Morphological and Functional Adaptation of Basal Ganglia Neurons after Cerebral Ischemia

Author(s):  
ISABELLA ZINI ◽  
EMILIO MERLO PICH ◽  
VALENTINA SOLFRINI ◽  
EVA VALENTE ◽  
ANDREA FRASOLDATI ◽  
...  
2018 ◽  
pp. 126-131

Ligadura de arteria carótida común y exsanguinación como modelo experimental de isquemia cerebral focal en ratas Ligation of common carotid artery and exsanguination as experimental model of focal cerebral ischemia in rats  Luz Marina Guerrero Espino Universidad Nacional de Trujillo, Calle Salaverry N⁰545-Trujillo, Perú DOI: https://doi.org/10.33017/RevECIPeru2013.0017/ Resumen La isquemia cerebral   es el proceso fisiopatológico caracterizado por la disfunción de una porción del tejido cerebral secundario a la disminución del flujo en una arteria cerebral concreta.  La mejor herramienta que se dispone hoy para el estudio de la fisiopatología de la isquemia cerebral son los modelos experimentales, que permiten abordar de manera simple el cuidado de esta afección que se caracteriza por su complejidad y heterogeneidad. Considerando que  las investigaciones sobre este campo es ilimitada, no solo por la trascendencia sino por el costo social de la enfermedad  ,el estudio de nuevas metodologías experimentales permitirán alcanzar mejores herramientas con rigor científico para el conocimiento fisiopatológico, terapéutico y preventivo de la enfermedad isquémica cerebral.Por lo que nos propusimos  determinar si la ligadura de arteria carótida común izquierda y exsanguinación     puede ser utilizada como modelo de isquemia cerebral focal en ratas. Se usaron 36 ratas, machos de 8 a 9 semanas de edad, con 200 ± 20gr de peso, que se mantuvieron en condiciones estándar de iluminación, ciclos diarios de 12 horas de luz y 12 de oscuridad, a una temperatura ambiente de 24-25°C, los especímenes fueron distribuidas en dos grupos: Grupo Sham o seudo operado y Grupo Experimental (operado). Se procedió a la sangría por punción cardiaca, y se extrajo de cada rata el 10% del volumen de sangre circulante. Bajo condiciones de asepsia y antisepsia, se realizó una incisión en la línea media del cuello y diseccionó la piel, tejido subcutáneo y músculo hasta lograr identificar la arteria carótida común (ACC) izquierda y se ligó con sutura Nylon 5-0 para interrumpir la circulación sanguínea de manera ipsilateral, se suturó la herida y se administró lidocaína, finalmente se dejó descansar al animal a una temperatura ligeramente mayor a la ambiental en cámaras previamente confeccionadas. Luego de 24 horas de aplicar la cirugía, se sacrificaron los especímenes por sobredosis de anestésico y se realizó la remoción del cerebro y el análisis histopatológico de secciones coronales en tres regiones neocortex, hipocampo y ganglios basales, por ser las áreas más vulnerables a daño en una lesión isquémica. Se encontró que las mayores características neuronales que indican daño celular se manifestaron en el Grupo Experimental con un p‹0.01 y la zona con mayor incidencia de daño es el ganglio basal, p ‹0.01. Se concluyó que la ligadura de arteria carótida común izquierda y ex sanguinación produce daños neuronales localizados y la zona más afectada es el ganglio basal. Descriptores:  isquemia cerebral focal, ligadura arteria carótida común ex sanguinación. Abstract Cerebral ischemia is the pathophysiological process characterized by dysfunction of a portion of brain tissue secondary to decreased flow in specific brain artery. The best tool available today to study the pathophysiology of cerebral ischemia are experimental models, which allow a simple way to address the care of the condition which is characterized by complexity and heterogeneity. Whereas research on this field is unlimited, not only because of the importance but by the social cost of the disease, the study of new experimental methodologies to achieve better tools for scientific rigor in pathophysiological knowledge, treatment and prevention of cerebral ischemic disease. So we set out to determine whether the common carotid artery ligation and ex sanguination left can be used as a model of focal cerebral ischemia in rats. We used 36 male rats of 8 to 9 weeks of age, with 200 ± 20g of weight, were maintained in standard lighting conditions, daily cycles of 12 hours of light and 12 of darkness were used at an ambient temperature of 24-25 ° C, the specimens were divided into two groups: Sham Groups and Experimental Group (operated). We proceeded to the bleeding via cardiac puncture, and extracted from each rat 10% of circulating blood volume. Under aseptic and antiseptic conditions, an incision was made in the midline of the neck and dissected the skin, subcutaneous tissue and muscle to be able to identify the left common carotid artery (CCA) and ligated with 5-0 nylon suture to interrupt circulation way of ipsilateral blood, the wound was sutured and received lidocaine finally allowed to rest the animal at a slightly higher environmental chambers previously made temperature. After 24 hours of surgery applied, the specimens were sacrificed by overdose of anesthetic and removal of the brain and histopathological analysis of coronal sections in three regions neocortex, hippocampus and basal ganglia was performed, being the most vulnerable areas to damage ischemic injury. It was found that the major characteristics that indicate neuronal cell damage demonstrated in the experimental group with p <0.01 and the area with the highest incidence of injury is the basal ganglia, p <0.01. It was concluded that ligation of left common carotid artery and ex sanguinación produces localized neuronal damage and the most affected area is the basal ganglia. Keywords: Focal cerebral ischemia, ligation of left common carotid artery and ex sanguination


