobula, all HAIR processes were seen in non-glomerular neuropil. The deuto- and tritocerebrum was devoid of HAIR processes. Double labeling experiments demonstrated that TH and HA immunoreactivity was not colocalized in any neuron. In some regions there wasm however, substantial superposition between the two systems. The morphology of the extensively arborizing aminergic neurons described suggests that they have modulatory functions in the brain and subesophageal ganglion.     

Early scale development in Heterodontus (Heterodontiformes; Chondrichthyes): a novel chondrichthyan scale pattern

In two species of Heterodontus, H. portusjacksoni and H. galeatus, the first scales to develop form two opposing rows along the caudal fin axis on both the left and right sides of the fin. The opposing rows originate from an initial scale located on either side of the posterior tip of the caudal fin, with subsequent scales erupting in a posterior to anterior direction along the tail axis. These scale rows may strengthen tail movements, providing aeration in the egg case, but are lost later in ontogeny. Development of subsequent body scales shows a more irregular origin and arrangement, from anterior to posterior, to cover the dorsal and ventral lobes of the caudal fin. Although the early developmental pattern of the scale associated with the Heterodontus caudal fin has not been previously described, several chondrichthyan taxa, including chimeroids, likewise possess ordered rows of flank scales early in ontogeny that are subsequently lost. These ordered scales contrast with previous suggestions that chondrichthyan scale development is entirely random. Instead, regulated and sequential development of scales may be a plesiomorphic character for both chondrichthyans and osteichthyans, with the less organized arrangement in later ontogenetic stages being a derived condition within Chondrichthyes.     

The nervous system of Loligo. II. Suboesophageal centres.

A well-marked hierarchy of centres can be recognized within the suboesophageal lobes and ganglia of the arms. The inputs and outputs of each lobe are described. There are sets of motoneurons and intermediate motor centres, which can be activated either from the periphery or from above. They mostly do not send fibres up to the optic or higher motor centres. However, there is a large set of fibres running from the magnocellular lobe to all the basal supraoesophageal lobes. The centre for control of the four eye-muscle nerves in the anterior lateral pedal lobe receives many fibres direct from the statocyst and from the peduncle and basal lobes, but none direct from the optic lobe. The posterior lateral pedal is a backward continuation of the oculomotor centre, containing large cells that may be concerned in initiating attacks by the tentacles. An intermediate motor centre in the posterior pedal lobe probably controls steering. It sends fibres to the funned and head retractors, and by both direct and interrupted pathways to the fin lobe. It receives fibres from the crista nerve and basal lobes, but none direct from the optic lobe. The jet control centre of the ventral magnocellular lobe receives fibres from the statocyst and skin and also from the optic and basal lobes. Some of these last also give extensive branches throughout the palliovisceral lobes. The branching patterns of the dendritic collaterals differ in the various lobes. Some estimates are given of the numbers of synaptic points. The dendritic collaterals of the motoneurons spread through large volumes of neuropil and they overlap. The incoming fibres spread widely and each presumably activates many motoneurons either together or serially. Many of the lobes contain numerous microneurons with short trunks restricted to the lobe, but there are none of these cells in the chromatophore lobes or fin lobes. The microneurons have only few dendritic collaterals, in contrast to the numerous ones on the nearby motoneurons.     

Analysis of neurotransmitter distribution in brain development of benthic and pelagic octopod cephalopods

