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Isolation, sequence, and bioactivity of PDVDHVFLRFamide and ADVGHVFLRFamide peptides from the locust central nervous system.

The brain and the retrocerebral complex of the locust, Locusta migratoria, were examined for the presence of FMRFamide-related peptides (FaRPs) using an RIA specific for -RFamide. RP-HPLC of these extracts using both C18 and phenyl columns revealed the presence of several FaRPs eluting at different percentages of acetonitrile. The sequences of two peptides were determined. One sequence is identical to the previously described SchistoFLRFamide (PDVDHVFLRFamide), whereas a second peptide is novel and differs from SchistoFLRFamide in positions 1 and 4 (ADVGHVFLRFamide). The bioactivity of these native and synthetic FaRPs on locust oviduct contractions has been examined. Both peptides showed inhibitory activity on the locust oviduct. Truncated versions of PDVDHVFLRFamide revealed that the essential features for inhibition lay in the sequence HVFLRFamide.

Amino Acid Sequence↗

Neuropeptide FF reduces food intake in rats.

The effect of neuropeptide FF (NPFF), a mammalian FMRFamide-like peptide with antiopioid activity, on food intake was investigated in food-deprived rat. The ICV administration of NPFF (5 or 10 micrograms/rat) reduced food intake during the first 60 min after administration. ICV injection of naloxone (10 or 100 micrograms/rat), an opioid antagonist, also decreased food intake. However, the combination of NPFF and naloxone showed no additivity in the anorexigenic effect, suggesting that NPFF and naloxone reduced food intake by the common mechanism. These results indicate that NPFF may function as an endogenous anorexigenic peptide with anitiopioid function.

Animals↗

Influence of methylation or substitution of the histidine of HVFLRFamide on biological activity and binding of locust oviduct.

The FMRFamide-related peptide PDVDHVFLRFamide (SchistoFLRFamide) is involved in the neural control of locust oviducts, where it acts as a potent inhibitor of spontaneous and induced contractions. Previous studies have shown that the His residue in the truncated analogue HVFLRFamide is critical for the retention of inhibitory biological activity, whereas VFLRFamide, in which inhibitory biological activity is lost, is the minimum sequence for binding of comparable affinity to the parent compound. In the present study we have used binding and bioassay to further investigate the properties of the His residue in determining the inhibitory biological activity of HVFLRFamide. Substitution of His by the D-isomer or Phe produced analogues with stimulatory rather than inhibitory activity, confirming the importance of the His moiety. In addition, inhibitory activity was retained when the His moiety was methylated at the N-3 position of the imidazole ring, but methylation of N-1 yielded a peptide that stimulated contractions. Inhibitory activity was further retained when Nn-methyl-L-His and D,L-1',2',4'-triazole-3-Ala were substituted for His. These results, along with binding studies, suggest that transfer of an imidazole proton is not responsible for inhibitory activity and further suggest that hydrogen bonding to the N-1 of the imidazole ring of histidine may be required to evoke the inhibitory response.

Amino Acid Sequence↗

Localization of octopaminergic neurones in insects.

This paper reviews data on the localization of octopaminergic neurons revealed by immunocytochemistry in insects, primarily the locusts Schistocerca gregaria and Locusta migratoria, cricket Gryllus bimaculatus, and cockroach Periplaneta americana. Supporting evidence for their octopaminergic nature is mentioned where available. In orthopteran ventral ganglia, the major classes of octopamine-like immunoreactive (-LI) neurones include: (1) efferent dorsal and ventral unpaired median (DUM, VUM) neurones; (2) several intersegmentally projecting DUM interneurones in the suboesophageal ganglion; other DUM interneurones are probably GABAergic; (3) a pair of anterior median cells in the prothoracic ganglion; (4) a single pair of ventral cells in most thoracic and some other ganglia; these appear to be plurisegmentally projecting interneurones. Eight categories of octopamine-LI neurones occur in the orthopteran brain. The basic projections of three types are described here: one class project to the optic lobes to form wide field projections. Another type descends to cross into the tritocerebral commissure and may invade the contralateral brain hemisphere. A further class is the median neurosecretory cells with axons in the nervi corpori cardiaci I. Available data for the honey bee Apis mellifera and moth Manduca sexta indicate that the octopamine-LI cell types found in orthopterans also occur in holometabolous insects. Immunocytochemical evidence suggests that some octopaminergic DUM cells contain an FMRFamide-related peptide and the amino acid taurine as putative cotransmitters.

