Octopamine-like immunoreactivity in the dorsal unpaired median (DUM) neurons innervating the accessory gland of the male cockroach Periplaneta americana

The musculature of the mushroom-shaped accessory gland receives innervation from trunks 5C1 of the phallic nerves, which arise from the posterior part of the terminal abdominal ganglion of the male cockroach Periplaneta americana. Anterograde cobalt filling through trunks 5C1 with the subsequent precipitating procedure has shown the fine innervation of the accessory gland. By retrograde cobalt filling through the same trunks, different types of cells have been mapped in the terminal abdominal ganglion. About 25 dorsal unpaired median (DUM) neurons have been identified among them. About 36 octopamine-like immunoreactive DUM neurons with large somata have been characterized in whole-mount preparations of the terminal abdominal ganglion. The combination of the cobalt-filling technique with immunohistochemical mapping of cells suggests an octopaminergic innervation of the musculature of the accessory gland by DUM neurons.     

The electrical activity of mechanoreceptive and neurosecretory neurons in the stick insectCarausim morosus

Summary Action potentials have been recorded from the neurosecretory cells which lie on the link nerve inCarausius morosus. The neurosecretory cells are spontaneously active in completely isolated preparations, firing with a regular but low frequency (<1 imp/s) or in small bursts (12 imp/s). The action potentials recorded extracellularly from the neurosecretory fibres are characteristically of long duration (2 to 10 ms), whereas those of motor or sensory fibres are of shorter duration (0.6 to 0.8 ms). The neurosecretory action potentials are also characterised by their slow conduction velocity (0.15 to 0.25 m/s) compared to those from motor and sensory fibres (0.54 to 0.7 m/s). The action potentials are propagated from the region of the cell body towards the terminals and have been recorded passing along all the major nerves in the periphery.Recordings from three of the non-neurosecretory cells which lie on peripheral nerves show that they respond to stretching of the nerves upon which they lie or of nerves which branch in the immediate vicinity. The action potentials are propagated away from the cell body towards the central nervous system. The neurons are termed peripheral nerve stretch receptors.We are grateful to the Science Research Council for financial support.     

Post-Tetanic Repetitive Activity in the Cat Soleus Nerve : Its origin, course, and mechanism of generation

Subsequent to conditioning by a high frequency stimulus axons of the cat soleus nerve respond to single stimuli with brief trains of repetitive action potentials. This phenomenon, post-tetanic repetitive activity (PTR), was studied in individual axons and single motor units of an in situ cat soleus nerve-muscle preparation. The occurrence, intensity, and duration of PTR are principally dependent on the frequency and duration of the conditioning tetanus. PTR occurs synchronously in the axon and muscles of single motor units. An analysis of the temporal relationships of the repetitive nerve and muscle potentials showed that PTR is generated in the motor nerve terminal. It is postulated that PTR is produced by a generator potential which is developed in the post-tetanic period between the unmyelinated nerve terminal and the last node of Ranvier.     

Monosynaptic connections mediate resistance reflex in crayfish (Procambarus clarkii) walking legs

Summary In crustacean walking legs, the coxo-basipodite chordotonal organ (CB) composed of about 50 sensory cells, evokes a resistance reflex in the levator (Lev) and depressor (Dep) muscles responsible for the movements of the coxo-basipodite joint where it is located. Mechanical stimulation of the CB strand and electrical stimulation of its sensory nerve have been performed along with systematic intracellular recordings from CB terminals (CB T) and levator (Lev) or depressor (Dep) motoneurons (MNs) in order to study their connections. Measurements of conduction times in the CB nerve demonstrated different pools of sensory fibres, the fastest of which reach the ganglion in 2.5 ms. During imposed movements to the CB strand, intracellularly recorded Lev or Dep MN display EPSPs that are correlated to spikes in the CB nerve, their delays are incompatible with a polysynaptic pathway. Systematic stimulation of the CB nerve demonstrates that about 4 to 8 CB fibres are connected with each Lev or Dep MN. Classical tests for monosynaptic connections indicate that EPSPs occurring between 3 ms and 6 ms correspond mainly to monosynaptic connections with CB T, whereas IPSPs (the latencies of which are above 12 ms) are polysynaptic. In spite of the high selectivity of the CB T onto MNs, eight simultaneous intracellular recordings of coupled CB T and MN (out of more than 300 MNs penetrated) have allowed a direct measurement of synaptic delays (less than 1 ms). The functional significance of these results is discussed in relation to the proprioceptive control of locomotor movements.Abbreviations CB Coxo-basipodite chordotonal organ - CB n CB sensory nerve - CB T CB sensory terminal - Dep depressor - Lev levator - MN motoneuron     

Some structural and functional properties of nerves and muscles in the oviduct of the pink bollworm moth,Pectinophora gossypiella (Saund.)

