Electrophysiological evidence of RML12 mosquito cell line towards neuronal differentiation by 20-hydroxyecdysdone

Abstract

Continuous cell lines from insect larval tissues are widely used in different research domains, such as virology, insect immunity, gene expression, and bio pharmacology. Previous study showed that introduction of 20-hydroxyecdysone to Spodoptera cell line induced a neuron-like morphology with neurite extensions. Despite some results suggesting potential presence of neuro-receptors, no study so far has shown that these neuron-induced cells were functional. Here, using microelectrode arrays, we showed that the mosquito cell line, RML12, differentiated with 20-hydroxyecdysone, displays spontaneous electrophysiological activity. Results showed that these cells can be stimulated by GABAergic antagonist as well as nicotinic agonist. These results provide new evidence of neuron-like functionality of 20-hydroxyecdysone induced differentiated mosquito cell line. Finally, we used this new model to test the effects of two insecticides, temephos and permethrin. Our analysis revealed significant changes in the spiking activity after the introduction of these insecticides with prolonged effect on the neuronal activity. We believe that this differentiated mosquito neuronal cell model can be used for high-throughput screening of new pesticides on insect nervous system instead of primary neurons or in vivo studies.

Spontaneous Cell Cluster Formation in Human iPSC-Derived Neuronal Spheroid Networks Influences Network Activity

“…Many different forms of neuronal cultures are observed to develop synchronous bursting behavior, including insect neurons such as locust ( Greenbaum et al, 2009 ) and mosquito ( Gaburro et al, 2018 ), and mammalian neurons such as rat ( Lam et al, 2019 ) and human ( Heikkilä et al, 2009 ). Although bursts are almost ubiquitous in neuronal culture systems, human neurons have burst characteristics that set them apart from, for example, rat neurons ( Hyvärinen et al, 2019 ).…”

Insect Cell-Based Models: Cell Line Establishment and Application in Insecticide Screening and Toxicology Research

“…The outcomes reveal that RML12 cells could be stimulated by GABAergic antagonist as well as nicotinic agonist. The finding provides new evidence of the neuron-like functionality of the 20-HE-induced differentiated cell line, and the cell model may be employed for HTS of candidate compounds in insect nervous systems [ 120 ].…”

Pharmaceutical technologies with potential application to insecticide discovery†

“…In some cases, it appears that voltage‐gated sodium, calcium, potassium or chloride channels have been induced concomitant with the morphological change 37–41 . More recently, it has been shown that 20‐OH E treatment of Aedes RML12 cells led to spontaneous neuroactivity as monitored by multiwell microelectrode array, and that this could be enhanced by treatment with the GABA antagonist gabazine and especially nicotine 42 …”