Vibrational sensitivity of the subgenual organ complex in female Sipyloidea sipylus stick insects in different experimental paradigms of stimulus direction, leg attachment, and ablation of a connective tibial sense organ

We document the sensitivity to sinusoidal vibrations for chordotonal organs in the stick insect tibia (Sipyloidea sipylus). In the tibia, the scolopidial subgenual organ (~ 40 scolopidial sensilla), distal organ (~ 20 scolopidial sensilla), and distal tibial chordotonal organ (~ 7 scolopidial sensilla) are present. We study the sensitivity of tibial sensory organs in all leg pairs to vibration stimuli as sensory thresholds by recording summed action potentials from Nervus cruris in the femur. The tibia was stimulated with a minishaker delivering vibrational stimuli. Because different experimental procedures may affect the vibration sensitivity, we here analysed possible effects of different experimental conditions: (1) the stimulus direction delivered in either horizontal or vertical direction to the leg; (2) recording responses only from the subgenual organ complex after ablation of the distal tibial chordotonal organ, and (3) the attachment of the leg to the minishaker by plastilin, beeswax-colophony, or freely standing legs. The tibial scolopidial organs give summed responses to vibration stimuli with highest sensitivity between 500 and 1000 Hz for all leg pairs. In the different experimental series, we find that (1) thresholds were influenced by stimulation direction with lower thresholds in response to vertical vibrations, (2) ablating the distal tibial chordotonal organ by cutting the distal-most tibia did not change the summed sensory thresholds significantly, and (3) the attachment material between legs and the minishaker (plastilin or beeswax-colophony mixture) did not significant influence the sensory thresholds against free-standing tarsi. The distal tibial chordotonal organ is a connective chordotonal organ attached to a tendon and is likely a proprioceptive organ. These results emphasise that vibrational thresholds are mainly direction-sensitive. Thus, the direction of stimulus delivery during electrophysiological recordings is relevant for comparisons of vibratory sensory thresholds.