Author(s):  
Fukhi Toshiya ◽  
Hasegawa YuKihiro ◽  
Seriyama Shinya ◽  
Takeuchi Toru ◽  
Sugita Koujiro ◽  
...  

ABSTRACT:Four patients presented with hemiballism-hemichorea as a clinical manifestation of white matter ischemia. These patients illustrate “positive” motor phenomena rather than limb weakness as a consequence of cerebral ischemia. In each patient, the involuntary movements disappeared following worsening of paresis. Subcortical white matter infarction in three patients and hemodynamic hypo-perfusion in the cerebral hemisphere contralateral to dyskinetic movements were possible causes. Neuroradiologically, none had pathological changes in the vicinity of the subthalamic nucleus. We presume from these observations that ischemia of the subcortical white matter, without involvement of the basal ganglia or the subthalamic nucleus, may cause hemiballism-hemichorea


2002 ◽  
Vol 22 (5) ◽  
pp. 526-533 ◽  
Author(s):  
Gerhard F. Hamann ◽  
Martin Liebetrau ◽  
Helge Martens ◽  
Dorothe Burggraf ◽  
Christian U. A. Kloss ◽  
...  

To define the location and extent of microvascular damage of the basal lamina after cerebral ischemia and reperfusion in rats, the authors subjected animals (n = 16) to 3 hours of focal cerebral ischemia and 24 hours of reperfusion using the suture middle cerebral artery occlusion model; sham-operated animals served as controls (n = 6). The Western blot technique was used to define the collagen type IV protein content in various brain regions, whereas immunohistochemistry identified microvascular basal lamina loss (anticollagen type IV staining). The extent of damage was quantified by automatic morphometric video-imaging analysis. Statistical analysis was based on the Mann-Whitney test and the paired Student's t-test. The ischemic hemisphere showed a reduction of the collagen type IV protein content after ischemia and reperfusion in the Western blot (reduction compared with the nonischemic side: total hemisphere, 33% ± 6%; basal ganglia, 49% ± 4%; cortex, 25% ± 7%; P < 0.01). There was also a decrease in the number of cerebral microvessels between the ischemic and nonischemic hemispheres (16% ± 3%), cortical (14% ± 3%), and basal ganglia areas (21% ± 4%) ( P < 0.01). Besides a reduction of the vessel number, there was also a loss in basal lamina antigen-positive stained area in ischemic areas (hemisphere, 20% ± 2%; cortex, 8% ± 3%; basal ganglia, 31% ± 3%; P < 0.001). Cortical areas had a less pronounced basal lamina loss than basal ganglia ( P < 0.05). For the first time, microvascular basal lamina damage, indicated by collagen type IV loss, is proven in rats by biochemical and morphometric analysis. These changes are comparable with those found in nonhuman primates. The authors report novel data regarding microvascular ischemic changes in the cortex. These data provide a basis for future experiments to determine the mechanisms of ischemic microvascular damage and to devise new therapeutic strategies.


