Frequency discrimination in the central nerve cords of locusts

The auditory mechanism in locusts, especially the central mechanism of frequency analysis, was examined electrophysiologically. The central auditory response had a sharp rise of threshold at a certain frequency range, and it showed a remarkable change when all peripheral fibres except the tympanic nerve were cut off. The response pattern was also changed by this procedure. These results indicate the presence of an inhibitory interaction mechanism of various auditory inputs. It can be concluded that locusts are able to discriminate sound frequency to some degree by this interactive mechanism.     

Structure-activity relationships of benzylidene anabaseines in nicotinic acetylcholine receptors of cockroach nerve cords

Ten analogues of 6'-chloro-3-benzylideneanabaseine (CBA) bearing substituents at the ortho- and the para-positions of the phenyl group were synthesized, together with two related compounds. The affinity of the synthesized compounds for nicotinic acetylcholine receptors (nAChRs) in the nerve cord of the American cockroach (Periplaneta americana L.) was examined by the radioligand binding assay using [(3)H]epibatidine (EPI), a nAChR agonist. All 12 tested compounds inhibited [(3)H]EPI binding, showing K(i) values ranging from 14.6 to 6830nM. The potency variation of para-substituted CBA analogues was explained by the steric (Delta B(1)) and electronic (sigma(p)) parameters of the para-substituents, or by the steric parameter and the charge of the N1 nitrogen atom (qN(1)). Among the CBA analogues, only two compounds containing a dimethylamino group and a methoxy group at the para-position showed high insecticidal activity against the German cockroach (Blattella germanica) when injected after pretreatment with metabolic inhibitors. High-affinity analogues of CBA might be suitable probes for use in classifying and characterizing insect nAChR subtypes.     

Selective 32P-labelling of individual species in a total tRNA population.

A simple procedure to label individual tRNA species in a total tRNA preparation has been developed. The principle of the method is as follows: total crude tRNA (from E. coli) is incubated in the presence of a crude aminoacyl-tRNA synthetase preparation, containing most aminoacyl-tRNA synthetases and only one specific amino acid corresponding to the tRNA species which is intended to be labelled. This achieves the purpose of charging the desired tRNA species thereby protecting its 3'OH-terminus; obviously all the other tRNA species will have a free 3'OH group. Periodate oxidation, followed by beta-elimination, destroys any free 3'OH. After deacylation of the specific aminoacylated tRNA at pH 8.8 the only free 3'OH group will be the one of the desired tRNA species. High specific activity (32P)-pCp is ligated to this 3'OH by means of T4-RNA ligase. Two-dimensional polyacrylamide gel electrophoresis (2D-PGE) and sequence analysis of the isolated tRNA show that the method is very specific. Individually labelled tRNA species can be used as probes for cloning tRNA genes.     

Phosphorylation reactions in bovine rod outer segments studied by 32P-labelling of intact retina

The protein phosphorylation pattern in the intact bovine retina has been investigated by labelling with 32P-phosphate under incubation conditions that preserve the electrical photoresponse of the photoreceptor cells. The phosphorylation of rod outer segment proteins was analysed after isolation of outer segments from the labelled retina. The global influence of light, Ca2+ and the phosphodiesterase inhibitor, isobutylmethylxanthine, on protein phosphorylation in rod outer segments was analysed. A 12 kDa protein is the most prominent phosphorylated species in the intact bovine retina. Its phosphorylation is increased by light and/or Ca2+. Evidence is presented that this strongly phosphorylated protein is not located in the outer segment, and we suggest that it may be a synaptic protein. Retinal rod outer segment membrane proteins with apparent molecular weights of 245, 226, 125, 110, 50, 46, 38 and 20 all show light-stimulated phosphorylation. Lowering the extracellular Ca2+ levels results in a decrease of the phosphorylation level of some of these proteins, viz. at 125, 50, 38 and probably at 20 kDa. Such proteins, whose phosphorylation level is influenced both by light and by elevated Ca2+, are candidates for mediators of phototransduction. The phosphorylated species at 245, 226, 110, 50 and 20 kDa are enriched in rod outer segment plasma membrane preparations. These protein species could participate in the light-regulated modulation of the Na+-conductance of the plasma membrane.     

Lysophosphatidic acid can activate platelets without increasing 32P-labelling of phosphatidic acid

Stimulation of platelets by thrombin produced a rise in [32P]phosphatidic acid labelling of platelets which was greater in medium without added calcium than in medium with 2.5 mM calcium. A rise in [32P]lysophosphatidic acid was also seen in platelets stimulated by thrombin in the presence of 2.5 mM extracellular calcium, though it was of lesser magnitude (average 35%) than the rise in phosphatidic acid. In platelets resuspended without added calcium no change in [32P]lysophosphatidic acid was seen in response to thrombin. Lysophosphatidic acid can itself induce platelet aggregation. Similarly to the calcium ionophore A23187, lysophosphatidic acid produced minimal change (in medium with no added calcium) to no change (in medium with 2.5 mM external calcium) in [32P]lysophosphatidic acid. The endoperoxide analog U46619 produced changes in 32P-labelling of platelet phosphatidic and lysophosphatidic acid similar to those produced by thrombin but of lesser magnitude. The results of these studies show that the action of lysophosphatidic acid on platelets differs from the action of thrombin, U46619 and platelet-activating factor, which produce a rapid rise in [32P]phosphatidic acid, and suggests that lysophosphatidic acid, like A23187, largely bypasses the initial receptor-coupled breakdown of phosphoinositides leading to formation of diacylglycerols and phosphatidic acid.     

