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Optical imaging of the spontaneous neuronal activities in the male rat major pelvic ganglion following denervation of the pelvic nerve.

Measurement of groups of neuronal activities following pelvic nerve transection to the major pelvic ganglion (MPG) of the rat was performed using voltage-sensitive dye (RH795) and an optical recording system. In control MPG, averaged neuron diameters were 32.0 +/- 0.6 x 22.6 +/- 0.4 microm. Application of KCl (10-50 mM) to the ganglia exhibited excitation which increased in a dose-dependent fashion. Fluctuating membrane potentials were not observed in control ganglion neurons. After the denervation of pelvic nerve chronically (2-6 weeks), the spontaneous neuronal activities were recorded in 91% of the experiments (n = 32). The activity was occurring somewhat periodically (2-15 s). Averaged neuron diameters were 41.3 +/- 1.3 x 24.7 +/- 0.9 microm in denervated MPG which is significantly larger than control. Since average neuron size increased 4 weeks after the denervation, the new excitatory activities could have influenced the change of the neuron size. The new activities might produce contraction of target organs in the pelvic viscera.

Animals↗

Optical imaging of spiral waves: pharmacological modification of spiral-type excitations in a 2-dimensional layer of ventricular myocardium.

Differential effects of sodium channel blockers, an I(Kr) blocker (nifekalant) and amiodarone on the spiral-type reentry, were investigated in rabbit hearts by using a high-resolution optical mapping system. Two-dimensional subepicardial layer of left ventricular myocardium with uniform anisotropy was prepared by endocardial cryoablation. During ventricular tachycardia (VT) elicited by cross-field stimulation, spiral-type excitations rotating around functional block lines (FBLs) were visualized. All the sodium channel blockers stabilized rotors; VT duration was prolonged in association with increases of FBLs and VT cycle length. The rotors in the presence of nifekalant were characterized by large meandering, long FBLs, and frequent front-tail interactions generating wave breaks. Amiodarone (acute application) increased FBLs and VT cycle length, but shortened the VT duration with minimal front-tail interaction. These results suggest that multifaceted drug action on both depolarization and repolarization may be required for the early termination of spiral-type reentry without causing breakup of rotors.

Amiodarone↗

Optical imaging reveals cation--Cl(-) cotransporter-mediated transient rapid decrease in intracellular Cl(-) concentration induced by oxygen--glucose deprivation in rat neocortical slices.

In brain slices from young (postnatal day (P) 10--15) rat somatosensory cortex, real-time neuronal intracellular Cl(-) concentration ([Cl(-)](i)) recordings were made by an optical technique measuring 6-methoxy-N-ethlquinolinium iodide (MEQ) fluorescence. Oxygen--glucose deprivation (in vitro model of ischemia) induced a long-lasting [Cl(-)](i) increase preceded by a rapid, transient [Cl(-)](i) decrease that could not be inhibited by blockers of Cl(-) pumps, Cl(-) channels, or Cl(-) antiporters, but was sensitive to cation-Cl(-) cotransporter inhibitors (bumetanide and furosemide). Use of low external Na(+) or high external K(+) revealed that the Na(+),K(+)-2Cl(-) cotransporter was inhibited by bumetanide and furosemide, whereas the K(+)-Cl(-) cotransporter was preferentially inhibited by furosemide under our experimental conditions. With a reduced inward driving force for Na(+) (reducing Na(+),K(+)-2Cl(-) cotransport), the transient [Cl(-)](i) decrease was only rarely induced by oxygen-glucose deprivation. In contrast, with a reduced outward driving force for K(+) (reducing K(+)-Cl(-) cotransport), the transient [Cl(-)](i) decrease still occurred. These results suggest that the transient [Cl(-)](i) decrease was primarily mediated by a rapid inhibition of the inwardly directed Na(+),K(+)-2Cl(-) cotransporter. Reverse transcriptase-polymerase chain reaction (RT-PCR) experiments suggested that the isoform involved is NKCC1. We hypothesize that the initial rapid Cl(-) efflux might effectively delay the irreversible Cl(-) influx that mediates neuronal injury.

