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Coupling of the cortical hemodynamic response to cortical and thalamic neuronal activity.

Accurate interpretation of functional MRI (fMRI) signals requires knowledge of the relationship between the hemodynamic response and the neuronal activity that underlies it. Here we address the question of coupling between pre- and postsynaptic neuronal activity and the hemodynamic response in rodent somatosensory (Barrel) cortex in response to single-whisker deflection. Using full-field multiwavelength optical imaging of hemoglobin oxygenation and electrophysiological recordings of spiking activity and local field potentials, we demonstrate that a point hemodynamic measure is influenced by neuronal activity across multiple cortical columns. We demonstrate that the hemodynamic response is a spatiotemporal convolution of the neuronal activation. Therefore, positive hemodynamic response in one cortical column might be explained by neuronal activity not only in that column but also in the neighboring columns. Thus, attempts at characterizing the neurovascular relationship based on point measurements of electrophysiology and hemodynamics may yield inconsistent results, depending on the spatial extent of neuronal activation. The finding that the hemodynamic signal observed at a given location is a function of electrophysiological activity over a broad spatial region helps explain a previously observed increase of local vascular response beyond the saturation of local neuronal activity. We also demonstrate that the oxy- and total-hemoglobin hemodynamic responses can be well approximated by space-time separable functions with an antagonistic center-surround spatial pattern extending over several millimeters. The surround "negative" hemodynamic activity did not correspond to observable changes in neuronal activity. The complex spatial integration of the hemodynamic response should be considered when interpreting fMRI data.

Animals↗

Downregulation of connexin 45 gene products during mouse heart development.

The electrical activity in heart is generated in the sinoatrial node and then propagates to the atrial and ventricular tissues. The gap junction channels that couple the myocytes are responsible for this propagation process. The gap junction channels are dodecamers of transmembrane proteins of the connexin (Cx) family. Three members of this family have been demonstrated to be synthesized in the cardiomyocytes: Cx40, Cx43, and Cx45. In addition, each of them has been shown to form channels with unique and specific electrophysiological properties. Understanding the conduction phenomenon requires detailed knowledge of the spatiotemporal expression pattern of these Cxs in heart. The expression patterns of Cx40 and Cx43 have been previously described in the adult heart and during its development. Here we report the expression of Cx45 gene products in mouse heart from the stage of the first contractions (8.5 days postcoitum [dpc]) to the adult stage. The Cx45 gene transcript was demonstrated by reverse transcriptase-polymerase chain reaction experiments to be present in heart at all stages investigated. Between 8.5 and 10.5 dpc it was shown by in situ hybridization to be expressed in low amounts in all cardiac compartments (including the inflow and outflow tracts and the atrioventricular canal) and then to be downregulated from 11 to 12 dpc onward. At subsequent fetal stages, the transcript was weakly detected in the ventricles, with the most distinct expression in the outflow tract. Cx45 protein was demonstrated by immunofluorescence microscopy to be expressed in the myocytes of young embryonic hearts (8.5 to 9.5 dpc). However, beyond 10.5 dpc the protein was no longer detected with this technique in the embryonic, fetal, or neonatal working myocardium, although it could be shown by immunoblotting that the protein was still synthesized in neonatal heart. In the major part of adult heart, Cx45 was undetectable. It was, however, clearly seen in the anterior regions of the interventricular septum and in trace amounts in some small foci dispersed in the ventricular free walls. Cx45 gene is the first Cx gene so far demonstrated to be activated in heart at the stage of the first contractions. The coordination of myocytes during the slow peristaltic contractions that occur at this stage would thus appear to be controlled by the Cx45 channels.

Amino Acid Sequence↗

Relative phase destabilization during interlimb coordination: the disruptive role of kinesthetic afferences induced by passive movement.

