The infrared receptor of Melanophila acuminata De Geer (Coleoptera: Buprestidae): ultrastructural study of a unique insect thermoreceptor and its possible descent from a hair mechanoreceptor

The paired infrared organs of Melanophila acuminata consist of 50-100 sensilla situated at the bottom of a pit next to the coxae of the mesothoracic legs, where no exocuticle is developed. Each sensillum is accompanied by a wax gland and has a cuticular lens-like spherule (diameter 12-15 mum) bulging out with its upper hemisphere above the surface, covered only by a thin cuticle of about 1 mum. Distal processes of two enveloping cells surround the entire spherule in the form of a flattened protoplasmatic layer with the exception of a small apical stalk connecting the spherule to the outer cuticle. The spherule is innervated by a single sensory neuron of the ciliary type which is anchored ventrally with the distal tip of its cylindrical and unbranched DOS in the spherule. The insertion of the dendrite, which contains a well-developed tubular body, is always eccentric like in a hair mechanoreceptor (sensillum trichodeum) and there is no evidence of any optical function of the spherule. Three enveloping cells exist, but only one - probably the trichogen cell - forms a relatively small outer receptor lymph cavity. In the posterior wall of the pit - where exocuticle is developed - so-called suppressed systems can be found which remain completely below the cuticle with their otherwise well-developed spherules. Additionally, there is a tendency towards basally flattening and longitudinally stretching of spherules which are situated more peripherally. They strongly resemble the basal regions of hair mechanoreceptors (sensilla trichodea) in their immediate neighbourhood which are also accompanied by wax glands. Because of the existence of these transitional stages and the great ultrastructural resemblance between infrared receptors and hair mechanoreceptors concerning the bauplan of the sensory neurons and their mode of innervating the cuticular apparatus, we conclude that the infrared sensilla are probably derived from hair mechanoreceptors. Based on these results and transmission measurements of infrared radiation through the cuticular components of the organ, a model of the possible function of the infrared receptor is presented.     

Sensory transduction in an insect mechanoreceptor: Linear and nonlinear properties

Mechanotransduction in the femoral tactile spine of the cockroach, Periplaneta americana, was examined as a function of displacement of the spine axially in its socket. Linear behaviour was analyzed by measurement of the frequency response function between displacement and action potential output using sinusoidal stimulation and random noise stimulation. The measured frequency response functions can be well fitted by a relationship which is a fractional power of complex frequency. This power was close to 0.5 for all experiments. To distinguish between the effects of nonlinearity and of inherent variability, the averaged responses of the preparation to repeated sequences of pseudorandom noise were compared to those from experiments in which continuous pseudorandom noise were used. The lack of sensitivity of the coherence function to these two methods of measurement suggests that mechanical stimuli are encoded into action potentials with a large signal-to-noise ratio. The low value of the coherence function which is characteristics of insect mechanoreceptors is therefore due to the strong non-linearity of their responses. To investigate the nonlinear properties of transduction, the second-order frequency response function of the tactile spine was measured for random noise stimulation experiments. Two models of the transduction process were considered in which a linear element with memory was cascaded with a nonlinear element without memory in the two possible configurations. Comparison of the experimental second-order frequency response functions with predictions based upon these two models and the measured first-order frequency response suggests that the transduction mechanism can be modelled by a linear element, which may be associated with the viscoelastic properties of the dendritic tubular body, and a zeromemory nonlinearity, which is most likely to be rectification by the dendritic membrane.     

Antisense inhibition of neuronal nicotinic receptors in the tobacco-feeding insect,Manduca sexta

Acetylcholine is the predominant excitatory transmitter in the insect central nervous system with many of its effects mediated by nicotinic acetylcholine receptors. These receptors are present at very high density and are structurally heterogeneous, although little is known about functional distinctions between them. An interesting system for examining these receptors is the larval stage of Manduca sexta, a nicotine-resistant tobacco-feeding insect. The nicotinic responses of cultured neurons were found to be blocked by mecamylamine and curare but highly resistant to alpha-bungarotoxin. The responses were also unaffected by the reducing agent dithiothreitol and the alkylating agent bromoacetylcholine suggesting that the alpha-subunit dicysteine agonist binding site is protected. To begin determining the functional roles of different subunits in these receptors, cultured neurons were treated with oligonucleotides based on the gene sequence of the alpha subunit, MARA1. Antisense DNA caused a significant downward shift in the amplitude distribution of nicotinic responses compared to sense or reverse antisense treatments. These treatments did not affect currents mediated by the application of GABA. The reduction in the nicotinic depolarization and inward currents did not affect the rate of current onset or recovery, suggesting that antisense MARA1 causes a partial block of all nicotinic responses in these neurons. These results demonstrate that receptor gene expression in insect neurons can be manipulated in a sequence-specific manner by antisense treatment and they provide evidence that MARA1 is important for normal nicotinic responses in Manduca.     

