视网膜单细胞成像技术研究

建立了一台基于37单元变形镜、Shack-hartman波前像差传感器和12位科研级CCD相机的自适应光学视网膜相机。采用一个中心波长为679nm的超辐射二极管(SLD)作为相机的光源,通过将超辐射二极管和多模光纤耦合,显著减小了SLD光源的空间相干性,从而消除了散斑噪声对成像的影响。多模光纤的输出提供了一种高亮度、均匀照明的光源,使人眼视网膜单细胞成像的速度达到4．8幅／秒。     

Development of the auditory sensitivity and formation of the acoustically guided defensive behavior in nestlings of the pied flycatcher Ficedula hypoleuca

Age dynamics of generation of the evoked potentials (EP) in the field L of caudal nidopallium (the higher integrative center of the avian auditory system) and development of the auditory-guided defensive behavior were studied in control and visually deprived pied flycatcher Ficedula hypoleuca nestlings. It was shown that the rhythmically organized monofrequency signals with sound frequency 3.5 kHz and higher produced the defensive behavior as the auditory sensitivity to these frequencies matured. After 9 days, the species-specific alarm signal produced more effectively the defensive behavior than the tonal signals. The rhythmically organized sound with filling frequency 0.5 kHz, occupying the less low-frequency diapason than the feeding signal, produced the effect opposite to the alarm signal to increase the nestling mobility. At the initial stage of the defensive behavior development the auditory threshold fell markedly in the frequency diapason corresponding to the frequency diapason of the alarm signal (5–6 kHz), which seemed to facilitate involvement of this diapason signals in the defensive integration. The auditory EP generation thresholds in the whole studied diapason were lower in the visually deprived nestlings than in the normally developing one; however, the ability of the acoustic signals to suppress alimentary reactions fell significantly.     

CyclinD2-mediated regulation of neurogenic output from the retinal ciliary margin is perturbed in albinism

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Linking phenotypic correlations from a diverse set of laboratory tests to field behaviors in the crayfish,Orconectes virilis

The presence of phenotypic behavioral correlations and their connection to fitness consequences of organisms have received considerable debate within the literature. Yet, little work has been carried out to connect any behavioral correlates found within a set of laboratory studies to natural behavior observed under complex environmental conditions. To help fill this gap, individual crayfish, collected from the same local population, completed five different behavioral assays in a laboratory setting in a random order. These data were used to reveal any possible correlations for behavioral scores across all of the laboratory tests. Subsequently, these same individuals were placed into the field and video recorded for 24 hr. A separate set of field behaviors, related to the laboratory assays, were quantified from the field videos. The normalized laboratory and field behaviors were used in three stepwise statistical analyses. First, normalized data were loaded into a PCA to generate a priori hypotheses on potential behavioral correlates. These hypotheses were subsequently tested using general multiple linear regression. Finally, structural equation modeling was performed to elucidate any behavioral modules from the laboratory assays that correlated with behavioral patterns present from the fieldwork. Three laboratory‐based behavioral modules were connected to three separate field assays: exploration–avoidance, bold–shy, and aggressiveness. Yet, some behaviors exhibited in the laboratory assays were uncorrelated with any behaviors found in the field and vice versa. Results from this study provide evidence that although many different behavioral correlates may exist within laboratory settings, these same modules may not translate directly into predicting behavior under natural settings.     

The Optimal Human Ventral Stream from Estimates of the Complexity of Visual Objects

The part of the primate visual cortex responsible for the recognition of objects is parcelled into about a dozen areas organized somewhat hierarchically (the region is called the ventral stream). Why are there approximately this many hierarchical levels? Here I put forth a generic information-processing hierarchical model, and show how the total number of neurons required depends on the number of hierarchical levels and on the complexity of visual objects that must be recognized. Because the recognition of written words appears to occur in a similar part of inferotemporal cortex as other visual objects, the complexity of written words may be similar to that of other visual objects for humans; for this reason, I measure the complexity of written words, and use it as an approximate estimate of the complexity more generally of visual objects. I then show that the information-processing hierarchy that accommodates visual objects of that complexity possesses the minimum number of neurons when the number of hierarchical levels is approximately 15.     

