Chavkin C, Bakhit C, Weber E, Bloom F E
Proc Natl Acad Sci U S A. 1983 Dec;80(24):7669-73. doi: 10.1073/pnas.80.24.7669.
The contents and molecular forms of five different prodynorphin-derived opioid peptides were compared in extracts of rat hippocampus by radioimmunoassay after C18-HPLC resolution. Dynorphin (Dyn) A(1-17) immunoreactivity (ir) and Dyn B-ir were heterogeneous in form; Dyn A(1-8)-ir, alpha-neoendorphin (alpha neo)-ir and beta-neoendorphin (beta neo)-ir each eluted as single homogeneous peaks of immunoreactivity. The fraction of immunoreactivity having the same retention as the appropriate synthetic standard was used to estimate the actual hippocampal content of each peptide. Comparison of these values showed that the concentrations of Dyn B, alpha neo, and Dyn A(1-8) were nearly equal, whereas both Dyn A(1-17) and beta neo were 1/5th to 1/10th the value of the other three. Calcium-dependent K+-stimulated release of these prodynorphin-derived opioids from hippocampal slices was detected. The stimulated rates of release were highest for Dyn B-ir followed by alpha neo-ir, then beta neo-ir and Dyn A(1-8)-ir with Dyn A(1-17)-ir lowest. The relative rates of stimulated release were in agreement with the relative proportions of peptide present within the tissue. This evidence of the presence and release of these opioid peptides considerably strengthens the hypothesis that this family of endogenous opioids plays a neurotransmitter role in the hippocampus.
采用C18高效液相色谱法分离后,通过放射免疫分析法比较了大鼠海马提取物中五种不同强啡肽衍生阿片肽的含量和分子形式。强啡肽(Dyn)A(1-17)免疫反应性(ir)和Dyn B-ir在形式上是异质的;Dyn A(1-8)-ir、α-新内啡肽(α neo)-ir和β-新内啡肽(β neo)-ir各自以单一的免疫反应性均匀峰洗脱。具有与相应合成标准品相同保留时间的免疫反应性部分用于估计每种肽在海马中的实际含量。这些值的比较表明,Dyn B、α neo和Dyn A(1-8)的浓度几乎相等,而Dyn A(1-17)和β neo的浓度是其他三种的1/5至1/10。检测到这些强啡肽衍生的阿片类物质从海马切片中钙依赖性钾刺激释放。刺激释放率最高的是Dyn B-ir,其次是α neo-ir,然后是β neo-ir和Dyn A(1-8)-ir,Dyn A(1-17)-ir最低。刺激释放的相对速率与组织中存在的肽的相对比例一致。这些阿片肽存在和释放的证据大大加强了这一假说,即这一家族的内源性阿片类物质在海马中发挥神经递质作用。