The disruption of three patterns of two-limb coordination, involving cyclical flexion-extension movements performed in the same or in different directions, was investigated through application of passive movement to a third limb by the experimenter. The three patterns referred to the homologous, homolateral, and heterolateral (diagonal) limb combinations which were performed in the sagittal plane. The passive movement involved a spatiotemporal trajectory that differed from the movements controlled actively. Even though subjects were instructed to completely ignore the passive limb movement, the findings of experiment 1 demonstrated a moderate to severe destabilization of the two-limb patterns, as revealed by analyses of power spectra, relative phase, cycle duration, and amplitude. This disruption was more pronounced in the homolateral and heterolateral than in the homologous effector combinations, suggesting stronger coupling between homologous than nonhomologous limb pairs. Moreover, passive mobilization affected antiphase (nonisodirectional) movements more than inphase (isodirectional) movements, pointing to the differential stability of these patterns. Experiment 2 focused on homolateral coordination and demonstrated that withdrawal of visual information did not alter the effects induced by passive movement. It was therefore hypothesized that the generation of extra kinesthetic afferences through passive limb motion was primarily responsible for the detriment in interlimb coordination, possibly conflicting with the sensory information accompanying active movement production. In addition, it was demonstrated that the active limbs were more affected by their homologous passive counterpart than by their nonhomologous counterpart, favoring the notion of "specific" interference. The findings are discussed in view of the potential role of kinesthetic afferences in human interlimb coordination, more specifically the preservance of relative phasing through a kinesthetic feedback loop.

Adolescent↗

Effect of neuritic cables on conductance estimates for remote electrical synapses.

The conductance of electrical synapses is usually estimated from voltage recordings at the neuronal somata under the assumption that each cell is isopotential. This approach neglects effects of intervening neurites. For a cell pair with unbranched neurites and an electrical synapse at their ends, we used cable theory to derive an analytical expression that relates the synaptic conductance to voltage recordings at the cell bodies and to the neurite properties. The equation implies that the conventional method significantly underestimates the actual synapse conductance if the neurite length is comparable to the electrotonic length constant and if the synaptic conductance is similar to the serial neurite conductance. For an experimental test, we cultured pairs of snail neurons on protein patterns, resulting in a geometry that matched the theoretical model. Using the isopotential theory, we estimated the synapse conductances and found them to be rather weak. To obtain the cable properties, we recorded spatiotemporal maps of signal propagation in the neurites using a voltage-sensitive dye. Fits of these maps to a passive cable model showed that the snail neurons are electrotonically rather compact. Given these features of our experimental system, the synaptic conductances derived with the nonisopotential model deviated from the estimates of the isopotential theory by about 13%. This discrepancy, although small, shows that even in electrotonically compact neurons coupled by weak synapses the impact of the neuritic cables on conductance estimates cannot be neglected. When applied to less compact and more strongly coupled cell pairs in vivo, our approach can supply the realistic estimates of synaptic conductances that are necessary for a better understanding of the role of electrical coupling in neural systems.

Animals↗

Synaptic delay learning in pulse-coupled neurons.

We present rules for the unsupervised learning of coincidence between excitatory postsynaptic potentials (EPSPs) by the adjustment of postsynaptic delays between the transmitter binding and the opening of ion channels. Starting from a gradient descent scheme, we develop a robust and more biological threshold rule by which EPSPs from different synapses can be gradually pulled into coincidence. The synaptic delay changes are determined from the summed potential--at the site where the coincidence is to be established--and from postulated synaptic learning functions that accompany the individual EPSPs. According to our scheme, templates for the detection of spatiotemporal patterns of synaptic activation can be learned, which is demonstrated by computer simulation. Finally, we discuss possible relations to biological mechanisms.

Excitatory Postsynaptic Potentials↗

Expression of genes (CAPN3, SGCA, SGCB, and TTN) involved in progressive muscular dystrophies during early human development.

