Comparative Neuroanatomy of the Mechanosensory Subgenual Organ Complex in the Peruvian Stick Insect, Oreophoetes peruana.

The subgenual organ complex in the leg of Polyneoptera (Insecta) consists of several chordotonal organs specialized to detect mechanical stimuli from substrate vibrations and airborne sound. In stick insects (Phasmatodea), the subgenual organ complex contains the subgenual organ and the distal organ located distally to the subgenual organ. The subgenual organ is a highly sensitive detector for substrate vibrations. The distal organ has a characteristic linear organization of sensilla and likely also responds to substrate vibrations. Despite its unique combination of sensory organs, the neuroanatomy of the subgenual organ complex of stick insects has been investigated for only very few species so far. Phylogenomic analysis has established for Phasmatodea the early branching of the sister groups Oriophasmata, the Old World phasmids, and Occidophasmata, the New World phasmids. The species studied for the sensory neuroanatomy, including the Indian stick insect Carausius morosus, belong to the Old World stick insects. Here, the neuroanatomy of the subgenual organ complex is presented for a first species of the New World stick insects, the Peruvian stick insect Oreophoetes peruana. To document the sensory organs in the subgenual organ complex and their innervation pattern, and to compare these between females and males of this species and also to the Old World stick insects, axonal tracing is used. This study documents the same sensory organs for O. peruana, subgenual organ and distal organ, as in other stick insects. Between the sexes of this species, there are no notable differences in the neuroanatomy of their sensory organs. The innervation pattern of tibial nerve branches in O. peruana is identical to other stick insect species, although the innervation pattern of the subgenual organ by a single tibial nerve branch is simpler. The shared organization of the organs in the subgenual organ complex in both groups of Neophasmatodea (Old World and New World stick insects) indicates the sensory importance of the subgenual organ but also of the distal organ. Some variation exists in the innervation of the chordotonal organs in O. peruana though a common innervation pattern can be identified. The findings raise the question for the ancestral neuroanatomical organization and innervation in stick insects.

Beyond the display: phase-only liquid crystal on Silicon devices and their applications in photonics [Invited].

Existing for almost four decades, liquid crystal on Silicon (LCOS) technology is rapidly growing into photonic applications. We review the basics of the technology, from the wafer to the driving solutions, the progress over the last decade and the future outlook. Furthermore we review the most exciting industrial and scientific applications of the LCOS technology.

Low-frequency vibration transmission and mechanosensory detection in the legs of cave crickets.

Vibrational communication is common in insects and often includes signals with prominent frequency components below 200 Hz, but the sensory adaptations for their detection are scarcely investigated. We performed an integrative study of the subgenual organ complex in Troglophilus cave crickets (Orthoptera: Rhaphidophoridae), a mechanosensory system of three scolopidial organs in the proximal tibia, for mechanical, anatomical and physiological aspects revealing matches to low frequency vibration detection. Microcomputed tomography shows that a part of the subgenual organ sensilla and especially the accessory organ posteriorly in this complex are placed closely underneath the cuticle, a position suited to evoke responses to low-frequency vibration via changes in the cuticular strain. Laser-Doppler vibrometry shows that in a narrow low-frequency range the posterior tibial surface reacts stronger to low frequency sinusoidal vibrations than the anterior tibial surface. This finding suggests that the posterior location of sensilla in tight connection to the cuticle, especially in the accessory organ, is adapted to improve detectability of low-frequency vibration signals. By electrophysiological recordings we identify a scolopidial receptor type tuned to 50-300 Hz vibrations, which projects into the central mechanosensory region specialised for processing low-frequency vibratory inputs, and most likely originates from the accessory organ or the posterior subgenual organ. Our findings contribute to understanding of the mechanical and neuronal basis of low-frequency vibration detection in insect legs and their highly differentiated sensory systems.

The mechanical leg response to vibration stimuli in cave crickets and implications for vibrosensory organ functions.

