CHARACTERISATION OF LARGE SECOND-ORDER OCELLAR NEURONES OF THE BLOWFLY CALLIPHORA ERYTHROCEPHALA

1. Blowflies have twelve large, second-order ocellar neurones (L-neurones) with axons in the single ocellar nerve. These neurones have fairly restricted arborizations in the posterior slope neuropile of the protocerebrum and cell bodies in the nerve, near to the fused ocellar retinae. 2. Like ocellar L-neurones of other insects, or large second-order neurones of the fly compound eye, blowfly L-neurones hyperpolarise in response to increases in light intensity and depolarise in response to decreases in light intensity. Both polarities of response have a strong phasic component. Adaptation to sustained illumination shifts the intensity­response curve, with little change in its gradient. 3. The maximum responses of blowfly L-neurones to sinusoidal changes in light intensity occur at stimulus frequencies of 5­10 Hz. 4. Hyperpolarising an L-neurone with small currents causes an increase in input resistance. Larger hyperpolarising currents cause oscillations in the membrane potential. The amplitude of the oscillations increases with current strength. Repolarisation generates brief rebound spikes of variable amplitude. 5. Injection of small hyperpolarising currents increases the amplitude of a response to a subsaturating pulse of light. This effect is not seen for saturating responses to light and is likely to be due to the increase in membrane resistance caused by hyperpolarisation.

Ultrastructural morphology of neurosecretory neurons in the barnacle Balanus amphitrite

Barnacles are very specialized Crustacea, with strongly reduced head and abdomen. Their nervous system is rather simple: the brain or supra-oesophageal ganglion (SG) is a small bilobed structure and the toracic ganglia are fused into a single ventral mass, the suboesophageal ganglion (VG). Neurosecretion was shown in barnacle nervous system by histochemical methods and numerous putative hormonal substances were extracted and tested. Recently six different types of dense-core granules were visualized in the median ocellar nerve of Balanus hameri and serotonin and FMRF-amide like substances were immunocytochemically detected in the nervous system of Balanus amphitrite. The aim of the present work is to localize and characterize at ultrastructural level, neurosecretory neuron cell bodies in the VG of Balanus amphitrite.Specimens of Balanus amphitrite were collected in the port of Genova. The central nervous system were Karnovsky fixed, osmium postfixed, ethanol dehydrated and Durcupan ACM embedded. Ultrathin sections were stained with uranyl acetate and lead citrate. Ultrastructural observations were made on a Philips M 202 and Zeiss 109 T electron microscopy.

Feedback synaptic interaction in the dragonfly ocellar retina.

The intracellular response of the ocellar nerve dendrite, the second order neuron in the retina of the dragonfly ocellus, has been modified by application of various drugs and a model developed to explain certain features of that response. Curare blocked the response completely. Both picrotoxin and bicuculline eliminated the "off" overshoot. Bicuculline also decreased the size of response and the sensitivity. gamma-Aminobutyric acid (GABA), however, increased the size of response. The evidence indicates the possibility that the receptor transmitter is acetylcholine and is inhibitory to the ocellar nerve dendrite whereas the feedback transmitter from the ocellar nerve dendrite may be GABA and is facilitory to receptor transmitter release. The model of synaptic feedback interaction developed to be consistent with these results has certain important features. It suggests that the feedback transmitter is released in the dark to increase input sensitivity from receptors in response to dim light. This implies that the dark potential of the ocellar nerve dendrite may be determined by a dynamic equilibrium established by synaptic interaction between it and the receptor terminals. Such a system is also well suited to signalling phasic information about changes in level of illumination over a wide range of intensities, a characteristic which appears to be a significant feature of the dragonfly median ocellar response.

