檳榔鹼對斑馬魚胚胎發育的影響

Abstract

Introduction:Arecoline, the major alkaloid in the betel nut, has been reported to be potent in inducing developmentally toxic effects by generally lowering the embryo weight and retarding development of the embryo. Zebrafish has been suggested as a model organism for vertebrate development, due to its rapid extracorporeal development, the transparent embryo, easy maintenance, and availability of gene markers. We explore the effect of arecoline on the skeletal muscle development of zebrafish. Material and Methods:Arecoline was administered to zebrafish embryos by incubation at concentrations ranging from 0.001–0.04 %, treating from 4 to 24 hpf (hours post fertilization). We recorded the morphological changes, survival rates, and the body length, and applied immunohistochemistry (IHC), confocal microscopy, and conventional light electron microscopy to investigate the morphological changes of zebrafish skeletal muscle tissue. Results:The survival rate of arecoline-treated embryos significantly declined as the arecoline concentration increased, the treated embryos also showed general growth retardation and the swimming (motor) ability impairments. Immunohistochemistry (IHC) and microscopic studies demonstrated that major phenotypical changes are the myosin heavy chain in a relatively looser arrangement. Ultrastructural observation revealed an alteration of myofibrils arrangement and the swelling of mitochondria. RT-PCR analyse of mitochondrial function genes showed that at 48 and 96 hpf, the expression of the genes including sdhb, coxI, cox4i1, atp5a1, and atp5f1 significantly declined as the arecoline concentration increased, but the expression of mt_cytb increased. Conclusions:We confirmed that arecoline causes general growth retardation, skeletal muscle impairment, mitochondrial structure damage and the alteration of mitochondria functional genes expression in zebrafish embryo.