Summary
The ultrastructural changes in the pyramidal neurons of the CA1 region of the hippocampus were studied 6 h, 24 h, 48 h, and 72 h following a transient 10 min period of cerebral ischemia induced by common carotid occlusion combined with hypotension. The pyramidal neurons showed delayed neuronal death (DND), i.e. at 24 h and 48 h postischemia few structural alterations were noted in the light microscope, while at 72 h extensive neuronal degeneration was apparent. The most prominent early ultrastructural changes were polysome disaggregation, and the appearance of electron-dense fluffy dark material associated with tubular saccules. Mitochondria and nuclear elements appeared intact until frank neuronal degeneration. The dark material accumulated with extended periods of recirculation in soma and in the main trunks of proximal dendrites, often beneath the plasma membrane, less frequently in the distal dendrites and seldom in spines. Protein synthesis inhibitors (anisomycin, cycloheximide) and an RNA synthesis inhibitor (actinomycin D), administered by intrahippocampal injections or subcutanously, did not mitigate neuronal damage. Therefore, DND is probably not apoptosis or a form of programmed cell death. We propose that the dark material accumulating in the postischemic period represents protein complexes, possibly aggregates of proteins or internalized plasma membrane fragments, which may disrupt vital cellular structure and functions, leading to cell death.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- DND:
-
delayed neuronal death
- ER:
-
endoplasmic reticulum
- GA:
-
Golgi apparatus
- HSP:
-
heat shock protein
- IR:
-
immunoreactivity
- PSD:
-
postsynaptic density
- RNA:
-
ribonucleic acid
- SER:
-
smooth endoplasmic reticulum
- UIR:
-
ubiquitin immunoreactivity
References
Auer RN, Olsson Y, Siesjö BK (1984) Hypoglycemic brain injury in the rat. Correlation of density of brain damage with the EEG isoelectric time: a quantitative study. Diabetes 33:1090–1098
Angeletti PU, Levi-Montalcini R, Caramia F (1971) Analysis of the effects of the antiserum to the nerve growth factor in adult mice. Brain Res 27:343–355
Aoki C, John TH, Pickel VM (1987) Ultrastructural localization of β-adrenergic receptor-like immunoreactivity in the cortex and neostriatum of rat brain. Brain Res 437:264–282
Blomqvist P, Lindvall O, Stenevi U, Wieloch T (1985) Cyclic AMP concentrations in rat neocortex and hippocampus following incomplete ischemia: effects of central noradrenergic neurons, prostaglandins and adenosine. J Neurochem 44:1345–1353
Brierley JB (1980) Hypoxic brain damage. In: Rose FC, Behan PO (eds) Animal models of neurological disease. Pitman Medical, Tunbridge Wells, pp 338–346
Brown AW, Brierley JB (1972) Anoxic-ischeamic cell change in rat brain light microscopic and fine-structural observations. J Neurol Sci 16:59–84
Brown AW, Levy DE, Kublik M, Harrow J, Plum F, Brierley JB (1979) Selective chromatolysis of neurons in the gerbil brain: A possible consequence of “epileptic” activity produced by common carotid artery occlusion. Ann Neurol 5:127–138
Bubis JJ, Fujimoto T, Ito U (1976) Experimental cerebral ischaemia in Mongolian gerbils. V. Ultrastructural changes in h 3 sector of the hippocampus. Acta Neuropathol (Berl) 36:285–294
Choi DW (1987) Ionic dependence of glutamate neurotoxicity. J Neurosci 7:369–379
Choi DW (1988) Glutamate neurotoxicity and diseases of the nervous system. Neuron 1:623–634
Finley D, Varshavsky A (1985) The ubiquitin system: functions and mechanisms. TIBS 9:343–346
Garcia JH, Lossinsky AS, Kauffman FC, Conger KA (1978) Neuronal ischemic injury: light microscopy, ultrastructure and biochemistry. Acta Neuropathol (Berl) 43:85–95
Gustafson I, Miyauchi Y, Wieloch T (1989) Postischemic administration of Idazoxan, an α-2 adrenergic receptor antagonist, decreases neuronal damage in the rat brain. J Cereb Blood Flow Metab 9:171–174
Haas AL, Bright PM (1985) The immunochemical detection and quantitation of intracellular ubiquitin-protein conjugates. J Biol Chem 260:12464–12473
