Osthole Antagonizes Microglial Activation in an NRF2-Dependent Manner.

Microglia are neuroglia in the brain with an innate immune function and participate in the progress of neurodegenerative diseases. Osthole (OST) is a coumarin derivative extracted from Cnidium monnieri and bears a microglia-antagonizing ability. However, the underlying mechanism of the antagonism is not clear. The lipopolysaccharides-induced microglial BV2 cell line and amyloid-overexpressing fruit fly were used as models to study OST treatment. We found that OST treatment is sufficient to evoke NRF2 cascade under an LPS-induced inflammatory environment, and silencing NRF2 is sufficient to abolish the process. Moreover, we found that OST is sufficient to antagonize microglial activation in both LPS-induced BV2 cells and Aß-overexpressing fruit flies, and silencing NRF2 abolishes OST's antagonism. Furthermore, OST treatment rescued survival, climbing, and the learning ability of Aß-overexpressing fruit flies and relieved oxidative stress. In conclusion, we proved that OST antagonizes microglial activation induced by either LPS or Aß and that NRF2 is necessary for OST's antagonism.

Alterations of RNA-binding protein found in neurons in Drosophila neurons and glia influence synaptic transmission and lifespan.

The found in neurons (fne), a paralog of the RNA-binding protein ELAV gene family in Drosophila, is required for post-transcriptional regulation of neuronal development and differentiation. Previous explorations into the functions of the FNE protein have been limited to neurons. The function of fne in Drosophila glia remains unclear. We induced the knockdown or overexpression of fne in Drosophila neurons and glia to determine how fne affects different types of behaviors, neuronal transmission and the lifespan. Our data indicate that changes in fne expression impair associative learning, thermal nociception, and phototransduction. Examination of synaptic transmission at presynaptic and postsynaptic terminals of the larval neuromuscular junction (NMJ) revealed that loss of fne in motor neurons and glia significantly decreased excitatory junction currents (EJCs) and quantal content, while flies with glial fne knockdown facilitated short-term synaptic plasticity. In muscle cells, overexpression of fne reduced both EJC and quantal content and increased short-term synaptic facilitation. In both genders, the lifespan could be extended by the knockdown of fne in neurons and glia; the overexpression of fne shortened the lifespan. Our results demonstrate that disturbances of fne in neurons and glia influence the function of the Drosophila nervous system. Further explorations into the physiological and molecular mechanisms underlying neuronal and glial fne and elucidation of how fne affects neuronal activity may clarify certain brain functions.

Imidacloprid Impairs Glutamatergic Synaptic Plasticity and Desensitizes Mechanosensitive, Nociceptive, and Photogenic Response of Drosophila melanogaster by Mediating Oxidative Stress, Which Could Be Rescued by Osthole.

BACKGROUND: Imidacloprid (IMD) is a widely used neonicotinoid-targeting insect nicotine acetylcholine receptors (nAChRs). However, off-target effects raise environmental concerns, including the IMD's impairment of the memory of honeybees and rodents. Although the down-regulation of inotropic glutamate receptor (iGluR) was proposed as the cause, whether IMD directly manipulates the activation or inhibition of iGluR is unknown. Using electrophysiological recording on fruit fly neuromuscular junction (NMJ), we found that IMD of 0.125 and 12.5 mg/L did not activate glutamate receptors nor inhibit the glutamate-triggered depolarization of the glutamatergic synapse. However, chronic IMD treatment attenuated short-term facilitation (STF) of NMJ by more than 20%. Moreover, by behavioral assays, we found that IMD desensitized the fruit flies' response to mechanosensitive, nociceptive, and photogenic stimuli. Finally, the treatment of the antioxidant osthole rescued the chronic IMD-induced phenotypes. We clarified that IMD is neither agonist nor antagonist of glutamate receptors, but chronic treatment with environmental-relevant concentrations impairs glutamatergic plasticity of the NMJ of fruit flies and interferes with the sensory response by mediating oxidative stress.

Neuromuscular, retinal, and reproductive impact of low-dose polystyrene microplastics on Drosophila.

Facing the challenge of global microplastics (MPs) pollution, full characterization of MPs biohazards is urgent. Recent intensive studies revealed that the toxicity depends on the material, size, and exposure concentration of MP. To better elucidate MPs biohazards, we investigated the impact of polystyrene-MPs of size 0.1 µm at a low dose of 50 µg/L on the neuromuscular, retinal, and reproductive phenotypes of fruit fly model, by voltage-clamped electrophysiology, electroretinogram, and reproductive assay, respectively. We found that MPs decreased the frequency of spontaneous junction currents of synapse and altered the receptor potential amplitude of the retina. Furthermore, MPs lowered the rate of embryo-laying of fruit flies. The differential gene expression of ligand-receptor interaction, endocytosis, phototransduction, and Toll/Imd signaling pathways might underlie these MPs-induced phenotypes. These findings call for further investigation on the potential biohazards of low-dose MPs.

DEHP toxicity on vision, neuromuscular junction, and courtship behaviors of Drosophila.

Bis(2-ethylhexyl) phthalate (DEHP) is the most common plasticizer. Previous studies have shown DEHP treatment accelerates neurological degeneration, suggesting that DEHP may impact retinal sensitivity to light, neurotransmission, and copulation behaviors. Although its neurotoxicity and antifertility properties have been studied, whether DEHP exposure disrupts vision and how DEHP influences neuromuscular junction (NMJ) have not been reported yet. Moreover, the impact of DEHP on insect courtship behavior is still elusive. Fruit flies (Drosophila melanogaster) were treated with series concentrations of DEHP and observed for lifespan, motor function, electroretinogram (ERG), electrophysiology of neuromuscular junction (NMJ), courtship behaviors, and relevant gene expression. Our results confirmed the DEHP toxicity on lifespan and capacity of motor function and updated its effect on copulation behaviors. Additionally, we report for the first time that DEHP exposure may harm vision by affecting the synaptic signaling between the photoreceptor and the laminar neurons. Further, DEHP treatment altered both spontaneous and evoked neurotransmission properties. Noteworthy, the effect of DEHP exposure on the copulation behavior is sex-dependent, and we proposed potential mechanisms for future investigation.