Mapping the Auditory Space of Culex pipiens Female Mosquitoes in 3D.

The task of directional hearing faces most animals that possess ears. They approach this task in different ways, but a common trait is the use of binaural cues to find the direction to the source of sound. In insects, the task is further complicated by their small size and, hence, minute temporal and level differences between two ears. A single symmetric flagellar particle velocity receiver, such as the antenna of a mosquito, should not be able to discriminate between the two opposite directions along the vector of the sound wave. Paired antennae of mosquitoes presume the usage of binaural hearing, but its mechanisms are expected to be significantly different from the ones typical for the pressure receivers. However, the directionality of flagellar auditory organs has received little attention. Here, we measured the in-flight orientation of antennae in female Culex pipiens pipiens mosquitoes and obtained a detailed physiological mapping of the Johnston's organ directionality at the level of individual sensory units. By combining these data, we created a three-dimensional model of the mosquito's auditory space. The orientation of the antennae was found to be coordinated with the neuronal asymmetry of the Johnston's organs to maintain a uniformly shaped auditory space, symmetric relative to a flying mosquito. The overlap of the directional characteristics of the left and right sensory units was found to be optimal for binaural hearing focused primarily in front of, above and below a flying mosquito.

Beyond the Limits of Light: An Application of Super-Resolution Confocal Microscopy (sCLSM) to Investigate Eocene Amber Microfossils.

Amber is known as one of the best sources of fossil organisms preserved with exceptional fidelity. Historically, different methods of imaging have been applied to amber, including optical microscopy and microtomography. These methods are sufficient to resolve millimeter-scaled fossils. However, microfossils, such as microarthropods, require another resolution. Here, we describe a non-destructive method of super resolution confocal microscopy (sCLSM) to study amber-preserved microfossils, using a novel astigmatid mite species (genus Histiogaster, Acaridae) from Eocene Rovno amber as a model. We show that the resolution obtained with sCLSM is comparable to that of scanning electron microscopy (SEM) routinely used to study modern mites. We compare sCLSM imaging to other methods that are used to study amber inclusions and emphasize its advantages in examination of unique fossil specimens. Furthermore, we show that the deterioration of amber, which manifests in its darkening, positively correlates with its increased fluorescence. Our results demonstrate a great potential of the sCLSM method for imaging of the tiniest organisms preserved in amber.

Sentinel lymph node detection by combining nonradioactive techniques with contrast agents: State of the art and prospects.

The status of sentinel lymph nodes (SLNs) has a substantial prognostic value because these nodes are the first place where cancer cells accumulate along their spreading route. Routine SLN biopsy ("gold standard") involves peritumoral injections of radiopharmaceuticals, such as technetium-99m, which has obvious disadvantages. This review examines the methods used as "gold standard" analogs to diagnose SLNs. Nonradioactive preoperative and intraoperative methods of SLN detection are analyzed. Promising photonic tools for SLNs detection are reviewed, including NIR-I/NIR-II fluorescence imaging, photoswitching dyes for SLN detection, in vivo photoacoustic detection, imaging and biopsy of SLNs. Also are discussed methods of SLN detection by magnetic resonance imaging, ultrasonic imaging systems including as combined with photoacoustic imaging, and methods based on the magnetometer-aided detection of superparamagnetic nanoparticles. The advantages and disadvantages of nonradioactive SLN-detection methods are shown. The review concludes with prospects for the use of conservative diagnostic methods in combination with photonic tools.

A transitional fossil mite (Astigmata: Levantoglyphidae fam. n.) from the early Cretaceous suggests gradual evolution of phoresy-related metamorphosis.

Metamorphosis is a key innovation allowing the same species to inhabit different environments and accomplish different functions, leading to evolutionary success in many animal groups. Astigmata is a megadiverse lineage of mites that expanded into a great number of habitats via associations with invertebrate and vertebrate hosts (human associates include stored food mites, house dust mites, and scabies). The evolutionary success of Astigmata is linked to phoresy-related metamorphosis, namely the origin of the heteromorphic deutonymph, which is highly specialized for phoresy (dispersal on hosts). The origin of this instar is enigmatic since it is morphologically divergent and no intermediate forms are known. Here we describe the heteromorphic deutonymph of Levantoglyphus sidorchukae n. gen. and sp. (Levantoglyphidae fam. n.) from early Cretaceous amber of Lebanon (129 Ma), which displays a transitional morphology. It is similar to extant phoretic deutonymphs in its modifications for phoresy but has the masticatory system and other parts of the gnathosoma well-developed. These aspects point to a gradual evolution of the astigmatid heteromorphic morphology and metamorphosis. The presence of well-developed presumably host-seeking sensory elements on the gnathosoma suggests that the deutonymph was not feeding either during phoretic or pre- or postphoretic periods.

Frequency tuning of swarming male mosquitoes (Aedes communis, Culicidae) and its neural mechanisms.

