Oxygen-Glucose Deprivation Models Ischemic Cell Death in CA1 Hippocampus
Mentor 1
James Moyer
Start Date
28-4-2023 12:00 AM
Description
In the United States, every 40 seconds someone suffers from stroke, and every 3.5 minutes someone dies from stroke. It has been shown that hippocampal CA1 neurons specifically are imperative for autonoetic consciousness, as well as retrieval of remote episodic memory. Areas important for learning and memory are nearly destroyed following ischemia (stroke), and overtime regain some but not all functionality. Calcium regulation is important for normal functioning of neuronal signaling cascades, but during ischemia too much calcium influx can trigger excitotoxic cell death. Therefore, regulation of intracellular Ca2+ is important for protection against ischemia, and may be useful in developing treatments that reduce ischemic brain damage and preserve cognitive functioning. In order to study the neuroprotective effects of treatment, baseline counts of cell death must be made for comparison. The present study used an in vitro rat brain slice model from 5 F344 rats to validate the hypothesis that cells that undergo oxygen-glucose deprivation (OGD) exhibit significantly more cell death than cells that did not undergo OGD. Dorsal hippocampal slices were prepared and subjected to 5 minutes of OGD, and cell death was assayed using the Trypan blue exclusion method. Dead and dying cells take up the Trypan blue while healthy neurons exclude the dye and are left unstained. The number of Trypan blue labeled neurons were then counted using the program ImageJ. The present study shows that just 5-min of OGD models an ischemic attack, resulting in a baseline control that can be used comparatively to examine effects of treatment on cell death. The general aim of this research is to produce a control value of cell death to demonstrate the effectiveness of this model for future neuroprotective research.
Oxygen-Glucose Deprivation Models Ischemic Cell Death in CA1 Hippocampus
In the United States, every 40 seconds someone suffers from stroke, and every 3.5 minutes someone dies from stroke. It has been shown that hippocampal CA1 neurons specifically are imperative for autonoetic consciousness, as well as retrieval of remote episodic memory. Areas important for learning and memory are nearly destroyed following ischemia (stroke), and overtime regain some but not all functionality. Calcium regulation is important for normal functioning of neuronal signaling cascades, but during ischemia too much calcium influx can trigger excitotoxic cell death. Therefore, regulation of intracellular Ca2+ is important for protection against ischemia, and may be useful in developing treatments that reduce ischemic brain damage and preserve cognitive functioning. In order to study the neuroprotective effects of treatment, baseline counts of cell death must be made for comparison. The present study used an in vitro rat brain slice model from 5 F344 rats to validate the hypothesis that cells that undergo oxygen-glucose deprivation (OGD) exhibit significantly more cell death than cells that did not undergo OGD. Dorsal hippocampal slices were prepared and subjected to 5 minutes of OGD, and cell death was assayed using the Trypan blue exclusion method. Dead and dying cells take up the Trypan blue while healthy neurons exclude the dye and are left unstained. The number of Trypan blue labeled neurons were then counted using the program ImageJ. The present study shows that just 5-min of OGD models an ischemic attack, resulting in a baseline control that can be used comparatively to examine effects of treatment on cell death. The general aim of this research is to produce a control value of cell death to demonstrate the effectiveness of this model for future neuroprotective research.
