Department of Chemistry, University of Miami, 1301 Memorial Drive, Coral Gables, FL 33124, USA.
Org Biomol Chem. 2011 Apr 7;9(7):2142-8. doi: 10.1039/c0ob00849d. Epub 2011 Feb 3.
A set of spectrally diverse stilbazolium dyes was identified in an uptake assay using cultured brainstem and cerebellum cells isolated from e19 chicks. Pretreatment of cells with indatraline, a monoamine reuptake inhibitor, allowed identification of dyes that may interact with monoamine transporters. Two structurally related, yet spectrally segregated, probes, (E)-1-methyl-4-[2-(2-naphthalenyl)ethenyl]-pyridinium iodide (NEP+, 3A) and (E)-4-[2-(6-hydroxy-2-naphthalenyl)ethenyl]-1-methyl-pyridinium iodide (HNEP+, 4A), were selected and further investigated using HEK-293 cells selectively expressing dopamine, norepinephrine or serotonin transporters. HNEP+ was selectively accumulated via catecholamine transporters, with the norepinephrine transporter (NET) giving the highest response; NEP+ was not transported, though possible binding was observed. The alternate modes of interaction enable the use of NEP+ and HNEP+ to image distinct cell populations in live brain tissue explants. The preference for HNEP+ accumulation via NET was confirmed by imaging uptake in the absence and presence of desipramine, a norepinephrine reuptake inhibitor.
采用从 e19 鸡胚分离培养的脑干和小脑细胞摄取测定法，鉴定了一组光谱多样的苯并噻唑鎓染料。细胞用单胺再摄取抑制剂 indatraline 预处理，可鉴定可能与单胺转运体相互作用的染料。选择了两种结构相关但光谱分离的探针，（E）-1-甲基-4-[2-（2-萘基）乙烯基]-吡啶翁碘化物（NEP+，3A）和（E）-4-[2-（6-羟基-2-萘基）乙烯基]-1-甲基-吡啶翁碘化物（HNEP+，4A），并用选择性表达多巴胺、去甲肾上腺素或 5-羟色胺转运体的 HEK-293 细胞进一步研究。HNEP+通过儿茶酚胺转运体选择性积累，去甲肾上腺素转运体（NET）给出最高的响应；NEP+不被转运，但观察到可能的结合。交替的相互作用模式使 NEP+和 HNEP+能够在活体脑组织外植体中成像不同的细胞群体。在不存在和存在去甲肾上腺素再摄取抑制剂去甲丙咪嗪的情况下，通过 NET 对 HNEP+积累的偏好通过成像得到证实。