Animals↗

Sex differences in the vocal motor pathway of the zebra finch revealed by real-time optical imaging technique.

Male zebra finches sing, whereas female zebra finches do not. To elucidate the neural mechanisms underlying sexual dimorphism in song behavior, the spatio-temporal properties of neural activity in the vocal motor pathway of the zebra finch were examined in sliced brain preparations using a real-time optical recording technique. Electrical stimulation to higher vocal center (HVC) fibers induced within 20 ms neural activities in the robust nucleus of the archistriatum (RA) of both male and female finches, although the amplitude was smaller and the latency was greater in females than in males. Bicuculline, a GABA(A) receptor antagonist, induced a robust activity in female RA, but had little effect in males. This suggests that neural connections from HVC to RA in female zebra finches are inhibited by GABAergic inputs. The results provide first evidence that an inhibitory neurotransmitter is involved in the sex difference in the motor vocal pathway of song birds.

Animals↗

Emerging patterns of cardiac conduction in the chick embryo: waveform analysis with photodiode array-based optical imaging.

Major difficulties investigating the developing cardiac conduction system stem from that the embryonic heart is extremely small (< 2 mm) and cardiac activation is relatively rapid (< 8 msec). The objective of this study was to investigate the electrophysiology of the embryonic chick cardiac conduction system at periseptation stages with a photodiode array-based detection method of optical mapping capable of high spatial and temporal resolution. Previous work indicated that, in chicken embryos, a switch occurs in ventricular activation pattern from immature base-to-apex to mature apex-to-base pattern at the time of ventricular septation. It was our aim to map activation in more detail to identify the active pathway or pathways of atrioventricular conduction at these particular stages. Analysis of preseptated hearts (n = 10) showed that the latest atrial activation took place just above the site of the earliest ventricular activation at the ventral left ventricular base. Analysis of postseptated hearts (n = 11) showed apex-to-base conduction consistent with activation through the maturing His-Purkinje system. Evaluation of hearts during septation revealed a gradual transition of ventricular activation patterns rather than an abrupt "switch." External pacing of preseptated hearts revealed significant slowing of interventricular conduction compared with spontaneous beats (spontaneous, 61.7 cm/sec +/- 9 cm/sec vs. paced, 36.5 cm/sec +/- 10 cm/sec). The more detailed mapping revealed that, before septation, the pattern of activation of the ventricular myocardium is consistent with direct atrial-ventricular myocardial connections at the left lateral atrioventricular junction; however, functional evidence for a preferential conduction pathway within the ventricles was present before septation.

Animals↗

Optical imaging of apoptosis as a biomarker of tumor response to chemotherapy.

A rapid and accurate assessment of the antitumor efficacy of new therapeutic drugs could speed up drug discovery and improve clinical decision making. Based on the hypothesis that most effective antitumor agents induce apoptosis, we developed a near-infrared fluorescent (NIRF) annexin V to be used for optical sensing of tumor environments. To demonstrate probe specificity, we developed both an active (i.e., apoptosis-recognizing) and an inactive form of annexin V with very similar properties (to account for nonspecific tumor accumulation), and tested the agents in nude mice each bearing a cyclophosphamide (CPA) chemosensitive (LLC) and a chemoresistant LLC (CR-LLC). After injection with active annexin V, the tumor-annexin V ratio (TAR; tumor NIRF/background NIRF) for untreated mice was 1.22+/-0.34 for LLC and 1.43+/-0.53 for CR-LLC (n=4). The LLC of CPA-treated mice had significant elevations of TAR (2.56+/-0.29, P=.001, n=4), but only a moderate increase was obtained for the CR-LLC (TAR=1.89+/-0.19, P=.183). The in vivo measurements correlated well with terminal deoxyribosyl transferase-mediated dUTP nick end labeling indexes. When inactive Cy-annexin V was used, with or without CPA treatment and in both CCL and CR-CCL tumors, tumor NIRF values ranged from 0.91 to 1.17 (i.e., tumor were equal to background). We conclude that active Cy-annexin V and surface reflectance fluorescence imaging provide a nonradioactive, semiquantitative method of determining chemosensitivity in LLC xenografts. The method maybe used to image pharmacologic responses in other animal models and, potentially, may permit the clinical imaging of apoptosis with noninvasive or minimally invasive instrumentation.

