Snail Hermissenda Research Articles

An artificial neural network system for temporal-spatial sequence processing

An artificial network is described that can learn, recognize, and generate higher-order temporal-spatial sequences. It consists of three parts: (1) comparator units, (2) a parallel array of artificial neural networks that are derived from the visual-vestibular networks of the snail Hermissenda, as well as hippocampal neuroanatomy, and (3) delayed feedback lines from the output of the system to the neural network layer. Its advantages include short training time, fast and accurate retrievals, toleration of spatial noise and temporal gaps in test sequences, and ability to store a large number of temporal sequences consisting of non-orthogonal spatial patterns.

Tumor Necrosis Factor‐α (TNF‐α), Interferon‐γ, and Interleukin‐6 but Not TNF‐β Induce Differentiation of Neuroblastoma Cells: The Role of Nitric Oxide

Previous observations have implicated GABA as a neurotransmitter released by the vestibular sensory neurons ("hair cells") of the snail Hermissenda onto visual sensory neurons, the type B cells, whose cell bodies are the sites of biophysical and biochemical changes during and following Pavlovian conditioning. Still other observations demonstrated that light-GABA pairings that simulate stimuli presented during Pavlovian conditioning cause prolonged elevation of intracellular Ca2+ and transformation of GABA-induced synaptic inhibition into excitation. Intracellular Ca2+ signals in response to GABA perfused onto the postsynaptic type B terminal branches are shown here to be prolonged on days after conditioning, but not after control paradigms. These and past results demonstrate two separate sites, i.e., the cell body and the terminal branches, for learning-induced changes after Pavlovian conditioning.

Associative learning in a network model of Hermissenda crassicornis. I. Theory.

A time-varying Resistance-Capacitance (RC) circuit computer model was constructed based on known membrane and synaptic properties of the visual-vestibular network of the marine snail Hermissenda crassicornis. Specific biophysical properties and synaptic connections of identified neurons are represented as lumped parameters (circuit elements) in the model; in the computer simulation, differential equations are approximated by difference equations. The model's output, membrane potential, an indirect measure of firing frequency, closely parallels the behavioral and electrophysiologic outputs of Hermissenda in response to the same input stimuli presented during and after associative learning. The parallelism of the computer modeled and the biologic outputs suggests that the model captures the features necessary and sufficient for associative learning.

Classical conditioning and protein kinase C activation regulate the same single potassium channel in Hermissenda crassicornis photoreceptors.

The patch-clamp technique was used to study the effects of classical conditioning and protein kinase C (PKC) activation on K+ channels of identified neurons in the snail Hermissenda crassicornis. Here we present evidence that classical conditioning and PKC activation similarly modify the same K+ channel. K+ channels were recorded in cells from animals with different training experience. The 64-pS K+ channel appeared with significantly lower frequency in the conditioned group compared to the frequencies in control animals (naive and unpaired). In addition, when present, the 64-pS channel exhibited a lower percentage of open time and an increased interval between opening bursts in cells from conditioned animals. The 42-pS K+ channel was observed with about the same frequency in all three groups, and its percentage of open time was invariant, regardless of the animal's experience. Incubation of the photoreceptor with the PKC activator phorbol 12,13-dibutyrate (PDBu) led to a profound decrease in the percentage of open time of the 64-pS K+ channel, from 35.7% in the control group to 2.5% in the PDBu-treated group. The inactive phorbol 4 alpha-phorbol 12-myristate 13-acetate had no effect. The use of the PKC inhibitor H-7 significantly blocked the phorbol effect. Inside-out patches obtained from phorbol preincubated cells likewise showed the same effect of PDBu on K+ channels, but the effect was not observed when phorbol was added after the cell-free patches were obtained from nontreated cells. By contrast, the percentage of open time of the 42-pS K+ channel remained unchanged after phorbol treatment.