Location and dynamic properties of the spike generator in an insect mechanosensory neuron

1.The cereal bristle hairs of the cockroach, Periplaneta americana, are each innervated by one mechanosensory cell and 1–5 chemosensory cells. In transepithelial recordings, chemo- and mechanosensory spikes could be discriminated from each other by their relative amplitude. 2. When current steps were applied via the sensory hair, trains of impulses were triggered whatever the polarity of the current. 3. All responses adapted to the current, but the time course of adaptation was fitted by a power law for outward currents and an exponential law for inward currents. 4. During application of outward currents, the spikes showed a negative initial phase on which a small positive component was superimposed; strong polarizations produced purely negative spikes. More classical spikes with a positive initial phase were induced by inward currents. 5. The present work supports the hypothesis of a direct excitability of the apical dendrite in cereal bristle mechanoreceptors and confirms previous results suggesting that spikes are normally triggered within that region during mechanical stimulations. It is also established, for the first time, that adaptation to currents may be different in the apical dendrite and in more basal regions of the same mechanosensory neuron.Abbreviation RP receptor potential - TEV transepithelial voltage - TTX tetrodotoxin     

昆虫神经生物学研究技术:细胞内记录

细胞内记录是昆虫神经生物学研究中的常用技术。它用来获得神经元兴奋和抑制过程及神经脉冲产生机制的信息。该技术的特点是把一根微电极的顶尖插入到神经细胞内进行电生理记录 ,这根电极还能用于向膜内输入电流。作者以对蝗虫Schistocercagregaria后胸神经节内的 2个运动神经元的活性记录为例介绍了这一技术     

昆虫神经生物学研究技术:细胞外记录

细胞外记录是昆虫神经生物学研究中最常用的技术之一。它用来获取神经元细胞是否处于激发状态以及激发的强度等信息。该技术的特点是把电极的顶尖放在神经或神经元的附近进行电生理记录。本文以对与蝗虫Schistocerca gregaria成虫后足胫节钟形感器有关的神经N5B2的记录为例介绍这一技术。     

细胞电生理技术在昆虫抗药性研究中的应用

害虫几乎对所有化学农药及Bt等生物农药都产生了抗性。离子通道是多种杀虫剂的作用靶,因此作为研究离子通道基本手段的电压钳与膜片钳技术在害虫抗性检测与抗性机理研究中越来越受到重视。该文综述了细胞电生理技术在害虫抗性机理、杀虫剂作用机理以及药物筛选中的应用。     

New field of insect science: Research on the use of insect properties

In recent years, research on “insect properties” has been attracting much attention for industrial applications of insect technology. This is a new field of research that attempts to analyze specific physiological properties of insects to develop technology for helping humankind. The term “insect properties” has been used to refer to “specific biological functions of insects” since around 1986, and it is now widely accepted in Japan. From 1996, the National Institute of Sericultural and Entomological Science (NISES) promoted “Research for utilization of insect properties” as a “Center of Excellence (COE)” project funded by the Science and Technology Agency. At this point, a new research field called “Research for utilization of insect properties” was initiated, and this led to the recognition of this field by the academic community. In the 21st century, remarkable results, including the development of transgenic silkworms and the full decoding of the silkworm genome, have been achieved. It is expected that this advanced technology will be a powerful tool for progress of research on the use of insect properties. This review presents an overview of the current state of research on use of insect properties as a new technology.     

KCl transport across an insect epithelium: II. Electrochemical potentials and electrophysiology

Summary The cellular mechanism of K-stimulated Cl transport in locust hindgut was studied using double-barrelled ionsensitive microelectrodes and electrophysiological techniques. Steady-state net electrochemical potentials for Cl and K and the conductances of apical and basal membranes and paracellular pathway were determined under control conditions, during exposure to 1mm cAMP, and following ion substitutions. Under control open-circuit conditions, intracellular Cl activity (a _Cl ^c ) was 3.5 times that predicted for passive equilibrium across the apical membrane. The net electrochemical potential opposing Cl entry from the mucosal side increased by 50% during cAMP stimulation of transepithelial Cl absorption whereas the net electrochemical potential favoring Cl exit across the basal membrane was unchanged. No correlation was observed between and the net electrochemical potential across the apical membrane for Na. The net electrochemical potential favoring K entry across the apical membrane was negligible underI _sc conditions when Cl transport rate was approximately 10 eq cm^–2 hr^–1. Locust rectal cells showed electrical and dye coupling. The results also indicate that most transepithelial diffusion of ions is transcellular and that epithelial tightness effectively increases during exposure to cAMP becauseR _a andR _b both decrease, by 80% whileR _j is unchanged. The cAMP-induced R _b was abolished in Cl-free saline whereas R _a was insensitive to Cl removal, but was blocked by removing K from the saline. Based on these findings, our model for Cl absorption in locust hindgut features i) an active entry step for Cl at the apical membrane which is stimulated by cAMP and by low levels of K on the mucosal side, but is not energized by or a large cAMP-stimulated Cl conductance in the basal membrane and a similar cAMP-stimulated K conductance in the apical membrane. cAMP dose-response curves are similar for the stimulation of active Cl absorption and Cl-independent (i. e. K) conductance, indicating that cAMP exerts dual control over active Cl transport and counter-ion permeability.     