The database on neurotransmitter distribution during central nervous system development of cephalopod mollusks is still scarce. We describe the ontogeny of serotonergic (5‐HT‐ir) and FMRFamide‐like immunoreactive (Fa‐lir) neurons in the central nervous system of the benthic Octopus vulgaris and Fa‐lir distribution in the pelagic Argonauta hians. Comparing our data to previous studies, we aim at revealing shared immunochemical domains among coleoid cephalopods, i.e., all cephalopods except nautiluses. During development of O. vulgaris, 5‐HT‐ir and Fa‐lir elements occur relatively late, namely during stage XII, when the brain neuropils are already highly differentiated. In stage XII‐XX individuals, Fa‐lir cell somata are located in the middle and posterior subesophageal mass and in the optic, posterior basal, and superior buccal lobes. 5‐HT is predominately expressed in cell somata of the superior buccal, anterior basal, and optic lobes, as well as in the subesophageal mass. The overall population of Fa‐lir neurons is larger than the one expressing 5‐HT. Fa‐lir elements are distributed throughout homologous brain areas of A. hians and O. vulgaris. We identified neuronal subsets with similar cell number and immunochemical phenotype in coleoids. These are located in corresponding brain regions of developmental stages and adults of O. vulgaris, A. hians, and the decapod squid Idiosepius notoides. O. vulgaris and I. notoides exhibit numerous 5‐HT‐ir cell somata in the superior buccal lobes but none or very few in the inferior buccal lobes. The latter have previously been homologized to the gastropod buccal ganglia, which also lack 5‐HT‐ir cell somata in euthyneuran gastropods. Among coleoids, 5‐HT‐ir neuronal subsets, which are located ventrally to the lateral anterior basal lobes and in the anterior middle subesophageal mass, are candidates for homologous subsets. Contrary to I. notoides, octopods exhibit Fa‐lir cell somata ventrally to the brachial lobes and 5‐HT‐ir cell somata close to the stellate ganglia. J. Morphol., 2012. © 2012 Wiley Periodicals, Inc.     

Molecular analysis of a novel FMRFamide-related peptide gene (SOFaRP(2)) and its expression pattern in the brain of the European cuttlefish Sepia officinalis

FMRFamide-related peptides (FaRPs) are among several neurotransmitters known to regulate the chromatophore function in the European cuttlefish Sepia officinalis. Here we report the cloning and sequencing of a novel S. officinalis FaRP gene (SOFaRP(2)). The complete 835-base pair cDNA sequence of the SOFaRP(2) gene contains an open reading frame of 567 base pairs encoding 188 amino acids and four putative FaRPs, NSLFRFamide, GNLFRFamide, TIFRFamide and PHTPFRFamide. All except TIFRFamide cause chromatophore expansion when assayed in an in vitro chromatophore bioassay. To investigate the expression pattern of SOFaRP(2) gene in the cuttlefish brain, in situ hybridization was performed using a full length RNA probe. The SOFaRP(2) gene was expressed primarily in the posterior chromatophore, anterior chromatophore, lateral basal and optic lobes among other brain locations. The SOFaRP(2) gene appears to be expressed in all brain regions involved in chromatophore regulation. These data suggests that some or all of the four FaRPs encoded by SOFaRP(2) might be involved in controlling chromatophore activity in cuttlefish.     

Central projections of first-order ocellar interneurons in the bug Triatoma infestans (Heteroptera: Reduviidae)

The projections of first-order ocellar interneurons were analyzed in the hematophagous bug Triatoma infestans by cobalt filling. The axons run between the calyces of the mushroom bodies and dorsal of the central body to different regions of the brain and the subesophageal and thoracic ganglia. The interneurons can be grouped into large L cells and small S cells. The L cells have cell bodies ranging from 11.5 to 25 μm and axons ranging from 8 to 25 μm diameter (measured in the ocellar nerve); the S cells have smaller cell bodies of 9 μm or less and axon diameters less than 5 μm. The projections of ten L cells are described in detail; they project to the protocerebral posterior slope (PS), the other ocellus (O), the optic neuropile, and the subesophageal, pro-, meso-, and metathoracic ganglia, either to ipsi- (PS I, II), or contra- (PS IV, V), or bilateral areas. In this case projections occur to the same areas (PSO, PS III) or different areas at each side (PSOE; E = eye). Large-descending (LD) first-order interneurons project to the contralateral posterior slope of the protocerebrum, the deutocerebrum, and subesophageal, pro-, mesa-, and metathoracic areas (LD I-III). Cell bodies are located in the dorsal protocerebral lobes and pars intercerebralis, except the PS II neuron and three LD cells, which are located in the ipsilateral posterior protocerebrum. This is the first report about ocellar pathways in Hemiptera. Their adaptive function is discussed with reference to the bugs' behavior as Chagas disease vectors. © 1996 Wiley-Liss, Inc.     