We investigate the influence of leg mechanics on the vibration input and function of vibrosensitive organs in the legs of the cave cricket Troglophilus neglectus, using laser Doppler vibrometry. By varying leg attachment, leg flexion, and body posture, we identify important influences on the amplitude and frequency parameters of transmitted vibrations. The legs respond best to relatively high-frequency vibration (200-2000 Hz), but in strong dependence on the leg position; the response peak shifts progressively over 500-1400 Hz towards higher frequencies following leg flexion. The response is amplified most strongly on the tibia, where specialised vibrosensory organs occur, and the response amplitude increases with the increasing frequency. Leg responses peaking at 800 and 1400 Hz closely resemble the tuning of the intermediate organ receptors in the proximal tibia of T. neglectus, which may be highly sensitive to positional change. The legs of free-standing animals with the abdomen touching the vibrating substrate show a secondary response peak below 150 Hz, induced by body vibration. Such responses may significantly increase the sensitivity of low-frequency receptors in the tibial accessory organ and the femoral chordotonal organ. The cave cricket legs appear suitable especially for detection of high-frequency vibration.

Neuronal Regression of Internal Leg Vibroreceptor Organs in a Cave-Dwelling Insect (Orthoptera: Rhaphidophoridae: Dolichopoda araneiformis).

Animals' adaptations to cave habitats generally include elaboration of extraoptic senses, and in insects the receptor structures located on the legs are supposed to become more prominent in response to constant darkness. The receptors for detecting substrate vibrations are often highly sensitive scolopidial sensilla localized within the legs or the body. For troglobitic insects the evolutionary changes in vibroreceptor organs have not been studied. Since rock is an extremely unfavorable medium for vibration transmission, selection on vibration receptors may be weakened in caves, and these sensory organs may undergo regressive evolution. We investigated the anatomy of the most elaborate internal vibration detection system in orthopteroid insects, the scolopidial subgenual organ complex in the cave cricket Dolichopoda araneiformis (Orthoptera: Ensifera: Rhaphidophoridae). This is a suitable model species which shows high levels of adaptation to cave life in terms of both phenotypic and life cycle characteristics. We compared our data with data on the anatomy and physiology of the subgenual organ complex from the related troglophilic species Troglophilus neglectus. In D. araneiformis, the subgenual organ complex contains three scolopidial organs: the subgenual organ, the intermediate organ, and the accessory organ. The presence of individual organs and their innervation pattern are identical to those found in T. neglectus, while the subgenual organ and the accessory organ of D. araneiformis contain about 50% fewer scolopidial sensilla than in T. neglectus. This suggests neuronal regression of these organs in D. araneiformis, which may reflect a relaxed selection pressure for vibration detection in caves. At the same time, a high level of overall neuroanatomical conservation of the intermediate organ in this species suggests persistence of the selection pressure maintaining this particular organ. While regressive evolution of chordotonal organs has been documented for insect auditory organs, this study shows for the first time that internal vibroreceptors can also be affected.

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 1000Hz 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.

Selective forces on origin, adaptation and reduction of tympanal ears in insects.

Insect ears evolved many times independently. As a consequence, a striking diversity exists in the location, construction and behavioural implementation of ears. In this review, we first summarise what is known about the evolutionary origin of ears and the presumed precursor organs in the various insect groups. Thereafter, we focus on selective forces for making and keeping an ear: we discuss detecting and localising predators and conspecifics, including establishing new "private" channels for intraspecific communication. More advanced aspects involve judging the distance of conspecifics, or assessing individual quality from songs which makes auditory processing a means for exerting sexual selection on mating partners. We try to identify negative selective forces, mainly in the context of energy expenditure for developing and keeping an ear, but also in conjunction with acoustic communication, which incorporates risks like eavesdropping by predators and parasitoids. We then discuss balancing pressures, which might oppose optimising an ear for a specific task (when it serves different functions, for example). Subsequently, we describe various scenarios that might have led to a reduction or complete loss of ears in evolution. Finally, we describe cases of sex differences in ears and potential reasons for their appearance.

Neurons of self-defence: neuronal innervation of the exocrine defence glands in stick insects.