Animals↗

Homologies between the amino acid sequences of some vertebrate peptide hormones and peptides isolated from invertebrate sources.

1. The 4K-prothoracicotropic hormone (PTTH) or bombyxin and the melanization-reddish coloration hormone of the silkworm Bombyx mori resemble insulin and insulin-like growth factors. 2. The family of adipokinetic/red pigment concentrating hormones has some similarity with glucagon. 3. Members of the FMRFamide family are found in vertebrates as well as in invertebrates. 4. In Locusta, a molecule immunologically and biologically related to amphibian melanophore stimulating hormone has been partially characterized. 5. Enkephalins and enkephalin-related peptides occur in insects and other invertebrates. 6. Peptides belonging to the tachykinin family have been isolated from molluscan (Octopus) salivary glands and from insect nervous tissue (Locusta migratoria). 7. Invertebrate arginine-vasotocin homologs have been isolated from an insect (Locusta migratoria) and from a mollusc (Conus). 8. In Leucophaea, Locusta and Drosophila, peptides resembling those of the vertebrate gastrin/cholecystokinin family have been identified. 9. As the number of different neuro-/gut peptides with possible function(s) as hormone, neurotransmitter or neuromodulator is now estimated to be of the order of a few hundred, more similarities will probably show up in the near future.

Amino Acid Sequence↗

Further mapping of the Achatina giant neurone types sensitive to the neuroactive peptides isolated from invertebrates.

1. The effects of the 10 synthetic neuroactive peptides originally isolated from invertebrates, applied locally to the neurone tested by the brief pneumatic pressure ejection on the identifiable neurone types of Achatina fulica Ferussac were examined. 2. Achatin-1 (Gly-D-Phe-Ala-Asp), an Achatina endogenous tetrapeptide having a D-phenylalanine residue, ejected locally, showed the depolarizing effects on nearly half of the number of neurone types tested. 3. ACEP-1 (Ser-Gly-Gln-Ser-Trp-Arg-Pro-Gln-Gly-Arg-Phe-NH2), isolated originally from Achatina atria, and pedal peptide (Pro-Leu-Asp-Ser-Val-Tyr-Gly-Thr-His-Gly-Met-Ser-Gly-Phe-Ala) and buccalin (Gly-Met-Asp-Ser-Leu-Ala-Phe-Ser-Gly-Gly-Leu-NH2), found in Aplysia neurones, showed excitatory effects on some Achatina neurone types. 4. Myomodulin (Pro-Met-Ser-Met-Leu-Arg-Leu-NH2), found in Aplysia neurones, produced a hyperpolarization on nearly half of the number of Achatina neurone types tested. The two FMRFamide-like peptides, <EDPFLRFamide (<Glu-Asp-Pro-Phe-Leu-Arg-Phe-NH2), isolated from Helix heart, and AF1 (Lys-Asn-Glu-Phe-Ile-Arg-Phe-NH2), from Ascaris head, also showed hyperpolarizing effects on more than half of the number of Achatina neurone types. 5. SALMFamide 1 (Gly-Phe-Asn-Ser-Ala-Leu-Met-Phe-NH2), isolated from Asterias nervous system, CCAP (Pro-Phe-Cys-Asn-Ala-Phe-Thr-Gly-Cys-NH2), from Carcinus pericardial organ, and corazonin (<Glu-Thr-Phe-Gln-Tyr-Ser-Arg-Gly-Trp-Thr-Asn-NH2), from Periplaneta cardiac corpus, had no effect on Achatina neurones.

Amino Acid Sequence↗

Identifiable Achatina giant neurones: their localizations in ganglia, axonal pathways and pharmacological features.

