Garcia Veronica J, Daur Nelly, Temporal Simone, Schulz David J, Bucher Dirk
Whitney Laboratory for Marine Bioscience, University of Florida, St. Augustine, Florida 32080, Department of Neuroscience, College of Medicine, University of Florida, Gainesville, Florida 32610.
Federated Department of Biological Sciences, New Jersey Institute of Technology and Rutgers University, Newark, New Jersey 07102, and.
J Neurosci. 2015 Apr 29;35(17):6786-800. doi: 10.1523/JNEUROSCI.0171-15.2015.
We studied the relationship between neuropeptide receptor transcript expression and current responses in the stomatogastric ganglion (STG) of the crab, Cancer borealis. We identified a transcript with high sequence similarity to crustacean cardioactive peptide (CCAP) receptors in insects and mammalian neuropeptide S receptors. This transcript was expressed throughout the nervous system, consistent with the role of CCAP in a range of different behaviors. In the STG, single-cell qPCR showed expression in only a subset of neurons. This subset had previously been shown to respond to CCAP with the activation of a modulator-activated inward current (IMI), with one exception. In the one cell type that showed expression but no IMI responses, we found CCAP modulation of synaptic currents. Expression levels within STG neuron types were fairly variable, but significantly different between some neuron types. We tested the magnitude and concentration dependence of IMI responses to CCAP application in two identified neurons, the lateral pyloric (LP) and the inferior cardiac (IC) neurons. LP had several-fold higher expression and showed larger current responses. It also was more sensitive to low CCAP concentrations and showed saturation at lower concentrations, as sigmoid fits showed smaller EC50 values and steeper slopes. In addition, occlusion experiments with proctolin, a different neuropeptide converging onto IMI, showed that saturating concentrations of CCAP activated all available IMI in LP, but only approximately two-thirds in IC, the neuron with lower receptor transcript expression. The implications of these findings for comodulation are discussed.
我们研究了北方黄道蟹口胃神经节(STG)中神经肽受体转录本表达与电流反应之间的关系。我们鉴定出一种转录本,它与昆虫中的甲壳类心脏活性肽(CCAP)受体以及哺乳动物神经肽S受体具有高度的序列相似性。该转录本在整个神经系统中均有表达,这与CCAP在一系列不同行为中的作用相一致。在STG中,单细胞定量PCR显示仅在一部分神经元中有表达。先前已表明,除了一个例外,这部分神经元会通过调制激活内向电流(IMI)的激活对CCAP产生反应。在一种显示有表达但无IMI反应的细胞类型中,我们发现了CCAP对突触电流的调制作用。STG神经元类型中的表达水平差异较大,但某些神经元类型之间存在显著差异。我们在两种已鉴定的神经元——外侧幽门(LP)神经元和下心(IC)神经元中测试了IMI对CCAP应用的反应幅度和浓度依赖性。LP神经元的表达高出几倍,并且显示出更大的电流反应。它对低浓度CCAP也更敏感,并且在较低浓度下就表现出饱和,因为S形拟合显示其半最大有效浓度(EC50)值更小且斜率更陡。此外,用促胃液素(一种作用于IMI的不同神经肽)进行的阻塞实验表明,饱和浓度的CCAP激活了LP神经元中所有可用的IMI,但在受体转录本表达较低的IC神经元中仅激活了约三分之二。我们讨论了这些发现对共同调制的意义。