BACKGROUND: Stick insects (Phasmatodea) use repellent chemical substances (allomones) for defence which are released from so-called defence glands in the prothorax. These glands differ in size between species, and are under neuronal control from the CNS. The detailed neural innervation and possible differences between species are not studied so far. Using axonal tracing, the neuronal innervation is investigated comparing four species. The aim is to document the complexity of defence gland innervation in peripheral nerves and central motoneurons in stick insects. RESULTS: In the species studied here, the defence gland is innervated by the intersegmental nerve complex (ISN) which is formed by three nerves from the prothoracic (T1) and suboesophageal ganglion (SOG), as well as a distinct suboesophageal nerve (Nervus anterior of the suboesophageal ganglion). In Carausius morosus and Sipyloidea sipylus, axonal tracing confirmed an innervation of the defence glands by this N. anterior SOG as well as N. anterior T1 and N. posterior SOG from the intersegmental nerve complex. In Peruphasma schultei, which has rather large defence glands, only the innervation by the N. anterior SOG was documented by axonal tracing. In the central nervous system of all species, 3-4 neuron types are identified by axonal tracing which send axons in the N. anterior SOG likely innervating the defence gland as well as adjacent muscles. These neurons are mainly suboesophageal neurons with one intersegmental neuron located in the prothoracic ganglion. The neuron types are conserved in the species studied, but the combination of neuron types is not identical. In addition, the central nervous system in S. sipylus contains one suboesophageal and one prothoracic neuron type with axons in the intersegmental nerve complex contacting the defence gland. CONCLUSIONS: Axonal tracing shows a very complex innervation pattern of the defence glands of Phasmatodea which contains different neurons in different nerves from two adjacent body segments. The gland size correlates to the size of a neuron soma in the suboesophageal ganglion, which likely controls gland contraction. In P. schultei, the innervation pattern appears simplified to the anterior suboesophageal nerve. Hence, some evolutionary changes are notable in a conserved neuronal network.

Functional Morphology of Leg Mechanosensory Organs in Early Postembryonic Development in the Stick Insect (Sipyloidea chlorotica).

The subgenual organ complex of stick insects has a unique neuroanatomical organisation with two elaborate chordotonal organs, the subgenual organ and the distal organ. These organs are present in all leg pairs and are already developed in newly hatched stick insects. The present study analyses for the first time the morphology of sensory organs in the subgenual organ complex for a membrane connecting the two sensory organs in newly hatched insects (Sipyloidea chlorotica (Audinet-Serville 1838)). The stick insect legs were analysed following hatching by axonal tracing and light microscopy. The subgenual organ complex in first juvenile instars shows the sensory organs and a thin membrane connecting the sensory organs resembling the morphology of adult animals. Rarely was this membrane not detected, where it is assumed as not developed during embryogenesis. The connection appears to influence the shape of the subgenual organ, with one end extending towards the distal organ as under tension. These findings are discussed for the following functional implications: (1) the physiological responses of the subgenual organ complex to mechanical stimuli after hatching, (2) the influence of the membrane on the displacement of the sensory organs, and (3) the connection between the subgenual organ and distal organ as a possible functional coupling.

Vibration receptor organs in the insect leg: neuroanatomical diversity and functional principles.

Detecting substrate vibrations is essential for insects in different behavioural contexts. These vibrational behaviours are mediated by mechanoreceptor organs detecting and processing vibrational stimuli transmitted in the environment. We discuss recently gained insights about the functional principles of insect vibration receptors, mainly leg chordotonal organs highly sensitive to vibrational stimuli, and the mechanisms of their diversification in neuroanatomy and functional morphology, in relation to the attachment structures and mechanical coupling. The two main input pathways for vibration stimuli transferred by the insect legs to vibrosensory organs via the cuticle and via the hemolymph are fundamental for explaining sensory specialisations. The vibroreceptor organs can diversify in their neuroanatomy and morphology in several key aspects. This provides the structural basis for complex adaptations in sensory evolution.

Laser focus positioning method with submicrometer accuracy.

Accurate positioning of a sample is one of the primary challenges in laser micromanufacturing. There are a number of methods that allow detection of the surface position; however, only a few of them use the beam of the processing laser as a basis for the measurement. Those methods have an advantage that any changes in the processing laser beam can be inherently accommodated. This work describes a direct, contact-free method to accurately determine workpiece position with respect to the structuring laser beam focal plane based on nonlinear harmonic generation. The method makes workpiece alignment precise and time efficient due to ease of automation and provides the repeatability and accuracy of the surface detection of less than 1 µm.