The oviduct of the pink bollworm is innervated by an intrinsic neural network arising from 4 nerves from the terminal ganglion. The nerve tracts in this network often contained elliptical swellings, each with a central nucleus.

A distinct surface topography was evident in the muscular sheath of the common and lateral oviduct. A very thin muscular envelope consisting of an inner band of circular fibers and an outer layer of longitudinal fibers was also found in the ovarioles. Although conventional A and I bands were recognized, the z-disk was composed of an irregular and loose meshwork, suggesting that the visceral muscles of the reproductive tract possess super-contracting properties. Even when the oviduct and the ovarioles were isolated from the central nervous system, an endogenous rhythmic activity was evident.

Two types of spontaneous excitatory postsynaptic potentials were detected in the oviduct. The type most frequently observed had a complex of multiple spikes with a duration of 18–32 msec. The other type had a saw-tooth shape and a duration of 80–160 msec. Spontaneous action potentials with a plateau-type configuration and a duration of 280–320 msec were also observed. After the removal of the terminal ganglion, endogenous electrical activity distinct from the events just described was found in the midand upper common oviduct. Such discharges seem to originate from the intrinsic neural network and had durations similar to those found for neurosecretory cells.     

The Sense Organs and Ventral Ganglion of Sagitta (Chaetognatha)

This paper describes some features of the chaetognath nervous system from ultrastructural observations and observations on material stained with specific techniques for nervous tissue, and from records of the activity of the locomotor muscles and ventral ganglion. Sensory cells grouped on the ventral surface of the head bear ciliary processes (some with multiple tubules), and are probably in connexion with the central nervous system by their own axons, unlike the sensory cells of the hair fan vibration receptors of head and body. The ventral ganglion is motor to the locomotor muscles of the body, and controls the rhythmic locomotor activity of the animal. Electrical events associated with contraction of these muscles are compound non-overshooting spike-like potentials. The ventral ganglion contains several large nerve fibres constant in position and connexions in different individuals. Some of these arise from cells in the ganglia of the head, and pass to the ventral ganglion, others from cells within the ventral ganglion, and probably supply the ciliary hair fan receptors of the body, whilst the motor axons issuing from the ventral ganglion are smaller in diameter. The ganglion is arranged on a ladder-like plan, and axons of the lateral cell bodies cross the central neuropil transversely before they contribute to the longitudinal tracts or pass out in the radial nerves. Synapses in the neuropil contain 30–40 nm electron lucent vesicles; the transmitter is unknown, but is unlikely to be either acetylcholine or l -glutamate. Occasional larger electron dense vesicles up to 70 nm in diameter are also found within nerve fibres of the neuropil. It is concluded that the arrangement of the peripheral nervous system is unlike that of several groups which have been suggested as related to chaetognaths.     

Effects of high calcium solutions on glutamate sensitivity of crayfish muscle fibres

Crayfish neuromuscular preparations were studied after 18--36 h exposure to high calcium solutions. As previously reported for frog neuromuscular preparations the treatment damaged the nerve terminals and decreased junctional potentials. The resting potentials and input resistances of the muscle fibres were not affected; but their sensitivity to glutamate was significantly decreased when compared to that of control muscles. After exposure to high calcium, the sensitivity to gamma-aminobutyric acid, the putative transmitter at inhibitory synapses, was increased. Apparently normal twitches were elicited by direct stimulation, and calcium spikes could still be observed in the fibres. A decreased sensitivity to glutamate was also noted in experiments carried out on denervated muscles 8 months after section of the motor axons. Possible relations between nerve terminal damage and the decrease in sensitivity to glutamate are discussed.     

The supply of oxygen to the flight muscles of insects: A Theory of tracheole physiology

In the flight muscles of insects, virtually every mitochondrion is in contact with or is encircled by terminal tracheoles which reach them by following the channels formed by the invaginated plasma membrane of the muscle fibres, the T-system tubules. In Musca, Calliphora and Drosophila (Diptera), Apis (Hymenoptera) and Tenebrio (Coleoptera) the terminal tracheoles are smooth-surfaced tubes with a lumen of about 50 nm. In Pieris (Lepidoptera) the terminal tracheoles occupy the regular transverse tubular system which runs between the mitochondria and across the fibrils on either side of the H zone. They are smooth tubules of 80–200 nm diameter. Preliminary observations suggest the same arrangement in Ischnura (Odonata). In Rhodnius and other Hemiptera the transverse T-tubule system forms large cavities among the mitochondria: these cavities in Rhodnius are occupied by smooth-walled tracheole endings. In the mature adult of Schistocerca (Orthoptera) T-tubules of varying size are utilized by terminal tracheoles (diameter 50–100 nm). The terminal tracheoles of the flight muscles are highly permeable to myrcene and kerosine. They commonly fill with liquid during rest and this liquid is resorbed during activity. It is suggested that these adaptations increase the efficiency of respiration in the flight muscles by ensuring that, when it is most needed, gaseous oxygen extends to the surface of the mitochondria, from which it is separated by a very permeable barrier.     