Insulin and progesterone increase 32PO4-labeling of phospholipids and inositol 1,4,5-trisphosphate mass in Xenopus oocytes.

After a 4-6 h induction period, insulin or progesterone induces Xenopus oocytes to enter prophase of meiosis. During the period of induction, both insulin and progesterone induced an increase in 32PO4 labeling of phosphatidylcholine and phosphatidylinositol. Through a mass assay, we found that insulin and progesterone increase inositol 1,4,5-trisphosphate (IP3) at about 15-30 s, 15 min and at about 2-3 h (0.5 GVBD50) after hormone addition. Since IP3 increases were small (from a basal of 66 to 104 nM), the results agree with prior conclusions that progesterone does not induce a large, cytosolic calcium elevation. Insulin is probably acting through the insulin-like growth factor-1 receptor as insulin concentrations greater than about 50 nM are required to increase IP3.     

Studies on the effects of acetylcholine and antiepileptic drugs on32Pi incorporation into phospholipids of rat brain synaptosomes

Studies were conducted on the effects of antiepileptic drugs on the acetylcholine-stimulated³²P labeling of phospholipids in rat brain synaptosomes. Of the four antiepileptic drugs investigated in the present study, namely phenytoin, carbamazepine, phenobarbital, and valproate, only phenytoin blocked the acetylcholine-stimulated³²P labeling of phosphatidylinositol and phosphatidic acid, and the acetylcholine-stimulated breakdown of polyphosphoinositides. Phenytoin alone, like atropine alone, had no effect on the³²P labeling of phospholipids nor on the specific radioactivity of [³²P]ATP. Omission of Na⁺ drastically reduced both the³²P labeling of synaptosomal phospholipids and the specific radioactivity of [³²P]ATP and furthermore it significantly decreased the phosphoinositide effect. It was concluded that certain antiepileptic drugs, such as phenytoin, could exert their pharmacological actions through their antimuscarinic effects. In addition the finding that phenytoin, which acts to regulate Na⁺ and Ca²⁺ permeability of neuronal membranes, also inhibited the phosphoinositide effects in synaptosomes, support the conclusions that Ca²⁺ and Na⁺ are probably involved in the molecular mechanism underlying this phenomenon in excitable tissues.Abbreviations used ACh Acetylcholine - PA phosphatidic acid - PI phosphatidylinositol - poly PI polyphosphoinositides (diphosphoinositide and triphosphoinositide) - PC phosphatidylcholine - PE phosphatidylethanolamine - PS phosphatidylserine - S.A. specific radioactivity     

Swelling of cockroach nerve cords in incubating solutions.

Cockroach thoracic ganglia and nerve-cord connectives were incubated for 30 min in several standard electrolyte solutions. Tissue swelling was calculated from specific gravities measured in a continuous gradient column. Ganglia and connectives swelled in all solutions tested, except those containing sucrose (80 mM). Oxygenation and addition of glucose (5 mM) to the media did not prevent swelling.     

The significance of the radial nerve cords in asteroids and echinoids

Summary The radial nerve cords of a number of different species of echinoids and asteroids have been examined with the electron microscope. They have been shown to consist of tracts of small naked axons with discrete regions of neuropile. Synapses within these areas of neuropile have been described and their significance discussed. A careful examination has failed to reveal any connections between the ectoneural nerve and the mesodermal nerves and muscles. Synapses across the connective tissue sheet separating these regions have been proposed to account for known functional connection. A hypothetical model of the nervous system has been proposed to account for known structure and function.The author wishes to acknowledge the help of Prof. M. S. Laverack during this study. Part of this work was carried out while the author was in receipt of a Queen Elizabeth Fellowship from the Commonwealth Government of Australia.     