Animals↗

Verbal creativity and schizotypal personality in relation to prefrontal hemispheric laterality: a behavioral and near-infrared optical imaging study.

Although anecdotal and correlational results have suggested a reliable relationship between creativity and psychosis, few studies have examined this relationship using empirical methods. In addition, little is known about the neural substrates of creative thinking. We investigated the creative thinking process in relation to schizotypal personality, schizophrenia and prefrontal hemispheric laterality using behavioral and near-infrared optical spectroscopy (NIRS) methods. Schizophrenic, psychometrically ascertained schizotypal, and healthy control subjects (all right-handed) participated in a novel "alternate uses" task designed to assess divergent thinking (DT) ability. The DT task required subjects to generate "uses" for conventional and ambiguous objects. Prefrontal activity was measured using NIRS while subjects were engaged in DT vs. a cognitive control task in a subset of the subjects. Behavioral data indicated that schizotypes had enhanced DT ability compared with schizophrenic and control subjects, who showed similar performance overall. NIRS data showed that DT was associated with bilateral prefrontal cortex (PFC) activation, but the right PFC particularly contributed to the enhanced creative thinking in psychometric schizotypes compared with the other two groups. Thus, creative thinking seems to robustly recruit bilateral PFC, but it is the right PFC that is preferentially activated in schizotypes in relation to their enhanced DT.

Adult↗

Optical imaging of turbid media using independent component analysis: theory and simulation.

A new imaging approach for 3-D localization and characterization of objects in a turbid medium using independent component analysis (ICA) from information theory is developed and demonstrated using simulated data. This approach uses a multisource and multidetector signal acquisition scheme. ICA of the perturbations in the spatial intensity distribution measured on the medium boundary sorts out the embedded objects. The locations and optical characteristics of the embedded objects are obtained from a Green's function analysis based on any appropriate model for light propagation in the background medium. This approach is shown to locate and characterize absorptive and scattering inhomogeneities within highly scattering medium to a high degree of accuracy. In particular, we show this approach can discriminate between absorptive and scattering inhomogeneities, and can locate and characterize complex inhomogeneities, which are both absorptive and scattering. The influence of noise and uncertainty in background absorption or scattering on the performance of this approach is investigated.

Biopolymers↗

Separation of voltage- and ligand-gated calcium influx in locust neurons by optical imaging.

Calcium ions can enter neurons through either ionotropic transmitter receptors or through voltage-gated calcium channels. Thus, an observed rise in intracellular calcium concentration upon synaptic stimulation can be due to either one of these mechanisms or to both of them. We analyzed the individual contribution of transmitter- and voltage-gated calcium entry in non-spiking somata, acutely dissociated from thoracic ganglia of the locust Locusta migratoria. By optically recording the calcium signal following different stimulation protocols, we isolated the voltage- and the transmitter-gated component and found that these components indeed summate to the total rise in calcium observed under control conditions.

Action Potentials↗

Intrinsic optical imaging reveals regionally different manifestation of spreading depression in hippocampal and entorhinal structures in vitro.

The spatiotemporal features of spreading depression (SD) were analyzed in vitro by using combined hippocampal-entorhinal cortex slices. SDs were induced by microinjection of 1 M KCl in the stratum radiatum of the CA1 region of the hippocampus. Measurements of extracellular field potentials, extracellular space (ECS) volume changes and intrinsic optical signal changes were combined to study SD features in different regions of the slice. Each SD was associated with a pronounced shrinkage of the extracellular space (ECS) volume and a decrease in light transmittance. The beginning of the optical signal change occurred simultaneously with the electrographic onset as measured with extracellular microelectrodes but outlasted the dc shift for tens of seconds. The amplitude of the intrinsic optical signal change displayed marked regional variations with greatest changes of 12% in cortical regions. The signal amplitudes were considerably lower in hippocampal regions. The analysis of spread patterns revealed two types of waves: fully propagated waves spreading from CA1 all the way to the temporal neocortex and abortive waves that ceased earlier. The spread velocities displayed pronounced regional differences with highest velocities of 5.4 +/- 0.3 mm/min in the area CA3 of the hippocampal formation and lowest velocities of 2.7 +/- 0.1 mm/min in cortical regions.

