Relation between the Severity of the Sensorimotor Cortical Edema with Cell Swelling and the Duration of Common Carotid Artery Occlusion in Rats (Morphometric Study)

The aim of the study. To examine the changes in structure and morphometry in sensorimotor cortical edema with cell swelling in mature white rats after common carotid artery occlusion of various durations.
Material and methods. Acute ischemia was modeled on white adult Wistar rats by 20-, 30- and 40-min occlusion of the common carotid arteries (CCA). Histological (hematoxylin-eosin and Nissl staining), immunohistochemical (NSE, MAP-2, GFAP) and morphometric methods were used. Morphometry was assessed on hematoxylin and eosin-stained specimens using ImageJ 1.53 plug-ins (Find Maxima, Find Foci). Statistical hypothesis testing (nonparametric criteria) was performed using Statistica 8.0 software.
Results. In the sensorimotor cortex (SMC) of white rats after 20, 30 and 40 minutes of CCA occlusion the signs of cytotoxic brain edema appeared, focal destructive and adaptive changes of neurons and astroglia evolved. The edema persisted throughout the observation period (7 days). The increase in the relative area, the number of cell swelling zones and their hydration (pixel brightness) was significant. On days 1 and 3 after CCA occlusion, some of the SMC astrocyte processes underwent destruction. Subpial and perivascular zones suffered to a greater extent. Mild and moderate (after unilateral 30-min CCA occlusion) to moderate and severe (after bilateral 40-min CCA occlusion) scattered structural and functional changes of the SMC with large areas of clearing in the «porous» neuropil, severe perivascular and perineuronal edema of the astrocyte processes developed. The latter was associated with a moderate reduction of the total neuronal density.
Conclusion. After occlusion of CCA, signs of edema with cellular swelling appeared in the SMC amid dystrophic and necrotic pyramidal neurons and activated neuroglial cells. To a greater extent, the signs of brain swelling were evident three days after bilateral 40-min occlusion of CCA.

1. Bashirova A.R., Sundukov D.V., Babkina A.S., Golubev M.A., Telipov I.N. Morphometry of cortical neurons in acute clozapine and ethanol poisoning. Obshchaya Reanimatologiya=General Reanimatology. 2020, 16 (6): 19-30 [In Russ.]. DOI: 10.15360/1813-9779-2020-6-19-30

2. Bailey D.M., Bartsch P., Knauth M., Baumgartner R.W. Emerging concepts in acute mountain sickness and high-altitude cerebral edema: from the molecular to the morphological. Cell Mol Life Sci. 2009, 66 (22): 3583-3594. DOI: 10.1007/s00018-009-0145-9.

3. Adeva M.M., Souto G., Donapetry C., Portals M., Rodriguez A., Lamas D. Brain edema in diseases of different etiology. Neurochem Int. 2012, 61 (2): 166-174. DOI: 10.1016/j.neuint.2012.05.007

4. Wang Y.F., Parpura V. Astroglial modulation of hydromineral balance and cerebral edema. Front Mol Neurosci. 2018, 11: 204. DOI: 10.3389/fnmol.2018.00204.

5. Slivka A., Murphy E., Horrocks L. Cerebral edema after temporary and permanent middle cerebral artery occlusion in the rat. Stroke. 1995, 26 (6): 1061-1065. DOI: 10.1161/01.str.26.6.1061.

6. Keep R.F., Jones H.C., Drewes L.R. This was the year that was: brain barriers and brain fluid research in 2019. Fluids Barriers CNS. 2020, 17(1): 20. DOI: 10.1186/s12987-020-00181-9.

7. Krueger M., Mages B., Hobusch C., Michalski D. Endothelial edema precedes blood-brain barrier breakdown in early time points after experimental focal cerebral ischemia. Acta Neuropathologica Communications. 2019, 7: 17. https: //doi.org/10.1186/s40478-019-0671-0

8. Ding Y., Liu J., Xu Y., Dong X., Shao B. Evolutionary adaptation of Aquaporin-4 in Yak (Bos grunniens) brain to high-altitude hypoxia of Qinghai-tibetan plateau. High altitude medicine & biology. 2020, 21 (2). DOI: 10.1089/ham.2019.0076

9. Hinzman J.M., DiNapoli V.A., Mahoney E.J., Gerhardt G.A., Hartings J.A. Spreading depolarizations mediate excitotoxicity in the development of acute cortical lesions. Exp Neurol. 2015, 267: 243-253. DOI: 10.1016/j.expneurol.2015.03.014.

10. Dreier J.P., Lemale C.L., Kola V., Friedman A., Schoknecht K. Spreading depolarization is not an epiphenomenon but the principal mechanism of the cytotoxic edema in various gray matter structures of the brain during stroke. Neuropharmacol. 2018, 15 (134): 189-207. DOI: 10.1016/j.neuropharm.2017.09.027.