The developmental expression pattern of four human genes, three of which are involved in progressive muscular dystrophies, was investigated. The rationale for these experiments is that these patterns might provide useful information on the pathophysiology underlying these myopathies. Despite the presence of overlapping clinical signs, the spatiotemporal expression profiles of the corresponding genes differed widely. Transcripts of alpha-sarcoglycan (SGCA) were visible as soon as myotomes were formed, and constitute, together with titin transcripts, precocious muscular system landmarks. beta-sarcoglycan (SGCB) was initially transcribed in a ubiquitous manner, and, toward the second part of the embryonic period, became specific to striated muscle, heart, and the central nervous system. Whereas titin (TTN) transcription and translation seem to be coupled, for the sarcoglycans, translation seemed restricted to skeletal muscle. Calpain3 (CAPN3) RNA was found in only skeletal muscles during the fetal period. It was, however, present earlier in the whole heart, where it selectively disappeared. Finally, evidence for differentially spliced calpain3 variants in smooth muscles was also seen. The expression profiles of these genes is suggestive of their having a role during myogenesis, knowledge of which could be pertinent to the understanding of the pathophysiology of the associated diseases.

Adult↗

Particulate adenylate cyclase plays a key role in human sperm olfactory receptor-mediated chemotaxis.

Human sperm chemotaxis is a critical component of the fertilization process, but the molecular basis for this behavior remains unclear. Recent evidence shows that chemotactic responses depend on activation of the sperm olfactory receptor, hOR17-4. Certain floral scents, including bourgeonal, activate hOR17-4, trigger pronounced Ca(2+) fluxes, and evoke chemotaxis. Here, we provide evidence that hOR17-4 activation is coupled to a cAMP-mediated signaling cascade. Multidimensional protein identification technology was used to identify potential components of a G-protein-coupled cAMP transduction pathway in human sperm. These products included various membrane-associated adenylate cyclase (mAC) isoforms and the G(olf)-subunit. Using immunocytochemistry, specific mAC isoforms were localized to particular cell regions. Whereas mAC III occurred in the sperm head and midpiece, mAC VIII was distributed predominantly in the flagellum. In contrast, G(olf) was found mostly in the flagellum and midpiece. The observed spatial distribution patterns largely correspond to the spatiotemporal character of hOR17-4-induced Ca(2+) changes. Behavioral and Ca(2+) signaling responses of human sperm to bourgeonal were bioassayed in the presence, or absence, of the adenylate cyclase antagonist SQ22536. This specific agent inhibits particulate AC, but not soluble AC, activation. Upon incubation with SQ22536, cells ceased to exhibit Ca(2+) signaling, chemotaxis, and hyperactivation (faster swim speed and flagellar beat rate) in response to bourgeonal. Particulate AC is therefore required for induction of hOR17-4-mediated human sperm behavior and represents a promising target for future design of contraceptive drugs.

Adenine↗

Visualizing signals moving in cells.

Cells display a highly complex spatiotemporal organization, required to exert a wide variety of different functions, for example, detection, processing, and propagation of nerve impulses by neurons; contraction and relaxation by muscle cells; movement by leukocytes; and adsorption and secretion of nutrients and metabolites by epithelial cells lining the gut. Successful execution of these complex processes requires highly dynamic information transfer between different regions and compartments within cells. Through the development of fluorescent sensors for intracellular signaling molecules coupled with improved microscopic imaging techniques, it has now become possible to investigate signal propagation in cells with high spatial and temporal resolution.

Animals↗

Identification and characterization of G protein-coupled receptors in the nocturnal halictid bee Megalopta genalis.

G protein-coupled receptors (GPCRs) are one of the largest families of membrane proteins in insects, regulating vision, neural signal transduction, and various physiological behaviors. Megalopta genalis exhibits a unique facultatively eusocial lifestyle and possesses adaptations for nocturnal activity; however, its GPCR family has not yet been systematically characterized. In this study, we performed genome-wide identification, phylogenetic analysis, and expression profiling of GPCRs in M. genalis by integrating genomic annotation and transcriptomic analysis. The results showed that a total of 99 GPCRs were identified in the genome of M. genalis, which were classified into four major families. Here, we show that M. genalis has undergone lineage-specific GPCR repertoire remodeling, marked by the expansion of novel orphan receptors and the systematic loss of multiple receptor subtypes, such as the neuropeptide receptors MIP-R and NPFR. Moreover, opsins have formed a diverse array of combinations and non-GPCR odorant receptors have undergone significant expansion via tandem duplication. Together, these features may represent part of the molecular repertoire associated with the adaptation of M. genalis to a nocturnal lifestyle. Furthermore, transcriptomic analysis revealed distinct spatiotemporal expression divergence within each of the Mth/Mthl and Fz GPCR families, suggesting functional specialization across development and adult tissues. This study provides the first systematic identification and initial functional characterization of GPCRs in M. genalis, revealing an evolutionary pattern characterized by the coexistence of contraction and expansion within the GPCR family. These findings lay a foundation for further studies aimed at elucidating the roles of these GPCRs in regulating M. genalis physiology and behavior.