1. An African giant snail (Achatina fulica Férussac), originally from East Africa, is now found abundantly in tropical and subtropical regions of Asia, including Okinawa in Japan. This is one of the largest land snail species in the world. The Achatina central nervous system is composed of the buccal, cerebral and suboesophageal ganglia. The 37 giant neurones were identified in these ganglia by the series of studies conducted over about 20 years. The identifications were made by the localization of these neurones in the ganglia, their axonal pathways and their pharmacological features. 2. In the left buccal ganglion, the four giant neurones, d-LBAN, d-LBMB, d-LBCN and d-LBPN, were identified. In the left and right cerebral ganglia, d-LCDN, d-RCDN, v-LCDN and v-RCDN were identified. The suboesophageal ganglia are further composed of the left and right parietal, the visceral, the left and right pleural, and the left and right pedal ganglia. In the right parietal ganglion, PON, TAN, TAN-2, TAN-3, RAPN, d-RPLN, BAPN, LPPN, LBPN, LAPN and v-RPLN were identified. In the visceral ganglion, VIN, FAN, INN, d-VLN, v-VLN, v-VAN, LVMN, RVMN and v-VNAN were identified. In the left parietal ganglion, v-LPSN was identified. In the left and right pedal ganglia, LPeNLN, RPeNLN, d-LPeLN, d-LPeCN, d-RPeAN, d-LPeDN, d-LPeMN and d-LPeEN were identified. 3. Of the small molecule compounds tested, dopamine, 5-hydroxytryptamine, GABA, L-glutamic acid, threo- or erythro-beta-hydroxy-L-glutamic acid were effective on the Achatina giant neurones. We suppose that these compounds act as the neurotransmitters for these neurones. 4. Of the neuroactive peptides, achatin-I(Gly-D-Phe-Ala-Asp). APGW-amide(Ala-Pro-Gly-Trp-NH2) and Achatina cardioexcitatory peptide (ACEP-1)(Ser-Gly-Gln-Ser-Trp-Arg-Pro-Gln-Gly-Arg-Phe-NH2) were proposed as neurotransmitters, because these were effective on the Achatina giant neurones and their presence was demonstrated in the Achatina ganglia. Further, myomodulin (Pro-Met-Ser-Met-Leu-Arg-Leu-NH2), buccalin (Gly-Met-Asp-Ser-Leu-Ala-Phe-Ser-Gly-Gly-Leu-NH2), FMRFamide (Phe-Met-Arg-Phe-NH2). [Ser2]-Mytilus inhibitory peptide ([Ser2]-MIP) (Gly-Ser-Pro-Met-Phe-Val-NH2), catch-relaxing peptide (CARP) (Ala-Met-Pro-Met-Leu-Arg-Leu-NH2), oxytocin (Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH2) and small cardioactive peptideB (SCPB) (Met-Asn-Tyr-Leu-Ala-Phe-Pro-Arg-Met-NH2) could also be neurotransmitters because these peptides were also effective on the Achatina giant neurones, though their presence in the ganglia of this animal has not yet been demonstrated. 5. Calcium current (ICa) was recorded from Achatina giant neurones in the Na(+)-free solution containing K(+)-channel blockers under voltage clamp. The Ca2+ antagonistic effects of brovincamine, verapamil, eperisone, diltiazem, monatepil, etc., were compared using the ICa of the Achatina neurones. 6. Almost all of the mammalian small molecule neurotransmitters were effective on the Achatina giant neurones, suggesting that these compounds are acting on the neurones of a wide variety of animal species. However, the pharmacological features of the Achatina neurone receptors to these compounds were not fully comparable to those of the mammalian receptors. For example, we proposed that beta-hydroxy-L-glutamic acid (either threo- or erythro-) could be an inhibitory neurotransmitter for an Achatina neurone. 7. In contrast, the Achatina giant neurones appear to have no receptor for the mammalian neuroactive peptides, except for oxytocin and Arg-vasotocin. On the other hand, many neuroactive peptides were isolated from invertebrate nervous tissues, including achatin-I, a neuroexcitatory tetrapeptide having a D-phenylalanine residue.

Animals↗

Autoradiographic distribution of receptors to FLFQPQRFamide, a morphine-modulating peptide, in rat central nervous system.

