A GABAergic and peptidergic sleep neuron as a locomotion stop neuron with compartmentalized Ca2+ dynamics

Animals must slow or halt locomotion to integrate sensory inputs or to change direction. In Caenorhabditis elegans, the GABAergic and peptidergic neuron RIS mediates developmentally timed quiescence. Here, we show RIS functions additionally as a locomotion stop neuron. RIS optogenetic stimulation caused acute and persistent inhibition of locomotion and pharyngeal pumping, phenotypes requiring FLP-11 neuropeptides and GABA. RIS photoactivation allows the animal to maintain its body posture by sustaining muscle tone, yet inactivating motor neuron oscillatory activity. During locomotion, RIS axonal Ca2+ signals revealed functional compartmentalization: Activity in the nerve ring process correlated with locomotion stop, while activity in a branch correlated with induced reversals. GABA was required to induce, and FLP-11 neuropeptides were required to sustain locomotion stop. RIS attenuates neuronal activity and inhibits movement, possibly enabling sensory integration and decision making, and exemplifies dual use of one cell across development in a compact nervous system.

Neuroendocrine modulation sustains the C. elegans forward motor state

Neuromodulators shape neural circuit dynamics. Combining electron microscopy, genetics, transcriptome profiling, calcium imaging, and optogenetics, we discovered a peptidergic neuron that modulates C. elegans motor circuit dynamics. The Six/SO-family homeobox transcription factor UNC-39 governs lineage-specific neurogenesis to give rise to a neuron RID. RID bears the anatomic hallmarks of a specialized endocrine neuron: it harbors near-exclusive dense core vesicles that cluster periodically along the axon, and expresses multiple neuropeptides, including the FMRF-amide-related FLP-14. RID activity increases during forward movement. Ablating RID reduces the sustainability of forward movement, a phenotype partially recapitulated by removing FLP-14. Optogenetic depolarization of RID prolongs forward movement, an effect reduced in the absence of FLP-14. Together, these results establish the role of a neuroendocrine cell RID in sustaining a specific behavioral state in C. elegans.

Neuroendocrine Modulation Sustains the C. elegans Forward Motor State

Neuromodulators shape neural circuit dynamics. Combining electron microscopy, genetics, transcriptome profiling, calcium imaging, and optogenetics, we discovered a peptidergic neuron that modulates C. elegans motor circuit dynamics. The Six/SO-family homeobox transcription factor UNC-39 governs lineage-specific neurogenesis to give rise to a neuron RID. RID bears the anatomic hallmarks of a specialized endocrine neuron: it harbors near-exclusive dense core vesicles that cluster periodically along the axon, and expresses multiple neuropeptides, including the FMRF-amide-related FLP-14. RID activity increases during forward movement. Ablating RID reduces the sustainability of forward movement, a phenotype partially recapitulated by removing FLP-14. Optogenetic depolarization of RID prolongs forward movement, an effect reduced in the absence of FLP-14. Together, these results establish the role of a neuroendocrine cell RID in sustaining a specific behavioral state in C. elegans.

Distribution of ionic currents in the soma and growing region of an identified peptidergic neuron in defined culture

1. Somata and lamellipodia of a distinct type of crustacean peptidergic neuron were isolated by severing the connecting neurite. The whole-cell variation of the patch-clamp technique was then used to study the electrical activity and ionic currents of each part. Neurons were enzymatically isolated from the X-organ of Cardisoma carnifex and cultured in defined medium. The neurons studied were recognizable by their large ovoid somata (approximately 35 microns minor diam) and broad, flat lamellipodium regrown from the remaining neurite. These cells are immunopositive against crustacean hyperglycemic hormone (CHH) antisera. Recordings were made 18-30 h after plating. 2. In current-clamp recordings, 10 of 12 lamellipodia fired overshooting action potentials (mean half width = 8.2 +/- 2.9 ms, mean +/- SD), whereas only two of seven somata did so. Spontaneous activity in isolated somata and lamellipodia was rarely encountered. The action potential in isolated lamellipodia has both Na+ and Ca2+ components, whereas the regenerative activity recorded in isolated somata was predominantly Ca(2+)-based. 3. The inward currents examined under voltage clamp consisted of Na current (INa) and Ca current (ICa). Both currents could be resolved in isolated somata and lamellipodia. INa was completely blocked by tetrodotoxin [TTX (1 microM)]. The INa density in lamellipodia was approximately 4-19 times greater than that of the somata from which they had been separated. In contrast, ICa density in lamellipodia was two to five times smaller than that of somata. The properties of ICa were similar in both somata and lamellipodia, with the exception that ICa in lamellipodia did not recover after large depolarizing prepulses. 4. Two types of outward current were readily identified under voltage clamp. These were the transient 4-aminopyridine-sensitive current and the delayed outward current that was partially sensitive to tetraethylammonium ions. The peak potassium current-density ratio for lamellipodia-somata pairs ranged from 0.5 to 1.7 (mean = 1.2, SD +/- 0.6), suggesting that both channel types are distributed relatively evenly over the cell surface. 5. The differences in current density in the different regions did not correlate with the length of the neurite remaining after separation of the soma and lamellipodium. 6. It is concluded that CHH neurons in the early phase of regeneration in primary culture show a distinct pattern of ion channel distribution. These ion channels are present in new membrane < 18 h after formation.(ABSTRACT TRUNCATED AT 400 WORDS)