Animals↗

Dynamics of yielding observed in a three-dimensional aqueous dry foam.

We study the onset of yielding in stable three-dimensional dry foams following the start up of steady shear flow. By means of a charge-coupled device camera equipped with a small depth-of-field objective, we visualize the Plateau border network in the bulk of the foam. The onset of yielding is identified with the deformation gamma(c) for which shear induced rearrangements start occurring. We show that gamma(c) is independent of shear rate gamma; in a quasistatic regime whereas at high strain rates, a rapid increase of gamma(c) with gamma; is observed, in qualitative agreement with theoretical models. Moreover, spatiotemporal image analyses are used to determine the velocity profile in the gap. We find that this profile remains linear up to strains far beyond gamma(c). Moreover, we have studied the strain history dependence of gamma(c).

Journal Article↗

Spatiotemporal coordination of rod and cone photoreceptor differentiation in goldfish retina.

In this study, we have compared spatial and temporal aspects of development of new rods and cones in the adult goldfish by using a combination of bromodeoxyuridine immunocytochemistry and opsin in situ hybridization to determine the intervals between terminal mitosis (cell "birth") and expression of opsin mRNA for each photoreceptor cell type. The goldfish opsins include rod opsin and four different cone opsins: red, green, blue, and ultraviolet. In a cohort of photoreceptors born at the same time, rods expressed opsin mRNA within 3 days of cell birth, while expression of cone opsin mRNA required at least 7 days. This temporal discrepancy in differentiation, coupled with a discordance in the site of cell genesis of rods and cones, allowed opsin expression to commence in both cell types in approximately the same retinal location. Commitment to the generic cone phenotype occurred within approximately 6 days throughout the cone cohort, as indicated by expression of interphotoreceptor retinoid-binding protein (IRBP) mRNA, but expression of a specific spectral phenotype was delayed until rods differentiated nearby. Onset of expression of cone opsin mRNA followed a phenotype-specific sequence: red, then green, then blue, and finally ultraviolet; in situ hybridization with two opsin probes confirmed that individual photoreceptors expressed only one type of opsin as they differentiated. This stepwise process of cone differentiation is consistent with the hypothesis that cell-cell interactions among developing photoreceptors may coordinate selection of specific photoreceptor phenotypes.

Animals↗

Dynamics of signaling between Ca(2+) sparks and Ca(2+)- activated K(+) channels studied with a novel image-based method for direct intracellular measurement of ryanodine receptor Ca(2+) current.

Ca(2+) sparks are highly localized cytosolic Ca(2+) transients caused by a release of Ca(2+) from the sarcoplasmic reticulum via ryanodine receptors (RyRs); they are the elementary events underlying global changes in Ca(2+) in skeletal and cardiac muscle. In smooth muscle and some neurons, Ca(2+) sparks activate large conductance Ca(2+)-activated K(+) channels (BK channels) in the spark microdomain, causing spontaneous transient outward currents (STOCs) that regulate membrane potential and, hence, voltage-gated channels. Using the fluorescent Ca(2+) indicator fluo-3 and a high speed widefield digital imaging system, it was possible to capture the total increase in fluorescence (i.e., the signal mass) during a spark in smooth muscle cells, which is the first time such a direct approach has been used in any system. The signal mass is proportional to the total quantity of Ca(2+) released into the cytosol, and its rate of rise is proportional to the Ca(2+) current flowing through the RyRs during a spark (I(Ca(spark))). Thus, Ca(2+) currents through RyRs can be monitored inside the cell under physiological conditions. Since the magnitude of I(Ca(spark)) in different sparks varies more than fivefold, Ca(2+) sparks appear to be caused by the concerted opening of a number of RyRs. Sparks with the same underlying Ca(2+) current cause STOCs, whose amplitudes vary more than threefold, a finding that is best explained by variability in coupling ratio (i.e., the ratio of RyRs to BK channels in the spark microdomain). The time course of STOC decay is approximated by a single exponential that is independent of the magnitude of signal mass and has a time constant close to the value of the mean open time of the BK channels, suggesting that STOC decay reflects BK channel kinetics, rather than the time course of [Ca(2+)] decline at the membrane. Computer simulations were carried out to determine the spatiotemporal distribution of the Ca(2+) concentration resulting from the measured range of I(Ca(spark)). At the onset of a spark, the Ca(2+) concentration within 200 nm of the release site reaches a plateau or exceeds the [Ca(2+)](EC50) for the BK channels rapidly in comparison to the rate of rise of STOCs. These findings suggest a model in which the BK channels lie close to the release site and are exposed to a saturating [Ca(2+)] with the rise and fall of the STOCs determined by BK channel kinetics. The mechanism of signaling between RyRs and BK channels may provide a model for Ca(2+) action on a variety of molecular targets within cellular microdomains.