11. Kvitnitskiy-Ryzhov Yu.N. Swelling and swelling of the brain. Kiev: «Zdorov'ya», 1978: 184 [In Russ.].

12. Ito U., Hakamata Y., Kawakami E., Oyanagi K. Degeneration of astrocytic processes and their mitochondria in cerebral cortical regions peripheral to the cortical infarction: heterogeneity of their disintegration is closely associated with disseminated selective neuronal necrosis and maturation of injury. Stroke. 2009, 40 (6): 2173-2181. DOI: 10.1161/STROKEAHA.108.534990.

13. Morgun A.V., Malinovskaya N.A., Komleva Yu.K., Lopatina O.L., Kuvacheva N.V., Panina Yu.A., Taranushenko T.E., Solonchuk R.Yu., Salmina A.B. Structural and functional heterogeneity of astrocytes in the brain: role in neurodegeneration and neuroinflammation. Byulleten' sibirskoy meditsiny. 2014, 13 (5): 138-148 [In Russ.]. DOI: 10.20538/1682-0363-2014-5-138-148

14. Stepanov A.S., Akulinin V.A., Mysik A.V., Stepanov S.S., Avdeev D.B. Neuro-Glio-Vascular Complexes of the Brain After Acute Ischemia. Obshchaya Reanimatologiya=General Reanimatology. 2017, 13 (6): 6-17. [In Russ.]. DOI: 10.15360/1813-9779-2017-6-6-17

15. Pekny M., Pekna M. Astrocyte reactivity and reactive astrogliosis: costs and benefits. Physiol Rev. 2014, 94: 1077-1098. DOI: 10.1152/physrev.00041.2013

16. Bai J., Lyden P.D. Revisiting cerebral postischemic reperfusion injury: new insights in understanding reperfusion failure, hemorrhage, and edema. Internation. J. Stroke. 2015, 10 (2): 143-152. DOI: 10.1111/ijs.12434

17. Von Kummer R., Dzialowski I. Imaging of cerebral ischemic edema and neuronal death. Neuroradiology. 2017; 59 (6): 545-553. DOI: 10.1007/s00234-017-1847-6.

18. Babkina A.S., Baeva A.A., Bashirova A.R., Blagonravov M.L., Golubev A.M., Grebenchikov O.A., Grechko A.V., Ershov A.V., Zakharchenko V.E., KuzovlevA.N., Muslimov B.G., Ostrova I.V., rerepelitsa S.A., retrova M.V., Romanova O.L., Saidov Sh.Kh., Silachev D.N., Sundukov D.V., Telipov I.N., Usmanov E.Sh., Khadzhieva M.B., Churilov A.A., Shabanov A.K., Shevelev O.A. Biological markers of damage and regeneration of the central nervous system. Moscow: OOO «VTsi»; 2021: 432. ISBN 9785604415900 [In Russ.].

19. Shi K., Tian De-Cai, Li Zhi-Guo, Ducruet A.F., Lawton M.T., Shi Fu-Dong. Global brain inflammation in stroke. Lancet Neurol. 2019; 18 (11): 1058-1066. DOI: 10.1016/S1474-4422(19)30078-X.

20. Paxinos G., Watson C. The Rat Brain in Stereotaxic Coordinates. 5th ed. San Diego: Elsevier Academic Press. 2005.

21. Herbert A.D., Carr A.M., Hoffmann E. FindFoci: a focus detection algorithm with automated parameter training that closely matches human assignments, reduces human inconsistencies and increases speed of analysis. PLoS One. 2014; 9 (12): e114749. DOI: 10.1371/journal.pone.0114749.

22. Bolon B., Garman R., Jensen K., Krinke G., Stuart B. Best practices approach to neuropathologic assessment in developmental neurotoxicity testing. Toxicol. rathol. 2006; 34 (3): 296-313. DOI: 10.1080/01926230600713269

23. Ooigawa H., Nawashiro H., Fukui S., Otani N, Osumi A, Toyooka T, Shima K. The fate of Nissl-stained dark neurons following traumatic brain injury in rats: difference between neocortex and hippocampus regarding survival rate. Acta Neuropathol. 2006; 112 (4): 471-481. DOI: 10.1007/s00401-006-0108-2

24. Borovikov V. Statistica. The art of analyzing data on a computer. Publishing House «Piter»; 2003: 2nd ed.: 688 [In Russ.].

25. Volkova D.A., Kositsyn N.S., Goloborod'ko E.V., Loginova N.A., Svinov M.M. Electrophysiological correlations of morphological restructuring in experimental local ischemia of different severity in the rat sensorimotor cortex. Byulleten' eksperimental'noy biologii i meditsiny= Bulletin of Experimental Biology and Medicine. 2013; 155 (2): 233236 [In Russ.]. DOI: 10.1007/s10517-013-2128-y

26. Stepanov S.S., Akulinin V.A., Avdeev D.B., Stepanov A.S., Gorbunova A.V. Reorganization of astrocytes of the neocortex of white rats after 20-minute occlusion of the common carotid arteries Rossijskij fiziologicheskij zhurnal im. I.M. Sechenova. 2019; 105 (5): 578-590 [In Russ.]. DOI: 10.1134/S086981391905011X

27. Semchenko V.V., Stepanov S.S., Bogolepov N.N. Brain synaptic plasticity (fundamental and applied aspects). M.: Direct Media; 2014 [In Russ.].