Regeneration of motorneurones to a cricket muscle after partial or complete denervation

The metathoracic extensor tibiae muscle of the cricket Teleogryllus oceanicus is innervated by two excitatory axons; one of which leaves the metathoracic ganglion through nerve 5, the other through nerve 3. Axons in nerve 5 frequently regenerate to reinnervate the extensor tibiae if the nerve is sectioned in a late nymphal stage; functional reinnervation is rare if the nerve is sectioned in young adults. The muscle may become reinnervated by several axons regenerating through nerve 5, and individual muscle fibres may receive inputs from two regenerated axons. Axons regrowing through nerve 5 to a partially-denervated extensor tibiae preferentially innervate fibres in the central portion of the muscle, which is the normal innervation field of nerve 5. If the muscle is totally denervated by transection of both nerve 5 and nerve 3b, reinnervation is less specific and fibres throughout the muscle may be reinnervated by axons in either nerve. Reinnervation by regenerating axons is progressive. The proportion of muscles which are functionally reinnervated by regenerated axons increases with survival time as does the proportion of fibres within a muscle with reinnervation. The amplitude of excitatory junctional potentials and of muscle contraction evoked by regenerated axons both increase with survival time.     

Substance P immunoreactivity in rat von Ebner's gland

Summary The present immunohistochemical study revealed substance P-immunoreactive neuronal elements in the von Ebner's gland of rats. Immunoreactive ganglion cells were observed as single cells or groups of several immunoreactive ganglion cells among intra-lingual muscles, at the base of the vallate papillae and near the von Ebner's gland. Very numerous substance P-immunoreactive varicose nerve fibres ran closely associated with the serous cells and excretory duct cells, and were seen to run along blood vessels in the gland. Since substance P-immunoreactive ganglion cells were present near the glands, the immunoreactive varicose nerve fibres in the von Ebner's gland may be partly derived from the intra-lingual ganglion cells. These substance P-immunoreactive varicose nerve fibres may have an effect on the secretory activity of the serous cells and duct cells, and on the vasodilation of blood vessels of the von Ebner's gland. Actin immunoreactivity was seen in numerous myoepithelial cells embracing serous cells and duct cells, and in the smooth muscle cells of blood vessels of the gland. By using a double immunolabelling technique with anti-substance P and anti-actin sera, substance P-immunoreactive varicose nerve fibres were found to be in close contact with myoepithelial cells.     

Histology,histochemistry, and emptying mechanism of the venom glands of some elapid snakes

The venom glands of several species of elapid snakes are described. The main venom gland consists of many tubules which usually contain large amounts of secretion product. The accessory gland surrounds the entire venom duct and is usually composed of uniform mucous epithelium. The epithelium lining the tubules of the accessory gland of Naja naja is composed of two distinct types of cells. Histochemical tests indicate that the main venom gland reacts with mercury bromphenol blue and PAS but not with alcian blue. The accessory gland reacts with PAS and alcian blue, and not with mercury bromphenol blue. Treatment of sections with sialidase demonstrates the presence of a sialomucin in the accessory gland. Stimulation of the muscles associated with the venom gland offers an indication of the venom expulsion mechanism of Bungarus caeruleus. A comparison of the venom apparatus of elapid and viperid snakes emphasizes marked differences in the internal anatomy of the venom glands, muscles associated with the gland, and arrangement of glandular components. The morphological differences and dissimilar venom expulsion mechanisms support the recent view of the polyphyletic origin of venomous snakes.     

Reinnervation of adult muscle in organ culture restores tetrodotoxin sensitivity in the absence of electrical activity.

Strips of denervated adult mouse diaphragm muscle maintained in organ culture were reinnervated by nerve processes growing out from explants of embryonic mouse spinal cord. In vivo, following denervation, the action potential loses its sensitivity to tetrodotoxin; this sensitivity is regained upon reinnervation. Similarly, action potentials in cultured muscle fibres were insensitive to tetrodotoxin, and sensitivity was restored in muscle fibres that became reinnervated in vitro. Tetrodotoxin sensitivity was also restored in cultured muscle fibres reinnervated in the continuous presence of d-tubocurarine, but it was not induced by 4 days of direct electrical stimulation of noninnervated muscles. We conclude that developing nerve terminals can exert a trophic action on adult muscle fibres that is independent of electrical activity in the muscle.     