2000 ◽  
Vol 20 (2) ◽  
pp. 337-342 ◽  
Author(s):  
T. Pfefferkorn ◽  
B. Staufer ◽  
M. Liebetrau ◽  
G. Bültemeier ◽  
M. R. Vosko ◽  
...  

In focal cerebral ischemia the plasminogen-plasmin system plays a role in the fibrinolysis of vessel-occluding clots and also in the proteolysis of extracellular matrix components, which potentially contributes to brain edema and bleeding complications. The authors investigated the plasminogen activation after middle cerebral artery occlusion with and without reperfusion (reperfusion intervals 9 and 24 hours) in rats by histologic zymography and compared areas of increased plasminogen activation to areas of structural injury, which were detected immunohistochemically. After 3 hours of ischemia, increased plasminogen activation was observed in the ischemic hemisphere. The affected area measured 5.2% ± 8.5% and 19.4% ± 30.1% of the total basal ganglia and cortex area, respectively. Reperfusion for 9 hours after 3 hours of ischemia led to a significant expansion of plasminogen activation in the basal ganglia (68.8% ± 42.2%, P < 0.05) but not in the cortex (43.0% ± 34.6%, P = 0.394). In the basal ganglia, areas of increased plasminogen activation were related to areas of structural injury ( r = 0.873, P < 0.001). No such correlation was found in the cortex ( r = 0.299, P = 0.228). In this study, increased plasminogen activation was demonstrated early in focal cerebral ischemia. This activation may promote early secondary edema formation and also secondary hemorrhage after ischemic stroke.


2003 ◽  
Vol 89 (06) ◽  
pp. 1072-1080 ◽  
Author(s):  
Dorothe Burggraf ◽  
Helge Martens ◽  
Gabriele Jäger ◽  
Gerhard Hamann

SummaryWhile recombinant tissue plasminogen activator (rt-PA) is successfully used in human ischemic stroke, it may also cause hemorrhagic complications. Animal experiments have shown that hemorrhages are related to microvascular basal lamina damage. We investigated the effects of different doses of rt-PA on the brain microvasculature. Experimental cerebral ischemia in rats was induced for 3 h and followed by 24 h reperfusion (suture model). Each group of rats (n = 6) received either treatment (0.9, 9, or 18 mg rt-PA/kg body weight) or saline (control group) at the end of ischemia. The loss of microvascular basal lamina antigen collagen type IV was measured by Western blot of the ischemic and non-ischemic basal ganglia and cortex. Compared with the contralateral non-ischemic area, collagen type IV was significantly reduced in the ischemic area: (basal ganglia/cortex) 43% +/- 9% / 64% +/- 4 %. Low/moderate doses of rt-PA had a protective effect: 0.9 mg 79% +/- 3% / 89% +/-6%, 9 mg 72% +/- 9%/ 81% +/- 12% (p < 0.05). Higher doses of rt-PA (18 mg) had a similar effect as seen in untreated controls: 57% +/- 11% / 59% +/- 9% (p < 0.05, Anova). MMP-9 and MMP-2, measured by gelatine zymography, steadily increased over higher doses of rt-PA: MMP-9 (basal ganglia/cortex): control 115% +/- 4% / 123% +/- 3% compared with 18 mg rt-PA 146% +/- 5%/ 162% +/- 6% (p < 0.05) and MMP-2: control 109% +/-4%/ 116% +/- 5% and 18 mg rt-PA 222% +/- 15%/ 252% +/- 2% (p < 0.05). Low to moderate doses of rt-PA protect the microvascular basal lamina, whereas high doses of rt-PA have the opposite effect, probably due to increased coactivation of MMP-2 and MMP-9.


