Friedrich-Wilhelm Schürmann

Dopamine-like immunoreactivity in the bee brain

SummaryThe distribution of dopamine-like immunoreactive neurons is described for the brain of the bee, Apis mellifera L., following the application of a pre-embedding technique on Vibratome sections. Immunoreactive somata are grouped into seven clusters, mainly situated in the protocerebrum. Immunoreactive interneurons have been detected in the different neuropilar compartments, except for the optic lobe neuropils. Strong immunoreactivity is found in the upper division of the central body, in parts of the stalk and in the α-lobe layers of the mushroom bodies. A dense network of many immunoreactive fibres surrounds the mushroom bodies and the central body. It forms a number of interhemispheric commissures/chiasmata, projecting partly into the contralateral mushroom body and central body. The lateral protocerebral neuropil contains some large wide-field-neurons. The antennal-lobe glomeruli receive fine projections of multiglomerular dopamine-like immunoreactive interneurons.

F-actin at identified synapses in the mushroom body neuropil of the insect brain

The distribution of f‐actin stained by fluorescent phalloidin was investigated in the brain of several insect species, with a special focus on the mushroom body. For localizing f‐actin in identified neurons and at synapses, additional staining with fluorescent dextrans and anti‐synapsin I immunostaining was employed. Intense f‐actin staining was consistently found in synaptic complexes of the mushroom body calyces (calycal microglomeruli [MG]). These MG contain a central core of presynaptic boutons, predominantly belonging to deutocerebral cholinergic excitatory projection neurons, which are surrounded by a shell of numerous Kenyon cell (KC) dendritic tips. In the cricket Gryllus bimaculatus, high‐resolution confocal laser scanning imaging revealed colocalization of f‐actin with KC dendritic spine parts within MG. Although presynaptic boutons appear to be mainly devoid of f‐actin‐phalloidin fluorescence, there appears to be an accumulation of f‐actin in KC dendritic spines synaptically contacting the boutons. Electron microscopy of boutons and dextran‐stained KC dendrites revealed their pre‐ and postsynaptic sites, with KCs being strictly postsynaptic elements. Their subsynaptic membrane appositions are considered to be associated with f‐actin. Focal accumulation of f‐actin in the dendritic tips of KCs was found to be a general feature of MG, with either spheroidal or indented boutons of different sizes, as encountered in the mushroom bodies of the cricket, honey bee, ant, and fruit fly. The structural similarities of calycal MG and f‐actin accumulation in KC dendrites with cerebellar microglomeruli are considered comparatively. The accumulation of f‐actin in KC dendrites is discussed in view of mushroom body plasticity and its potential role in learning and memory formation. J. Comp. Neurol. 475:303–314, 2004.

Octopamine-immunoreactive neurons in the central nervous system of the cricket, Gryllus bimaculatus

SummaryThe distribution of octopamine-immunoreactive neurons is described using whole-mount preparations of all central ganglia of the cricket, Gryllus bimaculatus. Up to 160 octopamine-immunoreactive somata were mapped per animal. Medial unpaired octopamine-immunoreactive neurons occur in all but the cerebral ganglia and show segment-specific differences in number. The position and form of these cells are in accordance with well-known, segmentally-organized clusters of large dorsal and ventral unpaired medial neurons demonstrated by other techniques. In addition, bilaterally arranged groups of immunoreactive somata have been labelled in the cerebral, suboesophageal and terminal ganglia. A detailed histological description of octopamine-immunoreactive elements in the prothoracic ganglion is given. Octopamine-immunoreactive somata and axons correspond to the different dorsal unpaired medial cell types identified by intracellular single-cell staining. In the prothoracic ganglion, all efferent neurons whose primary neurites are found in the fibre bundle of dorsal unpaired cells are immunoreactive. Intersegmental octopamine-immunoreactive neurons are also present. Collaterals originating from dorsal intersegmental fibres terminate in different neuropils and fibre tracts. Fine varicose fibres have been located in several fibre tracts, motor and sensory neuropils. Peripheral varicose octopamine-immunoreactive fibres found on several nerves are discussed in terms of possible neurohemal releasing sites for octopamine.

