Olson J E, Li G Z
Department of Emergency Medicine, Wright State University School of Medicine, Dayton, Ohio, USA.
Adv Exp Med Biol. 2000;483:213-8. doi: 10.1007/0-306-46838-7_23.
Taurine transport is important for volume regulation of cultured neurons and astroglial cells. Both cell types utilize similar mechanisms for taurine accumulation and efflux. However, taurine lost from cerebellar Purkinje cells in vivo is accumulated by adjacent astrocytes during hypoosmotic hyponatremia. To examine mechanisms for transfer of taurine between cell types, we measured relative sensitivities of taurine loss from cultured neurons and astrocytes. Primary cultures of hippocampal neurons and astrocytes were grown from embryonic and neonatal rat brain, respectively. Neurons were used after 10-14 days in culture. Astrocytes were used after 14 days in culture and were grown in the same culture medium used for neurons for 3 days prior to experimentation. Cells were incubated at 37 degrees C for 30 min in isoosmotic (290 mOsm) phosphate-buffered saline (PBS). The PBS was then changed to fresh isoosmotic or to hypoosmotic PBS (270 mOsm or 250 mOsm), made by reducing the NaCl concentration. Cell volume and taurine content were determined immediately before hypoosmotic exposure or 3, 15, or 30 min later. In isoosmotic PBS, astrocytes contained 162 +/- 18 nmol taurine/mg protein (mean +/- SEM), equivalent to an intracellular concentration of 30.2 +/- 2.1 mM. No taurine loss was detectable after 3 or 15 min in either hypoosmotic PBS, but after 30 min in 270 or 250 mOsm PBS, astrocyte taurine was reduced by 8.0% or 22.2%, respectively. Neurons initially contained 114 +/- 13 nmol taurine/mg protein, equivalent to an intracellular taurine concentration of 22.2 +/- 2.5 mM. After 3 min of exposure to 270 or 250 mOsm PBS, the cells had lost 17 +/- 5% or 25 +/- 4% of their taurine content, respectively. Cell volumes of each cell type were similarly affected by hypoosmotic exposure. We conclude that taurine loss from cultured hippocampal neurons is more sensitive to osmotic swelling than taurine loss from cultured hippocampal astrocytes. This characteristic, if present in cells of the hippocampus in vivo, could lead to net transfer of taurine from neurons to glial cells during pathological conditions which cause cell swelling.
牛磺酸转运对于培养的神经元和星形胶质细胞的容积调节很重要。这两种细胞类型利用相似的机制进行牛磺酸的积累和流出。然而,在低渗性低钠血症期间,体内小脑浦肯野细胞丢失的牛磺酸会被相邻的星形胶质细胞积累。为了研究牛磺酸在不同细胞类型之间转移的机制,我们测量了培养的神经元和星形胶质细胞中牛磺酸丢失的相对敏感性。海马神经元和星形胶质细胞的原代培养物分别从胚胎和新生大鼠脑分离培养而来。培养10 - 14天后使用神经元。培养14天后使用星形胶质细胞,在实验前3天,将其置于与神经元相同的培养基中培养。细胞在37℃下于等渗(290 mOsm)磷酸盐缓冲盐水(PBS)中孵育30分钟。然后将PBS更换为新鲜的等渗或低渗PBS(270 mOsm或250 mOsm,通过降低NaCl浓度制成)。在低渗暴露前或暴露3、15或30分钟后立即测定细胞容积和牛磺酸含量。在等渗PBS中,星形胶质细胞含有162±18 nmol牛磺酸/毫克蛋白(平均值±标准误),相当于细胞内浓度为30.2±2.1 mM。在任何一种低渗PBS中,3或15分钟后均未检测到牛磺酸丢失,但在270或250 mOsm PBS中30分钟后,星形胶质细胞中的牛磺酸分别减少了8.0%或22.2%。神经元最初含有114±13 nmol牛磺酸/毫克蛋白,相当于细胞内牛磺酸浓度为22.2±2.5 mM。暴露于270或250 mOsm PBS 3分钟后,细胞分别丢失了17±5%或25±4%的牛磺酸含量。每种细胞类型的细胞容积受到低渗暴露的影响相似。我们得出结论,培养的海马神经元中牛磺酸的丢失比培养的海马星形胶质细胞中牛磺酸的丢失对渗透性肿胀更敏感。如果在体内海马细胞中存在这种特性,那么在导致细胞肿胀的病理条件下,可能会导致牛磺酸从神经元向神经胶质细胞的净转移。