In this study, we used tissue from 10 adult normal monkeys (Table 2 ) that had been kept in cryoprotection in Dr. Rausell′s tissue bank in the Department of Anatomy, Histology, and Neuroscience, Medical School, Autonomous University of Madrid (Madrid, Spain).
The tissue preparation has been described in detail in previous studies [49 (link)]. Briefly, monkeys were anesthetized with intramuscular ketamine and given an overdose of intravenous Nembutal. They were then perfused through the ascending aorta with normal saline followed by a solution of 4% paraformaldehyde and 1% glutaraldehyde in phosphate buffer (PB, 0.1 M, pH 7.4). The brain was removed and blocked. All the blocks were subsequently postfixed in 4% paraformaldehyde for 4 h, infiltrated with 30% sucrose in 0.1 M PB at 4 °C with gentle agitation, frozen in dry ice, and stored at −80 °C. The frozen blocks of the flattened cortex were cut tangential to the pia mater into 25–30 μm thick sections in a freezing sliding microtome. Alternate series of sections were collected in a sterile cryoprotectant solution. These series would later be processed for Nissl staining, immunohistochemistry and double immunofluorescence for MCT8/OATP1C1 and a number of cell/vessel markers.
The tissue preparation has been described in detail in previous studies [49 (link)]. Briefly, monkeys were anesthetized with intramuscular ketamine and given an overdose of intravenous Nembutal. They were then perfused through the ascending aorta with normal saline followed by a solution of 4% paraformaldehyde and 1% glutaraldehyde in phosphate buffer (PB, 0.1 M, pH 7.4). The brain was removed and blocked. All the blocks were subsequently postfixed in 4% paraformaldehyde for 4 h, infiltrated with 30% sucrose in 0.1 M PB at 4 °C with gentle agitation, frozen in dry ice, and stored at −80 °C. The frozen blocks of the flattened cortex were cut tangential to the pia mater into 25–30 μm thick sections in a freezing sliding microtome. Alternate series of sections were collected in a sterile cryoprotectant solution. These series would later be processed for Nissl staining, immunohistochemistry and double immunofluorescence for MCT8/OATP1C1 and a number of cell/vessel markers.
Free full text: Click here
