ANALYSIS OF INTRINSIC CONNECTIVITY IN A MULTIFUNCTIONAL CENTRAL PACEMAKER NUCLEUS IN VERTEBRATES.

Abstract

Gymnotiform fish emit electric organ discharges (EODs) for both active electroreception and electrocommunication. EOD waveform and rhythm can be modified to cope with diverse environmental challenges. In pulse-type species, EODs are generated by a hierarchical electromotor network controlled by a medullary pacemaker nucleus (PN), which comprises intrinsic pacemaker cells (PM-cells) and projecting relay cells (R-cells). Active electroreception requires the emission of stereotyped EODs, an electromotor output that implies a functional PN configuration in which PM-cells rhythmically time EODs and R-cells transmit coordinated commands to downstream components of the electromotor system. To test whether electrical coupling (EC) between PN neurons supports this functional organization, intrinsic connectivity of the PN in Gymnotus omarorum was examined in brainstem slices using electrophysiology, immunohistochemistry, and dye-coupling analysis. Homotypic connections (PM-PM and R-R) exhibited low-magnitude, bidirectional EC with symmetrical, low-pass filter properties, supporting synchronous yet adaptable pacemaker activity and coordinated descending commands. Heterotypic connections (PM-R) also displayed bidirectional, symmetrical coupling but revealed direction-dependent filtering: an apparent high-pass behavior from PM- to R-cells and a low-pass behavior in the opposite direction. Together with precise PM-to-R discharge timing, direction-dependent filtering suggests a role of PM-cell axons in shaping signal flow. Dye coupling and immunohistochemical evidence further indicate that PN neurons are interconnected via gap junctions, likely formed by connexin 35. Thus, EC-based connectivity endows the PN with crucial functional attributes of its exploration mode of operation while preserving the capacity to organize communication signals under the influence of descending inputs, revealing remarkable functional versatility. Summary statementGap junction-mediated intrinsic connections within the electromotor nucleus of electric fish may sustain the emission of signals essential for sensory sampling as well as those supporting communication.