The neuropeptide FLFQPQRFamide is a structure related to FMRFamide which is able to inhibit the effects of both endogenous and exogenous opiates. This morphine-modulating activity is mediated via the stimulation of specific FLFQPQRFamide receptors, different from opiate receptors. In vitro quantitative receptor autoradiography was performed on frozen sections of rat central nervous system to characterize binding properties and visualize FLFQPQRFamide receptors using the specific ligand [125I]YLFQPQRFamide, a radio-iodinated analogue of FLFQPQRFamide. [125I]YLFQPQRFamide appeared to interact reversibly with a single class of binding sites (KD = 0.2 nM). The specific binding represented 80% of the total binding at 0.05 nM, the FLFQPQRFamide concentration used in this mapping study. Sites labelled with [125I]YLFQPQRFamide were distributed heterogeneously within the brain and spinal cord. A high density of FLFQPQRFamide binding sites was detected in the most external layers of the dorsal horn of spinal cord and various nuclei of pons and medulla including trigeminal, dorsal tegmental and reticular nuclei. Nucleus of solitary tract, parabrachial, ambiguous and facial nuclei are also intensively labelled. Some structures of mesencephalon and diencephalon exhibited a high density of FLFQPQRFamide binding sites: central gray, raphe nuclei and thalamic nuclei such as parafascicular, laterodorsal, central median, paratenial and paraventricular nuclei. Suprachiasmatic and mammillary nuclei, lateral, posterior and anterior areas of hypothalamus and medial preoptic area exhibited high labelling. FLFQPQRFamide binding sites were also seen in some structures of the dopaminergic meso-cortico-limbic system including ventral tegmental area, cingulate cortex, lateral septum and the head of the caudate-putamen. Dense labelling appeared in the presubiculum of hippocampus. The dissimilar mapping of FLFQPQRFamide and opiate brain receptors confirms our previous pharmacological findings in FLFQPQRFamide binding studies on rat spinal cord membranes, showing that FLFQPQRFamide receptors are different from opiate receptors. There was a good correspondence between localization of binding sites and that of the putative endogenous peptide. Both occur in brain areas previously associated with analgesic action of opiates. However, the mapping of FLFQPQRFamide receptors in the central nervous system suggests that the FLFQPQRFamide system could be implicated in other physiological functions.

Animals↗

Neuropeptide FLFQRFamide receptors within the ventral mesenchephalon and dopaminergic terminal areas: localization and functional antiopioid involvement.

The neuropeptide FLFQPQRF amide is a FMRFamide-like peptide with some anti-opiate properties. Neuropeptide FLFQPQRFamide receptors are present in the mammalian central nervous system and have been clearly identified as different from opiate receptors. Autoradiography has revealed neuropeptide FLFQPQRFamide receptor localization within the ventral mesencephalon, where opiate receptors are also located. In order to delineate anatomical localization of neuropeptide FLFQPQRFamide receptors, we used selective chemical lesions of dopaminergic cells and intrinsic perikarya of the ventral mesencephalon, coupled with in vitro autoradiographic techniques. We show that: (i) unilateral lesions of dopaminergic perikarya produced by 6-hydroxydopamine did not affect either ipsi or contralateral neuropeptide FLFQPQRFamide receptor density within the mesencephalon; (ii) unilateral lesions of intrinsic perikarya by ibotenic acid injected into the ventral tegmental area produced a significant reduction of neuropeptide FLFQPQRFamide receptors (40%) in this region; (iii) in the substantia nigra pars compacta, ibotenic acid unilateral lesions did not affect the density of neuropeptide FLFQPQRF-amide receptors; (iv) unilateral 6-hydroxydopamine or ibotenic acid lesions failed to affect neuropeptide FLFQPQRFamide binding in the dopaminergic projection areas. These results suggest that, like opiate receptors, the neuropeptide FLFQPQRFamide binding sites are not localized on dopaminergic neurons but are distributed on both soma of non dopaminergic cells in the ventral tegmental area and on fibers afferent to the ventral tegmental area and substantia nigra pars compacta. To evaluate the possibility that the stimulation of neuropeptide FLFQPQRFamide receptors may affect the opioid modulation of mesocorticolimbic dopaminergic neuron activity, we tested the effects of neuropeptide FLFQPQRFamide ventral tegmental area infusion (0.25-10 micrograms) on the behavioral activation induced by intra-ventral tegmental area morphine infusion. We observed that in the ventral tegmental area, the stimulation of neuropeptide FLFQPQRFamide receptors inhibits morphine-induced locomotor hyperactivity. These results suggest that in the ventral tegmental area, neuropeptide FLFQPQRFamide may participate, via an indirect mechanism, to the control of the mesocorticolimbic dopaminergic system activity by counteracting the effect of opioids.

