Dick R. Nässel

Histamine-like immunoreactivity in photoreceptors of the compound eyes and ocelli of the flies Calliphora erythrocephala and Musca domestica

SummaryAntibodies to histamine were used for immunocytochemical studies of the visual system in the flies Calliphora erythrocephala and Musca domestica. Specific immunolabeling of photoreceptors was found both in the compound eyes and ocelli of both species. In the compound eyes histamine-like immunoreactivity (HA-IR) was found in all the short visual fibers (photoreceptors R1–6) and one type of long visual fiber (photoreceptor R8). In addition, the ocellar photoreceptors also show HA-IR. In view of earlier biochemical and pharmacological/physiological findings by Elias and Evans (1983) and Hardie (1987) it thus seems likely that histamine is a neurotransmitter in insect photoreceptors. Interestingly, the second type of long visual fiber (photoreceptor R7) has recently been found to be GABA-immunoreactive (Datum et al. 1986). The two types of long visual fibers may hence use different transmitters which act on different receptors of the postsynaptic neurons in the second visual neuropil, the medulla. In addition to the photoreceptors in the retina and ocelli, we found processes of HA-IR neurons in one of the optic lobe neuropils, the lobula. This finding indicates that histamine may also be a transmitter in certain interneurons in the visual system.

Neurons reactive to antibodies against serotonin in the stomatogastric nervous system and in the alimentary canal of locust and crickets (orthoptera, insecta)

Immunoreactive neurons in the stomatogastric nervous system and in the alimentary tract of the locust Schistocerca gregaria and the crickets Gryllus bimaculatus and Acheta domesticus have been examined using antibodies against serotonin (5-hydroxytryptamine; 5-HT). For comparative anatomical analysis cobalt chloride infusion was applied. The innervation of the visceral muscles was studied electron microscopically. In all three species the majority of the 5-HT-immunoreactive cell bodies of the stomatogastric nervous system occur in the frontal ganglion in which 30-40% of the total number of cell bodies react with anti-5-HT. In the occipital ganglion only two to four cell bodies (1-2%) are 5-HT-immunoreactive. Single immunoreactive cell bodies were observed in the ventricular ganglia in only a few preparations. The 5-HT-immunoreactive neurons in the frontal ganglion are pseudounipolar or multipolar. The main process of the 5-HT-immunoreactive neurons of the frontal ganglion descend along the entire stomatogastric nervous system. Some arborizations of these processes ascend into the brain and others supply the neuropil of all stomatogastric ganglia. Side branches leave the stomatogastric nervous system and form a plexus along the surface of the entire intestinal tract from where 5-HT-immunoreactive fibers supply: all muscle layers of the muscularis; the external dilator muscles of the foregut and probably some somatic muscles, neurohaemal organs and Malpighian tubules (excretory system). Serotonin-immunoreactive fibers further proceed into salivary gland and the retrocerebral complex (corpora cardiaca and corpora allata). The retrocerebral glandular complex and the hindgut receive additional immunoreactive neurons from the central nervous system. Electron microscopic analysis of nerves innervating the muscle layers of the alimentary tract revealed one type of 5-HT-immunoreactive and one or two types of non-5-HT-immunoreactive fibers. All fiber types contact the sarcolemma of muscle fibers without any obvious synaptic membrane specializations. The 5-HT-immunoreactive fibers are in some regions in close contact with the haemolymph. These regions can be regarded as sites of neurohormonal release. The distribution of serotonin-immunoreactive neurons suggests that 5-HT acts as a neurotransmitter and/or modulator on intestinal muscles and some somatic muscles and glandular cells, and as a neurohormone released from neurohaemal sites into the body fluid.

Ultrastructural demonstration of serotonin-immunoreactivity in the nervous system of an insect (Calliphora erythrocephala)

Serotonin (5-HT) immunocytochemistry, was performed on the whole dissected nervous system of the blowfly. Employing the peroxidase-antiperoxidase technique and osmium postfixation, it was possible to examine 5-HT-immunoreactive neuronal elements first light microscopically in 25 microns sections, and, after re-embedding, to analyze the same sections electron microscopically in ultrathin sections. We describe the ultrastructure of 5-HT-positive terminals in the neural sheath of peripheral nerves and in the optic lobes. The immunoreactivity was observed in large (100 nm) granular vesicles, on membranes of clear vesicles, along neurotubules, and along the internal surface of the plasmalemma.