Preserved particulate organic carbon is likely derived from the subsurface sulfidic photic zone of the Proterozoic Ocean: evidence from a modern,oxygen-deficient lake

Biological processes in the Proterozoic Ocean are often inferred from modern oxygen-deficient environments (MODEs) or from stable isotopes in preserved sediment. To date, few MODE studies have simultaneously quantified carbon fixation genes and attendant stable isotopic signatures. Consequently, how carbon isotope patterns reflect these pathways has not been thoroughly vetted. Addressing this, we profiled planktonic productivity and quantified carbon fixation pathway genes and associated organic carbon isotope values (δ¹³C_POC) of size-fractionated (0.2–2.7 and >2.7 μm) particulate matter from meromictic Fayetteville Green Lake, NY, USA. The high-O₂ Calvin-Benson-Bassham (CBB) gene (cbbL) was most abundant in the <2.7 μm size fraction in shallow oxic and deep hypoxic waters, corresponding with cyanobacterial and eukaryote algal populations. The low-O₂ CBB gene (cbbM) was most abundant near the lower oxycline boundary in the larger size fraction, coincident with purple sulfur bacteria populations. The reverse citric acid cycle gene (aclB) was equally abundant in both size fractions in the deepest photic zone, coinciding with green sulfur bacteria populations. Methane coenzyme reductase A (mcrA), of anaerobic methane cyclers, was most abundant at the lower oxycline boundary in both size fractions, coinciding with Methanoregula populations. δ¹³C_POC values overlapped with the high-O₂ CBB fixation range except for two negative excursions near the lower oxycline boundary, likely reflecting assimilation of isotopically-depleted groundwater-derived carbon by autotrophs and sulfate-reducers. Throughout aphotic waters, δ¹³C_POC values of the large size fraction became ¹³C-enriched, likely reflecting abundant purple sulfur bacterial aggregates. Eukaryote algae- or cyanobacteria-like isotopic signatures corresponded with increases in cbbL, cbbM, and aclB, and enrichment of exopolymer-rich prokaryotic photoautotrophs aggregates. Results suggest that δ¹³C_POC values of preserved sediments from areas of the Proterozoic Ocean with sulfidic photic zones may reflect a mixture of alternate carbon-fixing populations exported from the deep photic zone, challenging the paradigm that sedimentary stable carbon isotope values predominantly reflect oxygenic photosynthesis from surface waters.     

Topology is the Principal Determinant in the Folding of a Complex All-alpha Greek Key Death Domain from Human FADD

In order to elucidate the relative importance of secondary structure and topology in determining folding mechanism, we have carried out a phi-value analysis of the death domain (DD) from human FADD. FADD DD is a 100 amino acid domain consisting of six anti-parallel alpha helices arranged in a Greek key structure. We asked how does the folding of this domain compare with that of (a) other all-alpha-helical proteins and (b) other Greek key proteins? Is the folding pathway determined mainly by secondary structure or is topology the principal determinant? Our Φ-value analysis reveals a striking resemblance to the all-beta Greek key immunoglobulin-like domains. Both fold via diffuse transition states and, importantly, long-range interactions between the four central elements of secondary structure are established in the transition state. The elements of secondary structure that are less tightly associated with the central core are less well packed in both cases. Topology appears to be the dominant factor in determining the pathway of folding in all Greek key domains.     

Chemical reduction of the diferric/radical center in protein R2 from mouse ribonucleotide reductase is independent of the proposed radical transfer pathway

The rates of reduction of the diferric/radical center in mouse ribonucleotide reductase protein R2 were studied by light absorption and EPR in the native protein and in three point mutants of conserved residues involved in the proposed radical transfer pathway (D266A, W103Y) or in the unstructured C terminal domain (Y370W). The pseudo-first order rate constants for chemical reduction of the tyrosyl radical and diferric center by hydroxyurea, sodium dithionite or the dihydro form of flavin adenine dinucleotide, were comparable with or higher (particularly D266A, by dithionite) than in native R2. Molecular modeling of the D266A mutant showed that the iron/radical site should be more accessible for external reductants in the mutant than in native R2. The results indicate that no specific pathway is required for the reduction. The dihydro form of flavin adenine dinucleotide was found to be a very efficient reductant in the studied proteins compared to dithionite alone. The EPR spectra of the mixed-valent Fe(II)Fe(III) sites formed by chemical reduction in the D266A and W103Y mutants were clearly different from the spectrum observed in the native protein, indicating that the structure of the diferric site was affected by the mutations, as also suggested by the modeling study. No difference was observed between the mixed-valent EPR spectra generated by chemical reduction in Y370W mutant and native mouse R2 protein.     

