Simple Dark-Field Microscopy with Nanometer Spatial Precision and Microsecond Temporal Resolution

Molecular motors such as kinesin, myosin, and F₁-ATPase are responsible for many important cellular processes. These motor proteins exhibit nanometer-scale, stepwise movements on micro- to millisecond timescales. So far, methods developed to measure these small and fast movements with high spatial and temporal resolution require relatively complicated experimental systems. Here, we describe a simple dark-field imaging system that employs objective-type evanescent illumination to selectively illuminate a thin layer on the coverslip and thus yield images with high signal/noise ratios. Only by substituting the dichroic mirror in conventional objective-type total internal reflection fluorescence microscope with a perforated mirror, were nanometer spatial precision and microsecond temporal resolution simultaneously achieved. This system was applied to the study of the rotary mechanism of F₁-ATPase. The fluctuation of a gold nanoparticle attached to the γ-subunit during catalytic dwell and the stepping motion during torque generation were successfully visualized with 9.1-μs temporal resolution. Because of the simple optics, this system will be applicable to various biophysical studies requiring high spatial and temporal resolution in vitro and also in vivo.     

Odors and olfaction

In this review, we discuss some of the neural processes involved in the perception of odors that, together with audition and vision, provide essential information for analyzing our surroundings. We shall see how odor detection and learning induce substantial structural and functional changes at the first relay of the olfactory system, i.e., the main olfactory bulb. Among the mechanisms that participate in these modifications is the persistence of a high level of interneuron neurogenesis within the adult olfactory bulb. Our goal is to present some observations related to the neurogenesis that may aid in understanding the neural mechanisms of sensory perception and shed light on the cellular basis of olfactory learning. We summarize the current ideas concerning the molecular mechanisms and organizational strategies used by the olfactory system to transduce, encode, and process information at various levels in the olfactory sensory pathway. Due to space constraints, this review focuses exclusively on the olfactory systems of vertebrates and primarily those of mammals.     

昆虫对气味分子的感受机制

昆虫对外界气味的感受作用是一个庞大而复杂的体系,多种蛋白参与了这一过程。其中包括气味结合蛋白,气味结合蛋白受体,气味降解酶等多种蛋白。昆虫不仅可以通过外界气味分子携带的信息来识别配偶,天敌,还可以通过对外界环境特征的识别来寻找食物来源,产卵等。明确昆虫的化学感受机制不仅可以帮助我们理解昆虫的行为,还有助于深入了解动物的行为机制。文章综述昆虫对气味分子的识别、气味分子在昆虫体内的运输以及电化学信号传导机制等方面的进展。     

A Method for Activation of Endogenous Acid-sensing Ion Channel 1a (ASIC1a) in the Nervous System with High Spatial and Temporal Precision

Protons activate acid-sensing ion channel 1a (ASIC1a) in the central nervous system (CNS) although the impact of such activation on brain outputs remains elusive. Progress elucidating the functional roles of ASIC1a in the CNS has been hindered by technical difficulties of achieving acidification with spatial and temporal precision. We have implemented a method to control optically the opening of ASIC1a in brain slices and also in awake animals. The light-driven H⁺ pump ArchT was expressed in astrocytes of mouse cortex by injection of adenoviral vectors containing a strong and astrocyte-specific promoter. Illumination with amber light acidified the surrounding interstitium and led to activation of endogenous ASIC1a channels and firing of action potentials in neurons localized in close proximity to ArchT-expressing astrocytes. We conclude that this optogenetic method offers a minimally invasive approach that enables examining the biological consequences of ASIC1a currents in any structure of the CNS and in the modulation of animal behaviors.     

Sensitivity-dependent Hierarchical Receptor Codes for Odors

Chemical Senses, 28, 87–104 Regrettably, an error occurred in the abstract of this article.The third sentence was incorrect and should have read ‘The chiral-non-discriminatingreceptors were newly     

High Precision Zinc Isotopic Measurements Applied to Mouse Organs

We present a procedure to measure with high precision zinc isotope ratios in mouse organs. Zinc is composed of 5 stable isotopes (⁶⁴Zn, ⁶⁶Zn, ⁶⁷Zn, ⁶⁸Zn and ⁷⁰Zn) which are naturally fractionated between mouse organs. We first show how to dissolve the different organs in order to free the Zn atoms; this step is realized by a mixture of HNO₃ and H₂O₂. We then purify the zinc atoms from all the other elements, in particular from isobaric interferences (e.g., Ni), by anion-exchange chromatography in a dilute HBr/HNO₃ medium. These first two steps are performed in a clean laboratory using high purity chemicals. Finally, the isotope ratios are measured by using a multi-collector inductively-coupled-plasma mass-spectrometer, in low resolution. The samples are injected using a spray chamber and the isotopic fractionation induced by the mass-spectrometer is corrected by comparing the ratio of the samples to the ratio of a standard (standard bracketing technique). This full typical procedure produces an isotope ratio with a 50 ppm (2 s.d.) reproducibility.     

