Acute cocaine induces fast activation of D1 receptor and progressive deactivation of D2 receptor striatal neurons: in vivo optical microprobe [Ca2+]i imaging.

Cocaine induces fast dopamine increases in brain striatal regions, which are recognized to underlie its rewarding effects. Both dopamine D1 and D2 receptors are involved in cocaine's reward but the dynamic downstream consequences of cocaine effects in striatum are not fully understood. Here we used transgenic mice expressing EGFP under the control of either the D1 receptor (D1R) or the D2 receptor (D2R) gene and microprobe optical imaging to assess the dynamic changes in intracellular calcium ([Ca(2+)](i)) responses (used as marker of neuronal activation) to acute cocaine in vivo separately for D1R- versus D2R-expressing neurons in striatum. Acute cocaine (8 mg/kg, i.p.) rapidly increased [Ca(2+)](i) in D1R-expressing neurons (10.6 ± 3.2%) in striatum within 8.3 ± 2.3 min after cocaine administration after which the increases plateaued; these fast [Ca(2+)](i) increases were blocked by pretreatment with a D1R antagonist (SCH23390). In contrast, cocaine induced progressive decreases in [Ca(2+)](i) in D2R-expressing neurons (10.4 ± 5.8%) continuously throughout the 30 min that followed cocaine administration; these slower [Ca(2+)](i) decreases were blocked by pretreatment with a D2R antagonist (raclopride). Since activation of striatal D1R-expressing neurons (direct-pathway) enhances cocaine reward, whereas activation of D2R-expressing neurons suppresses it (indirect-pathway) (Lobo et al., 2010), this suggests that cocaine's rewarding effects entail both its fast stimulation of D1R (resulting in abrupt activation of direct-pathway neurons) and a slower stimulation of D2R (resulting in longer-lasting deactivation of indirect-pathway neurons). We also provide direct in vivo evidence of D2R and D1R interactions in the striatal responses to acute cocaine administration.

Imaging separation of neuronal from vascular effects of cocaine on rat cortical brain in vivo.

MRI techniques to study brain function assume coupling between neuronal activity, metabolism and flow. However, recent evidence of physiological uncoupling between neuronal and cerebrovascular events highlights the need for methods to simultaneously measure these three properties. We report a multimodality optical approach that integrates dual-wavelength laser speckle imaging (measures changes in blood flow, blood volume and hemoglobin oxygenation), digital-frequency-ramping optical coherence tomography (images quantitative 3D vascular network) and Rhod(2) fluorescence (images intracellular calcium for measure of neuronal activity) at high spatiotemporal resolutions (30 µm, 10 Hz) and over a large field of view (3×5 mm(2)). We apply it to assess cocaine's effects in rat cortical brain and show an immediate decrease (3.5±0.9 min, phase 1) in the oxygen content of hemoglobin and the cerebral blood flow followed by an overshoot (7.1±0.2 min, phase 2) lasting over 20 min whereas Ca(2+) increased immediately (peaked at t=4.1±0.4 min) and remained elevated. This enabled us to identify a delay (2.9±0.5 min) between peak neuronal and vascular responses in phase 2. The ability of this multimodality optical approach for simultaneous imaging at high spatiotemporal resolutions permits us to distinguish the vascular versus cellular changes of the brain, thus complimenting other neuroimaging modalities for brain functional studies (e. g., PET, fMRI).

Differential effects of anesthetics on cocaine's pharmacokinetic and pharmacodynamic effects in brain.

Most studies of the effect of cocaine on brain activity in laboratory animals are preformed under anesthesia, which could potentially affect the physiological responses to cocaine. Here we assessed the effects of two commonly used anesthetics [alpha-chloralose (alpha-CHLOR) and isofluorane (ISO)] on the effects of acute cocaine (1 mg/kg i.v.) on cerebral blood flow (CBF), cerebral blood volume (CBV), and tissue hemoglobin oxygenation (S(t)O(2)) using optical techniques and cocaine's pharmacokinetics (PK) and binding in the rat brain using (PET) and [(11)C]cocaine. We showed that acute cocaine at a dose abused by cocaine abusers decreased CBF, CBV and S(t)O(2) in rats anesthetized with ISO, whereas it increased these parameters in rats anesthetized with alpha-CHLOR. Importantly, in ISO-anesthetized animals cocaine-induced changes in CBF and S(t)O(2) were coupled, whereas for alpha-CHLOR these measures were uncoupled. Moreover, the clearance of [(11)C]cocaine from the brain was faster for ISO (peak half-clearance 15.8 +/- 2.8 min) than for alpha-CHLOR (27.5 +/- 0.6 min), and the ratio of specific to non-specific binding of [(11)C]cocaine in the brain was higher for ISO- (3.37 +/- 0.32) than for alpha-CHLOR-anesthetized rats (2.24 +/- 0.4). For both anesthetics, cocaine-induced changes in CBF followed the fast uptake of [(11)C]cocaine in the brain (peaking at approximately 2.5-4 min), but only for ISO did the duration of the CBV and S(t)O(2) changes correspond to the rate of [(11)C]cocaine's clearance from the brain. These results demonstrate that anesthetics influence cocaine's hemodynamic and metabolic changes in the brain, and its binding and PK, which highlights the need to better understand the interactions between anesthetics and pharmacological challenges in brain functional imaging studies.

