Black I B, Chikaraishi D M, Lewis E J
Brain Res. 1985 Jul 22;339(1):151-3. doi: 10.1016/0006-8993(85)90635-3.
To begin examining molecular mechanisms underlying trans-synaptic regulation, tyrosine hydroxylase (TH) and its messenger RNA (mRNA) were examined in the superior cervical sympathetic ganglion (SCG) of adult rats. Basal levels of TH mRNA were detectable in control ganglia by RNA dot hybridization, using the 32P nick translated PstI-KpnI restriction fragment of pTH.4 as a probe. Reserpine induced a 3-fold rise in TH activity per microgram protein, and a simultaneous 3-fold increase in ganglion TH mRNA. As expected, ganglion decentralization (denervation) prevented the trans-synaptic induction of TH. In addition, decentralization prevented the increase in TH mRNA, suggesting that the increase in message was dependent on trans-synaptic stimulation. Northern blot analysis indicated that the cDNA (complementary DNA) probe hybridized to a single band of approximately 1900 nucleotides, which was the same size in all ganglia. Our observations indicate that induction of TH is associated with a trans-synaptic increase in mRNA coding for the enzyme. Consequently, trans-synaptic increases in impulse activity may induce TH by increasing neuronal levels of TH mRNA in the SCG.
为了开始研究跨突触调节的分子机制,我们检测了成年大鼠颈上神经节(SCG)中的酪氨酸羟化酶(TH)及其信使核糖核酸(mRNA)。使用pTH.4的32P缺口平移PstI - KpnI限制性片段作为探针,通过RNA斑点杂交在对照神经节中可检测到TH mRNA的基础水平。利血平使每微克蛋白质的TH活性升高3倍,同时神经节TH mRNA也同步增加3倍。正如预期的那样,神经节去中枢化(去神经支配)阻止了TH的跨突触诱导。此外,去中枢化阻止了TH mRNA的增加,这表明信息的增加依赖于跨突触刺激。Northern印迹分析表明,cDNA(互补DNA)探针与一条约1900个核苷酸的单带杂交,所有神经节中的该条带大小相同。我们的观察结果表明,TH的诱导与编码该酶的mRNA的跨突触增加有关。因此,冲动活动的跨突触增加可能通过增加SCG中神经元TH mRNA的水平来诱导TH。