Paedogenesis in the dipteran insect Heteropeza pygmaea: an interpretation

A comparison of the modified oocyte development in paedogenetic reproduction with ‘regular’ oogenesis in imaginal reproduction of H. pygmaea shows that the paedogenetically developing eggs start embryonic development long before the usual processes of oocyte growth have been terminated or even initiated. Under certain experimental conditions, uncoupling of meiotic and growth processes could be observed in eggs cultured in vitro and showing aberrant development. We, therefore, assume that in paedogenetic egg development a meiotic block is not properly established or removed precociously. An interpretation of the evolution of paedogenetic reproduction is given.     

Mimosa pudica: Electrical and mechanical stimulation of plant movements

Thigmonastic movements in the sensitive plant Mimosa pudica L., associated with fast responses to environmental stimuli, appear to be regulated through electrical and chemical signal transductions. The thigmonastic responses of M. pudica can be considered in three stages: stimulus perception, electrical signal transmission and induction of mechanical, hydrodynamical and biochemical responses. We investigated the mechanical movements of the pinnae and petioles in M. pudica induced by the electrical stimulation of a pulvinus, petiole, secondary pulvinus or pinna by a low electrical voltage and charge. The threshold value was 1.3–1.5 V of applied voltage and 2 to 10 µC of charge for the closing of the pinnules. Both voltage and electrical charge are responsible for the electro‐stimulated closing of a leaf. The mechanism behind closing the leaf in M. pudica is discussed. The hydroelastic curvature mechanism closely describes the kinetics of M. pudica leaf movements.     

Phentolamine selectively affects the fast sodium component of sensory adaptation in an insect mechanoreceptor

Phentolamine and related compounds have several different actions on nervous tissues in vertebrates and invertebrates, including a local anesthetic effect. However, recent work suggests that phentolamine can interfere with sensory transduction in insect mechanoreceptors at significantly lower concentrations than are required for conduction block. We tested the actions of phentolamine on sensory transduction and encoding in an insect mechanoreceptor, the cockroach tactile spine neuron and found that 500 microM phentolamine increased the action potential threshold by 50%. The passive membrane properties of the neuron were not affected, but one component of dynamic threshold change was strongly and selectively reduced. This component has previously been attributed to slowly inactivating sodium channels in the action potential initiating region, suggesting that these channels are the most phentolamine-sensitive sites.     

Variation in the electrical properties of cultured human proximal tubule cells

Summary Monolayers of human proximal tubule (HPT) cells, when grown on permeable supports and mounted in Ussing chambers, spontaneously display a transepithelial potential difference (PD), short-circuit current (Isc), and transepithelial specific resistance (R_T). These electrical parameters were used to determine the degree of heterogeneity among independent isolates of human proximal tubule cell cultures. Seventeen independent isolates of cells were assessed, totaling 260 individual determinations of spontaneous electrical properties. On average, these cell monolayers displayed an apicalnegative PD of 1.5 ± 0.1 mV, an Isc of 2.7 ± 0.2 μA/cm², and an R_T of 480 ± 19 ohms × cm². Each independent cell isolate, however, displayed electrical values within a narrow range, in some cases allowing isolates to be distinguished from one another. The individual isolates were also assessed for Na-coupled glucose transport, Na⁺,K⁺-ATPase activity, cAMP stimulation by parathyroid hormone (PTH), forskolin stimulation of Isc, and ouabain inhibition. With the exception of a strong correlation between Na⁺,K⁺-ATPase activity and Isc, these parameters, in contrast to electrical properties, were found to be consistent and did not reveal distinctions among the isolates. HPT cell cultures seem to consistently retain important features of proximal tubule differentiation while maintaining the variability, as demonstrated by electrical properties, that might be expected of cells isolated from a random population.     

