Nörenberg W, Illes P, Gebicke-Haerter P J
Department of Pharmacology, University of Freiburg, Germany.
Glia. 1994 Mar;10(3):165-72. doi: 10.1002/glia.440100303.
Human brain macrophages (microglia) have been isolated from mixed brain cell cultures initiated from explants of neurosurgical adult human tissue in one step according to a method developed for rat microglia. Cells were characterized enzyme-histochemically (NDPase) in mixed and immunocytochemically (anti-CD 14) in mixed and isolated cultures. Purified cells were used to investigate in more detail membrane currents by the patch clamp technique. In 14 cells microdialyzed with a standard, K(+)-containing intracellular solution there was no indication for a hyperpolarization-induced K(+)-inward current characteristic for newborn rat microglia. However, in 12 cells depolarizing pulses initiated a rapidly inactivating inward current which was followed by an outward current (in 4 cells). The outward current appeared to be carried by K+, since it was absent in another 18 cells, recorded by micropipettes containing Cs+ instead of K+ as the main intracellular cation. The depolarization-induced inward current persisted under these conditions. This current was inhibited by tetrodotoxin (5 microM) and by substitution of Na+ by choline in the bath solution. It is suggested that this Na(+)-current is specifically expressed in macrophages derived from adult brain.
人类脑巨噬细胞(小胶质细胞)已根据一种为大鼠小胶质细胞开发的方法,从神经外科成人脑组织外植体起始的混合脑细胞培养物中一步分离得到。细胞在混合培养物中通过酶组织化学(NDPase)进行表征,在混合培养物和分离培养物中通过免疫细胞化学(抗CD14)进行表征。纯化的细胞用于通过膜片钳技术更详细地研究膜电流。在用含钾标准细胞内溶液进行微透析的14个细胞中,没有迹象表明存在新生大鼠小胶质细胞特有的超极化诱导钾内向电流。然而,在12个细胞中,去极化脉冲引发了一种快速失活的内向电流,随后是外向电流(4个细胞中)。外向电流似乎由钾携带,因为在另外18个细胞中未出现这种电流,这些细胞是用含铯而非钾作为主要细胞内阳离子的微吸管记录的。在这些条件下去极化诱导的内向电流持续存在。该电流被河豚毒素(5微摩尔)以及在浴液中用胆碱替代钠所抑制。提示这种钠电流在源自成人大脑的巨噬细胞中特异性表达。