The role of calcium in prolonged modification of a GABAergic synapse

Caudal hair cell impulses cause postsynaptic inhibition of ipsilateral type B photoreceptors in the snail Hermissenda. This inhibition is shown to be GABAergic according to a number of criteria. HPLC, mass spectrophotometric, and immunocytochemical techniques demonstrated the presence of GABA in the hair cells and their axons. GABA agonists and antagonists mimic and block the synaptic effect in a manner consistent with endogenous GABAergic transmission. Other properties, including I-V relations, conductance changes and reversal potentials, are comparable for exogenous GABA responses and endogenous effects of the hair cell impulses. This inhibitory synapse has been found to undergo a long-lasting transformation into an excitatory synapse if GABA release is paired with post-synaptic depolarization. GABA, via GABAA and GABAB receptors in the B cell, causes the opening of calcium sensitive chloride and potassium channels that leads to the post-synaptic hyperpolarization. GABA also induces a long-lasting intracellular calcium elevation at the terminal branches of the B cell that greatly outlasts the voltage responses. Synaptic transformation induced by pairings is caused by a decrease in both GABA induced chloride and potassium conductances in the post-synaptic B cell, as well as a significant prolongation of the intracellular calcium accumulation in the B cell's terminal axonal branches.

Biophysical and behavioral correlates of memory storage, degradation, and reactivation.

Neural correlates of associative memory and "forgetting" were observed 1, 6, and 14 days after acquisition of a conditioned response (CR) in the marine snail Hermissenda. Behavioral expression of a light-rotation association, as indexed by contraction of the animal's foot in response to light, dissipated throughout the 14-day interval such that a CR was observed 1 and 6 days after conditioning but was absent 14 days later. In relation to naive or pseudoconditioned animals, membrane resistance (inversely related to neuronal membrane conductance and directly related to excitability) of the isolated Type B photoreceptor (B cell) was elevated in conditioned animals on Days 1 and 6, whereas no elevation was detectable on Day 14. However, both the behavioral response and the elevated membrane resistance in conditioned animals were hypersensitive to light-rotation pairings (i.e., exhibited "savings") on Day 14, which is indicative of a latent memory trace. In a second experiment, a current-induced depolarization of the B cell after 14 retention days resulted in an increase in input resistance of the B cell membrane in previously conditioned animals but a weaker, transient rise in resistance in B cells from animals exposed to the nonassociative control procedure 14 days earlier. This effect was Ca(2+)-dependent because no rise in resistance was observed if Ca2+ was removed from the extracellular bath. These results indicate that modification of membrane conductance (i.e., elevated resistance), although apparently critical for the behavioral expression of the memory, is not essential for the maintenance of the latent memory trace, whereas Ca2+ hypersensitivity may be a principal contributor to the storage of a latent memory trace and memory reactivation.

Early Enhancement of Calcium Currents by H- ras Oncoproteins Injected into Hermissenda Neurons

Influx of calcium through membrane channels is an important initial step in signal transduction of growth signals. Therefore, the effects of Ras protein injection on calcium currents across the soma membrane of an identified neuron of the snail Hermissenda were examined. With the use of these post-mitotic cells, a voltage-sensitive, inward calcium current was increased 10 to 20 minutes after Harvey-ras oncoproteins were injected. The effects of oncogenic Harvey ras p21 protein (v-Ras) occurred quickly and were sustained, whereas the effects of proto-oncogenic ras protein (c-Ras) were transient. This relative potency is consistent with the activities of these oncoproteins in stimulating cell proliferation. Thus, this calcium channel may be a target for Ras action.

Discrimination learning alters the distribution of protein kinase C in the hippocampus of rats

Protein kinase C (PKC), an enzyme that plays an essential role in eukaryotic cell regulation (Nishizuka, 1988; Huang et al., 1989), is critical to memory storage processes both in the marine snail Hermissenda crassicornis and in the rabbit (Alkon et al., 1988; Bank et al., 1988; Olds et al., 1989). Specifically, activation of PKC mimics neurobiological correlates of classical conditioning in both Hermissenda and the rabbit, and the distribution of the enzyme within the rabbit hippocampus changes after Pavlovian conditioning. Here, we report that the amount of PKC, as assayed by specific binding of 3H-phorbol-12,13-dibutyrate (3H-PDBU), decreased significantly within the hippocampal CA3 cell region in rats trained to solve a water maze task either by cognitive mapping or by visual discrimination strategies, but not in control rats. Furthermore, hippocampal lesions interfered with acquisition of both of these tasks. We interpret these findings to support the conclusion that distributional changes of PKC within the mammalian hippocampus play a crucial role in memory storage processes.