Glutamate‐like immunoreactivity marks compartments of the mushroom bodies in the brain of the cricket

In the mushroom bodies of the brain of the cricket Gryllus bimaculatus, the distribution of glutamate‐like immunoreactivity is shown by using several immunocytochemical staining protocols and confocal and conventional microscopy. Glutamate‐like staining of intrinsic cells of mushroom bodies (Kenyon cells), their axons and projections, is demonstrated for the first time. Two types of Kenyon cells constituting distinct, separated populations within the perikaryal layer and in prominent neuropilar subcompartments exhibit strong (type III cells) or medium (type II cells) glutamate‐like immunoreactivity, whereas the small neurons of a central population (type I cells) lack staining above background. Type III Kenyon cells display a strong immunoreactivity similarly found in some giant neurons and in identified antennal motorneurons by using glutamate as an excitatory transmitter, indicating that also distinct populations of the Kenyon cells use glutamate as a putative transmitter. The pattern of glutamate‐like immunoreactivity in the mushroom bodies and in other parts of the brain is different from γ‐aminobutyric acid (GABA)‐like immunoreactivity (investigated for comparison). GABA‐like immunostaining is particularly prominent in the mushroom body calyces where Kenyon cells have their dendritic branchings. Differences in glutamate‐like immunostaining in Kenyon cell subpopulations, together with differences in their arborization and axonal projection patterns, indicate a functional diversity of these neurons. J. Comp. Neurol. 418:227–239, 2000.

Prothoracic DUM neurons of the cricket Gryllus bimaculatus - responses to natural stimuli and activity in walking behavior

SummaryResponses to sensory stimuli and spike activity uring walking were investigated in bilaterally symmetrical dorsal unpaired median (DUM) neurons of the cricket. Intracellular recordings within the prothoracic ganglion were made either in restrained animals or in stationary walking specimens whilst parameters of their intended locomotion were measured. Three types of DUM cells were distinguished morphologically and physiologically. DUMa neurons send axons through segmental nerves. They often generated spontaneously large action potentials with low frequencies. Most DUMa neurons showed multimodal sensitivity, preferentially to cereal wind puffs and 15 kHz sound. Mean latencies ranged from 25 to 349 ms. Their large intraindividual variability could be correlated with behavioral modes during walking. Generally, the spike frequency increased with increased forward speed, while it was not related to turning. DUMb neurons projected either through the anterior or posterior connectives, but seemed physiologically similar to DUMa neurons. DUMc neurons were H-shaped with axons in both pairs of connectives. No external stimulus led to discrete spikes, but the regular spontaneous activity was modulated following cereal wind puffs to a restrained animal. During wind evoked escape the spike activity of another DUMc cell was modulated in phase with the rhythmic running behavior. The possibly different functions of DUMa and DUMc neurons during walking are discussed.

Effects of Serotonergic and Opioidergic Drugs on Escape Behaviors and Social Status of Male Crickets

Abstract We examined the effects of selective serotonin depletion and opioid ligands on social rank and related escape behavior of the cricket Gryllus bimaculatus. Establishment of social rank in a pair of males affected their escape reactions. Losers showed a lower and dominants a higher percentage of jumps in response to tactile cercal stimulation than before a fight. The serotonin-depleting drug α-methyltryptophan (AMTP) caused an activation of the escape reactivity in socially naive crickets. AMTP-treated animals also showed a lower ability to become dominants. With an initial 51.6±3.6% of wins in the AMTP group, the percentage decreased to 26±1.6% on day 5 after injection. The opiate receptor antagonist naloxone affected fight and escape similarly as AMTP. In contrast to naloxone, the opioid agonist [d-Ala2, N-Me-Phe4, Gly5-ol]-enkephalin decreased escape responsiveness to cercal stimulation in naive and subordinate crickets. We suggest that serotonergic and opioid systems are involved in the dominance induced depression of escape behavior.

Dense-core vesicles and non-synaptic exocytosis in the central body of the crayfish brain

SummaryThe central body in the median protocerebrum of the brain of the crayfish Cherax destructor is a distinctive area of dense neuropile, the nerve fibres of which contain three main types of vesicles: electronlucent vesicles (diameter 35 nm), dense-core vesicles (diameter 64 nm), and large structured dense-core vesicles (diameter 98 nm, maximum 170 nm). Different vesicle types were found together in the same neurons. Electronlucent vesicles were seen at presynaptic sites and rarely observed in the state of exocytosis. Exocytosis of densecore and structured dense-core vesicles was a regular feature on non-synaptic release sites either close to, or at some distance from pre- and subsynaptic sites. Non-synaptic exocytotic sites are more often observed than chemical synapses. Different forms of exocytosis seen at non-synaptic sites included the release of single densecore vesicles, packets of dense-core vesicles, and rows of dense-core vesicles lined up along cell membranes and around fibre invaginations. Swelling and the enhanced electron density of extracellular non-synaptic spaces may mark the positions of prior exocytotic events. In vitro treatment of the brain with tannic acid buffer solution followed by conventional double fixation resulted in the augmentation of non-synaptic exocytosis. Electron microscopy of proctolin- and serotonin-immunoreactive nerve fibres shows them to contain dense-core and electron-lucent vesicles and to be surrounded by many unlabelled profiles similarly laden with dense-core vesicles and electron-lucent vesicles, indicating the presence of other, not yet identified, neuroactive compounds.

