Ventral tegmental area/substantia nigra and prefrontal cortex rodent organotypic brain slices as an integrated model to study the cellular changes induced by oxygen/glucose deprivation and reperfusion: Effect of neuroprotective agents

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• 作者：Laura Colombo^a ; ¹ ; ^{laura.colombo982@gmail.com" class="auth_mail} ; Chiara Parravicini^a ; ¹ ; ^{chiara.parravicini@unimi.it" class="auth_mail} ; Davide Lecca^a ; ¹ ; ^{davide.lecca@unimi.it" class="auth_mail} ; Elena Dossi^b ; ^{elenadossi@gmail.com" class="auth_mail} ; Claudia Heine^c ; ^d ; ^{Claudia.Heine@medizin.uni-leipzig.de" class="auth_mail} ; Mauro Cimino^e ; ^{mauro.cimino@uniurb.it" class="auth_mail} ; Enzo Wanke^b ; ^{enzo.wanke@unimib.it" class="auth_mail} ; Peter Illes^c ; ^{Peter.Illes@medizin.uni-leipzig.de" class="auth_mail} ; Heike Franke^c ; ^{Heike.Franke@medizin.uni-leipzig.de" class="auth_mail} ; Maria P. Abbracchio^a ; ^{mariapia.abbracchio@unimi.it" class="auth_mail}
• 关键词：CTR ; control ; DIV ; days in vitro ; FJB ; Fluoro-Jade B ; LDH ; lactate dehydrogenase ; Iba1 ; ionized calcium binding adaptor molecule 1 ; IB4 ; Isolectin B4 ; MTT ; 3-(4 ; 5-dimethylthiazol-2-yl)-2 ; 5-diphenyltetrazolium bromide ; M/M ; macrophages/microglia ; OD ; optical density ; OGD ; oxygen and glucose deprivation ; OGD/R ; oxygen and glucose deprivation and reperfusion ; PI ; propidium iodide ; PPADS ; pyridoxalphosphate-6-azo-benzene-2&rsquo ; 4&rsquo ; -disulfonic acid ; RT ; room temperature ; PBS ; phosphate buffer
• 刊名：Neurochemistry International
• 出版年：January, 2014
• 年：2014
• 卷：66
• 期：Complete
• 页码：43-54
• 全文大小：3254 K

文摘

Unveiling the roles of distinct cell types in brain response to insults is a partially unsolved challenge and a key issue for new neuroreparative approaches. In vivo models are not able to dissect the contribution of residential microglia and infiltrating blood-borne monocytes/macrophages, which are fundamentally undistinguishable; conversely, cultured cells lack original tissue anatomical and functional complexity, which profoundly alters reactivity. Here, we tested whether rodent organotypic co-cultures from mesencephalic ventral tegmental area/substantia nigra and prefrontal cortex (VTA/SN-PFC) represent a suitable model to study changes induced by oxygen/glucose deprivation and reperfusion (OGD/R). OGD/R induced cytotoxicity to both VTA/SN and PFC slices, with higher VTA/SN susceptibility. Neurons were highly affected, with astrocytes and oligodendrocytes undergoing very mild damage. Marked reactive astrogliosis was also evident. Notably, OGD/R triggered the activation of CD68-expressing microglia and increased expression of Ym1 and Arg1, two markers of 鈥渁lternatively鈥?activated beneficial microglia. Treatment with two well-known neuroprotective drugs, the anticonvulsant agent valproic acid and the purinergic P2-antagonist PPADS, prevented neuronal damage. Thus, VTA/SN-PFC cultures are an integrated model to investigate OGD/R-induced effects on distinct cells and easily screen neuroprotective agents. The model is particularly adequate to dissect the microglia phenotypic shift in the lack of a functional vascular compartment.