Animals↗

Neuropeptide FF-containing efferent projections from the medial hypothalamus of rat: a Phaseolus vulgaris leucoagglutinin study.

Neuropeptide FF (FMRFamide-like peptide, morphine-modulating peptide) is an octapeptide isolated from the bovine brain. There is evidence that neuropeptide FF participates in the modulation of nociceptive mechanisms. Neuropeptide FF acts through its own receptors which are distinct from the opiate receptors. In the rat brain neuropeptide FF is found in two major cell populations. We have studied the efferent connections of the hypothalamic neuropeptide FF-containing cell group, which is located in the medial hypothalamus between the dorsomedial, ventromedial and periventricular hypothalamic nuclei. By using an anterograde tracing method (Phaseolus vulgaris leucoagglutinin) combined with double-staining immunohistochemistry we characterized the connections of this cell group with the limbic system, certain hypothalamic nuclei, periaqueductal gray and with the solitary tract nucleus. In the limbic system, the major targets were the lateral septal nucleus, bed nucleus of stria terminalis and certain subnuclei in the amygdala. These connections suggest that neuropeptide FF may act, in addition to its well-characterized action in the sensory system, in limbic functions. Efferent connections to the periaqueductal gray suggest that neuropeptide FF may modulate the opiate mediated analgesia at this site. Good correlation between our results and receptor autoradiography support the idea that the terminal areas which our results show are target areas of the neuropeptide FF-containing system.

Animals↗

Neuromodulatory effects of acetylcholine and serotonin on the sensitivity of leech mechanoreceptors.

1. Each segmental ganglion of the leech Hirudo medicinalis contains 6 touch (T) cells, 4 pressure (P) cells and 4 nociceptive (N) cells. The receptive terminals of these cells innervate the skin in discrete areas. These cells are known to have extrasynaptic receptors. 2. We tested the effect of transmitter substances present in leech CNS on the sensitivity of T and P cells to mechanical stimuli. Substances tested included octopamine, FMRFamide, proctolin, substance P, glutamate, GABA, acetylcholine and serotonin. 3. Only acetylcholine and serotonin had consistent effects. Serotonin (1 x 10(-3) M) increased the number of action potentials of T cells elicited by a standard stimulus. Serotonin (1 x 10(-4) M) and acetylcholine (1 x 10(-3) M) increased the number and frequency of action potentials in P cells.

Acetylcholine↗

Isolation and primary structure of two sulfakinin-like peptides from the fleshfly, Neobellieria bullata.

1. Two novel insect myotropic peptides termed neosulfakinin-I (Neb-SK-I) and neosulfakinin-II (Neb-SK-II) were isolated from the heads of 42 thousand fleshflies, Neobellieria bullata (Diptera, Sarcophagidae). 2. A series of four, high-performance liquid chromatographic (HPLC), fractionations performed on columns with different characteristic features yielded two purified biologically active, hindgut motility stimulating fractions, suitable for amino acid sequence analysis. 3. The proposed sequences for the two peptides are: Phe-Asp-Asp-Tyr-Gly-His-Met-Arg-Phe-(NH2), (Neb-SK-I) and X-X-Glu-Glu-Gln-Phe-Asp-Asp-Tyr-Gly-His-Met-Arg-Phe-(NH2), (Neb-SK-II). 4. These sulfakinins exhibit very high homology to putative drosulfakinin sequences which, however, have not yet been isolated, but were deduced from a cloned Drosophila gene encoding these peptides. 5. Here we provide the first evidence for the expression of such peptides present in Dipterans. 6. Insect sulfakinins show structural identities with the hormonally-active portion of vertebrate gastrin II-, cholecystokinin- and caerulin-related peptides and they share common carboxy terminal sequences with invertebrate/vertebrate peptides of the FMRFamide peptide family.