Animals↗

A foot-pointing task and spatiotemporal gait parameters during walking in sportsmen.

Unlike many sports, karate imposes pointing to a target with the lower limb. The nature of each sport could influence gait variables of sportsmen. The aim of this study was to examine the pointing accuracy of limb swing and the spatial and temporal parameters of the walking cycle during walking in karate experts (Group K, n=6) compared to accuracy of nonkarate sportsmen (Group S, n = 11). The second aim of this study was to compare the influence of this skill on the spatial and temporal parameters of the walking cycle between the two groups. The analysis was performed with a device composed of a gait analysis system (locometer) coupled to a lighted walkway. This system provides a measurement of the motor skill and the spatial and temporal gait during walking. Analysis for the pointing task indicated no differences between the Karate and the Sportsmen Groups during walking. Moreover, neither the spatial nor temporal characteristics of walking differed between the two groups. Karate does not therefore seem to develop motor programs specific to enhanced performance on this pointing task during walking or alter the walking cycle as compared to some other competitive sports.

Adult↗

Screening of ion channel receptor agonists using capillary electrophoresis-patch clamp detection with resensitized detector cells.

Efficient techniques for identifying endogenous and synthetic ligands of ion channels are important in understanding neuronal communication and for screening drug libraries. This paper describes a technique based on capillary electrophoresis (CE) separation coupled to patch-clamp (PC) detection where a pulsed-flow superfusion scheme was implemented for improved detection. The nicotinic acetylcholine receptor (nAChr) agonists acetylcholine, carbachol, and (-)-nicotine were fractionated and detected by patch-clamped pheochromcytoma detector cells. The high-conductance state of the nAChr during CE-PC detection was maintained and repetitively resensitized using pulsed-flow superfusion with agonist-free buffer. In this way, each agonist evoked an ensemble of peak currents that reflected the spatiotemporal distribution for the ligand at the cell surface. The technique takes advantage of the intrinsic high selectivity and sensitivity of membrane-expressed receptors and allowed for resolution and identification of closely migrating ligands. The method was employed for determination of acetylcholine content in cell lysates.

Acetylcholine↗

Hierarchical Bayesian inference in the visual cortex.

Traditional views of visual processing suggest that early visual neurons in areas V1 and V2 are static spatiotemporal filters that extract local features from a visual scene. The extracted information is then channeled through a feedforward chain of modules in successively higher visual areas for further analysis. Recent electrophysiological recordings from early visual neurons in awake behaving monkeys reveal that there are many levels of complexity in the information processing of the early visual cortex, as seen in the long-latency responses of its neurons. These new findings suggest that activity in the early visual cortex is tightly coupled and highly interactive with the rest of the visual system. They lead us to propose a new theoretical setting based on the mathematical framework of hierarchical Bayesian inference for reasoning about the visual system. In this framework, the recurrent feedforward/feedback loops in the cortex serve to integrate top-down contextual priors and bottom-up observations so as to implement concurrent probabilistic inference along the visual hierarchy. We suggest that the algorithms of particle filtering and Bayesian-belief propagation might model these interactive cortical computations. We review some recent neurophysiological evidences that support the plausibility of these ideas.