Glomerular bypass shunts and distribution of glomeruli in the kidney of the lesser spotted dogfish,Scyliorhinus caniculus

Summary Scanning electron microscopy of corroded resin casts of the renal vasculature of Scyliorhinus caniculus has revealed a novel vascular pathway arising from the afferent arteriole and bypassing the glomerulus. This glomerulus bypass shunt occurred in 36% of the glomerular casts examined. The shunt ran to join a peritubular network of capillaries and thereby offers the potential to vary the degree of glomerular perfusion and control the proportion of active glomeruli. In 29% of glomeruli two efferent arterioles drained the capillary knot. Glomeruli were located close to the dorsal margin of the posterior mass of the kidney, and towards the lateral edge of the anterior lobes of the kidney of female dogfish. In male dogfish, glomeruli were evenly distributed through the posterior mass of kidney, while in female dogfish 89% of glomeruli occurred in the posterior mass and 11% of glomeruli were located within the small anterior lobes.     

Crustacean cardioactive peptide-immunoreactive neurons innervating brain neuropils,retrocerebral complex and stomatogastric nervous system of the locust,Locusta migratoria

The distribution and morphology of crustacean cardioactive peptide-immunoreactive neurons in the brain of the locust Locusta migratoria has been determined. Of more than 500 immunoreactive neurons in total, about 380 are interneurons in the optic lobes. These neurons invade several layers of the medulla and distal parts of the lobula. In addition, a small group of neurons projects into the accessory medulla, the lamina, and to several areas in the median protocerebrum. In the midbrain, 12 groups or individual neurons have been reconstructed. Four groups innervate areas of the superior lateral and ventral lateral protocerebrum and the lateral horn. Two cell groups have bilateral arborizations anterior and posterior to the central body or in the superior median protocerebrum. Ramifications in subunits of the central body and in the lateral and the median accessory lobes arise from four additional cell groups. Two local interneurons innervate the antennal lobe. A tritocerebral cell projects contralaterally into the frontal ganglion and appears to give rise to fibers in the recurrent nerve, and in the hypocerebral and ingluvial ganglia. Varicose fibers in the nervi corporis cardiaci III and the corpora cardiaca, and terminals on pharyngeal dilator muscles arise from two subesophageal neurons. Some of the locust neurons closely resemble immunopositive neurons in a beetle and a moth. Our results suggest that the peptide may be (1) a modulatory substance produced by many brain interneurons, and (2) a neurohormone released from subesophageal neurosecretory cells.     

Cephalopod chromatophores: neurobiology and natural history

The chromatophores of cephalopods differ fundamentally from those of other animals: they are neuromuscular organs rather than cells and are not controlled hormonally. They constitute a unique motor system that operates upon the environment without applying any force to it. Each chromatophore organ comprises an elastic sacculus containing pigment, to which is attached a set of obliquely striated radial muscles, each with its nerves and glia. When excited the muscles contract, expanding the chromatophore; when they relax, energy stored in the elastic sacculus retracts it. The physiology and pharmacology of the chromatophore nerves and muscles of loliginid squids are discussed in detail. Attention is drawn to the multiple innervation of dorsal mantle chromatophores, of crucial importance in pattern generation. The size and density of the chromatophores varies according to habit and lifestyle. Differently coloured chromatophores are distributed precisely with respect to each other, and to reflecting structures beneath them. Some of the rules for establishing this exact arrangement have been elucidated by ontogenetic studies. The chromatophores are not innervated uniformly: specific nerve fibres innervate groups of chromatophores within the fixed, morphological array, producing 'physiological units' expressed as visible 'chromatomotor fields'. The chromatophores are controlled by a set of lobes in the brain organized hierarchically. At the highest level, the optic lobes, acting largely on visual information, select specific motor programmes (i.e. body patterns); at the lowest level, motoneurons in the chromatophore lobes execute the programmes, their activity or inactivity producing the patterning seen in the skin. In Octopus vulgaris there are over half a million neurons in the chromatophore lobes, and receptors for all the classical neurotransmitters are present, different transmitters being used to activate (or inhibit) the different colour classes of chromatophore motoneurons. A detailed understanding of the way in which the brain controls body patterning still eludes us: the entire system apparently operates without feedback, visual or proprioceptive. The gross appearance of a cephalopod is termed its body pattern. This comprises a number of components, made up of several units, which in turn contains many elements: the chromatophores themselves and also reflecting cells and skin muscles. Neural control of the chromatophores enables a cephalopod to change its appearance almost instantaneously, a key feature in some escape behaviours and during agonistic signalling. Equally important, it also enables them to generate the discrete patterns so essential for camouflage or for signalling. The primary function of the chromatophores is camouflage. They are used to match the brightness of the background and to produce components that help the animal achieve general resemblance to the substrate or break up the body's outline. Because the chromatophores are neurally controlled an individual can, at any moment, select and exhibit one particular body pattern out of many. Such rapid neural polymorphism ('polyphenism') may hinder search-image formation by predators. Another function of the chromatophores is communication. Intraspecific signalling is well documented in several inshore species, and interspecific signalling, using ancient, highly conserved patterns, is also widespread. Neurally controlled chromatophores lend themselves supremely well to communication, allowing rapid, finely graded and bilateral signalling.     