Amino Acid Sequence↗

Distribution and immunochemical characteristics of neuropeptide F (NPF) (Moniezia expansa) - immunoreactivity in Proteocephalus pollanicola (Cestoda: Proteocephalidea).

1. Using immunocytochemical techniques and confocal scanning laser microscopy, the proteocephalidean cestode, Proteocephalus pollanicola from Lough Neagh pollan (Coregomus autumnalis) was examined for the presence of the native platyhelminth neuropeptide, neuropeptide F (NPF). 2. An antiserum specific for whole-molecule NPF (1-39) (Moniezia expansa) did not immunostain nerve processes in P. pollanicola. A C-terminally-directed NPF (30-39) (M. expansa) antiserum immunostained nerve fibres and cell bodies of both the central and peripheral nervous systems, including innervation associated with the female reproductive system. 3. The pattern of immunoreactivity was identical to that obtained using antisera to the C-terminal region of mammalian NPY-superfamily peptides and the invertebrate neuropeptide, FMRFamide. 4. Under radioimmunoassay conditions, only the C-terminally-directed NPF antiserum cross-reacted with the P. pollanicola peptide and detected 58.74 ng/g wet weight of NPF-1 R in extracts of the worm. 5. Chromatographic characterisation of P. pollanicola NPF-immunoreactivity indicated an apparent molecular weight of 4400-4700 Da, similar to that of NPF (M. expansa). 6. Further analytical HPLC characterisation identified two molecular forms of P. pallanicoda NPF-immunoreactivity, both of which had different retention times from those of NPF (M. expansa) and NPF (Artioposthia triangulata). 7. These data suggest that P. pollanicola possesses a neuropeptide which is homologous in its C-terminal region to NPF (M. expansa) but differs in its mid- to N-terminal region.

Amino Acid Sequence↗

A Mytilus peptide related to the small cardioactive peptides (SCPs): structure determination and pharmacological characterization.

1. An SCP-related peptide, Ala-Pro-Asn-Phe-Leu-Ala-Tyr-Pro-Arg-Leu-NH2, was isolated from the ABRMs of Mytilus edulis. The peptide was designated Mytilus SCP. 2. At 10(-10) M or higher, Mytilus SCP showed a potentiating effect on phasic contraction of the ABRM in response to repetitive electrical stimulation. In contrast, the peptide did not show any potentiating effect on contractures in response to ACh and the FMRFamide-related Mytilus decapeptide, suggesting that the potentiating effect on phasic contraction was brought about by an action of the peptide on the nerve elements in the ABRM. 3. At 10(-8) M or higher, Mytilus SCP showed a catch-relaxing effect in addition to the potentiating effect. The relaxing effect was blocked by mersalyl, suggesting that it was also brought about by a presynaptic action. 4. Various analogues of Mytilus SCP were examined on the ABRM. Leu-Ala-Tyr-Pro-Arg-Leu-NH2 was suggested to be the minimum structure required for both potentiating and relaxing activities on the ABRM. Leu-D-Ala-Tyr-Pro-Arg-D-Leu-NH2 and D-Leu-Ala-Tyr-Pro-Arg-D-Leu-NH2 were found to show strong potentiating and relaxing activities, though they were less potent than the Mytilus SCP.

Amino Acid Sequence↗

The actions of RFamide neuroactive peptides on the isolated heart of the giant African snail, Achatina fulica.