Animals↗

Pattern formation in Taylor-Couette flow of dilute polymer solutions: dynamical simulations and mechanism.

We report spatiotemporal pattern formation in Taylor-Couette flow (i.e., flow between rotating cylinders) of viscoelastic dilute polymer solutions obtained for the first time from first-principles dynamical simulations. Solution structures with varying spatial and temporal symmetries, such as rotating standing waves, flames, disordered oscillations, and solitary vortex solutions which include diwhirls (stationary and axisymmetric) and oscillatory strips (axisymmetric or nonaxisymmetric), are observed, depending on the ratio of fluid relaxation time to the time period of inner cylinder rotation. The flow-microstructure coupling mechanisms underlying the pattern formation process are also discussed.

Journal Article↗

Role of Xrx1 in Xenopus eye and anterior brain development.

The anteriormost part of the neural plate is fated to give rise to the retina and anterior brain regions. In Xenopus, this territory is initially included within the expression domain of the bicoid-class homeobox gene Xotx2 but very soon, at the beginning of neurulation, it becomes devoid of Xotx2 transcripts in spatiotemporal concomitance with the transcriptional activation of the paired-like homeobox gene Xrx1. By use of gain- and loss-of-function approaches, we have studied the role played by Xrx1 in the anterior neural plate and its interactions with other anterior homeobox genes. We find that, at early neurula stage Xrx1 is able to repress Xotx2 expression, thus first defining the retina-diencephalon territory in the anterior neural plate. Overexpression studies indicate that Xrx1 possesses a proliferative activity that is coupled with the specification of anterior fate. Expression of a Xrx1 dominant repressor construct (Xrx1-EnR) results in a severe impairment of eye and anterior brain development. Analysis of several brain markers in early Xrx1-EnR-injected embryos reveals that anterior deletions are preceded by a reduction of anterior gene expression domains in the neural plate. Accordingly, expression of anterior markers is abolished or decreased in animal caps coinjected with the neural inducer chordin and the Xrx1-EnR construct. The lack of expansion of mid-hindbrain markers, and the increase of apoptosis in the anterior neural plate after Xrx1-EnR injection, indicate that anterior deletions result from an early loss of anterior neural plate territories rather than posteriorization of the neuroectoderm. Altogether, these data suggest that Xrx1 plays a role in assigning anterior and proliferative properties to the rostralmost part of the neural plate, thus being required for eye and anterior brain development.

Animals↗

CREB and Elk-1 phosphorylation by metabotropic glutamate receptors in striatal neurons (review).

Metabotropic glutamate receptors (mGluRs) are implicated in the regulation of diverse neuronal plasticity and neuropathological processes in the central nervous system. Activation of mGluRs couples glutamatergic signals to second messengers in a subtype-specific manner: activation of group I mGluRs upregulates Ca2+ cascades, while group II/III downregulates the adenylate cyclase and cAMP cascades. Dominant presynaptic inhibitory actions of group II/III mGluRs on the glutamate release, extensive cross-talk between kinases by various second messengers downstream to the group I mGluRs, and desensitization of mGluRs in response to prolonged stimulation of glutamate input have been documented in the regulation of glutamatergic transmission. In addition to the spatiotemporal processes, interactions with ionotropic glutamate receptors, and protein phosphatase activity against kinase actions further regulate glutamatergic signals. These overall activities in medium spiny neurons contribute to modifying striatal outflow in striatopallidal and striatonigral neurons. Thus, characterization of the roles of mGluRs in the regulation of intracellular effectors is crucial for the understanding of diverse neuronal plasticity implicated with the receptors including long-term potentiation and long-term depression, neurotoxicity, actions of abused drugs, and neurodegenerative diseases. In this review we attempted to provide a broad spectrum on how mGluRs regulate the phosphorylation of cAMP response element-binding protein and Elk-1, well known inducible transcription factors by extracellular stimuli, by emphasizing major kinase interactions in medium spiny neurons.

Animals↗