The neuronal innervation pattern of the subgenual organ complex in Peruphasma schultei (Insecta: Phasmatodea).

The proximal tibia of orthopteroid insects contains sensory organs, the subgenual organ complex, detecting mechanical stimuli including substrate vibration. In stick insects, two chordotonal organs occur in close proximity, the subgenual organ and the distal organ, which likely detect substrate vibrations. In most stick insects, both organs are innervated by separate nerve branches. To obtain more data on the neuroanatomy of the subgenual organ complex from the New World phasmids (Occidophasmata), the present study documents the neuronal innervation of sensory organs in the subgenual organ complex of Peruphasma schultei, the first species from Pseudophasmatinae investigated for this sensory complex. The innervation pattern shows a distinct nerve branch for the subgenual organ and for the distal organ in most cases. Some variability in the innervation, which generally occurs for these chordotonal organs, was noted for both organs in P. schultei. The most common innervation for both organs was by a single nerve branch for each organ. The innervation of the subgenual organ resembled the nerve pattern of another New World phasmid, but was simpler than in the Old World phasmids (Oriophasmata) studied so far. Therefore, the peripheral neuronal innervation of sensory organs could reflect phylogenetic relationships and provide phylogenetic information, while the overall neuroanatomy of the subgenual organ complex is similar in stick insects.

Pigment-dispersing hormone in Daphnia interneurons, one type homologous to insect clock neurons displaying circadian rhythmicity.

We report identification of a beta-type pigment-dispersing hormone (PDH) identical in two water flea species, Daphnia magna and Daphnia pulex. It has been identified by cloning of precursors, chromatographic isolation from tissue extracts followed by immunoassays and de novo-mass spectrometric sequencing. The peptide is restricted to a complex system of distinct interneurons in the brain and visual ganglia, but does not occur in neurosecretory cells projecting to neurohemal organs as in decapod crustaceans. Thirteen neuron types individually identified and reconstructed by immunohistochemistry were almost identical in terms of positions and projection patterns in both species. Several neurons invade and form plexuses in visual ganglia and major brain neuropils including the central body. Five neuron types show contralateral pathways and form plexuses in the lateral, dorsal, or postlateral brain neuropils. Others are local interneurons, and a tritocerebral neuron connects the protocerebrum with the neuropil of the locomotory second antenna. Two visual ganglia neuron types lateral to the medulla closely resemble insect medulla lateral circadian clock neurons containing pigment-dispersing factor based upon positional and projectional criteria. Experiments under 12:12 h light/dark cycles and constant light or darkness conditions showed significant circadian changes in numbers and activities of one type of medulla lateral PDH neuron with an acrophase in the evening. This simple PDH system shows striking homologies to PDH systems in decapod crustaceans and well-known clock neurons in several insects, which suggests evolutionary conservation of an ancient peptidergic interneuronal system that is part of biological clocks.

Vibration detection in arthropods: Signal transfer, biomechanics and sensory adaptations.

In arthropods, the detection of vibrational signals and stimuli is essential in several behaviours, including mate recognition and pair formation, prey detection, and predator evasion. These behaviours have been studied in several species of insects, arachnids, and crustaceans for vibration production and propagation in the environment. Vibration stimuli are transferred over the animals' appendages and the body to vibrosensory organs. Ultimately, the stimuli are transferred to act on the dendrites of the mechanosensitive sensilla. We refer to these two different levels of transfer as macromechanics and micromechanics, respectively. These biomechanical processes have important roles in filtering and pre-processing of stimuli, which are not carried out by neuronal components of sensory organs. Also, the macromechanical transfer is posture-dependent and enables behavioural control of vibration detection. Diverse sensory organs respond to vibrations, including cuticular sensilla (slit sensilla, campaniform sensilla) and internal chordotonal organs. These organs provide various adaptations, as they occur at diverse body positions with different mechanical couplings as input pathways. Macromechanics likely facilitated evolution of vibrosensory organs at specific body locations. Thus, vibration detection is a highly complex sensory capacity, which employs body and sensory mechanics for signal filtering, amplification, and analysis of frequency, intensity and directionality.

