Short-Term Exposure of Zebrafish Embryos to Arecoline Leads to Retarded Growth, Motor Impairment, and Somite Muscle Fiber Changes

Abstract

The areca nut-chewing habit is common in Southeast Asia, India, and Taiwan, and arecoline is the most abundant and potent component in the areca nut. The effects of arecoline on birth defects have been explored in many species, including chicken, mice, and zebrafish. The effects of arecoline on embryos after long-term exposure are well established; however, the effects of short-term embryo exposure to arecoline are not understood. Using zebrafish, we study the effects of short-term exposure of arecoline on embryos to model the human habit of areca nut-chewing during early pregnancy. Arecoline, at concentrations from 0.001% to 0.04%, was administered to zebrafish embryos from 4 to 24 hours post fertilization. The morphological changes, survival rates, body length, and skeletal muscle fiber structure were then investigated by immunohistochemistry, confocal microscopy, and conventional electron microscopy. With exposure of embryos to increasing arecoline concentrations, we observed a significant decline in the hatching and survival rates, general growth retardation, lower locomotor activity, and swimming ability impairment. Immunofluorescent staining demonstrated a loose arrangement of myosin heavy chains, and ultrastructural observations revealed altered myofibril arrangement and swelling of the mitochondria. In addition, the results of flow-cytometry and JC-1 staining to assay mitochondria activity, as well as reverse transcription-polymerase chain reaction analyses of functional gene expression, revealed mitochondrial dysfunctions after exposure to arecoline. We confirmed that short-term arecoline exposure resulted in retarded embryonic development and decreased locomotor activity due to defective somitic skeletal muscle development and mitochondrial dysfunction.

Association of betel nut chewing with exercise performance in a military male cohort: the CHIEF study

“…The betel nut metabolite, arecoline, may lead to muscular mitochondria dysfunction and affect either the arrangement or the integrity of muscle fibres or a reduction of locomotor activity 22. With regard to the cardiovascular system, current betel nut chewing would increase HR, irrespective of the frequency of usage, due to central sympathetic response, but the effect on BP may vary by the peripheral cholinergic effect, resulting in a fall of the diastolic component 4.…”

Pharmaceutical Assessment Suggests Locomotion Hyperactivity in Zebrafish Triggered by Arecoline Might Be Associated with Multiple Muscarinic Acetylcholine Receptors Activation

“…Betel nut chewing becomes popular since they elicit a psychoactive effect in humans [ 2 ]. In addition, betel nut is also listed as the most addictive substance in the world along with tobacco, alcohol, and caffeine [ 3 ]. The psychoactive effect in betel nut is associated with arecoline (methyl 1,2,5,6-tetrahydro-1-methyl nicotinate) [ 4 ], which is the most abundant nicotinic acid-based alkaloid in betel nut [ 5 ].…”

“…Furthermore, arecoline alkaloid is able to promote peristalsis and glandular secretion by acting on both muscarinic and nicotinic receptors [ 1 , 7 ]. International Agency for Research on Cancer (IARC) has found evidence demonstrating that chewing betel nut could cause oral cancer in humans [ 3 ]. However, arecoline can be used for medicinal purposes such as anti-helminthic, mental illness drugs, digestive agents, as well as psychosomatic medicine [ 1 ].…”

“…Meanwhile, in zebrafish studies, it has been proved that 0.01–0.04% (100–400 ppm) of arecoline caused general growth retardation and a lower heart rate [ 10 ]. Another experiment with zebrafish larvae that used 0.001–0.04% (10–400 ppm) arecoline also generate developmental retardation and morphological deformities [ 3 ]. All these results showed that arecoline in a higher concentration (>1 ppm) has a potentially adverse effect on the animal, especially in the developmental stage.…”

Nanomaterial híbrido (TiO2-MWCNT): Síntese, Caracterização e Ecotoxicidade aquática

“…Peng et al (2015) correlacionaram alterações em fibras musculares de embriões de zebrafish, expostos a Arecolina, com a diminuição no crescimento das larvas. Além disso, estudos in vitro e in vivo realizados porSavi et al (2014) Gráficos representativos de corte transversal.…”