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.
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 "alternatively" 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.
Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan 20133, Italy. Electronic address: laura.colombo982@gmail.com.
Laura
L
Colombo
Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan 20133, Italy. Electronic address: chiara.parravicini@unimi.it.
Chiara
C
Parravicini
Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan 20133, Italy. Electronic address: davide.lecca@unimi.it.
Davide
D
Lecca
Department of Biotechnologies and Biosciences, University of Milano-Bicocca, Milano 20126, Italy. Electronic address: elenadossi@gmail.com.
Elena
E
Dossi
Universität Leipzig, Rudolf-Boehm-Institut für Pharmakologie und Toxikologie, Leipzig 04107, Germany; Translational Centre for Regenerative Medicine (TRM), University of Leipzig, Leipzig 04103, Germany. Electronic address: Claudia.Heine@medizin.uni-leipzig.de.
Claudia
C
Heine
Department of Biomolecular Sciences, University of Urbino, Urbino 61029, Italy. Electronic address: mauro.cimino@uniurb.it.
Mauro
M
Cimino
Department of Biotechnologies and Biosciences, University of Milano-Bicocca, Milano 20126, Italy. Electronic address: enzo.wanke@unimib.it.
Enzo
E
Wanke
Universität Leipzig, Rudolf-Boehm-Institut für Pharmakologie und Toxikologie, Leipzig 04107, Germany. Electronic address: Peter.Illes@medizin.uni-leipzig.de.
Peter
P
Illes
Universität Leipzig, Rudolf-Boehm-Institut für Pharmakologie und Toxikologie, Leipzig 04107, Germany. Electronic address: Heike.Franke@medizin.uni-leipzig.de.
Heike
H
Franke
Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, Milan 20133, Italy. Electronic address: mariapia.abbracchio@unimi.it.
Maria P
MP
Abbracchio
D000818:Animals; D000831:Animals, Newborn; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology; D018920:Coculture Techniques; D005947:Glucose / Q000378:metabolism*; D051379:Mice; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009924:Organ Culture Techniques; D010100:Oxygen / Q000378:metabolism*; D017397:Prefrontal Cortex / Q000187:drug effects / Q000378:metabolism*; D015424:Reperfusion / Q000379:methods; D013378:Substantia Nigra / Q000187:drug effects / Q000378:metabolism*; D017557:Ventral Tegmental Area / Q000187:drug effects / Q000378:metabolism*
D018696:Neuroprotective Agents; D005947:Glucose; D010100:Oxygen
Inflammation; Microglia; Neuroprotection; Neurotoxicity; Oxygen and glucose deprivation/reperfusion; PPADS