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氨基糖苷类抗生素与氯化钴对巨骨舌鱼侧线系统消融的比较

Comparison of Aminoglycoside Antibiotics and Cobalt Chloride for Ablation of the Lateral Line System in Giant Danios.

作者信息

Mekdara P J, Tirmizi S, Schwalbe M A B, Tytell E D

机构信息

Department of Biology, Tufts University, 200 Boston Avenue, Ste 4700, Medford, MA 02155, USA.

出版信息

Integr Org Biol. 2022 Mar 21;4(1):obac012. doi: 10.1093/iob/obac012. eCollection 2022.

Abstract

The mechanoreceptive lateral line system in fish is composed of neuromasts containing hair cells, which can be temporarily ablated by aminoglycoside antibiotics and heavy metal ions. These chemicals have been used for some time in studies exploring the functional role of the lateral line system in many fish species. However, little information on the relative effectiveness and rate of action of these chemicals for ablation is available. In particular, aminoglycoside antibiotics are thought to affect canal neuromasts, which sit in bony or trunk canals, differently from superficial neuromasts, which sit directly on the skin. This assumed ablation pattern has not been fully quantified for commonly used lateral line ablation agents. This study provides a detailed characterization of the effects of two aminoglycoside antibiotics, streptomycin sulfate and neomycin sulfate, and a heavy metal salt, cobalt (II) chloride hexahydrate (CoCl), on the ablation of hair cells in canal and superficial neuromasts in the giant danio () lateral line system, as a model for adult teleost fishes. We also quantified the regeneration of hair cells after ablation using CoCl and gentamycin sulfate to verify the time course to full recovery, and whether the ablation method affects the recovery time. Using a fluorescence stain, 4-Di-2-ASP, we verified the effectiveness of each chemical by counting the number of fluorescing canal and superficial neuromasts present throughout the time course of ablation and regeneration of hair cells. We found that streptomycin and neomycin were comparably effective at ablating all neuromasts in less than 12 h using a 250 μM dosage and in less than 8 h using a 500 μM dosage. The 500 μM dosage of either streptomycin or neomycin can ablate hair cells in superficial neuromasts within 2-4 h, while leaving those in canal neuromasts mostly intact. CoCl (0.1 mM) worked the fastest, ablating all of the hair cells in less than 6 h. Complete regeneration of the neuromasts in the lateral line system took 7 days regardless of chemicals used to ablate the hair cells. This study adds to the growing knowledge in hearing research about how effective specific chemicals are at ablating hair cells in the acoustic system of vertebrates.

摘要

鱼类的机械感受侧线系统由包含毛细胞的神经丘组成,氨基糖苷类抗生素和重金属离子可使这些毛细胞暂时失活。一段时间以来,这些化学物质一直被用于探索侧线系统在许多鱼类物种中的功能作用的研究中。然而,关于这些化学物质用于失活的相对有效性和作用速率的信息却很少。特别是,人们认为氨基糖苷类抗生素对位于骨管或躯干部位管道中的管神经丘的影响,与直接位于皮肤上的表面神经丘不同。对于常用的侧线失活剂,这种假定的失活模式尚未得到充分量化。本研究详细描述了两种氨基糖苷类抗生素硫酸链霉素和硫酸新霉素,以及一种重金属盐六水合氯化钴(CoCl)对巨暹罗鲤侧线系统中管神经丘和表面神经丘毛细胞失活的影响,以此作为成年硬骨鱼类的模型。我们还对使用CoCl和硫酸庆大霉素失活后毛细胞的再生情况进行了量化,以验证完全恢复的时间进程,以及失活方法是否会影响恢复时间。使用荧光染料4-Di-2-ASP,我们通过计算在毛细胞失活和再生的整个时间进程中出现的荧光管神经丘和表面神经丘的数量,来验证每种化学物质的有效性。我们发现,使用250μM的剂量,链霉素和新霉素在不到12小时内就能有效失活所有神经丘,使用500μM的剂量则在不到8小时内就能做到。500μM剂量的链霉素或新霉素可在2-4小时内使表面神经丘中的毛细胞失活,而管神经丘中的毛细胞大多保持完整。CoCl(0.1 mM)起效最快,在不到6小时内就能使所有毛细胞失活。无论使用何种化学物质使毛细胞失活,侧线系统中神经丘的完全再生都需要7天。这项研究增加了听力研究中关于特定化学物质在脊椎动物听觉系统中使毛细胞失活的有效性的知识。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d27d/8964175/8bd9293a2684/obac012fig1.jpg

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