Are all auditory sensilla of bushcrickets bimodal? Comment on: R. D. Zhantiev and O. S. Korsunovskaya, Functions of chordotonal sensilla in bushcrickets (Orthoptera, Tettigoniidae); Entomological Review, 2021, vol. 101 (6), pp. 755-766.

Detection of sound and substrate vibration is crucial for the survival and reproduction of many animals, particularly insects. Bushcrickets (Orthoptera, Tettigoniidae), developed a large mechanosensory organ complex in their legs to detect such stimuli. As demonstrated by various studies in the past, sensilla in distinct functional groups form specialized vibratory organs (the subgenual organ and the accessory organ), respond sensitively to both vibration and sound (in the intermediate organ [IO]), or mediate hearing (in the crista acustica [CA]; the tympanal hearing organ). In their recent publication, Zhantiev and Korsunovskaya addressed auditory and vibratory sensitivity in the IO and the CA in two species of bushcrickets, using single-cell recording and staining of sensory neurons from their soma in an isolated foreleg. Their main finding was that not only the IO but also the complete CA contains bimodal sensilla responding with high sensitivity to both sound and vibration, which would be a true change in the paradigm of how the auditory/vibratory sense in Orthoptera works. In addition, they revealed vibratory tuning of the IO sensilla, which differs largely from that in previous studies. We propose three major experimental causes of such discrepancies: calibration, experiments with isolated legs, and differences in the sites of recording. To judge the causes of these discrepancies more adequately, a detailed comparison of methods and a number of control experiments are needed. This will deepen our understanding of sensory adaptations and specialization of insect mechanosensory organs to stimuli entering the system by different input pathways.

Genomics, transcriptomics, and peptidomics of Daphnia pulex neuropeptides and protein hormones.

We report 43 novel genes in the water flea Daphnia pulex encoding 73 predicted neuropeptide and protein hormones as partly confirmed by RT-PCR. MALDI-TOF mass spectrometry identified 40 neuropeptides by mass matches and 30 neuropeptides by fragmentation sequencing. Single genes encode adipokinetic hormone, allatostatin-A, allatostatin-B, allatotropin, Ala(7)-CCAP, CCHamide, Arg(7)-corazonin, DENamides, CRF-like (DH52) and calcitonin-like (DH31) diuretic hormones, two ecdysis-triggering hormones, two FIRFamides, one insulin, two alternative splice forms of ion transport peptide (ITP), myosuppressin, neuroparsin, two neuropeptide-F splice forms, three periviscerokinins (but no pyrokinins), pigment dispersing hormone, proctolin, Met(4)-proctolin, short neuropeptide-F, three RYamides, SIFamide, two sulfakinins, and three tachykinins. There are two genes for a preprohormone containing orcomyotropin-like peptides and orcokinins, two genes for N-terminally elongated ITPs, two genes (clustered) for eclosion hormones, two genes (clustered) for bursicons alpha, beta, and two genes (clustered) for glycoproteins GPA2, GPB5, three genes for different allatostatins-C (two of them clustered) and three genes for IGF-related peptides. Detailed comparisons of genes or their products with those from insects and decapod crustaceans revealed that the D. pulex peptides are often closer related to their insect than to their decapod crustacean homologues, confirming that branchiopods, to which Daphnia belongs, are the ancestor group of insects.

The tracheal system in the stick insect prothorax and prothoracic legs: Homologies to Orthoptera and relations to mechanosensory functions.

Arthropod respiration depends on the tracheal system running from spiracles at the body surface through the body and appendages. Here, three species of stick insects (Carausius morosus, Ramulus artemis, Sipyloidea sipylus) are investigated for the tracheae in the prothorax and foreleg. The origin of the tracheae from the mesothoracic spiracle that enter the foreleg is identified: five tracheae originate from the mesothoracic spiracle, of which two enter the foreleg (supraventral trachea, trachea pedalis anterior). These two tracheae run separately through the leg to the femur-tibia joint where they fuse, but in the proximal tibia split again into two tracheae. The leg tracheae in stick insects are homologous to those in Tettigoniidae (bushcrickets). Stick insects have two chordotonal organs in the proximal tibia (subgenual organ and distal organ) which locate dorsally of the leg trachea. The tracheal system shows no adaptation specific to the propagation of airborne sound, like enlarged spiracles or tracheal volumes. Tracheal vesicles form in the tibia proximally to the mechanosensory organs, but no tracheal sacks or expansions occur at the level of the sensory organs that could mediate the detection of airborne sound or amplify substrate vibrations transmitted in the hemolymph fluid. Rather, the morphological characteristics indicate a respiratory function.

