The eversible tentacle organs of Polyommatus caterpillars (Lepidoptera, Lycaenidae): Morphology, fine structure, sensory supply and functional aspects.

In their late (3rd and 4th) larval stages, caterpillars of the myrmecophilous lycaenid (Lepidoptera) species Polyommatus coridon and Polyommatus icarus, possess on their 8th abdominal segment two eversible so called tentacle organs (TOs). Previous histological and behavioural results have proposed that the TOs may release a volatile substance that elicits "excited runs" in attendant ants. In our study we investigated for the first time the temporal in- and eversion pattern of TOs. Using nerve tracing, Micro-CT, light- and electron microscopy techniques we studied (i) the histology of the 8th abdominal segment, (ii) the fine structure of the cuticular and cellular apparatus of the TOs, (iii) the attachment sites of the retractor muscle of each TO and (iv) the fine structure of the long slender tentacle hairs which are exposed to the outside, when the TOs are everted and fold back into the TO-sac during inversion. Our data show that the tentacle hairs are typical insect mechanoreceptors, each innervated by a small bipolar sensory cell with a tubular body in the tip of the outer dendritic segment. The latter is enclosed by a cuticular sheath previously called the "internal cuticular duct" and misinterpreted in earlier studies as the space, where the tentacle hairs actively secrete fluids. However, we found no glandular structures nearby or in the wall of the TO-sac. Also we did not reveal any conspicuous signs of secretory activity in one of the enveloping cells belonging to a tentacle hair. Although highly unusual features for an insect mechanoreceptor are: (a) the hair-shaft lumen of tentacle hairs contains flocculent material as well small vesicles and (b) the thin cuticular wall of the hair-shaft and its spines possess few tiny pores. Our data do not support the assumption of previous studies that volatile substances are released via the tentacle organs during their interactions with ants which in turn are supposed to cause excited runs in ants.

Stimulus transmission in the auditory receptor organs of the foreleg of bushcrickets (Tettigoniidae) I. The role of the tympana.

The auditory organs of the tettigoniid are located just below the femoral tibial joint in the forelegs. Structurally each auditory organ consists of a tonotopically organized crista acustica and intermediate organ and associated sound conducting structures; an acoustic trachea and two lateral tympanic membranes located at the level of the receptor complex. The receptor cells and associated satellite structures are located in a channel filled with hemolymph fluid. The vibratory response characteristics of the tympanic membranes generated by sound stimulation over the frequency range 2-40 kHz have been studied using laser vibrometry. The acoustic trachea was found to be the principal structure through which sound energy reached the tympana. The velocity of propagation down the trachea was observed to be independent of the frequency and appreciably lower than the velocity of sound in free space. Structurally the tympana are found to be partially in contact with the air in the trachea and with the hemolymph in the channel containing the receptor cells. The two tympana were found to oscillate in phase, with a broad band frequency response, have linear coherent response characteristics and small time constant. Higher modes of vibration were not observed. Measurements of the pattern of vibration of the tympana showed that these structures vibrate as hinged flaps rather than vibrating stretched membranes. These findings, together with the morphology of the organ and physiological data from the receptor cells, suggest the possibility of an impedance matching function for the tympana in the transmission of acoustic energy to the receptor cells in the tettigoniid ear.

Comparison of song frequency and receptor tuning in two closely related bushcricket species.

The songs and the structure and physiology of the auditory organs in the closely related bushcricket species Tettigonia viridissima and Tettigonia cantans were investigated comparatively using bioacoustical, histological and neurophysiological methods. The morphology of the crista acustica, the main auditory receptor organ, is very similar in the two species in respect to both the distribution of scolopidia along the length axis of the crista and the dimensions of corresponding scolopidia and attachment structures. The only obvious difference is that T. viridissima has one more scolopidium in the crista acustica and that the overall length of the crista is by about 50 microns larger than in T. cantans. In contrast, differences were found in the physiology of individual auditory receptor cells. Comparison of the threshold characteristics of all the receptor cells of the crista acustica in both species reveals a differential sensitivity of groups of auditory receptor cells at dominant frequencies of the song. In each species, the sensitivity of auditory receptor cells is method to the energy spectrum of the song. These differences in the physiology can partly be explained by differences in transmission characteristics of the acoustic trachea.

Specific differences in sound production and pattern recognition in tettigoniids.

A brief comparative description of the stridulatory songs of nine different tettigoniid species is given to introduce a set of four parameters (phase of sound production during opening and closing movement of the wings, syllable repetition mode, syllable similarity, and impulse pattern of the syllables) to characterize the temporal pattern of tettigoniid songs. The importance of different song parameters for female phonotaxis was investigated in two tettigoniid species (Ephippiger ephippiger and Tettigonia viridissima). Two-choice experiments revealed that the impulse pattern of the closing syllable is an important parameter for the phonotactic behaviour of E. ephippiger, whereas the syllable pattern is a decisive parameter for species discrimination in T. viridissima.

Comparison of the physiology of the auditory receptor organs in Gryllus bimaculatus and Ephippiger ephippiger: CSD recordings within the auditory neuropiles.

The syllables of the song of the tettigoniid Ephippiger ephippiger consist of a series of short sound impulses with a broad-banded frequency spectrum. Syllables of the song of the gryllid species Gryllus bimaculatus are nearly pure tones with sharply tuned frequency maxima. A comparison of the physiology of the auditory receptor organs of both species was carried out by using acoustical stimuli with different carrier frequencies and time-amplitude patterns. The neuronal ensemble activity of receptor cell groups of the tympanal organ was measured within the prothoracic ganglion using the CSD technique. In E. ephippiger, response maxima were found at carrier frequencies mirroring the broad frequency content of the conspecific song. The receptor cells of E. ephippiger are highly sensitive to transient sound impulses. In G. bimaculatus, the receptor cell population is more sharply tuned to the basic frequencies of the natural songs; pure tones represent more effective stimuli than transient sound signals. The causes for these species-specific differences are discussed with regard to probable adaptations of the receptor organs to the parameters of the conspecific songs.

Species-specific sound production in three ephippigerine bushcrickets.

Songs of three Ephippigerine species (Ephippiger ephippiger, E. discoidalis and E.perforatus) have been recorded and analysed. Manipulation experiments have been carried out by removing single teeth from the pars stridens. The songs of manipulated animals show characteristic gaps within the impulse structure of the opening and closing syllables. Morphological measurements were carried out by means of SEM-photographs of the pars stridens. Combining both the bioacoustic and morphological results reveals that only the lateral part of the pars stridens is used during stridulation. Furthermore it could be shown that the individual impulse interval pattern within one syllable is also highly constant between syllables. The impulse interval pattern correlates with the pattern of tooth spacing on the pars stridens.