Journal of neuroscience methods
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J. Neurosci. Methods · Sep 2012
Evaluation of transcranial Doppler flow velocity changes in intracerebral hemorrhage rats using ultrasonography.
This study investigated the blood flow velocity changes in seven major arteries in rat brain before and after intracerebral hemorrhage (ICH) using high frequency transcranial Doppler (TCD) ultrasonography (13-4 MHz). Eighteen adult Sprague Dawley rats received either the collagenase-injection surgery (ICH, n=12) or the saline-injection surgery (control, n=6) after baseline TCD flow velocity values were recorded. The TCD flow velocity changes were measured at 0.5h after the surgery and daily for the following 8 days in seven major cerebral arteries, including bilateral internal carotid arteries, bilateral middle cerebral arteries (MCAs), bilateral posterior cerebral arteries (PCAs), and basilar artery. ⋯ This study demonstrates the feasibility and reliability of monitoring TCD flow velocity in cerebral arteries using ultrasonography technique in a rat ICH model. The results of this study extend our knowledge in the cerebrovascular changes during intracranial hemorrhage and suggest a possibility of clinical application of TCD ultrasonography in studying the dynamic cerebral circulation after strokes. Moreover, this method could be extensively applied in further studies using potential neuroprotective treatments that affect the cerebral dynamics in the intracerebral hemorrhage.
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J. Neurosci. Methods · Sep 2012
Optogenetic approaches to characterize the long-range synaptic pathways from the hypothalamus to brain stem autonomic nuclei.
Recent advances in optogenetic methods demonstrate the feasibility of selective photoactivation at the soma of neurons that express channelrhodopsin-2 (ChR2), but a comprehensive evaluation of different methods to selectively evoke transmitter release from distant synapses using optogenetic approaches is needed. Here we compared different lentiviral vectors, with sub-population-specific and strong promoters, and transgenic methods to express and photostimulate ChR2 in the long-range projections of paraventricular nucleus of the hypothalamus (PVN) neurons to brain stem cardiac vagal neurons (CVNs). Using PVN subpopulation-specific promoters for vasopressin and oxytocin, we were able to depolarize the soma of these neurons upon photostimulation, but these promoters were not strong enough to drive sufficient expression for optogenetic stimulation and synaptic release from the distal axons. ⋯ However, neither was able to drive sufficient expression to observe and photostimulate the long-range projections to brainstem autonomic regions. We conclude that a viral vector approach with a strong promoter is required for successful optogenetic stimulation of distal axons to evoke transmitter release in pre-autonomic PVN neurons. This approach can be very useful to study important hypothalamus-brainstem connections, and can be easily modified to selectively activate other long-range projections within the brain.