Reduced Ca²⁺ transient amplitudes may signify increased or decreased depolarization depending on the neuromodulatory...

Neuromodulators regulate neuronal excitability and bias neural circuit outputs. Optical recording of neuronal Ca²⁺ transients is a powerful approach to study the impact of neuromodulators on neural circuit dynamics. We are investigating the polymodal nociceptor ASH in Caenorhabditis elegans to better understand the relationship between neuronal excitability and optically recorded Ca²⁺ transients. ASHs depolarize in response to the aversive olfactory stimulus 1-octanol (1-oct) with a concomitant rise in somal Ca²⁺, stimulating an aversive locomotory response. Serotonin (5-HT) potentiates 1-oct avoidance through Gα_q signaling, which inhibits L-type voltage-gated Ca²⁺ channels in ASH. Although Ca²⁺ signals in the ASH soma decrease, depolarization amplitudes increase because Ca²⁺ mediates inhibitory feedback control of membrane potential in this context. Here, we investigate octopamine (OA) signaling in ASH to assess whether this negative correlation between somal Ca²⁺ and depolarization amplitudes is a general phenomenon, or characteristic of certain neuromodulatory pathways. Like 5-HT, OA reduces somal Ca²⁺ transient amplitudes in ASH neurons. However, OA antagonizes 5-HT modulation of 1-oct avoidance behavior, suggesting that OA may signal through a different pathway. We further show that the pathway for OA diminution of ASH somal Ca²⁺ consists of the OCTR-1 receptor, the G_o heterotrimeric G-protein, and the G-protein activated inwardly rectifying channels IRK-2 and IRK-3, and this pathway reduces depolarization amplitudes in parallel with somal Ca²⁺ transient amplitudes. Therefore, even within a single neuron, somal Ca²⁺ signal reduction may indicate either increased or decreased depolarization amplitude, depending on which neuromodulatory signaling pathways are activated, underscoring the need for careful interpretation of Ca²⁺ imaging data in neuromodulatory studies.