A Study of the Afferent Pathways of the Dragonfly Lateral Ocellus from Extra-Cellularly Recorded Spike Discharges

1. The lateral ocellar nerve of adult dragonflies contains at least two kinds of afferent nerve fibres: the ‘giant’ afferent and small afferents. Efferent fibres are also present, but are not described here. 2. The afferent fibres and receptor axons were studied by extracellular recording of their spike discharges. Experiments using light and electrical (D.C.) stimulation and the application of magnesium were performed. 3. Responses of the giant afferent were analysed into a number of exponential components. The number could be altered experimentally. 4. The giant afferent shows the phenomenon of ‘delayed response’, a delayed onset of the off-response to brief light stimuli. A qualitative model for delayed responses, incorporating the exponential components, is described. 5. It is concluded that the small afferents behave in accordance with Ruck's model of ocellar of functioning but that the giant afferent does not. It is proposed that inhibion in the giant afferent fibre is produced electrotonically, but there may be a synaptic contribution also.

Neural Organization of the Median Ocellus of the Dragonfly

Intracellular responses from receptors and postsynaptic units have been recorded in the median ocellus of the dragonfly. The receptors respond to light with a graded, depolarizing potential and a single, tetrodotoxin-sensitive impulse at "on." The postsynaptic units (ocellar nerve dendrites) hyperpolarize during illumination and show a transient, depolarizing response at "off." The light-evoked slow potential responses of the postsynaptic units are not altered by the application of tetrodotoxin to the ocellus. It appears, therefore, that the graded receptor potential, which survives the application of tetrodotoxin, is responsible for mediating synaptic transmission in the ocellus. Comparison of pre- and postsynaptic slow potential activity shows (a) longer latencies in postsynaptic units by 5–20 msec, (b) enhanced photosensitivity in postsynaptic units by 1–2 log units, and (c) more transient responses in postsynaptic units. It is suggested that enhanced photosensitivity of postsynaptic activity is a result of summation of many receptors onto the postsynaptic elements, and that transients in the postsynaptic responses are related to the complex synaptic arrangements in the ocellar plexus to be described in the following paper.

Neural Organization of the Median Ocellus of the Dragonfly

Two types of presumed synaptic contacts have been recognized by electron microscopy in the synaptic plexus of the median ocellus of the dragonfly. The first type is characterized by an electron-opaque, button-like organelle in the presynaptic cytoplasm, surrounded by a cluster of synaptic vesicles. Two postsynaptic elements are associated with these junctions, which we have termed button synapses. The second synaptic type is characterized by a dense cluster of synaptic vesicles adjacent to the presumed presynaptic membrane. One postsynaptic element is observed at these junctions. The overwhelming majority of synapses seen in the plexus are button synapses. They are found most commonly in the receptor cell axons where they synaptically contact ocellar nerve dendrites and adjacent receptor cell axons. Button synapses are also seen in the ocellar nerve dendrites where they appear to make synapses back onto receptor axon terminals as well as onto adjacent ocellar nerve dendrites. Reciprocal and serial synaptic arrangements between receptor cell axon terminals, and between receptor cell axon terminals and ocellar nerve dendrites are occasionally seen. It is suggested that the lateral and feedback synapses in the median ocellus of the dragonfly play a role in enhancing transients in the postsynaptic responses.

Electrophysiology of the Insect Dorsal Ocellus

Dorsal ocelli are small cup-like organs containing a layer of photoreceptor cells, the short axons of which synapse at the base of the cup with dendritic terminals of ocellar nerve fibers. The ocellar ERG of dragonflies, recorded from the surface of the receptor cell layer and from the long lateral ocellar nerve, contains four components. Component 1 is a depolarizing sensory generator potential which originates in the distal ends of the receptor cells and evokes component 2. Component 2 is believed to be a depolarizing response of the receptor axons. It evokes a hyperpolarizing postsynaptic potential, component 3, which originates in the dendritic terminals of the ocellar nerve fibers. Ocellar nerve fibers in dragonflies are spontaneously active, discharging afferent nerve impulses (component 4) in the dark-adapted state. Component 3 inhibits this discharge. The ERG of the cockroach ocellus is similar. The main differences are that component 3 is not as conspicuous as in the dragonflies and that in most cases ocellar nerve impulses appear only as a brief burst at "off." In one preparation a spontaneous discharge of nerve impulses was observed. As in the dragonflies, this was inhibited by illumination.