Tests of search image and learning in the wild: Insights from sexual conflict in damselflies

Search image formation, a proximal mechanism to maintain genetic polymorphisms by negative frequency‐dependent selection, has rarely been tested under natural conditions. Females of many nonterritorial damselflies resemble either conspecific males or background vegetation. Mate‐searching males are assumed to form search images of the majority female type, sexually harassing it at rates higher than expected from its frequency, thus selectively favoring the less common morph. We tested this and how morph coloration and behavior influenced male perception and intersexual encounters by following marked Ischnura elegans and noting their reactions to conspecifics. Contrary to search image formation and associative learning hypotheses, although males encountered the minority, male‐like morph more often, sexual harassment and clutch size were similar for both morphs. Prior mating attempts or copula with morphs did not affect a male''s subsequent reaction to them; males rarely attempted matings with immature females or males. Females mated early in the day, reducing the opportunity for males to learn their identity beforehand. Once encountered, the male‐like morph was more readily noticed by males than the alternative morph, which once noticed was more likely to receive mating attempts. Flexible behavior gave morphs considerable control over their apparency to males, influencing intersexual encounters. Results suggested a more subtle proximal mechanism than male learning maintains these color polymorphisms and call for inferences of learning to be validated by behavior of wild receivers and their signalers.     

Cardiac neural crest is necessary for normal addition of the myocardium to the arterial pole from the secondary heart field

In cardiac neural-crest-ablated embryos, the secondary heart field fails to add myocardial cells to the outflow tract and elongation of the tube is deficient. Since that study, we have shown that the secondary heart field provides both myocardium and smooth muscle to the arterial pole. The present study was undertaken to determine whether addition of both cell types is disrupted after neural crest ablation. Marking experiments confirm that the myocardial component fails to be added to the outflow tract after neural crest ablation. The cells destined to go into the outflow myocardium fail to migrate and are left at the junction of the outflow myocardium with the nascent smooth muscle at the base of the arterial pole. In contrast, the vascular smooth muscle component is added to the arterial pole normally after neural crest ablation. When the myocardium is not added to the outflow tract, the point where the outflow joins the pharynx does not move caudally as it normally should, the aortic sac is smaller and fails to elongate resulting in abnormal connections of the outflow tract with the caudal aortic arch arteries.     

Projection from the ventral bed nucleus of the stria terminalis to the retrorubral field in rats and the effects of cells in these areas on mating in male rats versus gerbils

This research identified the rat counterpart of the lateral cell group of the sexually dimorphic area (SDA) found in medial preoptic area (MPOA) gerbil of gerbils. The lateral SDA (lSDA) is critical for mating in male gerbils and contains most of the SDA cells projecting to the retrorubral field (RRF), a projection that is also important for mating. Therefore, to locate the counterpart of the lateral SDA, we traced the inputs to the rat RRF, which were dense in the ventral part of the bed nucleus of the stria terminalis (BST). To determine if the ventral BST or its projection to the RRF affects mating in male rats, we disrupted them bilaterally by placing cell-body lesions bilaterally in the ventral BST or unilaterally there and in the contralateral RRF. We also studied the effects of RRF lesions in both rats and gerbils. Bilateral ventral BST lesions, which left the medial preoptic nucleus intact, produced persistent and severe mating deficits. Disconnecting the ventral BST from the RRF also had long-lasting, but less severe, consequences. RRF lesions produced only temporary mating deficits in rats, but virtually eliminated mating in gerbils. The recovery of mating in rats after RRF, but not ventral BST, lesions, and the intermediate effects of disconnecting these areas from each other suggest that the ventral BST may contain mating-related projection neurons other than those projecting to the RRF or that its RRF-projecting cells send collaterals to another site. In either case, the pedunculopontine tegmental nucleus or raphe nuclei may be involved.