Serotonin-like immunoreactivity in the optic lobes of three insect species

SummaryThe cellular localization of 5-HT in the optic lobes of three insect species was assayed with the use of antibodies raised against 5-HT. In Schistocerca, Periplaneta, and Calliphora all neuropil regions of the optic lobe, the lamina, medulla and lobula, contain 5-HT-immunoreactive varicose fibres in different patterns, like columns and layers. Such fibres also connect the lobula to neuropil in the lateral protocerebrum. In Calliphora also 5-HT-positive fibres of the medulla and lobula plate have projections to the lateral protocerebrum, whereas the origin of the lamina fibres is not certain.In all species the processes displaying 5-HT-like immunoreactivity appear to be derived from a relatively small number of cell bodies, each neuron thus having processes over a large volume of the neuropil of the optic lobe in different layers.

The retina and retinal projection on the lamina ganglionaris of the crayfish Pacifastacus leniusculus (Dana)

The retinular cell morphology and ommatidia arrangement in the crayfish retina are studied, Each ommatidium contains eight receptor cells (R1–R8). Seven of the them (R1–R7) contribute to a large spindle‐shaped rhabdom with orthogonal layers of microvilli.

Types and arrangements of neurons in the crayfish optic lamina

SummaryThe neural arrangements in the optic lamina of the crayfish Pacifastacus leniusculus Dana have been studied by light microscopy by means of silver impregnation techniques. The lamina is composed of columnar synaptic compartments (cartridges). Each cartridge is composed of seven receptor terminals distributed in two layers and second-order monopolar neurons connecting the lamina with the second synaptic region, the medulla externa.The neurons found in the lamina consist of five classes: monopolar neurons, centrifugal small-field neurons, tangential neurons, multipolar cells (possibly of a glial nature) and photoreceptor axons (Fig. 13).Among the monopolar cells, five types are classified (M1–M5) according to their lamina arborizations. Two types are stratified (M3 and M4) corresponding to the photoreceptor terminal strata. On this basis, the lamina plexiform layer is subdivided into two layers (epl1 and epl2). The remaining monopolar neurons have lateral processes in both layers, two of them within one cartridge (M1 and M2) and one over several cartridges (M5).There is one type of small-field centrifugal neuron (C1) and two types of tangential medulla to lamina neurons (Tan1 and Tan2), both having processes covering a large number of cartridges.Multipolar cells with cell bodies distal (MP1) or proximal (MP2) to the plexiform layer send processes to several cartridges.The receptor axons consist of three types. One has terminals in epl1 or epl2, the second has its terminal in epl1 and a thin process to epl2, and the third (corresponding to the 8th retinular cell) bypasses the lamina and has a terminal in the medulla externa.A brief comparison is made with the neural arrangements in the lamina of the Norway lobster Nephrops norvegicus L.

Quantitative determination of biogenic amines and DOPA in the CNS of adult and larval blowflies, Calliphora erythrocephala

Abstract 1. Using high performance liquid chromatography with electrochemical detection we have assayed serotonin (5-HT), octopamine (OA). dopamine (DA), noradrenaline (NA), adrenaline (A) and dihydroxyphenylalanine (DOPA) in the CNS of adult and larval blowflies, Calliphora erythrocephala . 2. The adult CNS was also partitioned and optic lobes, brain minus optic lobes and ventral ganglia were assayed separately. 3. In the adult and larval CNS all the assayed compounds except NA and A were detected. 4. In the adult flies the amines and DOPA occurred within the range of 0.29–4.0 pmol per tissue. 5. In the larval CNS DOPA was present in large amounts, 9.8 pmol per tissue, whereas the others were in the range of 0.51–1.1 pmol per tissue. 6. Measured in pmol per tissue, the adult optic lobes contained less of all the compounds than the remaining brain and the ventral ganglia less than the entire brain (except for DOPA). 7. The larval CNS (including the precursors of imaginai neurons) contained less 5-HT, OA and DA than the adult, but much more DOPA.