Animals↗

Chronic administration of dehydroepiandrosterone sulfate (DHEAS) primes for facilitated induction of long-term potentiation via sigma 1 (sigma1) receptor: optical imaging study in rat hippocampal slices.

Dehydroepiandrosterone sulfate (DHEAS), one of the most abundant neurosteroids synthesized de novo in the nervous system, has well characterized effects on memory and cognitive performances. However, little is known about the underlying synaptic mechanisms. In this study, we investigated the effects of chronic administration of DHEAS (20 mg/kg for 7 days) on the plasticity of Schaffer collateral-CA1 synapses by applying an optical recording technique on the hippocampal slices stained with voltage-sensitive dyes. We report here that chronically administered DHEAS significantly facilitated the induction of frequency-dependent LTP, termed DHEAS-facilitated LTP. While tetanus of at least 50 pulses (at 100 Hz) were required to induce LTP in control rats, only 20 pulses were needed in DHEAS-treated animals. In contrast DHEA, the non-sulfated form of DHEAS, had no facilitating effect on the induction of LTP. We found that chronically administered DHEAS did not alter the presynaptic glutamate release in response to both single pulse and tetanic stimulation, suggesting that certain alterations happened in postsynaptic neurons. Co-administration of the sigma 1 (sigma1) receptor antagonists, haloperidol or NE100, with DHEAS completely inhibited the DHEAS-facilitated LTP. However, acute administration of sigma1 receptor antagonists to the slices did not affect the induction of DHEAS-facilitated LTP, suggesting that sigma1 receptor is a key target of chronic actions of DHEAS but is not involved in the induction of DHEAS-facilitated LTP. Our findings provide evidence that chronically administered DHEAS plays a priming role in inducing a facilitated synaptic plasticity probably via a chronic activation of sigma1 receptor in rat hippocampal CA1 pyramidal cells.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Optical imaging of hydroxyapatite in the calcified vasculature of transgenic animals.

OBJECTIVE: To detect the hydroxyapatite component of vascular calcification in vivo so that the process of calcium deposition can be studied in transgenic model systems. METHODS AND RESULTS: We have previously developed a near-infrared fluorescent bisphosphonate derivative that binds with high affinity and specificity to hydroxyapatite, and an intraoperative near-infrared fluorescence imaging system for small animals. Using these tools, and a transgenic mouse strain with homozygous deletion of the matrix GLA protein (Mgp(-/-)), we demonstrate that the hydroxyapatite component of vascular calcification can be detected in vivo with high sensitivity, specificity, and resolution. CONCLUSIONS: The hydroxyapatite component of vascular calcification can be detected optically, in real-time, without sacrifice of the animal. It is now possible to study the earliest events associated with vascular mineralization, at the cell and organ level, and to monitor the process in living animals.

Animals↗

Glial nitric oxide-mediated long-term presynaptic facilitation revealed by optical imaging in rat spinal dorsal horn.

We investigated a presynaptic form of long-term potentiation (LTP) in horizontal slices of the rat spinal cord by visualizing presynaptic and postsynaptic excitation with a voltage-sensitive dye. To record presynaptic excitation, we stained primary afferent fibers anterogradely from the dorsal root. A single-pulse test stimulation of C fiber-activating strength to the dorsal root elicited action potential (AP)-like or compound AP-like optical signals throughout the superficial dorsal horn. After conditioning (240 pulses at 2 Hz for 2 min), the presynaptic excitation was augmented. Furthermore, new excitation was elicited in the areas that were silent before conditioning. For postsynaptic recording, projection neurons in spinal lamina I were stained retrogradely from the periaqueductal gray in the brain stem. The test stimulation elicited AP-like or EPSP-like optical signals in the stained neurons. After conditioning, the EPSP-like responses were augmented, and previously silent neurons were converted to active ones. Results obtained with a nitric oxide (NO) donor, NO synthase inhibitors, metabotropic glutamate receptor (mGluR) agonist and mGluR1 antagonist, and a glial metabolism inhibitor suggest that after conditioning, presynaptic excitation is facilitated by NO released from glial cells via the activation of mGluR1. The results also indicate the possible presence of additional presynaptic and postsynaptic mechanism(s) for the LTP induction. Activity-dependent LTP of nociceptive afferent synaptic transmission in the spinal cord is believed to underlie central sensitization after inflammation or nerve injury. This glial NO-mediated control of presynaptic excitation may contribute to the induction at least in part.

