不同培养条件下背根神经节源性永生细胞系(F-11)神经元离子通道的转录谱分析。

In vitro models Pub Date : 2022-11-01 Epub Date: 2022-11-03 DOI:10.1007/s44164-022-00036-7
Erick Orozco Morato, Brittany Knight, Lakshmi S Nair
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引用次数: 0

摘要

病理性疼痛是一种常见病,影响着大多数患有各种潜在疾病的成年人。目前可用的药物止痛治疗方法有几种负面副作用,长期使用可能危及生命。由于疼痛感知的异质性和导致疼痛的神经元机制的多样性,高通量筛选可能具有潜在镇痛特性的小分子对于确定既有效又安全的新镇痛疗法至关重要。F-11 杂交永生化细胞系是目前用于药物筛选的背根神经节(DRG)细胞系之一。虽然F-11细胞通常被用作原发性DRG感觉神经元的类似物,但它们在生理特性上存在很大差异。本研究调查了分化方案对 F-11 细胞中成熟神经元离子通道和受体表达的影响。我们使用了一个定制的基因阵列,其中包括电压门控离子通道、瞬时受体电位通道和大麻素受体等八十多种神经元离子通道和受体,对以下几组细胞进行了评估:对照组 F-11 细胞、在不同培养条件下培养的 F-11 细胞和小鼠 DRG 组织。结果发现,与小鼠原代DRG神经元相比,F-11细胞中大多数被研究的离子通道和受体的表达量都较低。与对照 F-11 细胞相比,在低血清(LSM)条件下培养的 F-11 细胞中电压门控离子通道和大麻素受体等几个研究目标的表达量有所增加。研究表明,培养条件会显著调节所研究离子通道和受体的转录表达,长期培养(21 天)可能会对许多研究靶点的表达产生不利影响。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Transcriptional profiling of neuronal ion channels in dorsal root ganglion-derived immortal cell line (F-11) under different culture conditions.

Pathological pain is a prevalent condition that affects majority of adults with a variety of underlying disease conditions. Current available pharmacological pain treatments have several negative and potentially life-threatening side effects associated with their long-term use. Due to the heterogeneity of pain perception and the diversity of neuronal mechanisms that contribute to pain, high-throughput screening of small molecules that may have underlying analgesic properties is essential for identifying new analgesic treatments that are both effective and safe. The F-11 hybrid immortalized cell line is one of the currently available dorsal root ganglion (DRG) cell lines used for drug screening. While F-11 cells are commonly used as analogs to primary DRG sensory neurons, they differ significantly in physiological properties. The present study investigated the impact of differentiation protocols on the expression of mature neuron ion channels and receptors in F-11 cells. Using a customized gene array of more than eighty neuronal ion channels and receptors including voltage-gated ion channels, transient receptor potential channels, and cannabinoid receptors, we assessed the following groups: control F-11 cells; F-11 cells cultured under different culture conditions, and murine DRG tissue. The expression profiles of majority of the investigated ion channels and receptors in F-11 cells were found to be lower compared to primary mouse DRG neurons. F-11 cells cultured under low serum (LSM) conditions had increased expression of several investigated targets including voltage-gated ion channels and cannabinoid receptors when compared to control F-11 cells. The study showed that the culture conditions significantly modulated the transcriptional expression of studied ion channels and receptors, and that long-term culture (21 days) may adversely affect the expression of many of the studied targets.

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