The effect of intravenous lidocaine on brain activation during non-noxious and acute noxious stimulation of the forepaw: a functional magnetic resonance imaging study in the rat.

BACKGROUND: Lidocaine can alleviate acute as well as chronic neuropathic pain at very low plasma concentrations in humans and laboratory animals. The mechanism(s) underlying lidocaine's analgesic effect when administered systemically is poorly understood but clearly not related to interruption of peripheral nerve conduction. Other targets for lidocaine's analgesic action(s) have been suggested, including sodium channels and other receptor sites in the central rather than peripheral nervous system. To our knowledge, the effect of lidocaine on the brain's functional response to pain has never been investigated. Here, we therefore characterized the effect of systemic lidocaine on the brain's response to innocuous and acute noxious stimulation in the rat using functional magnetic resonance imaging (fMRI). METHODS: Alpha-chloralose anesthetized rats underwent fMRI to quantify brain activation patterns in response to innocuous and noxious forepaw stimulation before and after IV administration of lidocaine. RESULTS: Innocuous forepaw stimulation elicited brain activation only in the contralateral primary somatosensory (S1) cortex. Acute noxious forepaw stimulation induced activation in additional brain areas associated with pain perception, including the secondary somatosensory cortex (S2), thalamus, insula and limbic regions. Lidocaine administered at IV doses of either 1 mg/kg, 4 mg/kg or 10 mg/kg did not abolish or diminish brain activation in response to innocuous or noxious stimulation. In fact, IV doses of 4 mg/kg and 10 mg/kg lidocaine enhanced S1 and S2 responses to acute nociceptive stimulation, increasing the activated cortical volume by 50%-60%. CONCLUSION: The analgesic action of systemic lidocaine in acute pain is not reflected in a straightforward interruption of pain-induced fMRI brain activation as has been observed with opioids. The enhancement of cortical fMRI responses to acute pain by lidocaine observed here has also been reported for cocaine. We recently showed that both lidocaine and cocaine increased intracellular calcium concentrations in cortex, suggesting that this pharmacological effect could account for the enhanced sensitivity to somatosensory stimulation. As our model only measured physiological acute pain, it will be important to also test the response of these same pathways to lidocaine in a model of neuropathic pain to further investigate lidocaine's analgesic mechanism of action.

Quantification of cocaine-induced cortical blood flow changes using laser speckle contrast imaging and Doppler optical coherence tomography.

We present a dual-imaging technique combining laser speckle contrast imaging and spectral-domain Doppler optical coherence tomography to enable quantitative characterization of local cerebral blood flow (CBF) changes in rat cortex in response to drug stimulus (e.g., cocaine) at high spatiotemporal resolutions. To examine the utility of this new technique, animal experiments were performed to study the influences of anesthetic regimes (e.g., isoflurane, alpha-chloralose) on the pharmadynamic effects of acute cocaine challenge. The results showed that cocaine-evoked CBF patterns (e.g., increases in alpha-chloralose and decreases in isoflurane regimes) were quantitatively characterized, thus rendering it a potentially useful tool for imaging studies of brain functions.

Estimating coronary artery lumen area with optimization-based contour detection.

A modified optimization-based contour detection method was presented to compute the lumen area of the coronary artery from intravascular ultrasound (IVUS) video images. First, the search range for the artery inner wall was determined based on the continuity of IVUS video frames. Next, the internal and external energy were calculated to describe the smoothness of the arterial wall and the grayscale variation of ultrasound images, respectively. Here, a novel form of the external energy which combines the gradient and variance of the intensity of image in the radial direction was used. Finally, the minimal energy path based on the optimum contour of the artery wall was obtained using circular dynamic programming (DP). By the comparison with the typical DP procedure using the traditional external energy form, based only on the image gradient, the reliability of this modified method is considerably improved in the measurement of coronary artery lumen area.

[Coronary resistance system in evaluation of microvascular dysfunction after intracoronary microembolization: an experimental study].