Putative neurohemal areas in the peripheral nervous system of an insect, Gryllus bimaculatus, revealed by immunocytochemistry

The morphology and position of putative neurohemal areas in the peripheral nervous system (ventral nerve cord and retrocerebral complex) of the cricket Gryllus bimaculatus are described. By using antisera to the amines dopamine, histamine, octopamine, and serotonin, and the neuropeptides crustacean cardioactive peptide, FMRFamide, leucokinin 1, and proctolin, an extensive system of varicose fibers has been detected throughout the nerves of all neuromeres, except for nerve 2 of the prothoracic ganglion. Immunoreactive varicose fibers occur mainly in a superficial position at the neurilemma, indicating neurosecretory storage and release of neuroactive compounds. The varicose fibers are projections from central or peripheral neurons that may extend over more than one segment. The peripheral fiber varicosities show segment-specific arrangements for each of the substances investigated. Immunoreactivity to histamine and octopamine is mainly found in the nerves of abdominal segments, whereas serotonin immunoreactivity is concentrated in subesophageal and terminal ganglion nerves. Immunoreactivity to FMRFamide and crustacean cardioactive peptide is widespread throughout all segments. Structures immunoreactive to leucokinin 1 are present in abdominal nerves, and proctolin immunostaining is found in the terminal ganglion and thoracic nerves. Codistribution of peripheral varicose fiber plexuses is regularly seen for amines and peptides, whereas the colocalization of substances in neurons has not been detected for any of the neuroactive compounds investigated. The varicose fiber system is regarded as complementary to the classical neurohemal organs.

Types, numbers and distribution of synapses on the dendritic tree of an identified visual interneuron in the brain of the locust.

Abstract The descending contralateral movement detector (DCMD), an identified descending interneuron in the brain of the locust Schistocerca gregaria has been investigated by using light and electron microscopy. We describe the fine structure, distribution and numbers of synapes that it receives from another identified brain neuron, the lobular giant movement detector (LGMD), and from unidentified neurons. The DCMD dendrites emerging from the integrative segment vary in form and number between individuals and sexes but always form a flattened dendritic domain. The arborizations and the integrative segment appear to be exclusively postsynaptic. Two types of synaptic contacts (Type 1 and 2) onto the DCMD can be discerned as having either round (Type 1) or pleiomorphic synaptic vesicles (Type 2) and by large (Type 1) or small (Type 2) subsynaptic appositions. Contact zones of Type 1 synapses are smaller than those of Type 2. LGMD-synapses are of Type 1 and occur intermingled with presynaptic sites of unidentified units. Some branches of the DCMD receiving input from unidentified units are devoid of contacting LGMD processes. Synapses of both types are randomly distributed over the DCMD integrative segment and at fibres with similar sizes.Type 1 synapses are much more frequent than Type 2 synapses and their number is negatively correlated with fibre diameter. For a whole DCMD dendritic arborization, a total of 8500 active zones of chemical synapses has been calculated, including a mininum of 2250 LGMD-synapses and about 1000 Type 2 synapses. The DCMD may thus receive a considerable amount of input from as yet unidentified neurons.

The distribution of neurones immunoreactive for β-tyrosine hydroxylase, dopamine and serotonin in the ventral nerve cord of the cricket, Gryllus bimaculatus

The cellular localization of the biogenic amines dopamine and serotonin was investigated in the ventral nerve cord of the cricket, Gryllus bimaculatus, using antisera raised against dopamine, β-tyrosine hydroxylase and serotonin. Dopamine-(n<-70) and serotonin-immunoreactive (n<-120) neurones showed a segmental arrangement in the ventral nerve cord. Some neuromeres, however, did not contain dopamine-immunoreactive cell bodies. The small number of stained cells allowed complete identification of brain and thoracic cells, including intersegmentally projecting axons and terminal arborizations. Dopamine-like immunostaining was found primarily in plurisegmental interneurones with axons descending to the soma-ipsilateral hemispheres of the thoracic and abdominal ganglia. In contrast, serotonin-immunostaining occurred predominantly in interneurones projecting via soma-contralaterally ascending axons to the thorax and brain. In addition, serotonin-immunoreactivity was also present in efferent cells and afferent elements. Serotonin-immunoreactive, but no dopamine-immunoreactive, varicose fibres were observed on the surface of some peripheral nerves. Varicose endings of both dopamine-and serotonin-immunoreactive neurones occurred in each neuromere and showed overlapping neuropilar projections in dorsal and medial regions of the thoracic ganglia. Ventral associative neuropiles lacked dopamine-like immunostaining but were innervated by serotonin-immunoreactive elements. A colocalization of the two amines was not observed. The topographic representation of neurone types immunoreactive for serotonin and dopamine is discussed with respect to possible modulatory functions of these biogenic amines in the central nervous system of the cricket.