Hagberg H, Lehmann A, Sandberg M, Nyström B, Jacobsson I, Hamberger A (1985) Ischemia induced shift of inhibitory and excitatory amino acids from intra- to extracellular compartments. J Cereb Blood Flow Metab 5:413–419
Ito U, Spatz M, Walker JT, Klatzo I, (1975) Experimental cerebral ischemia in mongolian gerbils. Acta Neuropathol (Berl) 32:209–223
Jørgensen MB, Johansen FF, Diemer NH (1987) Removal of the enthorhinal cortex protects hippocampal CA-1 neurons from ischemic damage. Acta Neuropath (Berl) 73:189–194
Kadota T, Kadota K (1979) Filamentous contacts containing subjunctional dense lattice and tubular smooth endoplasmic reticulum in cat lateral geniculate nuclei. Brain Res 177:49–59
Kirino T (1982) Delayed neuronal death in the gerbil hippocampus following ischemia. Brain Res 239:57–69
Kirino T, Sano K (1984) Fine structural nature of delayed neuronal death following ischemia in the gerbil hippocampus. Acta Neuropathol (Berl) 62:209–218
Kirino T, Tamura A, Sano K (1984) Delayed neuronal death in the rat hippocampus following transient forebrain ischemia. Acta Neuropathol (Berl) 64:139–147
Kiessling M, Xie Y, Ullrich B, Thilmann R (1991) Are the neuroprotective effects of the protein synthesis inhibitor cycloheximide due to prevention of apoptosis? J Cereb Blood Flow Metab 11:S357
Levi-Montalcini R, Caramia F, Angeletti PU (1969) Alterations in the fine structure of nucleoli in sympathetic neurons following NGF-antiserum treatment. Brain Res 12:54–73
Magnusson K, Wieloch TW (1989) Impairment of protein ubiquitination may cause delayed neuronal death. Neurosci Lett 96:264–270
Martin DP, Schmidt RE, DiStefano PS, Lowry OH, Carter JG, Johnson EM Jr (1988) Inhibitors of protein synthesis and RNA synthesis prevent neuronal death caused by nerve growth factor deprivation. J Cell Biol 106:829–844
Mattson MP, Murrain M, Guthrie PB, Kater SB (1989) Fibroblast growth factor and glutamate: Opposing roles in the generation and degeneration of hippocampal neuroarchitecture. J Neurosci 9:3728–3740
Mayer RJ, Arnold J, László L, Landon M, Lowe J (1991) Ubiquitin in health and disease. Biochem Biophys Acta 1089:141–157
McGee-Russell, Brown AW, Brierley JB (1970) A combined light and electron microscope study of early anoxic-ischemic cell change in rat brain. Brain Res 20:193–200
Meldrum BS, Swan JH (1989) Competitive and non-competitive NMDA antagonists as cerebroprotective agents. In: Krieglstein J (ed) Pharmacology of cerebral ischemia 1988. CRC Press Inc Boca Raton 157–163
Monaghan DT, Bridges RJ, Cotman CW (1989) The excitatory amino acid receptors: Their classes, pharmacology, and distinct properties in the function of the central nervous system. Annu Rev Pharmacol Toxicol 29:365–402
Munekata K, Hossmann K-A, Xie Y, Seo K, Oschlies U (1987) Selective vulnerability of hippocampus: Ribosomal aggregation, protein synthesis and tissue pH. In: Raichle ME, Powers WJ (eds) Cerebrovascular disease. Raven Press, New York, pp. 107–117
Nellgård B, Gustafson I, And Wieloch T (1991) Lack of protection by MK-801 (Dizoclipine maleate) on ischemic neuronal damage in the rat hippocampus following transient cerebral ischemia. Anesthesiology 75:279–287
Nellgård and Wieloch (1992) Postischemic blockade of AMPA but not NMDA receptors mitigates neuronal damage in the rat brain following transient cerebral ischemia. J Cereb Blood Flow Metabol (in press)
Onodera H, Sato G, Kogure K (1986) Lesions of the Schaffer collaterals prevent ischemic death of CA1 pyramidal cells. Neurosci Lett 68:169–174
Parag HA, Raboy B, Kulka RG (1987) Effect of heat shock protein degradation in mammalian cells: involvement of the ubiquitin system. EMBO J 6:55–61
Paxinos G, Watson C (1982) The rat brain in stereotaxic coordinates. Sidney, Academic Press
Peters A, Palay SL, Webster HF (1976) The fine structure of the nervous system: the neuron and supporting cells. WB Saunders Co, Philadelphia London Toronto
Petito CK, Pulsinelli WA (1984A) Delayed neuronal recovery and neuronal death in rat hippocampus following severe cerebral ischemia: possible relationship to abnormalities in neuronal processes. J Cereb Blood Flow Metab 4:194–205