OBJECTIVE: To assess the microvascular function of coronary artery after intracoronary microembolization using coronary resistance system. METHODS: The left anterior descending coronary artery (LAD) of 10 pigs weighing 21 kg-25 kg were embolized by repetitive injection of microspheres 45 micro m in diameter through a 2.8F Tracker catheter. Intra-vascular ultrasound (IVUS) images, intracoronary Doppler and pressure signals in the middle segment of LAD were acquired by use of intracoronary ultrasound imaging catheter, Doppler flow wire and pressure wire separately. Intracoronary bolus injection of 18 micro g adenosine was administered to maximally vasodilate the coronary arterial bed through the 2.8F Tracker catheter. The resting and hyperemic signals were acquired respectively before microembolization and in different levels of microembolization. Coronary resistance system reflecting the resistance to pulsatile coronary flow was established by a self-made software of PC system. The resting and hyperemic CR parameters included average resting coronary resistance (rCR) and average minimal coronary resistance (min-CR), the first-harmonic rCR and min-CR, the first-harmonic rCR orientation and min-CR orientation, and so on. Factor analysis was performed to extract the best coronary parameter from the coronary resistance parameters. RESULTS: Factor analysis showed that the first-harmonic rCR and first-harmonic min-CR were correlated better with the first component extracted from the resting and hyperemic CR parameters than rCR and min-CR, with the correlation coefficient being 0.913 and 0.950 in the first-harmonic CR and first-harmonic min-CR respectively. No significant difference in min-CR was found between the value at the dosage of 5 x 10(4) microspheres and that before microembolization. The min-CR value increased markedly from 271 mm Hg.ml(-1).s(-1) +/- 99 mm Hg.ml(-1).s(-1) at the dosage of injecting 5 x 10(4) microspheres to 361 mm Hg.ml(-1).s(-1) +/- 158 mm Hg.ml(-1).s(-1) at the dosage of injecting 10 x 10(4) microspheres (P < 0.05). The min-CR value remained almost unchanged from the dosage of 10 x 10(4) to 15 x 10(4) microspheres. There was no significant difference concerning the first-harmonic min-CR between the value at the dosage of 5 x 10(4) microspheres and that before microembolization. Along with the increase of number of microspheres injected the min-CR value increased gradually. The min-CR value was increased significantly than that before microembolization since the number of microspheres injected surpassed 14 x 10(4). CONCLUSION: The first-harmonic min-CR reflected the coronary microvascular dysfunction in different extents of microembolization better than min-CR. The extent of coronary microvascular dysfunction wasn't linearly related to the extent of microembolization.

A new look at glutamate and ischemia: NMDA agonist improves long-term functional outcome in a rat model of stroke.

Ischemic stroke triggers a massive, although transient, glutamate efflux and excessive activation of NMDA receptors (NMDARs), possibly leading to neuronal death. However, multiple clinical trials with NMDA antagonists failed to improve, or even worsened, stroke outcome. Recent findings of a persistent post-stroke decline in NMDAR density, which plays a pivotal role in plasticity and memory formation, suggest that NMDAR stimulation, rather than inhibition, may prove beneficial in the subacute period after stroke. AIM: This study aims to examine the effect of the NMDAR partial agonist d-cycloserine (DCS) on long-term structural, functional and behavioral outcomes in rats subjected to transient middle cerebral artery occlusion, an animal model of ischemic stroke. MATERIALS #ENTITYSTARTX00026; METHODS: Rats (n = 36) that were subjected to 90 min of middle cerebral artery occlusion were given a single injection of DCS (10 mg/kg) or vehicle (phosphate-buffered saline) 24 h after occlusion and followed up for 30 days. MRI (structural and functional) was used to measure infarction, atrophy and cortical activation due to electrical forepaw stimulation. Memory function was assessed on days 7, 21 and 30 postocclusion using the novel object recognition test. A total of 20 nonischemic controls were included for comparison. RESULTS: DCS treatment resulted in significant improvement of somatosensory and cognitive function relative to vehicle treatment. By day 30, cognitive performance of the DCS-treated animals was indistinguishable from nonischemic controls, while vehicle-treated animals demonstrated a stable memory deficit. DCS had no significant effect on infarction or atrophy. CONCLUSION: These results support a beneficial role for NMDAR stimulation during the recovery period after stroke, most likely due to enhanced neuroplasticity rather than neuroprotection.

Simultaneous imaging of cortical hemodynamics and blood oxygenation change during cerebral ischemia using dual-wavelength laser speckle contrast imaging.

A dual-wavelength laser speckle contrast imaging technique (DW-LSCI) is presented for simultaneous imaging of cerebral blood flow and hemoglobin oxygenation changes at high spatiotemporal resolutions. Experimental validation was performed using a rat transient forebrain ischemia model. The results showed that DW-LSCI was able to track detailed hemodynamic and metabolic changes induced by ischemia, i.e., decreased oxy- and total hemoglobin concentrations and blood flow as well as increased deoxy-hemoglobin concentration in the downstream regions, thus allowing us to distinguish cerebral arterial and venous flows. Simultaneous cerebral blood flow and oxygenation imaging at high spatiotemporal resolutions is crucial to the understanding of neural process and brain functions.

Optical coherence Doppler tomography quantifies laser speckle contrast imaging for blood flow imaging in the rat cerebral cortex.

A dual-imaging modality is demonstrated for high-resolution quantitative imaging of local cerebral blood flow in the rat cortex by combining simultaneous spectral-domain Doppler optical coherence tomography (SDOCT) and full-field laser speckle contrast imaging (LSCI). Preliminary studies in tissue flow phantom and cocaine-induced cerebral blood flow changes indicated that by correlating coregistered cortical arterial blood flow, the relative measurement of flow changes by LSCI could be accurately calibrated by the absolute flow imaging provided by SDOCT (least square fit, r(2) approximately 0.96). Quantitative LSCI of cerebral blood flow is crucial to the quantitative analyses of the spatiotemporal hemodynamics of functional brain activations and thus improved understanding of neural process.