DeForest Mellon

Identification of three classes of multiglomerular, broad-spectrum neurons in the crayfish olfactory midbrain by correlated patterns of electrical activity and dendritic arborization

We recorded electrical activity from three different classes of broad-spectrum, multiglomerular neurons in the crayfish (Procambarus clarkii, P. blandingi) olfactory midbrain. Responses were obtained to odorants and electrical stimuli applied to the antennules of isolated, perfused head preparations. All three neuronal types responded to a complex mixture of five amino acids as well as to solutions of a commercial fish food. At least two classes also responded to individual amino acids and to sugars. The response properties and the morphologies of the neurons were unique to each type. Responses of Type I cells were stimulus-dependent excitatory postsynaptic potentials and superimposed impulse trains; those in Type II were stimulus-dependent inhibitory postsynaptic potentials; those in Type III were compound responses consisting of short latency hyperpolarizations, followed by depolarizing post-synaptic potentials and impulses. All three cell types had extensive, multiglomerular dendritic arbors in the olfactory lobe, but each of their respective branching pattern morphologies was distinctive. Two types had additional dendrite branches in the lateral antennular neuropil and the olfactory-globular tract neuropil. We conclude that these broad-spectrum neurons are part of a parallel olfactory pathway that is separate from the putative quality coding circuitry in the crayfish olfactory system.

PHYSIOLOGICAL CHARACTERIZATION OF ANTENNULAR FLICKING REFLEXES IN THE CRAYFISH

Abstract The cellular substrates of antennular flicking behavior in the crayfish Procambarus clarkii were investigated. Flicking involves fast downward movements of the external filament of each biramous antennule (1st antenna), and is mediated by phasic contractions of a short muscle, the external filament depressor. Phasic contractions of the external filament depressor depend upon stereotyped impulse bursts in a single motorneuron (P1). These bursts have a characteristic impulse frequency profile that is consistent upon successive occurrences. The temporal characteristics of the impulse burst suggest that the central depolarizations generating each burst may be similar to driver potentials described for motor neurons in crustacean cardiac ganglia. Responses of the external filament to odorants have a long latency and are characterized by repetitive bursts and tonic activity in some external filament depressor fibers. Tonic activity in a slowly contracting muscle, the antennular depressor muscle, is also evoked by chemical stimulation. Flicking is consistently evoked only by mechanical or hydrodynamic stimulation of the cephalothorax, antennae and antennules. The sensitivity and short latency of the hydrodynamic antennule-generated flick reflex is consistent with the sensitivity of rapidly conducting, hydrodynamically activated mechanoreceptor neurons in both antennular filaments. I propose that antennular flicking, which has been shown to enhance the dynamic response characteristics of olfactory receptor neurons on the external antennular filament, has evolved as a response to the turbulence associated with fluid movement, within which chaotic odorant concentration fronts may be imbedded.The cellular substrates of antennular flicking behavior in the crayfish Procambarus clarkii were investigated. Flicking involves fast downward movements of the external filament of each biramous antennule (1st antenna), and is mediated by phasic contractions of a short muscle, the external filament depressor. Phasic contractions of the external filament depressor depend upon stereotyped impulse bursts in a single motorneuron (P1). These bursts have a characteristic impulse frequency profile that is consistent upon successive occurrences. The temporal characteristics of the impulse burst suggest that the central depolarizations generating each burst may be similar to driver potentials described for motor neurons in crustacean cardiac ganglia. Responses of the external filament to odorants have a long latency and are characterized by repetitive bursts and tonic activity in some external filament depressor fibers. Tonic activity in a slowly contracting muscle, the antennular depressor muscle, is also evoked by chemical stimulation. Flicking is consistently evoked only by mechanical or hydrodynamic stimulation of the cephalothorax, antennae and antennules. The sensitivity and short latency of the hydrodynamic antennule-generated flick reflex is consistent with the sensitivity of rapidly conducting, hydrodynamically activated mechanoreceptor neurons in both antennular filaments. I propose that antennular flicking, which has been shown to enhance the dynamic response characteristics of olfactory receptor neurons on the external antennular filament, has evolved as a response to the turbulence associated with fluid movement, within which chaotic odorant concentration fronts may be imbedded.

