Neuronal alterations in developing cortical infarction

An experimental study in monkeys

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✓ The sequential neuronal alterations that occur during the early phase of developing cortical infarction in the squirrel monkey were studied by light and electron microscopy. A technique used to select ischemic tissue based on spectrophotometry is described. Neuronal shrinkage, characterized by angularity, cytoplasmic eosinophilia, and nuclear pyknosis on light microscopy and by an increase in electron density of the cytoplasmic and nucleoplasmic matrix on electron microscopy, was the predominant reaction. The increased electron density of the cytoplasm and nucleoplasm suggested a diffuse alteration at the molecular level and the appearance of this abnormality between 3 and 6 hours corresponded with the development of an irreversible neurological deficit. In contrast, approximately 10% of the neurons became very swollen and pale. The pattern of perineuronal astrocytic alterations suggested that some form of interaction involving fluid transfer may exist between astrocyte and neuron and that shrinkage or swelling of neurons may depend in part upon the presence or absence of direct fluid exchange with astrocytes. Most terminal boutons became progressively shrunken and dense resembling the changes which occur in anterograde axonal degeneration. Swelling and fragmentation of large lysosomes occurred at 12 hours. Disruption of neural membranes was widespread by 24 hours and was more severe in swollen neurons.

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Address reprint requests to: John R. Little, M.D., Cerebrovascular Clinical Research Center, Mayo Clinic and Mayo Foundation, Rochester, Minnesota 55901.

© AANS, except where prohibited by US copyright law.

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    Light photomicrographs. Calibrations equal 25 µ in A through F. A. Representative neurons from the nonischemic cortex. H & E. B. At 90 minutes, the neurons have developed an angular contour and increased nuclear basophilia. H & E. C. At 3 hours, the two neurons in the center of the field demonstrate vacuolation of the cytoplasm; some kinking of the apical dendrites is present. Thionin. D. At 4½ hours, a swollen neuron is surrounded by shrunken angular neurons; clear spaces are present beside some of the shrunken cells. H & E. E. At 12 hours, shrunken angular neurons with pyknotic nuclei and adjacent clear spaces are seen. H & E. F. At 24 hours, “incrustations” have developed. Some of these basophilic structures (arrows) appear to be outlining clear vacuoles. Thionin.

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    Electron photomicrograph. Calibrations equal 5 µ. A. Representative neuron from the nonischemic cortex. B. At 3 hours of ischemia, an angular neuron is seen, with increased electron density of the nucleoplasmic and cytoplasmic matrix. The ribosomes have a more diffuse distribution and there is a mild distension of the endoplasmic reticulum (arrows). The neuropil has a loose appearance produced by an increase in the interstitial space. A few swollen astrocytic processes (a) can be identified.

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    Electron photomicrographs; calibrations equal 1 µ. A. Representative neuron from the nonischemic cortex. Note the tightly packed neuropil. B. At 90 minutes, note the swollen mitochondria (m), distended rough endoplasmic reticulum (er), and slight enlargement of the interstitial space (arrows). C. At 3 hours, the ribosomes are more uniformly dispersed and there is an increase in the electorn density of the cytoplasmic matrix; note further increase in the extracellular space (arrows) and the dense degenerating terminal bouton (b). D. At 6 hours, the cisterns of the Golgi apparatus (g) are engulfing a portion of the cytoplasm; apposition of the membranes (arrows) produces a multilaminated appearance. Numerous ribosomes stud the external surface of the nuclear membrane. E. At 12 hours, the membranes of this swollen lysosome (1) have ruptured (arrows); the interstitial space is greatly expanded and the neuronal processes are shrunken and dense. F. At 24 hours, “incrustations” (i) containing numerous tightly packed ribosomes are seen. Note the narrow stalk (arrow) connecting the proximal “incrustation” to the soma of the neuron.

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    Electron photomicrographs; calibration equals 5 µ. A. At 4½ hours, a swollen neuron is seen with numerous swollen mitochondria and distended cisternae of the endoplasmic reticulum dispersed throughout the expanded, low density cytoplasm. Nuclear and plasma membranes are intact No swollen astrocytic processes are in contact with the perikaryon; however, a process from a “dark” neuron (d) is present in the upper left corner. B. At 6 hours, the neuron is shrunken. The mitochondria (m) contain a homogeneous and moderately dense matrix in which few intact cristae can be identified Distension of the cisternae of the endoplasmic reticulum (er) is prominent. The neuron is surrounded by distended astrocytic processes (a). Two terminal boutons (b) can be identified in contact with the plasma membrane; an axo-dendntic synapse is present in the lower right corner (arrow)

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    Electron photomicrographs; calibrations equal 5 µ. A. At 12 hours, shrunken neuron. Note the marked irregularity of the soma membrane. Distension of the cytoplasmic vacuoles (er) has resulted in the segregation of some peripheral cytoplasm. The large dense structure on the right, which probably represents an “incrustation,” is attached to the soma by two narrow stalks (arrows). Numerous swollen astrocytic processes (a) surround the dark neuron. B. At 12 hours, swollen neuron. Irregular enlargement of the space between the membranes of the nuclear envelope is present and numerous ribosomes stud its external surface. The plasma membrane has become disrupted (arrows), allowing the cytoplasmic contents to mingle freely with the surrounding neuropil. Considerable enlargement of the interstitial space is evident and numerous shrunken dense neuronal processes can be identified. Part of a dense neuron (d) is present in the upper left corner.

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    Electron photomicrographs; calibrations equal 5 µ. A. At 24 hours, the nucleus, containing dense clumps of chromatin and a nucleolus, appears to be intact. Complete disruption of the plasma membrane has occurred. A normal-looking lysosome (l) can be identified. B. At 24 hours, shrunken neuron. A clump of condensed cytoplasm (double arrows) is being “pinched off” of the soma of the neuron by a greatly swollen vacuole. Numerous dense, angular structures (arrows) which probably represent “incrustations” can be identified in close proximity to the perikaryon. The surrounding astrocytic processes are enormously distended (a).

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