New techniques in female pudendal somatosensory evoked potential testing

Objective—The primary nerves innervating the female genitalia are the dorsal nerve of the clitoris (DNC) and the perineal nerve, which innervate the clitoris and the external genitalia/distal vagina, respectively. We describe two novel electrodiagnostic techniques for evaluating the integrity of these female genital somatosensory pathways.

Subjects and methods—Seventy-seven healthy women (mean age 29.3 years) were enrolled for this study. We performed DNC somatosensory evoked potentials (SEPs), stimulating through self-adhesive disk electrodes on either side of the clitoris. Perineal nerve SEPs were evoked through a vaginal probe. Cortical responses were measured through cup electrodes affixed to the scalp at Cpz and Fpz. Stimulus parameters were duration 0.1?ms, frequency 4.1?Hz, filters 5–5,000?Hz, at three times sensory threshold.

Results—DNC and perineal nerve SEPs from both the right and left sides were reproducible and easily discerned. The mean P1 latencies were: right DNC 39.4?ms (SD 2.8?ms), left DNC 39.3?ms (SD 3.3?ms), right perineal nerve 37.8?ms (SD 3.4?ms), left perineal nerve 37.6?ms (SD 3.1?ms). We recorded SEP responses from 90 to 92% of subjects for the DNC, and 69% for the perineal nerve.

Conclusions—We are able to evoke somatosensory potentials from the four primary somatic nerves that mediate female genital cutaneous sensation. In healthy subjects, the DNC responses are robust and maintain laterality. The perineal nerve responses are less consistently obtained, but when recorded, are easily discerned. These preliminary data provide a foundation from which to study female genital innervation, particularly as it applies to sexual function.     

Monoamines and acetyl-cholinesterase in the pineal gland and habenula of the ferret

Summary A histochemical method for demonstrating amines by fluorescence showed that the pinealocytes of the ferret contained a high concentration of a yellow fluorophore (probably 5-HT). Numerous green-fluorescent (noradrenaline-containing) nerve fibres occurred around intrapineal blood vessels, between pinealocytes and in the N. conarii (which entered the gland caudally). A collection of neuron-like cells (the pineal ganglion) lay, surrounded by a meshwork of nerve fibres, in the posterior part of the pineal. Neither the cells nor the fibres of the pineal ganglion contained monoamines, but both showed the presence of acetyl-cholinesterase which otherwise was found in the pineal only in fibres which stretched from the ganglion towards the cranial pole of the gland. The medial habenular nucleus showed a remarkable perivascular green fluorescence not seen in the lateral habenular nucleus nor anywhere else in the adjacent diencephalon and brain stem. The cells and fibres of this nucleus also contained much acetyl-cholinesterase.Bilateral superior cervical ganglionectomy, or treating animals with reserpine, removed the green fluorescence from both pineal nerve fibres and the habenula. Ganglionectomy also resulted in a progressive alteration in the colour of the parenchymal fluorescence from yellow to green; the original yellow colour was restored by treating ganglionectomised animals with nialamide (a monoamine oxidase inhibitor). L-Dopa, 5-hydroxytryptophan or nialamide, alone or in combination, had no effect on the fluorescence of the nerve fibres or cells of the pineal, or on the habenula.These results are related to previous findings that pinealectomy or ganglionectomy prevents the acceleration by artificial light of oestrus in ferrets.     

The action of iontophoretically applied L-glutamate on an insect visceral muscle

1) lontophoretic application of L-glutamate was employed to study the distribution of glutamate receptors in the superior longitudinal (SL) muscles of the locust (Locusta migratoria) hindgut, in which spontaneous activity was inhibited using normal saline containing 5 mM MgCl_2. 2) Junctional glutamate potentials with a rise time of 50–100 ms (peak) and a decay time of 250–400 ms were recorded at localized sites using ejection pulses in the range 5–10 nC. Most active sites were found in interfiber clefts and were spaced at about 250–300 μm intervals. 3) Desensitization of glutamate receptors occurred using ejection frequencies > 0.2 Hz. Desensitization could be irreversibly blocked using the lectin concanavalin A. 4) Depolarizing (D-) and biphasic depolarizing/hyperpofarizing (DH -) extrajunctional glutamate potentials were observed using ejection pulses > 15 nC. 5) δ-Philanthotoxin (δ-PTX) at concentrations > 0.3 Uml^?1 inhibited junctional glutamate potentials in a dose-dependent manner, 50% inhibition was achieved using 0.45 Uml^?1 δ-PTX. 6) Subthreshold concentrations of proctolin (up to 5 × 10^?10M) had no visible effect on glutamate potentials, suggesting that proctolin possibly does not act by modulating glutamate activity. 7) It is proposed that glutamate plays a transmitter role in SL muscles, while the role of proctolin is still unclear.     