2002 ◽  
Vol 22 (12) ◽  
pp. 1452-1452

In the article “Microvascular Basal Lamina Injury After Experimental Focal Cerebral Ischemia and Reperfusion in the Rat,” by Gerhard F. Hamann, Martin Liebetrau, Helge Martens, Dorothe Burggraf, Christian U. A. Kloss, Gundula Bültemeier, Natalie Wunderlich, Gabriele Jäger, and Thomas Pfefferkorn, published in the May 2002 issue (pp 526–533) of the Journal of Cerebral Blood Flow & Metabolism, an error was overlooked in the Abstract. The correct numbers regarding microvascular damage are given in the Results section, but the Abstract should read as follows: “The ischemic hemisphere showed a reduction of the collagen type IV protein content after ischemia and reperfusion in the Western blot (reduction compared with the nonischemic side: total hemisphere, 33% ± 6%; basal ganglia, 25% ± 7%; cortex, 49% ± 4%; P < 0.01). There was also a decrease in the number of cerebral microvessels in the ischemic and nonischemic hemispheres (20% ± 2%), cortical (8% ± 3%), and basal ganglia areas (31% ± 3%) ( P < 0.001). Besides a reduction of the vessel number, there was also a loss in basal lamina antigen-positive stained area in ischemic areas (hemisphere, 16% ± 3%; cortex, 14% ± 3%; basal ganglia, 21% ± 4%; P < 0.01). …” This change does not influence the interpretation or the conclusions of the paper: microvascular basal lamina damage occurs in rats after experimental cerebral ischemia and reperfusion and is comparable to that observed in nonhuman primates. In addition, the conclusion that microvessel walls in cortical areas are less damaged than those in basal ganglia remains unchanged.


1985 ◽  
Vol 5 (4) ◽  
pp. 529-536 ◽  
Author(s):  
Kazumi Yamamoto ◽  
Toshiki Yoshimine ◽  
Takehiko Yanagihara

Regional cerebral ischemia was produced in the rabbit by unilateral transorbital occlusion of the middle cerebral artery (procedure I); the middle cerebral and azygos anterior cerebral or anterior communicating artery (procedure II); or the middle cerebral, azygos anterior cerebral or anterior communicating, and internal carotid artery (procedure III). Evolution of ischemic lesions was examined with the immunohistochemical reaction for tubulin. With procedure I, ischemic lesions did not become constantly visible for 6 h in the basal ganglia and for 8 h in the frontoparietal region of the cerebral cortex. With procedure II, it was shortened to 3 h in the basal ganglia and to 6 h in the cerebral cortex. With procedure III, the ischemic lesions were observed in 1 h both in the basal ganglia and in the cerebral cortex as loss of the reaction for tubulin in the neuropil, nerve cell bodies, and dendrites. The evidence of neuronal damage became apparent in the same areas later by staining with hematoxylin–eosin. The experimental model presented here may be suitable for investigation of the mechanism that shifts reversible ischemia to cerebral infarction and for evaluation of the effectiveness of pharmacological intervention.


Author(s):  
R.V.W. Dimlich ◽  
M.H. Biros

In severe cerebral ischemia, Purkinje cells of the cerebellum are one of the cell types most vulnerable to anoxic damage. In the partial (forebrain) global ischemic (PGI) model of the rat, Paljärvi noted at the light microscopic level that cerebellar damage is inconsistant and when present, milder than in the telencephalon, diencephalon and rostral brain stem. Cerebellar injury was observed in 3 of 4 PGI rats following 5 minutes of reperfusion but in none of the rats after 90 min of reperfusion. To evaluate a time between these two extremes (5 and 90 min), the present investigation used the PGI model to study the effects of ischemia on the ultrastructure of cerebellar Purkinje cells in rats that were sacrificed after 30 min of reperfusion. This time also was chosen because lactic acid that is thought to contribute to ischemic cell changes in PGI is at a maximum after 30 min of reperfusion.


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