Astrocyte-neuron lactate shuttle plays a pivotal role in sensory-based neuroprotection in a rat model of permanent middle cerebral artery occlusion.

We have previously demonstrated protection from impending cortical ischemic stroke is achievable by sensory stimulation of the ischemic area in an adult rat model of permanent middle cerebral artery occlusion (pMCAo). We have further demonstrated that a major underpinning mechanism that is necessary for such protection is the system of collaterals among cerebral arteries that results in reperfusion of the MCA ischemic territory. However, since such collateral flow is weak, it may be necessary but not sufficient for protection and therefore we sought other complementary mechanisms that contribute to sensory-based protection. We hypothesized that astrocytes-neuron lactate shuttle (ANLS) activation could be another potential underpinning mechanism that complements collateral flow in the protection process. Supporting our hypothesis, using functional imaging, pharmacological treatments, and postmortem histology, we showed that ANLS played a pivotal role in sensory stimulation-based protection of cortex and therefore serves as the other supporting mechanism underpinning the protection process.

Astrocyte-neuron lactate shuttle plays a pivotal role in sensory-based neuroprotection in a rat model of permanent middle cerebral artery occlusion.

We have previously demonstrated protection from impending cortical stroke is achievable by sensory stimulation of the ischemic area in an adult rat model of permanent middle cerebral artery occlusion (pMCAo). We have further demonstrated that a major underpinning mechanism that is necessary for such protection is the system of collaterals among cerebral arteries that results in reperfusion of the MCA ischemic territory. However, since such collateral flow is weak, it may be necessary but not sufficient for protection and therefore we were seeking other complementary mechanisms that contribute to sensory-based protection. We hypothesized that astrocytes-to-neuron shuttle (ANLS) is another potential underpinning mechanism that could complement collateral flow in the protection process. Supporting our hypothesis, using functional imaging, pharmacological treatments, and postmortem histology, we show that ANLS has a pivotal role in sensory-based protection of cortex and therefor serves as the other supporting mechanism underpinning the protection process.

Development of highly sensitive, flexible dual L-glutamate and GABA microsensors for in vivo brain sensing.

Real-time tracking of neurotransmitter levels in vivo has been technically challenging due to the low spatiotemporal resolution of current methods. Since the imbalance of cortical excitation/inhibition (E:I) ratios are associated with a variety of neurological disorders, accurate monitoring of excitatory and inhibitory neurotransmitter levels is crucial for investigating the underlying neural mechanisms of these conditions. Specifically, levels of the excitatory neurotransmitter L-glutamate, and the inhibitory neurotransmitter GABA, are assumed to play critical roles in the E:I balance. Therefore, in this work, a flexible electrochemical microsensor is developed for real-time simultaneous detection of L-glutamate and GABA. The flexible polyimide substrate was used for easier handling during implantation and measurement, along with less brain damage. Further, by electrochemically depositing Pt-black nanostructures on the sensor's surface, the active surface area was enhanced for higher sensitivity. This dual neurotransmitter sensor probe was validated under various settings for its performance, including in vitro, ex vivo tests with glutamatergic neuronal cells and in vivo test with anesthetized rats. Additionally, the sensor's performance has been further investigated in terms of longevity and biocompatibility. Overall, our dual L-glutamate:GABA sensor microprobe has its unique features to enable accurate, real-time, and long-term monitoring of the E:I balance in vivo. Thus, this new tool should aid investigations of neural mechanisms of normal brain function and various neurological disorders.

Head Implants for the Neuroimaging of Awake, Head-Fixed Rats.

Anesthetics, commonly used in preclinical and fundamental scientific research, have a depressive influence on the metabolic, neuronal, and vascular functions of the brain and can adversely influence neurophysiological results. The use of awake animals for research studies is advantageous but poses the major challenge of keeping the animals calm and stationary to minimize motion artifacts throughout data acquisition. Awake imaging in smaller-sized rodents (e.g., mice) is very common but remains scant in rats as rats are bigger, stronger, and have a greater tendency to oppose movement restraints and head fixation over the long durations required for imaging. A new model of neuroimaging of awake, head-fixed rats using customized hand-sewn slings, 3D-printed head implants, head caps, and a headframe is described. The results acquired following a single trial of single-whisker stimulation suggest an increase in the intensity of the evoked functional response. The acquisition of the evoked functional response from awake, head-fixed rats is faster than that from anesthetized rats, reliable, reproducible, and can be used for repeated longitudinal studies.

Pulse waveform analysis on temporal changes in ocular blood flow due to caffeine intake: a comparative study between habitual and non-habitual groups.

