Neuropeptides in sensory structures of nematodes.

The peptidergic innervation of sensory structures in two species of nematodes, Ascaris suum (order Ascaridida, family Ascarididae) and Cystidicola farionis (order Spirurida, family Cystidicolidae) was studied. Immunocytochemical methods were used for localization of FMRF-amide-like neuropeptides in the nervous system. Immunoreactivity to FMRF-amide, RF-amide and SALMF-amide was detected in the central nervous system of the species studied, and also in the cephalic papillary nerves, in axons of the amphids and the deirids, and in nerves innervating caudal papillae.

Immunocytochemical localization of histamine in flatworms.

Specific antibodies against histamine were used to demonstrate the occurrence and cellular distribution of histamine-like immunoreactivity in three species of flatworms (phylum Platyhelminthes). In the parasitic cestode Diphyllobothrium dendriticum, histamine-reactivity was found in neurons of the main nerve cords, and in cells lining the central and peripheral excretory ducts. In the free-living microturbellarian Microstomum lineare and in the planarian Polycelis nigra, histamine-immunoreactivity was restricted to cells and fibres of the nervous system. The occurrence of histamine or a related substance in the nervous system of flatworms, which represent primary bilateria, indicates the importance of this neuroactive substance in the animal kingdom.

Development of immunoreactivity to the invertebrate neuropeptide small cardiac peptide B in the tapeworm Diphyllobothrium dendriticum.

Neurons immunoreactive to small cardiac peptide B (SCPb) occur in the scolex and neck region of adult Diphyllobothrium dendriticum. The localisation of the SCPb-IR neurons in the peripheral nervous system is very pronounced; they are closely associated to the bothridial musculature in the scolex. SCPb-IR neurons were not observed in plerocercoid larvae but appeared after cultivation in vitro at 37 degrees C for 30 h. Functional and developmental aspects of the SCPb-IR neurons are discussed.

Neuropeptides in a microturbellarian--whole mount immunocytochemistry.

A whole mount immunofluorescence method was used for the localization of immunoreactivity (IR) to four regulatory peptides and the bioamine serotonin in the nervous system of Stenostomum leucops (Turbellaria, Platyhelminthes). The flatworm S. leucops belongs to the taxon Catenulida which, according to the new phylogenetic system by Ax [2], forms a key group between the coelenterates and more advanced flatworm species. Positive IR was obtained using antisera against FMRF-amide, beta-endorphin, growth hormone releasing factor (GRF), substance P, and serotonin. The distribution patterns of these neuropeptide-like immunoreactivities differ significantly from each other. Antisera against Leu-enkephalin, bovine pancreatic polypeptide (BPP), bombesin, cholecystokinin (CCK-8), neurotensin, somatostatin, growth hormone (GH), secretin, and neurophysin II gave negative results. This primitive flatworm shows similarities with hydra in the lack of IR to anti-somatostatin, anti-Leu-enkephalin, and anti-BPP. These antisera give positive IR in more advanced flatworm species, indicating a later convergent evolution of vertebrate-like peptides within the phylum Platyhelminthes.

Immunocytochemical demonstration of neuropeptides and serotonin in the tapeworm Diphyllobothrium dendriticum.

The present immunocytochemical study concerns the distribution of four neuropeptides, FMRF-amide, vasotocin, leu-enkephalin and neurotensin, and of the bioamine serotonin in the plerocercoid larva of Diphyllobothrium dendriticum. Anti-FMRF-amide and vasotocin-reactivity occurs in perikarya and nerve fibres in the CNS and PNS of this worm. The peptide-containing fibres surround and seem to innervate the musculature and to terminate beneath the basal lamina of the tegument at the inner surface of the bothridia, suggesting a neurotransmitter function. Anti-leu-enkephalin reaction occurs in perikarya and fibres in the main nerve cords and in the PNS. Anti-neurotensin reactive fibres were observed in the neuropile of the nerve cords. Serotonin immunoreactivity was found in neurons in the ganglionic commissure of the brain and along the main nerve cords. This study is the first immunocytochemical identification of neuropeptides and serotonin in a parasitic flatworm and the information gained may be of importance for the development of new antihelminthics.

Release of neurosecretory material by protrusions of bounding membranes extending through the axolemma, in Diphyllobothrium dendriticum (Cestoda).

The release of neurosecretory material was studied in the cestode tapeworm Diphyllobothrium dendriticum. In plerocercoid larvae cultivated in vitro the neurosecretory vesicles release their content by way of protrusions extending through the axolemma into the intercellular space. The released material is directed predominantly towards the muscles. No omega figures were detected. The significance of this new type of release is discussed.

Ultrastructure of epidermal eyespots of Microstomum lineare (Turbellaria, Macrostomida).

The eyespots of Microstomum lineare were studied by electron microscopy, light microscopy, and fluorescence microscopy. Each eyespot consists of two ciliary photoreceptor cells shielded by pigment cells and additional sensory cells. The photoreceptor cells are characterized by a distal intracellular cavity lined with 50-100 interwoven cilia. The other sensory cells are of two ultrastructurally different types, one with long cilia predominating and the other with balloonlike cilia. The pigment cells, which envelop processes of the sensory cells, contain pigment vacuoles varying in size and content and give a bright red fluorescence by the Falck-Hillarp method. The eyespots are suggested to perform a dual function as photoreceptors and chemoreceptors. The evolutionary significance of ciliary photoreceptors in Turbellaria is discussed.

The nervous system of Microstromum lineare (Turbellaria, Macrostomida). I. A fluorescence and electron microscopic study.

The nervous system (NS) of Microstomum lineare (Turbellaria, Macrostomida) was studied by electron and light microscopy, combined with fluorescence histochemistry (Falck-Hillarp method for biogenic monoamines). The NS is primitively organized, with a bilobed brain, two lateral nerve cords lacking commissures, and peripheral nerve cells scattered along the nerve cords. The stomatogastric NS, with a pharyngeal nerve ring, is joined to the central NS by a pair of connective ganglia. A green fluorescence in all parts of the NS indicates catecholaminergic neurons as the dominant neuron type. Ultrastructurally, two types of neurons were identified on the basis of their vesicle content: 1. Aminergic (catecholaminergic) neurons containing dense-core vesicles of varying electron-density and size, i.e., small dense-core vesicles (diameter 50--100 nm), vesicles with a highly electron-dense core (60--140 nm), and vesicles with an eccentric dense-core. 2. Presumed peptidergic neurosecretory neurons containing large granular vesicles (diameter about 200 nm) in the stomatogastric NS and peripheral parts of the central NS. In light microscopy, paraldehyde-thionin stained neurons were observed in the same areas.