Early regulation of membrane excitability by ras oncogene proteins

Two electrode voltage clamp conditions were used to study the early effects on ionic membrane channels of the intracellularly injected proto-oncogenic form of c-Ha-ras (c-ras) and its oncogenic counterpart v-Ha-ras (v-ras). These experiments were conducted on isolated somata of identified fully differentiated neurons of the sea snail Hermissenda. 20 min after c-ras, and 10 min after v-ras intracellular injections into type B medial photoreceptors of Hermissenda, the peak amplitude of two outward potassium currents (IA and IC), across the isolated Type B soma membrane begin to decrease. These two currents have been previously isolated by differences in activation and inactivation kinetics and their response to pharmacological blockers. c- or v-ras injections did not have any effect on a voltage-dependent inward calcium current. Reduction of IA preceded that of IC. Current reductions due to c-ras, but not to v-ras injection reversed spontaneously after 40 min. The voltage dependence of the steady state inactivation of IA shifted toward more negative potentials with ras injections. Ras-mediated cell transformations therefore, could involve, perhaps as initial events, prolonged modification of membrane currents.

Specificity of molecular changes in neurons involved in memory storage

Evidence implicating molecular steps in memory storage is discussed, particularly with reference to molecular specificity and uniqueness and the possible relevance of these steps to other types of long-lasting transformations such as those of development, regeneration, and tumorigenesis. The role of protein kinase C-mediated phosphorylation of identified protein subtrates, such as a 20,000-dalton GTP-binding protein, is described for associative memory of the snail Hermissenda, associative conditioning of the rabbit, and long-term potentiation. Cyclic AMP-mediated phosphorylation during sensitization of the snail Aplysia is also examined.

Sequential changes of potassium currents in Hermissenda Type B photoreceptor during early stages of classical conditioning

Classical conditioning of the marine snail Hermissenda can be produced in a single session of 50 pairings of light and rotation stimuli. Voltage clamp measurements of two outward K + currents, I A and I Ca 2+/1 bK + were obtained from medial Type B photoreceptors that were isolated from the nervous system 1 day after animals were exposed to paired light and rotation stimuli or control procedures (Unpaired, or no exposure to light and rotation), I Ca 2+−K + was found to be unchanged 18–30 h after 50 training trials. This result is consistent with a previous study where I Ca 2+/−K + was found to be unchanged after 50 light and rotation trials, although significantly reduced by 100 trials. In the present study 50 pairings of light and rotation produced a significant reduction in I A, suggesting an important role for this current in the earliest stages of classical conditioning.

The interstimulus interval and classical conditioning in the marine snail Hermissenda crassicornis

We examined the role of the interstimulus interval for the conditioned association between light and rotation stimuli in the marine snail Hermissenda. This interval between the conditioned stimulus (CS) and the unconditioned stimulus (US) is an important and widespread property of vertebrate associative learning. We demonstrated that with a forward CS-US delay of 1.0 s we were able to produce significant 24-h retention of an associative memory after 50 training trials. Other paired treatments providing intervals of 1.5 s, 0.5 s, simultaneous, and backward arrangements did not support retention at 24 h.

The nature of memory

Memory, and its failings, are surely the most important factors in determining human behaviour. In humans and the higher animals the memory process is manifestly so complex as to defy direct experimental investigation. However, the snail Hermissenda crassicornis, with a nervous system many orders of magnitude simpler, has been shown to be capable of associative learning, and investigation of its cellular basis throws light on the mechanisms governing the memory process generally, and, more specifically, as now being revealed in the rabbit hippocampus and cerebellum.

Classical conditioning of Hermissenda: origin of a new response

Training of the marine snail Hermissenda crassicornis with paired light and rotation was previously shown to result in acquisition and retention of a behavioral change with many features characteristic of vertebrate associative learning. Here, this behavioral change is demonstrated to be classical, Pavlovian-like conditioning. A new response to light is formed (the CR) that is pairing-specific and resembles the unconditioned response (UCR) to rotation. The conditioned and unconditioned responses are relatively rapid, occurring within seconds of the onset of light or rotation stimuli, and correspond to pairing-specific reductions in speed during the same time period. Since the CR is independent of the presentation of rotation, and it is also expressed by the same effector system (the foot) responsible for the UCR, light stimulation has assumed some of the functional character of rotation.