Buccellato Francesca R, Foi Laura, Veber Daniela, Pravettoni Giulio, Scalabrino Giuseppe
Institute of General Pathology and Center of Excellence on Neurodegenerative Diseases, University of Milan, Milan, Italy.
Glia. 2004 Mar;45(4):406-11. doi: 10.1002/glia.10345.
In the present study, we hypothesized that cobalamin (Cbl) deficiency might affect astrocytes and oligodendrocytes of rat spinal cord (SC) differently. Radiolabeled Cbl ([Cyano-14C]cyano-Cbl) was used to investigate whether the in vitro uptake of Cbl is different in primary cultures of oligodendrocytes and astrocytes. In culture medium supplemented with serum that naturally contains Cbl, the time course of labeled Cbl uptake by neonatal oligodendrocytes had two peaks, at 8 h and 24 h. No uptake was observed when the same cells were cultured in a serum-free medium and consequently in the absence of Cbl. Oligodendrocytes isolated from adult rat SC showed no uptake under any of the tested conditions. Astrocytes isolated from adult Cbl-deficient and newborn rat SC, both cultured in a medium supplemented with serum, showed peak Cbl uptake at 8 h and 12 h, respectively, whereas those isolated from the SC of an adult normal rat cultured under the same conditions showed no uptake throughout the experimental period. Astrocytes isolated from normal, Cbl-deficient adult rats and newborns cultured in a serum-free medium not containing Cbl, showed a similar trend of Cbl uptake with a peak at 24 h. Oligodendrocytes isolated from Cbl-deficient rats showed no uptake when cultured in medium with or without serum. This study provides evidence for a difference in the uptake of labeled Cbl between rat SC astrocytes and oligodendrocytes in relation to (1) age of the donor SC, (2) Cbl status of the donor SC, and (3) Cbl deficiency in the incubation medium that facilitates Cbl uptake in neonatal and adult astrocytes.
在本研究中,我们假设钴胺素(Cbl)缺乏可能对大鼠脊髓(SC)的星形胶质细胞和少突胶质细胞产生不同影响。使用放射性标记的Cbl([氰基 - 14C]氰钴胺素)来研究少突胶质细胞和星形胶质细胞原代培养物中Cbl的体外摄取是否存在差异。在补充了天然含有Cbl的血清的培养基中,新生少突胶质细胞摄取标记Cbl的时间进程有两个峰值,分别在8小时和24小时。当相同细胞在无血清培养基中培养因而缺乏Cbl时,未观察到摄取现象。从成年大鼠脊髓分离的少突胶质细胞在任何测试条件下均未显示摄取。从成年Cbl缺乏大鼠和新生大鼠脊髓分离的星形胶质细胞,在补充血清的培养基中培养时,分别在8小时和12小时出现Cbl摄取峰值,而在相同条件下从成年正常大鼠脊髓分离的星形胶质细胞在整个实验期间均未显示摄取。在不含Cbl的无血清培养基中培养的正常、成年Cbl缺乏大鼠和新生大鼠的星形胶质细胞,显示出类似的Cbl摄取趋势,峰值在24小时。从Cbl缺乏大鼠分离的少突胶质细胞在含或不含血清的培养基中培养时均未显示摄取。本研究提供了证据,表明大鼠脊髓星形胶质细胞和少突胶质细胞在标记Cbl摄取方面存在差异,这与(1)供体脊髓的年龄、(2)供体脊髓的Cbl状态以及(3)孵育培养基中的Cbl缺乏有关,后者促进新生和成年星形胶质细胞摄取Cbl。