Attention Increases the Temporal Precision of Conscious Perception: Verifying the Neural-ST2 Model

What role does attention play in ensuring the temporal precision of visual perception? Behavioural studies have investigated feature selection and binding in time using fleeting sequences of stimuli in the Rapid Serial Visual Presentation (RSVP) paradigm, and found that temporal accuracy is reduced when attentional control is diminished. To reduce the efficacy of attentional deployment, these studies have employed the Attentional Blink (AB) phenomenon. In this article, we use electroencephalography (EEG) to directly investigate the temporal dynamics of conscious perception. Specifically, employing a combination of experimental analysis and neural network modelling, we test the hypothesis that the availability of attention reduces temporal jitter in the latency between a target''s visual onset and its consolidation into working memory. We perform time-frequency analysis on data from an AB study to compare the EEG trials underlying the P3 ERPs (Event-related Potential) evoked by targets seen outside vs. inside the AB time window. We find visual differences in phase-sorted ERPimages and statistical differences in the variance of the P3 phase distributions. These results argue for increased variation in the latency of conscious perception during the AB. This experimental analysis is complemented by a theoretical exploration of temporal attention and target processing. Using activation traces from the Neural-ST² model, we generate virtual ERPs and virtual ERPimages. These are compared to their human counterparts to propose an explanation of how target consolidation in the context of the AB influences the temporal variability of selective attention. The AB provides us with a suitable phenomenon with which to investigate the interplay between attention and perception. The combination of experimental and theoretical elucidation in this article contributes to converging evidence for the notion that the AB reflects a reduction in the temporal acuity of selective attention and the timeliness of perception.     

Distinguishing species

Given two organisms, how can one distinguish whether they belong to the same species or not? This might be straightforward for two divergent organisms, but can be extremely difficult and laborious for closely related ones. A molecular marker giving a clear distinction would therefore be of immense benefit. The internal transcribed spacer 2 (ITS2) has been widely used for low-level phylogenetic analyses. Case studies revealed that a compensatory base change (CBC) in the helix II or helix III ITS2 secondary structure between two organisms correlated with sexual incompatibility. We analyzed more than 1300 closely related species to test whether this correlation is generally applicable. In 93%, where a CBC was found between organisms classified within the same genus, they belong to different species. Thus, a CBC in an ITS2 sequence-structure alignment is a sufficient condition to distinguish even closely related species.     

High Precision U/Th Dating of First Polynesian Settlement

Previous studies document Nukuleka in the Kingdom of Tonga as a founder colony for first settlement of Polynesia by Lapita peoples. A limited number of radiocarbon dates are one line of evidence supporting this claim, but they cannot precisely establish when this event occurred, nor can they afford a detailed chronology for sequent occupation. High precision U/Th dates of Acropora coral files (abraders) from Nukuleka give unprecedented resolution, identifying the founder event by 2838±8 BP and documenting site development over the ensuing 250 years. The potential for dating error due to post depositional diagenetic alteration of ancient corals at Nukuleka also is addressed through sample preparation protocols and paired dates on spatially separated samples for individual specimens. Acropora coral files are widely distributed in Lapita sites across Oceania. U/Th dating of these artifacts provides unparalleled opportunities for greater precision and insight into the speed and timing of this final chapter in human settlement of the globe.     

Odors: implicit memory and performance effects.

In order to assess the influence of odors on human performance and implicit memory for odors, 108 subjects completed a variety of tests in weakly scented (jasmine, lavender or odorless) rooms without having been made aware of the odor. After a 30 min interval the subjects were shown slides of different surroundings, including the room they had been in, and were requested to rate how well a set of 12 odors, including a blank, would fit to these surroundings. Half of these contexts contained visual cues related to two of the presented odors (leather and coffee). After the rating of fit the subjects had to rate the odors for pleasantness, were asked to identify the odors with their correct names and to tell where and when they had last smelled these odors. One subject remembered smelling the odor (jasmine) in the room and was discarded from the analysis of the results for the rating of fit. None of the others reported recollection of the experimental odors. The results showed that in general jasmine had a negative and lavender a positive effect on test performance. If an odor-related visual cue was present in the context, the related odor was always rated highest in fit to that context. Furthermore, the subjects working in rooms with an odor subsequently assigned this odor to the visual context of that room to a significantly higher degree than subjects working in rooms with different odors. Since none of the subjects reported that they had smelled the odor in the rooms where performance testing took place, it was concluded that the memory for these odors was implicit. Further analysis showed that such memory was only found in subjects who were unable to supply the right name for the odor. The possible consequences of this latter finding for understanding the relationship between sensory (episodic) and semantic odor memory are discussed.     

Response of Bacterioplankton Communities to Cadmium Exposure in Coastal Water Microcosms with High Temporal Variability

Multiple anthropogenic disturbances to bacterial diversity have been investigated in coastal ecosystems, in which temporal variability in the bacterioplankton community has been considered a ubiquitous process. However, far less is known about the temporal dynamics of a bacterioplankton community responding to pollution disturbances such as toxic metals. We used coastal water microcosms perturbed with 0, 10, 100, and 1,000 μg liter⁻¹ of cadmium (Cd) for 2 weeks to investigate temporal variability, Cd-induced patterns, and their interaction in the coastal bacterioplankton community and to reveal whether the bacterial community structure would reflect the Cd gradient in a temporally varying system. Our results showed that the bacterioplankton community structure shifted along the Cd gradient consistently after a 4-day incubation, although it exhibited some resistance to Cd at low concentration (10 μg liter⁻¹). A process akin to an arms race between temporal variability and Cd exposure was observed, and the temporal variability overwhelmed Cd-induced patterns in the bacterial community. The temporal succession of the bacterial community was correlated with pH, dissolved oxygen, NO₃⁻-N, NO₂⁻-N, PO₄³⁻-P, dissolved organic carbon, and chlorophyll a, and each of these parameters contributed more to community variance than Cd did. However, elevated Cd levels did decrease the temporal turnover rate of community. Furthermore, key taxa, affiliated to the families Flavobacteriaceae, Rhodobacteraceae, Erythrobacteraceae, Piscirickettsiaceae, and Alteromonadaceae, showed a high frequency of being associated with Cd levels during 2 weeks. This study provides direct evidence that specific Cd-induced patterns in bacterioplankton communities exist in highly varying manipulated coastal systems. Future investigations on an ecosystem scale across longer temporal scales are needed to validate the observed pattern.