Jenson Lacey J, Paulson Sally L, Bloomquist Jeffrey R
Department of Entomology and Nematology, Neurotoxicology Laboratory, Emerging Pathogens Institute, University of Florida, 2055 Mowry Road, Gainesville, FL 32610-0006, USA.
Invert Neurosci. 2012 Dec;12(2):119-27. doi: 10.1007/s10158-012-0138-5. Epub 2012 Jul 14.
The goal of this research was to induce neuron-like properties in Sf21 cells, an insect ovarian cell line, which could lead to a new high-throughput insecticide screening method and a way to mass produce insect neuronal material for basic research. This study applied differentiation agents to produce viable neuron-like cells. In the presence of the molting hormone 20-hydroxyecdysone (20-HE), or insulin, in the growth medium, a maximum of ca. 30 % of Sf21 cells expressed an apparent neuronal morphology of unipolar, bipolar, or multipolar axon-like processes within 2-3 days. Maximal differentiation occurred after 2 days in the presence of 50 μM 20-HE or 3 days in 10 μM insulin. Both 20-HE and insulin displayed time- and concentration-dependent differentiation with biphasic curves, suggesting that two binding sites or processes were contributing to the observed effects. In addition, combinations of 20-HE and insulin produced apparent synergistic effects on differentiation. Caffeine, a central nervous system stimulant, inhibited induction of elongated processes by 20-HE and/or insulin, with an IC(50) of 9 nM for 20-HE, and the inhibition was incomplete, resulting in about one-quarter of the differentiated cells remaining, even at high concentrations (up to 1 mM). The ability to induce a neural phenotype simplifies the studies of insect cells, compared to either the use of primary nervous tissue or genetic engineering techniques. The presence of ion channels or receptors in the differentiated cells remains to be determined.
本研究的目标是在昆虫卵巢细胞系Sf21细胞中诱导出类神经元特性，这可能会带来一种新的高通量杀虫剂筛选方法，以及一种为基础研究大量生产昆虫神经元材料的途径。本研究应用分化剂来产生有活力的类神经元细胞。在生长培养基中存在蜕皮激素20-羟基蜕皮酮（20-HE）或胰岛素的情况下，最多约30%的Sf21细胞在2-3天内表现出单极、双极或多极轴突样突起的明显神经元形态。在50 μM 20-HE存在下2天后或10 μM胰岛素存在下3天后发生最大分化。20-HE和胰岛素均呈现出具有双相曲线的时间和浓度依赖性分化，这表明两个结合位点或过程促成了观察到的效应。此外，20-HE和胰岛素的组合对分化产生了明显的协同效应。咖啡因，一种中枢神经系统兴奋剂，抑制20-HE和/或胰岛素诱导的伸长突起，对20-HE的IC(50)为9 nM，且抑制不完全，即使在高浓度（高达1 mM）下仍有大约四分之一的分化细胞留存。与使用原代神经组织或基因工程技术相比，诱导神经表型的能力简化了昆虫细胞的研究。分化细胞中离子通道或受体的存在仍有待确定。