Modification of structure and synaptic physiology in transformed Shrimp muscle

Summary1.The gross anatomy of the nerves and muscles in the pincer and snapper claws ofAlpheus armillatus is described. In both claws there are three muscles: the opener muscle, the main closer muscle, Cl1, and the small closer muscle, Cl2.2.The number of fibers in each of the two closer muscles is similar for a pair of claws removed from the same animal. However, the size of individual closer muscle fibers in the snapper is about two times greater, both in length and diameter, than in the pincer.3.Sarcomeres in fibers of the opener muscle and the small closer muscle are similar in length for the pincer and the snapper. In contrast, sarcomeres in the main closer muscle of the pincer are shorter than those in the snapper Cl2 muscle.4.Excitatory junctional potentials (ejps) recorded from single pincer Clx muscle fibers at 1 Hz were always small in amplitude (≦12 mV) and facilitated poorly at 10 Hz. Snapper Cl1 ejps ranged up to about. 30 mV in amplitude and displayed a large degree of facilitation at 10 Hz.5.During the process of pincer transformation into a snapper claw, parallel changes occur in both the sarcomere length of Cl1 muscle fibers and the degree of ejp facilitation.

Reflex actions of the functional divisions in the crayfish oculomotor system

SummaryThe electrical responses of motor neurons in different anatomical subdivisions of the crayfish oculomotor system were examined during various kinds of experimentally manipulated sensory stimulation. Geotactic reflexes are effected by neurons in the anterior motor cluster and the medulla terminalis. Optokinetic and proprioceptive nystagmus are generated by neurons in the lateral motor cluster. This functional diversity in the major subdivisions contrasts with an intradivisional homogeneity of function, in that the different motor neurons of each all contribute to reflexes initiated by different kinds of sensory input.

Nitric oxide as a putative messenger molecule in the crayfish olfactory midbrain

NADPH-d histochemistry was used to investigate presumptive nitric oxide synthase (NOS)-containing neurons in the crayfish olfactory midbrain. Three anatomically different types of local olfactory interneurons exhibiting NADPH-d activity were observed: two pairs of large interneurons as well as positively stained globuli cells. Branches derived from the large interneurons were confined to the ipsilateral olfactory lobe and accessory lobe, but only a few branches innervated the olfactory lobe glomeruli. Local field potential recordings on the olfactory lobe showed that administration of SNP or SIN-1 (10-4 M) into the brain had reversible inhibitory effects on electrically-evoked responses of unidentified neuronal cell populations.

Integration of hydrodynamic and odorant inputs by local interneurons of the crayfish deutocerebrum

SUMMARY Intracellular electrodes were used to record from local interneurons in the olfactory lobes of the midbrain in the crayfish Procambarus clarkii. Cells that resembled previously studied central targets of olfactory receptor neurons on the lateral antennular flagellum were specifically examined for their responses to hydrodynamic stimuli. Initiation of water movement past the antennular flagellum, confined within an olfactometer, evoked a triphasic excitatory-inhibitory-excitatory postsynaptic potential lasting up to 2 s that generated spikes on depolarizing phases of the response sequence. Odorant pulses seamlessly imbedded in the water pulse past the antennule evoked purely excitatory, dose-dependent postsynaptic responses and associated spike trains. The latency of the initial phase of the response to water was approximately half as long as the latency of the response to odorant, suggesting that different afferent pathways are involved in responses to hydrodynamic and odorant stimuli, respectively. In some olfactory lobe interneurons that resembled previously described cells classified as Type I, conjoint stimulation of fluid onset and odorant evoked responses that were twice the amplitude of the summed response to either hydrodynamic or odorant stimulation alone, suggesting that the olfactory responses were potentiated by hydrodynamic input. Individuals of at least one other class of first-order interneuron that responded to both hydrodynamic and odorant stimulation were occasionally recorded from. These results indicate that multimodal integration of chemical and mechanical information occurs at the level of first-order sensory interneurons in the crayfish brain.

Parasol cells of the hemiellipsoid body in the crayfish Procambarus clarkii: Dendritic branching patterns and functional implications