Petito CK, Pulsinelli WA (1984B) Sequential development of reversible and irreversible neuronal damage following cerebral ischemia. J Neuropathol Exp Neurol 43:141–153
Petito C, Feldmann E, Pulsinelli W, Plum F (1987) Delayed hippocampal damage in humans following cardiac arrest. Neurology 37:1282–1286
Pratt RM, Greene RM (1976) Inhibition of palatal epithelial cell death by altered protein synthesis. Dev Biol 54:135–145
Pulsinelli WA, Duffy TE (1983) Regional energy balance in rat brain after transient forebrain ischemia. J Neurochem 40:1500–1503
Pulsinelli WA, Brierley JB, Plum F (1982) Temporal profile of neuronal damage in a model of transient forebrain ischemia. Ann Neurol 11:491–499
Rechsteiner M (1987) Ubiquitin-mediated pathways for intracellular proteolysis. Ann Rev Cell Biol 3:1–30
Rothman SM (1984) Synaptic release of excitatory amino acid neurotransmitters mediates anoxic neuronal death. J Neurosci 4:1884–1891
Seubert P, Lee K, Lynch G (1989) Ischemia triggers NMDA receptor-linked cytoskeletal proteolysis in hippocampus. Brain Res 492:366–370
Sheardown MJ, Nielson EØ, Hansen AJ, Jacobsen P, Honoré T (1990) 2,3-Dihydroxy-6-nitro-7-sulfamoyl-benzo(F)-quinoxaline: a neuroprotectant for cerebral ischemia. Science 247:571–574
Shigeno T, Yamasaki Y, Kato G, Kusaka K, Mima T, Takakura K, Graham D, Furukawa S (1991) Reduction of delayed neuronal death by inhibition of protein synthesis. Neurosci Lett 120:117–119
Siesjö BK, Bengtsson F (1989) Calcium fluxes, calcium antagonists, and calcium related pathology in brain ischemia, hypoglycemia, and spreading depression: a unifying hypothesis. J Cereb Blood Flow Metab 9:127–140
Siesjö BK (1985) Acid-base homeostasis in the brain: Physiology, chemistry, and neurochemical pathology. In: Kogure K, Hossmann K-A, Siesjö BK, Welsh FA (eds) Progress in Brain Research, Vol 63. Elsevier, Amsterdam, pp 121–154
Siman R, Noszek CJ, Kegerise C (1989) Calpain I activation is specifically related to excitatory amino acid induction of hippocampal damage. J Neurosci 9:1579–1590
Smith M-L, Auer RN, Siesjö BK (1984A) The density and distribution of ischemic brain injury in the rat following 2–10 min of forebrain ischemia. Acta Neuropathol (Berl) 64:319–332
Smith M-L, Bendek G, Dahlgren N, Rosen I, Wieloch T, Siesjö BK (1984B) Models for studying long-term recovery following forebrain ischemia in the rat. A 2-vessel occlusion model. Acta Neurol Scand 69:385–401
Tatsuoka H (1986) Ultrastructural changes of afferent nerve terminals in the cat medial geniculate body after destruction of the inferior colliculus. J Submicro Cytol 18:35–46
van Reempts J, Borgers M (1985) Ischemic brain injury and cell calcium: Morphologic and therapeutic aspects. Ann Emergency Med 14:736–742
Westerberg E, Monaghan DT, Kalimo H, Cotman CW, Wieloch TW (1989) Dynamic changes of excitatory amino acid receptors in the rat hippocampus following transient cerebral ischemia. J Neurosci 9:798–805
Widmann R, Kuroiwa T, Bonnekoh P, Hossmann K-A (1991) [14C]Leucine incorporation into brain proteins in gerbils after transient ischemia: Relationship to selective vulnerability of hippocampus. J Neurochem 56:789–796
Wieloch T (1985) Neurochemical correlates to selective neuronal vulnerability. In: Kogure K, Hossmann K-A, Siesjö BK, Welsh FA (eds) Progress in Brain Research, Vol. 63. Elsevier, Amsterdam, pp 69–85
Wieloch T, Lindvall O, Blomqvist P, Gage F (1985) Evidence for amelioration of ischemic neuronal damage in the hippocampal formation by lesions of the perforant path. Neurol Res 7:24–26
Wyllie AH, Kerr JFR, Currie AR (1980) Cell death: the significance of apoptosis. In: Bourne GH, Danielli JF, Jeon KW (eds) International Review of cytology, Vol 68, Academic Press pp 252–306
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Deshpande, J., Bergstedt, K., Lindén, T. et al. Ultrastructural changes in the hippocampal CA1 region following transient cerebral ischemia: evidence against programmed cell death. Exp Brain Res 88, 91–105 (1992). https://doi.org/10.1007/BF02259131
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02259131