1. The effects of a range of RFamide peptides were examined on the frequency, amplitude and tone of the isolated heart of Achatina fulica. 2. FMRFamide, FLRFamide and SDPNFLRFamide were potent excitants while LPLRFamide, KHEYLRFamide and GSFFRFamide were weak excitants. 3. DNFLRFamide and SSLFRFamide were potent inhibitory peptides while LFRFamide, GSLFRFamide and KNEFIRFamide were weak inhibitory peptides. 4. MRFamide, LRFamide and RFamide were inactive. 5. It is concluded that a tetrapeptide sequence is the minimal requirement for activity on the Achatina heart. With the exceptions of DNFLRFamide and in most preparations GSFFRFamide, LRFamide peptides are excitatory while FRFamide peptides are inhibitory. 6. Due to its inhibitory effect and high potency, the sequence DNFLRFamide warrants further investigation.

Amino Acid Sequence↗

Physiological and pharmacological studies on annelid and nematode body wall muscle.

1. This review covers the pharmacology and physiology of the body wall muscle systems of nematodes and annelids. 2. Both acetylcholine and gamma-aminobutyric acid (GABA) play important roles in the control of body wall muscle in both phyla. In annelids and nematodes, acetylcholine is the excitatory neuromuscular transmitter while GABA is the inhibitory neuromuscular transmitter. In addition, 5-hydroxytryptamine (5-HT) has a modulatory role at annelid body wall muscle but little if any effect on nematode body wall muscle. 3. The acetylcholine receptor of the body wall muscle can be classified as nicotinic-like in both phyla though the annelid receptor has not been analysed in detail. In nematodes, vertebrate ganglionic nicotinic agonists were the most effective of those so far examined while mecamylamine and benzoquinonium were the most effective antagonists. Both neuronal bungarotoxin and neosurugatoxin were potent antagonists of acetylcholine excitation at the nematode receptor. 4. The GABA receptor of the body wall muscle exhibits similarities with the vertebrate GABA-A receptor in both phyla. Picrotoxin is a very weak or inactive antagonist at leech and nematode GABA receptors, while bicuculline methiodide blocks leech GABA receptors but is inactive on nematode GABA receptors. Picrotoxin does block GABA responses of earthworm body wall muscle. All these GABA responses are chloride mediated. 5. Neuroactive peptides of the RFamide family occur in both phyla and FMRFamide has been identified in leeches. RFamides probably have an important role in heart regulation in leeches and in modulation of their body wall muscles. RFamides also modulate nematode body wall muscle activity with KNEFIRFamide raising muscle tone while SDPNFLRFamide relaxes the muscle. It is likely that this family and other neuroactive peptides play an important role in the physiology of body wall muscle throughout both phyla.

Amino Acid Sequence↗

AF1, a sequenced bioactive neuropeptide isolated from the nematode Ascaris suum.

An FMRFamide-like neuropeptide, named AF1, was isolated from head extracts of the nematode Ascaris suum using five steps of HPLC. AF1 is a heptapeptide with the amino acid sequence Lys-Asn-Glu-Phe-Ile-Arg-Phe-NH2. Synthetic AF1 (10(-9) to 10(-7) M) rapidly and reversibly abolished slow membrane potential oscillations of identified ventral and dorsal inhibitory motoneurons and selectively reduced their input resistances. Synaptic transmission was not blocked. In intact Ascaris, AF1 inhibited locomotory movements. This study indicates a potential physiological role for an endogenous neuropeptide in nematodes.

Action Potentials↗

Transdifferentiation from striated muscle of medusae in vitro.

We have established an in vitro transdifferentiation and regeneration system which is based entirely on mononucleated striated muscle cells. The muscle tissue is isolated from anthomedusae and activated by various means to undergo cell cycles and transdifferentiation to several new cell types. In all cases DNA-replication is initiated and the division products are smooth muscle cells, characterized by their ultrastructure and monoclonal antibodies, and nerve/sensory cells, characterized by their ultrastructure and FMRFamide-staining. Both cell types are found at a 1:1 ratio after the first division. The nerve cells stop to replicate, whereas the smooth muscle cells continue and keep producing in each successive division a smooth muscle cell and a nerve cell. The observed data indicate that smooth muscle cells behave like stem cells. Depending on the destabilization and culturing methods, some isolated muscle tissue will form a bilayered fragment and within only two cell cycles manubria (the feeding and sexual organ) or tentacles will regenerate. In this case six to eight new non-muscle cell types have been formed by transdifferentiation.

Animals↗