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Glia. 2001 Feb;33(2):169-79.

Astrocytic clasmatodendrosis in hippocampal organ culture.

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Department of Neurology, University of Chicago, 5841 S. Maryland, Chicago, IL 60637, USA.


Mechanisms by which astrocytes are irreversibly injured from ischemic brain injury remain incompletely defined. More than 90 years ago Alzheimer showed that astrocytes lose their distal processes (i.e., undergo "clasmatodendrosis") when irreversibly injured by a reduction in blood flow, a process shown by Friede and van Houten (1961) to be due to energy failure and acidosis. Such alterations in astrocytic morphology can relate directly to changes in cell function. However, astrocytic clasmatodendrosis has largely been lost to the modern literature, perhaps because of a inability to study it under controlled conditions. In the present study, novel four-dimensional (4D)and digital deblurring imaging of glial fibrillary acidic protein (GFAP) immunostaining changes in hippocampal organ cultures (HOTCs) were used to establish an in vitro model of astrocytic clasmatodendrosis. Also, astrocytes in primary culture were transfected with green fluorescent protein (GFP) to show the occurrence of clasmatodendrosis via a parallel and separate means. In HOTCs, a significant reduction in astrocytic process length occurred 15 min (and remained for 60 min) after exposure to acidic Ringer's and mitochondrial inhibition in the pyramidal cell body layer. Time-lapsed images of primary cultures showed thinning of cell processes within 15 min of exposure to acidic Ringer's and mitochondrial inhibition. Distal processes subsequently broke away but retained their fluorescence for minutes before disintegrating along with their parent cell bodies. This report shows the spatiotemporal occurrence of clasmatodendrosis in astrocytes of HOTCs closely parallels that seen in vivo. Thus, HOTCs, where microenvironmental conditions can be controlled and single, identified cells can be followed in space and time, can be applied to study the interrelations between energy metabolism and pH that result in clasmatodendrosis.

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