Die Zweiteiligkeit der Chromatophoren bei Cryptomonaden

In theChrysophyceae as well as in different species ofCryptomonas bilobed chromatophores are present. These chromatophores consist of two large parietal lobes closed to the lateral sides of the cell and joined by a narrow bridge on its dorsal part. A survey of all species with a single bilobed chromatophore is given. Besides, also species with two separate chromatophores have been found. The presence of several chromatophores inCryptomonas cells is doubtful. The morphology of the chromatophores has to be taken into consideration in the taxonomy ofCryptomonas.

     

Distribution of APGWamide-immunoreactivity in the brain and reproductive organs of adult Pygmy squid, Idiosepius pygmaeus

The neuropeptide APGWamide is involved in the control of the reproductive behavior in molluscs. Using immunocytochemistry, we investigated the distribution of APGWa-immunoreactive neurons in the brain and reproductive organs of adult male and female specimen of Idiosepius pygmaeus. The study showed that the APGWamide-immunoreactive neurons and fibers are localized in the dorsal basal and vertical lobes of the supraesophageal mass, the palliovisceral lobe of the posterior subesophageal mass and olfactory lobe of the optic tract in male brains, with the highest number of APGWamide-immunoreactive neurons in the palliovisceral and olfactory lobes. In females, only the palliovisceral and olfactory lobes contained APGWa-immunoreactive neurons. The number of APGWamide-immunoreactive neurons in male I. pygmaeus brain is significantly higher than in females. Furthermore, APGWamide-immunoreactive fibers are localized exclusively in male reproductive organs and mantle muscles. Together these data suggest a role for APGW-amide in the control of male reproduction.     

Serotonin-like immunoreactivity in the central nervous system of two ixodid tick species

Immunocytochemistry was used to describe the distribution of serotonin-like immunoreactive (5HT-IR) neurons and neuronal processes in the central nervous system (CNS), the synganglion, of two ixodid tick species; the winter tick, Dermacentor albipictus and the lone star tick, Amblyomma americanum. 5HT-IR neurons were identified in the synganglion of both tick species. D. albipictus had a significantly higher number of 5HT-IR neurons than A. americanum. The labeling pattern and number of 5HT-IR neurons were significantly different between sexes in D. albipictus, but were not significantly different between sexes in A. americanum. 5HT-IR neurons that were located in the cortex of the synganglion projected processes into the neuropils, invading neuromeres in the supraesophageal ganglion including the protocerebrum, postero-dorsal, antero-dorsal and cheliceral neuromeres. In the subesophageal ganglion, dense 5HT-IR neuronal processes were found in the olfactory lobes, pedal, and opisthosomal neuromeres. Double-labeling with neurobiotin backfilled from the first leg damaged at the Haller’s organ revealed serotoninergic neuronal processes surrounding the glomeruli in the olfactory lobes. The high number of the 5HT-IR neurons and the extensive neuronal processes present in various regions of the synganglion suggest that serotonin plays a significant role in tick physiology. This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or a recommendation by the USDA for its use. The U.S. Government’s right to retain a non-exclusive, royalty free license in and to any copyright is acknowledged.     