Action Potentials↗

Near-infrared-emissive polymersomes: self-assembled soft matter for in vivo optical imaging.

We demonstrate that synthetic soft materials can extend the utility of natural vesicles, from predominantly hydrophilic reservoirs to functional colloidal carriers that facilitate the biomedical application of large aqueous-insoluble compounds. Near-infrared (NIR)-emissive polymersomes (50-nm- to 50-microm-diameter polymer vesicles) were generated through cooperative self assembly of amphiphilic diblock copolymers and conjugated multi(porphyrin)-based NIR fluorophores (NIRFs). When compared with natural vesicles comprised of phospholipids, polymersomes were uniquely capable of incorporating and uniformly distributing numerous large hydrophobic NIRFs exclusively in their lamellar membranes. Within these sequestered compartments, long polymer chains regulate the mean fluorophore-fluorophore interspatial separation as well as the fluorophore-localized electronic environment. Porphyrin-based NIRFs manifest photophysical properties within the polymersomal matrix akin to those established for these high-emission dipole strength fluorophores in organic solvents, thereby yielding uniquely emissive vesicles. Furthermore, the total fluorescence emanating from the assemblies gives rise to a localized optical signal of sufficient intensity to penetrate through the dense tumor tissue of a live animal. Robust NIR-emissive polymersomes thus define a soft matter platform with exceptional potential to facilitate deep-tissue fluorescence-based imaging for in vivo diagnostic and drug-delivery applications.

Animals↗

Health information department reorganization resulting from the implementation of optical imaging.

Our experience clearly demonstrates that transitional periods which include significant change can be very difficult. Nonetheless, we have described our organizational issues not to discourage, but to educate our readers. We are confident that the staffing changes, departmental reorganization, and other changes generated by the imaging system will have a positive impact on everyone. The institution will enjoy improved customer service and enhanced patient care.

Hospital Bed Capacity, 500 and over↗

Detection of single and multiple targets in tissue phantoms with fluorescence-enhanced optical imaging: feasibility study.

PURPOSE: To evaluate the feasibility of reconstructing single and multiple targets by using fluorescence-enhanced tomography of a breast tissue phantom. MATERIALS AND METHODS: Frequency-domain fluorescence measurements were performed in cup-shaped tissue phantoms of clinically relevant size (diameter, 10 cm) in which single or multiple 0.5-1.0-cm(3) targets that contained micromolar concentrations of indocyanine green with 1:0 and 100:1 target-to-background (T:B) contrast ratios had been embedded. Rapid acquisition of time-dependent fluorescent light measurements was performed at the phantom surface in response to point illumination of excitation light by using a gain-modulated intensified charge-coupled device detection system. Boundary surface measurements were used to tomographically reconstruct the interior targets located with various experimental conditions. RESULTS: Single 1.0-cm(3) targets located between 1.43 and 2.82 cm deep from the phantom surface at a T:B contrast ratio of 100:1 and three approximately 0.55-cm(3) targets located about 1.30 cm deep at a T:B contrast ratio of 1:0 were reconstructed with minimal or no artifacts by using boundary surface fluorescence measurements and an approximate extended Kalman filter algorithm. CONCLUSION: It is feasible to detect single or multiple fluorescent targets in tissue phantoms of clinically relevant size by using fluorescence-enhanced optical tomography.

Algorithms↗