The primary function of hearing in mosquitoes is believed to be intraspecific communication. This view dictated the principle of many behavioral studies, namely, the attraction of male mosquitoes to the sounds that mimicked a female tone. However, after the avoidance response to certain frequencies of sound was demonstrated, it became clear that attraction tests cannot fully account for all the capabilities of the mosquito auditory system. In addition, the tuning curves obtained by electrophysiological measurements differ from the behavioral ones. We designed a simple but robust field test based on responses of swarming mosquitoes to sound stimulation, but not limited to the attraction response. Here we report the auditory thresholds over a wide range of sound frequencies measured in the field from swarms of Aedes communis mosquitoes. In parallel, the auditory sensitivity of male mosquitoes taken from the same swarms was measured electrophysiologically. Surprisingly, we found high acoustic sensitivity; 26 dBSPL on average, in the frequency range 180-220 Hz (ambient temperature 12 °C). In addition, responses were found in the high-frequency range, 500-700 Hz (the so-called 'mirror channel'). Two types of auditory units were recorded: more sensitive broadband neurons and less sensitive units with distinct narrow (quality factor Q6 = 7.4) frequency tunings in the range 180-350 Hz. We propose that the former provides the detection of signal while the latter are used for frequency identification in order to make a behavioral choice.

Directional and frequency characteristics of auditory neurons in Culex male mosquitoes.

The paired auditory organ of the mosquito, the Johnston's organ (JO), being the receiver of the particle velocity component of sound, is directional by its structure. However, to date almost no physiological measurements of its directionality have been made. In addition, the recent finding on the grouping of the JO auditory neurons into antiphase pairs demands confirmation by different methods. Using the vector superposition of the signals produced by two orthogonally oriented speakers, we measured the directional characteristics of individual units as well as their relationships in physiologically distinguishable groups - pairs or triplets. The feedback stimulation method allowed us to discriminate responses of the two simultaneously recorded units, and to show that they indeed responded in antiphase. Units of different frequency tuning as well as highly sensitive units (thresholds of 27 dB SPVL and below) were found in every angular sector of the JO, providing the mosquito with the ability to produce complex auditory behaviors.

Frequency organization of the Johnston's organ in male mosquitoes (Diptera, Culicidae).

The Johnston's organs (JO) of mosquitoes are the most complex mechanosensitive organs yet found in insects. Previous findings on the behavior of mosquitoes suggest that, together with exceptional sensitivity, their auditory system can discriminate frequencies. Analysis of compound responses of the JO did not provide unambiguous evidence of such discrimination, nor did it help to find its mechanism. Using the feedback stimulation method, we measured the tuning frequencies of the JO sensory neurons. Here we present electrophysiological evidence that male mosquitoes of Culex pipiens possess at least eight groups of auditory neurons that are distinct in their frequency tuning, with individual frequencies ranging from 85 to 470 Hz. Most of the neurons are tuned to 190-270 Hz, which corresponds to the difference between male and female flight tones. Axons of the JO sensory units propagate graded amplified receptor potentials rather than all-or-none action potentials, are grouped into pairs or triplets and often respond in anti-phase to each other. Some features of the mosquito auditory system suggest an analogy to the retinal mechanisms. Together with our previous findings on frequency tuning in female mosquitoes of different species, this study presents evidence in favor of sophisticated frequency analysis of sound in mosquitoes.

Previous motor activity affects the transition from uncertainty to decision making in snails.

One of the most widely accepted benefits of enhanced physical activity is an improvement in the symptoms of depression, including the facilitation of decision making. Up until now, these effects have been shown in rodents and humans only. Little is known about their evolutionary origin or biological basis, and the underlying cellular mechanisms also remain relatively elusive. Here, we demonstrate for the first time that preceding motor activity accelerates decision making in an invertebrate, the pond snail Lymnaea stagnalis To investigate decision making in a novel environment, snails, which normally live in water, were placed on a flat dry surface to simulate the potentially threatening consequence of being in an arid environment. This stimulus initiated two distinct phases in snail behaviour: slow circular movements, followed by intense locomotion in a chosen direction. The first phase was prolonged when the test arena was symmetrically lit, compared with one with an apparent gradient of light. However, forced muscular locomotion for 2 h prior to the test promoted the transition from random circular motions to a directional crawl, accompanied by an increase in crawling speed but with no effect on the choice of direction. Intense locomotion for 2 h also produced a strong excitatory effect on the activity of serotonergic neurons in L. stagnalis Our results suggest that the beneficial effects of physical exercise on cognitive performance in mammals might have deep roots in evolution, granting the opportunity to unravel the origins of such effects at the single-neuron and network levels.

Direct and decarboxylation-dependent effects of neurotransmitter precursors on firing of isolated monoaminergic neurons.

To elucidate mechanisms that underlie the profound physiological effects of the monoamine precursors 5-hydroxy-L-tryptophan (5-HTP) and L-3,4-dihydroxyphenylalanine (L-DOPA), we examined their action on single monoaminergic neurons isolated from the ganglia of the gastropod snail Lymnaea stagnalis. In isolated serotonergic PeA motoneurons, 5-HTP produced excitation. The effect was mimicked by serotonin at 0.5-1 microM, masked by pretreatment with serotonin at higher concentrations, and abolished by the inhibitor of aromatic amino acid decarboxylase (AAAD) m-hydroxybenzylhydrazine (NSD-1015), the inhibitor of the vesicular monoamine transporter reserpine or the serotonin receptor antagonist mianserin. Exposure of the dopaminergic interneurons RPeD1 to L-DOPA caused a biphasic effect composed of a depolarization followed by a hyperpolarization. AAAD inactivation with NSD-1015, as well as the blockade of dopamine receptors with sulpiride, resulted in the enhancement of the excitatory effect, and the abolition of the inhibitory effect. Dopamine caused hyperpolarization and masked the inhibitory phase of L-DOPA action. The results show that precursors affect the rate of firing of isolated monoaminergic neurons and that their effect is completely or partially mediated by the enhanced synthesis of the respective neurotransmitter, followed by extrasynaptic release of the latter and activation of extrasynaptic autoreceptors.