The giant neurone system in ophiuroids

Summary The nervous system of Ophiura texturata contains nerve fibres and cell bodies that are an order of magnitude larger than anything previously described in the Asteroidea and Echinoidea. These large nerve cells are designated giant fibres. Giant nerve cells are present in both the ectoneural and hyponeural nervous system. The layout of these nerve cells is described and it is shown that the organization is repeated in each segmental ganglion that makes up the radial nerve cord. The circumoral nerve ring is composed, in the main, of tracts of nerve fibres joining the radial nerves, and it contains only limited areas of neuropil associated with the alimentary canal and muscles of the disc and jaws. Degeneration studies have shown that each segmental ganglion of the radial nerve cords contains a discrete population of neurones separate from adjacent ganglion and that there are not anatomically continuous giant fibres along the whole length of the nerve cord.     

The anatomy and physiology of the stomatogastric nervous system ofSquilla

Summary The stomatogastric nervous system of a mantis shrimp,Squilla oratoria, is described. The motor nerves of the stomatogastric ganglion (STG) and their innervation of muscles of the posterior cardiac plate (pcp) and pyloric systems are detailed.The STG contains more than 25 neurons. It sends out one pair of major output nerves. The pcp-pyloric cycle recorded from the motor axons in this nerve consists of rhythmic bursts of several units which fire with a characteristic phase relationship to each other. The rhythm is intrinsic to the STG itself, but it is modifiable.Recordings from the peripheral nerves reveal that identifiable cardiac plate, pyloric dilator and pyloric neurons control sequential contractions of the pcp and pyloric muscles to constrict or dilate a number of their attached ossicles.Several modulatory input fibres in the stomatogastric nerve, activated via stimulation of the superior or inferior oesophageal nerve (son, ion), prime or trigger the cyclic motor outputs. The son inputs induce distinct effects on the cardiac and pcp-pyloric pattern generators, while the ion inputs, via the oesophageal ganglion, excite only the pcp-pyloric generator.On the basis of anatomical and physiological observations, the possible functions of motor neurons involved in the pcp-pyloric cycle are described with reference to opening of the pcp and pyloric channels.This stomatogastric nervous system inSquilla is compared to that in decapods which has been well analyzed.Abbreviations CG commissural ganglion - ion inferior oesophageal nerve - lvn lateral ventricular nerve - OG oesophageal ganglion - pep posterior cardiac plate - son superior oesophageal nerve - STG stomatogastric ganglion - stn stomatogastric nerve - ivn inferior ventricular nerve     

The radial decline of nerve impulses in a restricted cylindrical extracellular space

In order to increase the potentials recorded extracellularly from nerve fibres, peripheral nerves are often placed in restricted space with cylindrical geometry. Equations are derived for computing the potentials expected at the surface of the cylinder, based on the potentials at the external surface of a small nerve fibre located on the long axis of the cylinder. These equations are evaluated numerically, using two formulae for a nerve impulse given in the literature. In both cases there is little attenuation for cylinders with radii less than 0.5 mm, but the potential declines approximately as a power of radius b for 1<b<10 mm. Various factors which might affect these results under different experimental conditions are discussed.     

Simultaneous Recording of Input and Output of Lateral Geniculate Neurones

TO understand the way in which the cat dorsal lateral geniculate nucleus (LGN) processes visual information it would be useful to know the number and type of retinal inputs to individual LGN neurones. Using electrical stimulation of the optic nerve Bishop et al.¹concluded that an impulse in a single optic nerve fibre is sufficient to excite a single LGN neurone. From the appearance of excitatory postsynaptic potentials (EPSPs) recorded essentially intracellularly, Creutzfeldt suggested that LGN neurones are driven by perhaps one² or a few³ retinal ganglion cells. Hubel and Wiesel⁴ proposed models of convergence of several retinal inputs on single LGN neurones based on analyses of receptive fields. Guillery⁵ produced anatomical evidence that some types of LGN neurones receive inputs from several different retinal fibres. Now we report direct observations which were made by recording simultaneously from single LGN neurones and from individual retinal ganglion cells which provided excitatory input to them. We shall not consider inhibitory influences, which are currently under study.