Baines D, Mallon B S, Love S
Department of Pathology and Microbiology, University of Bristol, UK.
Brain Res Mol Brain Res. 1992 Dec;16(3-4):330-8. doi: 10.1016/0169-328x(92)90243-5.
We have studied the effects of sodium butyrate on cell morphology and the expression of mRNAs encoding voltage-gated sodium channels in five neuronal cell lines, B35, B50, B65, B103 and B104, all derived from the rat CNS. The cells were grown in medium supplemented with 2.5 mM sodium-n-butyrate and examined daily by phase-contrast microscopy. Sodium butyrate caused slowing of cell division and the formation of longer and more highly branched cytoplasmic processes than were present in untreated cells. Expression of sodium channel mRNA was analysed by PCR with primers that allow the transcripts encoding the different types of sodium channel to be distinguished according to the lengths of the PCR products. The identity of the PCR products was confirmed by restriction enzyme digestion. Southern blotting and hybridization with internal radiolabelled probes. Prior to sodium butyrate treatment, expression of sodium channel mRNA was largely restricted to B50 and B104 cells: B50 cells showed expression of rat brain types I and II sodium channel and B104 cells expressed rat brain type III sodium channel. After treatment for 5 days with sodium butyrate, sodium channel mRNA was detected in all five cell lines. In addition to type I and type II sodium channel, B50 cells expressed rat brain type III sodium channel. These three types of sodium channel were also expressed by B35, B65 and B103 cells. Even after butyrate treatment, B104 cells expressed only type III sodium channel. The treatment also induced expression of rat skeletal muscle SkM1 sodium channel in B35 cells but only trace amounts in the other neuronal cell lines.
我们研究了丁酸钠对五种神经元细胞系（B35、B50、B65、B103和B104，均源自大鼠中枢神经系统）的细胞形态以及编码电压门控钠通道的mRNA表达的影响。将细胞培养在添加了2.5 mM正丁酸钠的培养基中，每天通过相差显微镜进行检查。丁酸钠导致细胞分裂减缓，并形成比未处理细胞中更长且分支更多的细胞质突起。通过PCR分析钠通道mRNA的表达，所用引物可根据PCR产物的长度区分编码不同类型钠通道的转录本。通过限制性内切酶消化、Southern印迹以及与内部放射性标记探针杂交来确认PCR产物的身份。在丁酸钠处理之前，钠通道mRNA的表达主要局限于B50和B104细胞：B50细胞显示出大鼠脑I型和II型钠通道的表达，而B104细胞表达大鼠脑III型钠通道。用丁酸钠处理5天后，在所有五种细胞系中均检测到钠通道mRNA。除了I型和II型钠通道外，B50细胞还表达大鼠脑III型钠通道。B35、B65和B103细胞也表达这三种类型的钠通道。即使在丁酸钠处理后，B104细胞也仅表达III型钠通道。该处理还诱导B35细胞中大鼠骨骼肌SkM1钠通道的表达，但在其他神经元细胞系中仅诱导出微量表达。
