Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, NV, USA.
Neurogastroenterol Motil. 2013 Sep;25(9):e609-20. doi: 10.1111/nmo.12174. Epub 2013 Jun 30.
Purinergic signaling provides regulation of colonic motility. Smooth muscle cells (SMC), interstitial cells of Cajal (ICC), and platelet-derived growth factor receptor α-positive (PDGFRα(+) ) cells are electrically coupled and form a functional (SIP) syncytium that constitutes the receptive field for motor neurotransmitters in the tunica muscularis. Each cell type in the SIP syncytium has specialized functions in mediating motor neurotransmission. We compared gene transcripts for purinergic receptors and membrane-bound enzymes for purine degradation expressed by each cell type of the SIP syncytium.
Fluorescence-activated cell sorting (FACS) was used to purify SMC, ICC, and PDGFRα(+) cells from mixed cell populations of colonic muscles dispersed from reporter strains of mice with constitutive expression of green fluorescent proteins. Differential expression of functional groups of genes related to purinergic signaling was determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).
We detected marked phenotypic differences among SMC, ICC, and PDGFRα(+) cells. Substantial numbers of genes of importance in purinergic neurotransmission were enriched in PDGFRα(+) cells in relation to SMC and ICC. Notably, genes related to mediating effects and extracellular biotransformation of enteric purinergic inhibitory neurotransmitters were strongly expressed by PDGFRα(+) cells.
CONCLUSIONS & INFERENCES: Our results demonstrate differential expression of genes for proteins involved in purinergic signaling in the SIP syncytium. These results may further clarify the specific functions of each cell type, identify novel biomarkers for postjunctional cells, and provide hypotheses for further studies to understand the physiological roles of cells of the SIP syncytium.
嘌呤能信号提供了对结肠运动的调节。平滑肌细胞(SMC)、Cajal 间质细胞(ICC)和血小板衍生生长因子受体α阳性(PDGFRα(+))细胞电偶联并形成一个功能性(SIP)合胞体,构成了肌层中运动神经递质的感受野。SIP 合胞体中的每种细胞类型在介导运动神经传递中都具有特定的功能。我们比较了 SIP 合胞体中每种细胞类型表达的嘌呤能受体和嘌呤降解的膜结合酶的基因转录物。
荧光激活细胞分选(FACS)用于从具有组成型绿色荧光蛋白表达的报告株小鼠的结肠肌肉混合细胞群体中纯化 SMC、ICC 和 PDGFRα(+)细胞。通过定量逆转录聚合酶链反应(qRT-PCR)确定与嘌呤能信号相关的功能基因群的差异表达。
我们在 SMC、ICC 和 PDGFRα(+)细胞之间检测到明显的表型差异。在 PDGFRα(+)细胞中,与 SMC 和 ICC 相比,大量与嘌呤能神经传递有关的重要基因被富集。值得注意的是,与调节肠内嘌呤能抑制性神经递质的介导作用和细胞外生物转化有关的基因在 PDGFRα(+)细胞中强烈表达。
我们的研究结果表明,SIP 合胞体中参与嘌呤能信号的蛋白质的基因表达存在差异。这些结果可能进一步阐明每种细胞类型的特定功能,为后突触细胞鉴定新的生物标志物,并为进一步研究 SIP 合胞体细胞的生理功能提供假说。