Light- and electron-microscopic immunocytochemistry of peptidergic neurons innervating thoracico-abdominal neurohaemal areas in the blowfly

SummaryVentral thoracic neurosecretory cells (VTNCs) of the blowflies, Calliphora erythrocephala and C. vomitoria, innervating thoracic neuropil and the dorsal neural sheath of the thoracico-abdominal ganglion have been shown to be immunoreactive to a variety of mammalian peptide antisera. In the neural sheath the VTNC terminals form an extensive neurohaemal network that is especially dense over the abdominal ganglia. The same areas are invaded by separate, ut overlapping serotonin-immunoreactive (5-HT-IR) projections derived from neuronal cell bodies in the suboesophageal ganglion. Immunocytochemical studies with different antisera, applied to adjacent sections at the lightmicroscopic level, combined with extensive cross-absorption tests, suggest that the perikarya of the VTNCs contain co-localized peptides related to gastrin/cholecystokinin (CCK), bovine pancreatic polypeptide (PP), Met- and Leuenkephalin and Met-enk-Arg6-Phe7 (Met-enk-RF). Electron-microscopic immunogold-labeling shows that some of the terminals in the dorsal sheath react with several of the individual peptide antisera, whilst others with similar cytology are non-immunoreactive. In the same region, separate terminals with different cytological characteristics contain 5-HT-IR. Both 5-HT-IR and peptidergic terminals are localized outside the cellular perineurium beneath the acellular permeable sheath adjacent to the haemocoel. Hence, we propose that various bioactive substances may be released from thoracic neurosecretory neurons into the circulating haemolymph to act on peripheral targets. The same neurons may also interact by synaptic or modulatory action in the CNS in different neuropil regions of the thoracic ganglion.

Mapping of serotonin-immunoreactive neurons in the larval nervous system of the flies Calliphora erythrocephala and Sarcophaga bullata

SummarySerotonin-immunoreactive (5-HTi) neurons were mapped in the larval central nervous system (CNS) of the dipterous flies Calliphora erythrocephala and Sarcophaga bullata. Immunocytochemistry was performed on cryostat sections, paraffin sections, and on the entire CNS (whole mounts).The CNS of larvae displays 96–98 5-HTi cell bodies. The location of the cell bodies within the segmental cerebral and ventral ganglia is consistent among individuals. The pattern of immunoreactive fibers in tracts and within neuropil regions of the CNS was resolved in detail. Some 5-HTi neurons in the CNS possess axons that run through peripheral nerves (antenno-labro-frontal nerves).The suboesophagealand thoracico-abdominal ganglia of the adult blowflies were studied for a comparison with the larval ventral ganglia. In the thoracico-abdominal ganglia of adults the same number of 5-HTi cell bodies was found as in the larvae except in the metathoracic ganglion, which in the adult contains two cell bodies less than in the larva. The immunoreactive processes within the neuropil of the adult thoracico-abdominal ganglia form more elaborate patterns than those of the larvae, but the basic organization of major fiber tracts was similar in larval and adult ganglia. Some aspects of postembryonic development are discussed in relation to the transformation of the distribution of 5-HTi neurons and their processes into the adult pattern.

Neuronal connectivity patterns in the compound eyes of Artemia salina and Daphnia magna (Crustacea: Branchiopoda)

SummaryThe neuronal types and patterns in the visual system of the species Artemia salina and Daphina magna have been studied with the Golgi method and electron microscopy. The lamina contains five classes of neurons: photoreceptor axons, monopolar, centrifugal, tangential and amacrine neurons. The terminals of the receptor axons are distributed in two (A. salina) or three (D. magna) layers. The dilated terminals have an extensive and wide array of fine branches. One axon from each ommatidium bypasses the lamina and terminates in the medulla in A. salina. A. salina has four types of monopolar neurons, two of which are stratified, whereas in D. magna only two types are found, one of which is bistratified. Tangential T-neurons connect the lamina with the protocerebrum. D. magna has in addition one tangential T-neuron connecting both the lamina and the medulla with the protocerebrum. In both species monopolar-type centrifugal neurons connect the medulla and the lamina, whereas that of A. salina has a wide laminar distribution. Both species also have amacrine cells in the lamina. The medulla contains, besides those shared with the lamina, transmedullary neurons (two types in A. salina), amacrine cells and neurons originating in the protocerebrum.“Cartridge”-type synaptic compartments are lacking in the investigated species, although a periodic arrangement is discernible in the distal portion of the lamina of A. salina. The receptors from three types of specialized contacts in Artemia, one of which involves a dyad. D. magna has only one-to-one synapses. Neurosecretory fibres are absent in A. salina.