GABA-, histamine-, and FMRFamide-immunoreactivity in the visual, vestibular and central nervous systems of Hermissenda crassicornis.

Hermissenda crassicornis is a model for studying the molecular and cellular basis for classical conditioning, based on its ability to associate light with vestibular stimulation. We used confocal microscopy to map histamine (HA), FMRF-amide, and γ-aminobutyric acid (GABA) immunoreactivity in the central nervous system (CNS), eyes, optic ganglia and statocysts of the nudibranchs. For HA immunoreactivity, we documented both consistently and variably labeled CNS structures across individuals. We also noted minor differences in GABA immunoreactivity in the CNS compared to previous work on Hermissenda. Contrary to expectations, we found no evidence for GABA inside the visual or vestibular systems. Instead, we found only FMRFamide- and HA immunoreactivity (FMRFamide: 4 optic ganglion cells, 4-5 hair cells; HA: 3 optic ganglion cells, 8 hair cells). Overall, our results can act as basis for comparisons of nervous systems across nudibranchs, and suggest further exploration of intraspecific plasticity versus evolutionary changes in gastropod nervous systems.

The Model Organism Hermissenda crassicornis (Gastropoda: Heterobranchia) Is a Species Complex.

Hermissenda crassicornis is a model organism used in various fields of research including neurology, ecology, pharmacology, and toxicology. In order to investigate the systematics of this species and the presence of cryptic species in H. crassicornis, we conducted a comprehensive molecular and morphological analysis of this species covering its entire range across the North Pacific Ocean. We determined that H. crassicornis constitutes a species complex of three distinct species. The name Hermissensa crassicornis is retained for the northeast Pacific species, occurring from Alaska to Northern California. The name H. opalescens is reinstated for a species occurring from the Sea of Cortez to Northern California. Finally, the name H. emurai is maintained for the northwestern species, found in Japan and in the Russian Far East. These three species have consistent morphological and color pattern differences that can be used for identification in the field.

Neurochemical and neuroanatomical identification of central pattern generator neuron homologues in Nudipleura molluscs.

Certain invertebrate neurons can be identified by their behavioral functions. However, evolutionary divergence can cause some species to not display particular behaviors, thereby making it impossible to use physiological characteristics related to those behaviors for identifying homologous neurons across species. Therefore, to understand the neural basis of species-specific behavior, it is necessary to identify homologues using characteristics that are independent of physiology. In the Nudipleura mollusc Tritonia diomedea, Cerebral Neuron 2 (C2) was first described as being a member of the swim central pattern generator (CPG). Here we demonstrate that neurochemical markers, in conjunction with previously known neuroanatomical characteristics, allow C2 to be uniquely identified without the aid of electrophysiological measures. Specifically, C2 had three characteristics that, taken together, identified the neuron: 1) a white cell on the dorsal surface of the cerebral ganglion, 2) an axon that projected to the contralateral pedal ganglion and through the pedal commissure, and 3) immunoreactivity for the peptides FMRFamide and Small Cardioactive Peptide B. These same anatomical and neurochemical characteristics also uniquely identified the C2 homologue in Pleurobranchaea californica (called A1), which was previously identified by its analogous role in the Pleurobranchaea swim CPG. Furthermore, these characteristics were used to identify C2 homologues in Melibe leonina, Hermissenda crassicornis, and Flabellina iodinea, species that are phylogenetically closer to Tritonia than Pleurobranchaea, but do not display the same swimming behavior as Tritonia or Pleurobranchaea. These identifications will allow future studies comparing and contrasting the physiological properties of C2 across species that can and cannot produce the type of swimming behavior exhibited by Tritonia.

Relative spike timing in stochastic oscillator networks of the Hermissenda eye.

The role of relative spike timing on sensory coding and stochastic dynamics of small pulse-coupled oscillator networks is investigated physiologically and mathematically, based on the small biological eye network of the marine invertebrate Hermissenda. Without network interactions, the five inhibitory photoreceptors of the eye network exhibit quasi-regular rhythmic spiking; in contrast, within the active network, they display more irregular spiking but collective network rhythmicity. We investigate the source of this emergent network behavior first analyzing the role of relative input to spike-timing relationships in individual cells. We use a stochastic phase oscillator equation to model photoreceptor spike sequences in response to sequences of inhibitory current pulses. Although spike sequences can be complex and irregular in response to inputs, we show that spike timing is better predicted if relative timing of spikes to inputs is accounted for in the model. Further, we establish that greater noise levels in the model serve to destroy network phase-locked states that induce non-monotonic stimulus rate-coding, as predicted in Butson and Clark (J Neurophysiol 99:146-154, 2008a; J Neurophysiol 99:155-165, 2008b). Hence, rate-coding can function better in noisy spiking cells relative to non-noisy cells. We then study how relative input to spike-timing dynamics of single oscillators contribute to network-level dynamics. Relative timing interactions in the network sharpen the stimulus window that can trigger a spike, affecting stimulus encoding. Also, we derive analytical inter-spike interval distributions of cells in the model network, revealing that irregular Poisson-like spike emission and collective network rhythmicity are emergent properties of network dynamics, consistent with experimental observations. Our theoretical results generate experimental predictions about the nature of spike patterns in the Hermissenda eye.

Comparative study of visuo-vestibular conditioning in Lymnaea stagnalis.

In this review, we compare the current understanding of visuo-vestibular conditioning in Hermissenda crassicornis and Lymnaea stagnalis on the basis of behavioral, electrophysiologic, and morphologic studies. Paired presentation of a photic conditioned stimulus (CS) and an orbital rotation unconditioned stimulus (US) results in conditioned escape behavior in both species. In Hermissenda, changes in excitability of type B photoreceptors and morphologic modifications at the axon terminals follow conditioning. Caudal hair cells, which detect mechanical turbulence, have reciprocal inhibition with type B photoreceptors. In Lymnaea, the interaction between photoreceptors and hair cells is dependent on statocyst location. Furthermore, the organization of the Lymnaea eye is complex, with more than 100 photoreceptors distributed in a uniquely folded retina. Although the optimal conditions to produce long-term memory (memory persistent for >1 week) are almost identical in Hermissenda and Lymnaea, physiologic and morphologic differences suggest that the neuronal mechanisms underlying learning and memory are distinct.