Interneurons of the subesophageal ganglion of Sarcophaga bullata responding to gustatory and mechanosensory stimuli

Summary Intracellular recordings were made from interneurons in the subesophageal ganglion (SEG) of Sarcophaga bullata while stimulating the labellar lobes with solutions of sucrose, NaCl and with distilled water. Neurons that responded to sucrose did not respond to NaCl and vice versa, while sucrose-sensitive neurons often responded weakly to water. Several of the recorded neurons were filled with Lucifer Yellow, and their morphology was reconstructed. Most showed extensive arborizations within the SEG, suggesting that they were local interneurons involved in the early stages of gustatory processing. Some of the filled neurons had extensive projections to the brain, in addition to arborizations in the SEG. This is the first published record of gustatory interneurons in the higher flies.Abbreviations LY lucifer yellow - SEG subesophageal ganglion     

On the fine structure of the Octopus iris

Summary The Octopus iris is composed of five different layers: A, the external epithelium; B, the chromatophore layer; C, the iridocyte layer; D, the layer of muscles and collagen strands; E, the pigment epithelium. The nerves innervating the sphincter and the chromatophore muscles are identified and their neuromuscular junction is described. The motor endings of chromatophore nerves have an additional ending in presynaptic position which probably functions as a modifier of neuromuscular transmission. The chromatophores are naked and exhibit a tubular channel system between plasmalemma and pigment container which looks similar to the T-system of muscle cells.The financial support of this investigation by the Swiss National Foundation is gratefully acknowledged.     

Assembly of plasmid DNA and chromatophore inRhodospirillum rubrum

Summary Rhodospirillum rubrum, a photosynthetic bacterium, contains many photosynthetic vesicular membranous structures called chromatophores. The organism contains a 55 kb specific plasmid which is essential for photosynthesis, but the exact relationship between the chromatophore and the plasmid is uncertain. In this study we examined the precise localization of the plasmids, especially in relation to the chromatophores. Fluorescence in situ hybridization indicated that there are several copies of the plasmid per cell and that some plasmids are localized close to the cellular envelope. In situ hybridization at the electron-microscopic level further revealed that the plasmid localized to the periphery of the chromatophore close to the envelope. Moreover, when the chromatophore fraction was purified from cells, the plasmid DNA was observed as a cluster around the chromatophore vesicles. The assembly of the plasmid and chromatophore may be related to chromatophore formation by invagination of cell membrane.     

CNS microstructure in the wandering wolf spider Arctosa kwangreungensis (Araneae: Lycosidae)

The supraesophageal ganglion of the wolf spider Arctosa kwangreungensis is made up of a protocerebral and tritocerebral ganglion, whereas the subesophageal ganglionic mass is composed of a single pair of pedipalpal ganglia, four pairs of appendage ganglia, and a fused mass of abdominal neuromeres. In the supraesophageal ganglion, complex neuropile masses are located in the protocerebrum which include optic ganglia, the mushroom bodies, and the central body. Characteristically, the only nerves arising from the protocerebrum are the optic nerves, and the neuropiles of the principal eyes are the most thick and abundant in this wandering spider. The central body which is recognized as an important association center is isolated at the posterior of the protocerebrum and appears as a complex of highly condensed neurons. These cells give off fine parallel bundles of axons arranged in the mushroom bodies. The subesophageal nerve mass can be divided into two main tracts on the basis of direction of the neuropiles. The dorsal tracts are contributed to from the motor or interneurons of each ganglion, whereas the ventral tracts are from incoming sensory axons.     