PURPOSE: To evaluate and compare the temporal changes in pulse waveform parameters of ocular blood flow (OBF) between non-habitual and habitual groups due to caffeine intake. METHOD: This study was conducted on 19 healthy subjects (non-habitual 8; habitual 11), non-smoking and between 21 and 30 years of age. Using laser speckle flowgraphy (LSFG), three areas of optical nerve head were analyzed which are vessel, tissue, and overall, each with ten pulse waveform parameters, namely mean blur rate (MBR), fluctuation, skew, blowout score (BOS), blowout time (BOT), rising rate, falling rate, flow acceleration index (FAI), acceleration time index (ATI), and resistive index (RI). Two-way mixed ANOVA was used to determine the difference between every two groups where p < 0.05 is considered significant. RESULT: There were significant differences between the two groups in several ocular pulse waveform parameters, namely MBR (overall, vessel, tissue), BOT (overall), rising rate (overall), and falling rate (vessel), all with p < 0.05. In addition, the ocular pulse waveform parameters, i.e., MBR (overall), skew (tissue), and BOT (tissue) showed significant temporal changes within the non-habitual group, but not within the habitual group. The temporal changes in parameters MBR (vessel, tissue), skew (overall, vessel), BOT (overall, vessel), rising rate (overall), falling rate (overall, vessel), and FAI (tissue) were significant for both groups (habitual and non-habitual) in response to caffeine intake. CONCLUSION: The experiment results demonstrated caffeine does modulate OBF significantly and response differently in non-habitual and habitual groups. Among all ten parameters, MBR and BOT were identified as the suitable biomarkers to differentiate between the two groups.

Ocular Blood Flow in Rabbits under Deep Anesthesia: A Real-Time Measurement Technique and Its Application in Characterizing Retinal Ischemia.

In this study, we reported a new technique based on laser speckle flowgraphy to record the ocular blood flow in rabbits under deep anesthesia, and proposed parameters to characterize retinal ischemia. We applied the proposed technique to study the correlation of blood flow between the eyes of normal non-anesthetized animals, and to characterize the occlusion of the internal carotid artery (ICA) and external carotid artery (ECA). We established a correlation in blood flow between the eyes of non-anesthetized animals, and derived two new parameters, namely, the laterality index and vascular perfusion estimate (VPE). Our experimental results from 16 eyes (of 13 New Zealand white rabbits) showed a reduction in ocular blood flow with a significant decrease in the VPE after the occlusion of the ECA (p < 0.001). A low/minimal effect on blood flow was observed with the occlusion of the ICA. In conclusion, we demonstrated a means for the real-time measurement of the ocular blood flow in rabbits under deep anesthesia by using laser speckle flowgraphy and the VPE as an indicator of successful occlusion. The proposed technique might be applicable in quantifying the efficacy of new drugs and interventions for the treatment of retinal ischemia.

Effects of water drinking test on ocular blood flow waveform parameters: A laser speckle flowgraphy study.

The water-drinking test (WDT) is a provocative test used in glaucoma research to assess the effects of elevated intraocular pressure (IOP). Defective autoregulation due to changes in perfusion pressure may play a role in the pathophysiology of several ocular diseases. This study aims to examine the effects of WDT on ocular blood flow (in the form of pulse waveform parameters obtained using laser speckle flowgraphy) to gain insight into the physiology of ocular blood flow and its autoregulation in healthy individuals. Changes in pulse waveform parameters of mean blur rate (MBR) in the entire optic nerve head (ONH), the vasculature of the ONH, the tissue area of the ONH, and the avascular tissue area located outside of the ONH were monitored over time. Significant increases in the falling rate of MBR over the entire ONH and its tissue area and decreases in blowout time (BOT) of the tissue area were observed only at 10 minutes after water intake. Significant increases in the skew of the waveform and the falling rate were observed in the vasculature of the ONH at 40 and 50 minutes after water intake, respectively. In the avascular region of the choroid, the average MBR increased significantly up to 30 minutes after water intake. Furthermore, the rising rate in this region increased significantly at 20 and 40 minutes, and the falling rate and acceleration-time index were both significantly increased at 40 minutes after water intake. Our results indicate the presence of effective autoregulation of blood flow at the ONH after WDT. However, in the choroidal region, outside of the ONH, effective autoregulation was not observed until 30 minutes after water intake in healthy study participants. These pulse waveform parameters could potentially be used in the diagnosis and/or monitoring of patients with glaucoma.

Rats exhibit reference-dependent choice behavior.

Human preferences depend on whether a chosen outcome appears to be a loss or a gain compared with what had been expected, i.e., in comparison to a reference point. Because reference dependence has such a strong influence on human decision-making, it is important to uncover its origins, which will in turn help delineate the underlying mechanisms. It remains unknown whether rats use reference points in decision-making, and yet, the study of rats could help address the question of whether reference dependence is evolutionarily conserved among mammals and could provide a nonhuman animal model to investigate the neural mechanisms underlying this important cognitive process. The aim of the current study was to determine whether rats show reference-dependent choice behavior. We developed a novel paradigm by modifying the "T" maze by installing "pockets" to the left and right of the "T" stem that held reward pellets so rats would potentially develop reference values for each option prior to choice. We found that the rats were indeed sensitive to the way alternatives were presented. That is, they exhibited reference-dependent choice behavior by avoiding the choice option framed as a loss (e.g., having four reward pellets in the pocket, but receiving only one), at least under conditions with certain outcomes and clear differences between the reference and outcome quantities. Despite the small number of rats in this study, this species-level capacity suggests that reference dependence in general and loss aversion in particular may be conserved traits that evolved at or before the emergence of mammals.