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The Indirect Neuron-astrocyte Coculture Assay: An In Vitro Set-up for the Detailed Investigation of Neuron-glia Interactions
Published on: November 14, 2016
Astrocytes and radial glia-like cells, but not neurons, display a nonapoptotic increase in caspase-3 expression
Morgan E Stevenson1, Nicole A Lensmire1, Rodney A Swain1
1Department of Psychology, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin.
Background:
Exercise induces plasticity in the hippocampus, which includes increases in neurogenesis, the proliferation of new neurons, and angiogenesis, the sprouting of new capillaries from preexisting blood vessels. Following exercise, astrocytes also undergo morphological changes that parallel the events occurring in the neurovascular system. Interestingly, there have also been reports of apoptosis in the hippocampus following aerobic exercise. This experiment aimed to identify which population of hippocampal cells undergoes apoptosis after an acute bout of exercise.
Methods:
Cleaved caspase-3, a terminal protein in the apoptotic cascade, was initially used to identify apoptotic cells in the hippocampus after rats completed an acute bout of exercise. Next, the proportion of immature neurons, adult neurons, astrocytes, or radial glia-like cells expressing cleaved caspase-3 was quantified. TUNEL staining was completed as a second measure of apoptosis.
Results:
Following exercise, cleaved caspase-3 expression was increased in the CA1 and DG regions of the hippocampus. Cleaved caspase-3 was not highly expressed in neuronal populations, and expression was not increased in these cells postexercise. Instead, cleaved caspase-3 was predominantly expressed in astrocytes. Following exercise, there was an increased number of cleaved caspase-3 positive astrocytes in DG and CA1, and cleaved caspase-3 positive radial glia-like cells located in the subgranular zone. To determine whether cleaved caspase-3 expression in these glial cells was associated with apoptosis, a TUNEL assay was completed. TUNEL staining was negligible in all groups and did not mirror the pattern of caspase-3 labeling.
Conclusions:
Cleaved caspase-3 expression was detected largely in non-neuronal cell populations, and the pattern of cleaved caspase-3 expression did not match that of TUNEL. This suggests that after exercise, cleaved caspase-3 expression may serve a nonapoptotic role in these hippocampal astrocytes and radial glia-like cells. It will be important to identify the function of exercise-induced cleaved caspase-3 expression in the future experiments.
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