Evolution of the arthropod neuromuscular system. 2. Inhibitory innervation of the walking legs of a scorpion: Vaejovis spinigerus (Wood, 1863), Vaejovidae, Scorpiones, Arachnida

Inhibitory motoneurons which supply the leg musculature are identified and characterized in the scorpion, Vaejovis spinigerus (Wood, 1863) (Vaejovidae, Scorpiones, Arachnida). (1) Successive intracellular muscle fiber recordings from antagonists, and correlation of the monitored inhibitory postsynaptic potentials with spikes in motor nerves, suggest supply of the scorpion leg musculature by common inhibitory motoneurons. (2) Anti-GABA immunohistochemistry is combined with transmission electron microscopy to estimate the number of inhibitory motor axons present in the main leg nerve. The number of immunoreactive axons decreases toward more distal leg segments, from 14 to 18 in the basis to 6-8 in the tibia. No immunoreactive axons are detected beyond the tibia. (3) The distribution of putative inhibitory neurons in the subesophageal ganglion mass is determined by anti-GABA immunohistochemistry, revealing notable similarities to the situation in pterygote insects. This provides a framework for the characterization of the inhibitory motoneurons. (4) Backfills from leg nerves are combined with anti-GABA immunocytochemistry to identify inhibitory motoneurons in the central nervous system. Putative inhibitory motoneurons occur in three clusters per hemi-segment. Two clusters are located near the posterior edge of the neuromere, one lateral, the other more medial, and both contain ca. 8-10 cell bodies. The third cluster consists of two somata located contralaterally, just off the ganglion midline.     

Liopropoma dorsoluteum sp. nov., a new serranid fish from Okinawa, Japan

A new serranid fish,Liopropoma dorsoluteum sp. nov., is described on the basis of two specimens from Yaeyama Is., Okinawa, Japan. The new species is most similar toL. erythaeum Randall & Taylor, 1988, in having the following characters: Dorsal fin rays VIII, 12; anal fin rays III, 9; pored lateral line scales 52–53; anterior nostril situated midway between posterior nostril and anterior tip of snout; slightly forked caudal fin with both lobes rounded. It differs from the latter species in having a shorter pectoral fin (23.4–23.8% SL vs. 26.9–29.0% SL), greater preanus length (65.6–68.0% SL vs. 63.3–65.1% SL), fewer gill rakers (6+12 vs. 6–7+14–15) and yellow coloration on the back (vs. light red on head and body) in fresh specimens.     

Distribution of the octopamine receptor AmOA1 in the honey bee brain

Octopamine plays an important role in many behaviors in invertebrates. It acts via binding to G protein coupled receptors located on the plasma membrane of responsive cells. Several distinct subtypes of octopamine receptors have been found in invertebrates, yet little is known about the expression pattern of these different receptor subtypes and how each subtype may contribute to different behaviors. One honey bee (Apis mellifera) octopamine receptor, AmOA1, was recently cloned and characterized. Here we continue to characterize the AmOA1 receptor by investigating its distribution in the honey bee brain. We used two independent antibodies produced against two distinct peptides in the carboxyl-terminus to study the distribution of the AmOA1 receptor in the honey bee brain. We found that both anti-AmOA1 antibodies revealed labeling of cell body clusters throughout the brain and within the following brain neuropils: the antennal lobes; the calyces, pedunculus, vertical (alpha, gamma) and medial (beta) lobes of the mushroom body; the optic lobes; the subesophageal ganglion; and the central complex. Double immunofluorescence staining using anti-GABA and anti-AmOA1 receptor antibodies revealed that a population of inhibitory GABAergic local interneurons in the antennal lobes express the AmOA1 receptor in the cell bodies, axons and their endings in the glomeruli. In the mushroom bodies, AmOA1 receptors are expressed in a subpopulation of inhibitory GABAergic feedback neurons that ends in the visual (outer half of basal ring and collar regions) and olfactory (lip and inner basal ring region) calyx neuropils, as well as in the collar and lip zones of the vertical and medial lobes. The data suggest that one effect of octopamine via AmOA1 in the antennal lobe and mushroom body is to modulate inhibitory neurons.