Multimodal, higher‐order sensory integration in decapod crustaceans occurs in local interneurons (parasol cells) within a structure in the lateral protocerebrum, the hemiellipsoid body, which is located dorsal to the terminal medulla. The hemiellipsoid body is targeted by projection neuron inputs by means of the olfactory globular tract from bilateral deutocerebral neuropils, the accessory lobes, which receive secondary visual, mechanosensory, and olfactory inputs. Parasol cell dendrites arborize extensively within the two neuropils of the hemiellipsoid body and possibly have some neurites within another neuropil at its base. The two neuropils of the hemiellipsoid body, neuropils I and II, are known to receive asymmetrical inputs from the contralateral and ipsilateral accessory lobes, and our current study addresses the question of the distribution of parasol cells within these two neuropils. Three anatomic methods were used to analyze this distribution: intracellular filling of cells with neurobiotin and visualization of the cells by using either a fluorescent or a peroxidase avidin conjugate, or placement of a fluorescent lipophilic tracer within a lobe of the hemiellipsoid body. All of these methods demonstrated that single parasol cells exclusively arborize within one of the two lobes of the hemiellipsoid body, but not in both lobes. Electrophysiological recordings from pairs of parasol cells with dendrites in the same or different lobes confirm a functional separation between neuropils I and II. Comparisons are made between insect and crustacean systems, emphasizing the inputs to the hemiellipsoid body and the mushroom body and similarities between extrinsic cells in insects and parasol cells in decapod crustaceans. J. Comp. Neurol. 462:168–179, 2003.

Combining Dissimilar Senses: Central Processing of Hydrodynamic and Chemosensory Inputs in Aquatic Crustaceans

Aquatic environments are by their nature dynamic and dominated by fluid movements driven by lunar tides, temperature and salinity density gradients, wind-driven currents, and currents generated by the earths rotation. Accordingly, animals within the aquatic realm must be able to sense and respond to both large-scale (advection) and small-scale (eddy turbulence) fluid dynamics, for chemical signals critically important for their survival are embedded within such movements. Aquatic crustaceans possess many types of near-field fluid-flow detectors and two general classes of chemoreceptors on their body appendages: high-threshold, near-field receptors that may be somewhat equated with the sense of taste, and low-threshold far-field receptors that can be considered as olfactory. This review briefly summarizes the distribution of hydrodynamic and high-threshold chemoreceptors in aquatic crustaceans and the physiological characteristics of olfactory receptors in lobsters; it also examines recent physiological evidence for the central nervous integration of inputs from olfactory receptors and hydrodynamic detectors, two dissimilar senses that must be combined within the brain for survival. Marine crustaceans have provided valuable insights about mechanisms of primary olfactory sensory physiology; their additional sensitivity to hydrodynamic stimulation makes them a potentially useful model for examining how these two critical sensory inputs are combined within the brain to enhance foraging behavior. Multimodal sensory processing is critically important to all animals, and the principles and concepts derived from these crustacean studies may provide generalities about neuronal processing across taxa.

RESPONSE PROPERTIES OF HIGHER LEVEL NEURONS IN THE CENTRAL OLFACTORY PATHWAY OF THE CRAYFISH

Abstract We have examined the electrical activity of interneurons within the higher levels of the crayfish olfactory system. In unstimulated isolated crayfish head preparations, local protocerebral interneurons (LPI) of the hemiellipsoid bodies generate periodic, low-frequency membrane depolarizations. The most reasonable explanation for these baseline fluctuations, which were exhibited by all of the LPIs examined and which were reversibly abolished by either tetrodotoxin or low-calcium saline solution, is that they reflect periodic synaptic drive from the axon terminals of olfactory projection neurons. One-third of tested LPIs generated impulses in response to the odor stimuli we applied to the antennules. Those cells that did respond exhibited a brief excitatory postsynaptic potential and one or two action potentials, even during prolonged odor pulses. Many of the responding neurons also exhibited a delayed impulse burst 1 or 2 s following the stimulus pulse. Most of the responding cells recovered their sensitivity to odors very slowly, exhibiting disadaptation periods of several minutes. The apparent refractory nature of individual LPIs to olfactory stimulation is attributed in part to a hypothesized selectivity of connections between projection neurons and protocerebral targets and in part to the electrical isolation of the recording electrode from many regions of the extensive LPI dendritic tree.

Changes in myofibrillar gene expression during fiber-type transformation in the claw closer muscles of the snapping shrimp, Alpheus heterochelis.

Isotopes of a number of crustacean myofibrillar proteins have been identified with sodium dodecyl sulfate-polyacrylamide electrophoresis, and their distribution in muscles of the snapping shrimp has been examined. Fast-slow differences in distribution have been observed for myosin light chains and tropomyosin. In contrast, three troponin T subunits have been resolved, each specific to one of the three muscles examined. This result suggests that expression of crustacean contractile proteins is not accomplished by a simple coexpression of a battery of slow or fast isotopes. In addition, the expression of these proteins was examined during the quasi-developmental fiber-type transition of the main claw closer muscle during the reversal of claw asymmetry in response to the loss of the large snapper appendage. The changes observed appear similar to the cross-innervation induced changes in gene expression of vertebrate muscle.