The auditory system of non-calling grasshoppers (Melanoplinae: Podismini) and the evolutionary regression of their tympanal ears.

Reduction of tympanal hearing organs is repeatedly found amongst insects and is associated with weakened selection for hearing. There is also an associated wing reduction, since flight is no longer required to evade bats. Wing reduction may also affect sound production. Here, the auditory system in four silent grasshopper species belonging to the Podismini is investigated. In this group, tympanal ears occur but sound signalling does not. The tympanal organs range from fully developed to remarkably reduced tympana. To evaluate the effects of tympanal regression on neuronal organisation and auditory sensitivity, the size of wings and tympana, sensory thresholds and sensory central projections are compared. Reduced tympanal size correlates with a higher auditory threshold. The threshold curves of all four species are tuned to low frequencies with a maximal sensitivity at 3-5 kHz. Central projections of the tympanal nerve show characteristics known from fully tympanate acridid species, so neural elements for tympanal hearing have been strongly conserved across these species. The results also confirm the correlation between reduction in auditory sensitivity and wing reduction. It is concluded that the auditory sensitivity of all four species may be maintained by stabilising selective forces, such as predation.

Neuronal Innervation of the Subgenual Organ Complex and the Tibial Campaniform Sensilla in the Stick Insect Midleg.

Mechanosensory organs in legs play are crucial receptors in the feedback control of walking and in the detection of substrate-borne vibrations. Stick insects serve as a model for the physiological role of chordotonal organs and campaniform sensilla. This study documents, by axonal tracing, the neural innervation of the complex chordotonal organs and groups of campaniform sensilla in the proximal tibia of the midleg in Sipyloidea sipylus. In total, 6 nerve branches innervate the different sensory structures, and the innervation pattern associates different sensilla types by their position. Sensilla on the anterior and posterior tibia are innervated from distinct nerve branches. In addition, the variation in innervation is studied for five anatomical branching points. The most common variation is the innervation of the subgenual organ sensilla by two nerve branches rather than a single one. The fusion of commonly separated nerve branches also occurred. However, a common innervation pattern can be demonstrated, which is found in >75% of preparations. The variation did not include crossings of nerves between the anterior and posterior side of the leg. The study corrects the innervation of the posterior subgenual organ reported previously. The sensory neuroanatomy and innervation pattern can guide further physiological studies of mechanoreceptor organs and allow evolutionary comparisons to related insect groups.

Early postembryogenic development of the subgenual organ complex in the stick insect Sipyloidea sipylus.

Stick insects have elaborate mechanosensory organs in their subgenual organ complex in the proximal tibia, particularly the distal organ with scolopidial sensilla in linear arrangement. For early postembryonic developmental stages of Sipyloidea sipylus (Phasmatodea: Necrosciinae), the neuroanatomy of the scolopidial organs in the subgenual organ complex and the campaniform sensilla is documented by retrograde axonal tracing, and compared to the adult neuroanatomy. Already after hatching of the first larval instars are the sensory structures of subgenual organ and distal organ as well as tibial campaniform sensilla differentiated. In the distal organ, the full set of sensilla is shown in all larval stages examined. This finding indicates that the sensory organs differentiate during embryogenesis, and are already functional by the time of hatching. The constancy of distal organ sensilla over postembryonic stages allows investigation of the representative number of sensilla in adult animals as well as in larval instars. Some anatomical changes occur by postembryogenic length increase of the distal organ, and grouping of the anterior subgenual sensilla. The embryonic development of scolopidial sensilla is similar for auditory sensilla in hemimetabolous Orthoptera (locusts, bushcrickets, crickets) where tympanal membranes develop during postembryogenic stages, conferring a successive gain of sensitivity with larval moults.