Excitable properties of insect neurones in culture: A developmental study

The passive and excitable electrical properties of cockroach neurones growing in vitro have been investigated using intracellular recording techniques. The resting membrane potentials of the neurones are similar to those of their in vivo counterparts but the input resistances and membrane capacitive properties are more typical of embryonic insect neurones. During the first 12 days of growth in vitro the neurones exhibit delayed rectification in response to the injection of depolarising current steps. After this period “all or none” action potentials can be evoked by depolarising pulses in approximately half of the neurones tested. These spikes are abolised by 1 μM tetrodotoxin but are unaffected by 5 mM Co²⁺. Spontaneous excitatory activity develops in approx 25% of the neurones after 3 weeks in culture.     

Effects of intraoral mechanoreceptor stimulation on reticular formation neurones, in the rabbit

The effects of intraoral mechanoreceptor stimulation on the firing rate of single neurones of the brain stem reticular formation (RF) were investigated in rabbits. 30% of RF neurones responded to periodontal mechanoreceptor stimulation; 16% to mucosal mechanoreceptor stimulation and 6% to both types of stimuli. Periodontal stimulation induced mainly inhibitory effects localized within the mesencephalic and rostral pontine RF. Among periodontal afferents incisors were the most widely represented. The effects of mucosal mechanoreceptor stimulation were predominant in the medullary and pontine RF and they were mainly excitatory. The present results support the hypothesis that brain stem RF neurones can be recruited into regulating mastication and biting also by stimulation of intraoral mechanoreceptors.     

Effects of 20-hydroxyecdysone and tunicamycin on glycoprotein synthesis in an insect cell line

Treatment of CH-MRRL cells with either 20-hydroxyecdysone or tunicamycin resulted in a decrease in the incorporation of labeled sugars into glycoproteins. This change appears to be largely quantitative, as few qualitative changes in protein bands were apparent as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Tunicamycin caused a greater change in the amount of labeled sugar incorporated into specific glycoproteins than did 20-hydroxyecdysone. This was more apparent in [¹⁴C]-mannose-labeled than in [¹⁴C]-N-acetylglucosamine-labeled glycoproteins. Both compounds caused changes in cell surface glycoproteins. These changes are discussed in relation to previous work on binding of lectins to the cell surface and on the mode of action of tunicamycin.     

A look into the invisible: ultraviolet-B sensitivity in an insect (Caliothrips phaseoli) revealed through a behavioural action spectrum

Caliothrips phaseoli, a phytophagous insect, detects and responds to solar ultraviolet-B radiation (UV-B; λ ≤ 315 nm) under field conditions. A highly specific mechanism must be present in the thrips visual system in order to detect this narrow band of solar radiation, which is at least 30 times less abundant than the UV-A (315–400 nm), to which many insects are sensitive. We constructed an action spectrum of thrips responses to light by studying their behavioural reactions to monochromatic irradiation under confinement conditions. Thrips were maximally sensitive to wavelengths between 290 and 330 nm; human-visible wavelengths (λ ≥ 400 nm) failed to elicit any response. All but six ommatidia of the thrips compound eye were highly fluorescent when exposed to UV-A of wavelengths longer than 330 nm. We hypothesized that the fluorescent compound acts as an internal filter, preventing radiation with λ > 330 nm from reaching the photoreceptor cells. Calculations based on the putative filter transmittance and a visual pigment template of λ_max = 360 nm produced a sensitivity spectrum that was strikingly similar to the action spectrum of UV-induced behavioural response. These results suggest that specific UV-B vision in thrips is achieved by a standard UV-A photoreceptor and a sharp cut-off internal filter that blocks longer UV wavelengths in the majority of the ommatidia.     

Reduction of mechanical sensitivity in an insect mechanoreceptor correlated with destruction of its tubular body

Summary The modified cilium (dendrite) of epithelial mechanoreceptors of insects contains microtubules in different arrangements: (1) microtubules distributed over the entire receptor and not fixed in a special configuration, therefore called free microtubules, (2) densely packed, interconnected microtubules called the tubular body, and (3) 9 doublet microtubules. These groups of microtubules have been discussed in relation to mechanotransduction.In a preceding paper the free microtubules were proved to be not involved in mechanotransduction. In this paper the hypothesis is examined that the tubular body may be essential to mechanotransduction. For this purpose the effect of the microtubule-disassembling drug vinblastine on both the tubular body and the sensitivity is examined in a femoral mechanoreceptor of the cricket Acheta domesticus.After 6- to 26-h exposure to vinblastine the tubular body is partially or totally destroyed. Simultaneously, mechanical sensitivity decays to zero. In contrast, the pacemaker property for nerve impulses of the apical dendritic segment is only slightly altered. We conclude from these results that the tubular body is essential to mechanotransduction. Three experiments in which a (small) response persisted, despite a totally destroyed tubular body, suggest that receptor potentials can in principle be evoked without an intact tubular body.In addition to the irreversible reduction of receptor sensitivity, vinblastine causes a reversible reduction during repetitive stimulation. This adaptation is supposed to be the consequence of altered properties of the tubular body.