Donor-recipient interconnections between giant nerve fibers in transplanted ventral nerve cords of earthworms

Twelve segments of ventral nerve cord (VNC) from donor earthworms, Eisenia foetida, were transplanted into recipient worms from which a comparable length of VNC had been removed. Within the first few days after transplantation, bud-like formations, containing outgrowths of the giant nerve fibers, were evident at the ends of transplanted and recipient VNC. Morphological and electrophysiological evidence indicated that by 4-10 days after transplantation, medial (MGF) and lateral (LGF) giant fibers within the transplanted VNC formed cell-specific connections with their counterparts in the recipient VNC. Although the diameters of the giant fiber connections in the transplant-recipient junctions were often larger than normal, spike conduction across the junction was initially slow (approximately 1.0 m/s) but gradually increased over the next 2-3 weeks. Within the transplant, giant fibers were initially normal in appearance, but spike conduction was slow (1-2 m/s). During the next few weeks velocities increased by as much as fourfold and then stabilized for the next several months. However, by 4-5 weeks after transplantation, giant fiber morphology within the transplant was altered significantly, as indicated by the formation of numerous branch-like extensions along the length of each giant fiber. By 9-10 months there were further morphological changes in the transplant, as indicated by decreased branching of the giant fibers and altered neuropile. Despite these morphological changes, through-conduction of giant fiber spikes remained reliable.     

Ultrastructure of the circumoral nerve ring and the radial nerve cords in holothurians (Echinodermata)

The circumoral nerve ring and the radial nerve cords (RNCs) of Eupentacta fraudatrix and Pseudocnus lubricus (Holothuroidea) were examined as an example of holothurian nervous tissue. The RNC is composed of outer ectoneural and inner hyponeural layers, which are interconnected with one another via short neural bridges. The circumoral nerve ring is purely ectoneural. Both ectoneural and hyponeural components are epithelial tubes with a thick neuroepithelium at one side. A thin ciliated non-neuronal epithelium complements the neuroepithelium to form a tube, thereby enclosing the epineural and hyponeural canals. The whole of the ectoneural and hyponeural subsystems is separated from the surrounding tissue by a continuous basal lamina. The nerve ring and the ectoneural and hyponeural parts of the radial nerves are all neuroepithelia composed of supporting cells and neurons. Supporting cells are interpreted as being glial cells. Based on ultrastructural characters, three types of neurons can be distinguished: (1) putative primary sensory neurons, whose cilium protrudes into the epineural or hyponeural canal; (2) non-ciliated neurons with swollen rough endoplasmic reticulum cisternae; (3) monociliated neurons that are embedded in the trunk of nerve fibers. Different types of synapses occur in the neuropile area. They meet all morphological criteria of classical chemical synapses. Vacuolated cells occur in the neuroepithelium of E. fraudatrix, but are absent in P. lubricus; their function is unknown. The cells of the non-neuronal epithelia that overlie the ectoneural and hyponeural canals are hypothesized to belong to the same cell type as the supporting cells of the neuroepithelium.     

Sodium and lithium movements and axonal function in cockroach nerve cords.

Exposure to sodium-deficient (tris) saline caused an appreciable decline in the sodium content of intact connectives in the absence of equivalent reduction in the amplitude of the recorded action potentials. Return of sodium-depleted connectives to normal saline resulted in a rapid recovery of axonal function despite only a partial (less than 70%) recovery in sodium content. Replacement of sodium ions by those of lithium in the bathing medium resulted in a substantial accumulation of this cation. Lithium movements exhibited a marked asymetry, no significant decline in concentration being observed upon return to normal saline. These results are tentatively interpreted in terms of an exchangeable glial sodium fraction and are discussed in relation to extra-axonal sodium regulation.     

Intersegmental coordination of swimmeret rhythms in isolated nerve cords of crayfish

Summary In crayfish,Pacifastacus leniusculus, abdominal ganglia that can generate the motor pattern normally associated with swimmeret beating continue to do so when the number of connected ganglia is reduced from six to two. The period and phase of the rhythm produced by these shortened chains of ganglia are the same as those produced by the full abdominal nerve cord. These results demonstrate that interactions between any two neighboring ganglia suffice to establish the metachronal phase-lag characteristic of the swimmeret rhythm.Several kinds of interganglionic interneurons that are part of the swimmeret system originate in each abdominal ganglion. These premotor interneurons receive synaptic input in the ganglion of origin and project to other ganglia. Axons from interganglionic neurons also terminate in each ganglion, and some of these terminals receive PSPs from the swimmeret pattern generators in the ganglion where they terminate. Currents injected into these interneurons and axon terminals can reset the swimmeret rhythm. These results demonstrate that premotor interganglionic interneurons exist that have the properties required to coordinate adjacent ganglia. The structures and physiological properties of these interneurons are described and discussed in the context of Stein's model of intersegmental coordination in the swimmeret system.     

The structure of Archiannelid Dinophilus gyrociliatus ventral nerve cords

Dinophilus gyrociliatus is a typical representative of the synthetic group Archiannelida. Here we describe the structure of Dinophilus young adults ventral nerve cords. Five nerve cords, irregular commissures and intersegmental nerves are revealed by tubulin and FMRFamide immunostaining. 5-HT-stainning marks the complex varicose nerve plexus with numerous irregularly distributed small perikarya. Four nerve cords and solitary cell bodies occur after clyoxylic-induced reaction for catecholamines visualization. Altogether, our results indicated that morphology of Dinophilus ventral nerve cords differs from the Polychaetes archetype in many respects.