An Assay for Lateral Line Regeneration in Adult Zebrafish

Abstract

Due to the clinical importance of hearing and balance disorders in man, model organisms such as the zebrafish have been used to study lateral line development and regeneration. The zebrafish is particularly attractive for such studies because of its rapid development time and its high regenerative capacity. To date, zebrafish studies of lateral line regeneration have mainly utilized fish of the embryonic and larval stages because of the lower number of neuromasts at these stages. This has made quantitative analysis of lateral line regeneration/and or development easier in the earlier developmental stages. Because many zebrafish models of neurological and non-neurological diseases are studied in the adult fish and not in the embryo/larvae, we focused on developing a quantitative lateral line regenerative assay in adult zebrafish so that an assay was available that could be applied to current adult zebrafish disease models. Building on previous studies by Van Trump et al. 17 that described procedures for ablation of hair cells in adult Mexican blind cave fish and zebrafish (Danio rerio), our assay was designed to allow quantitative comparison between control and experimental groups. This was accomplished by developing a regenerative neuromast standard curve based on the percent of neuromast reappearance over a 24 hr time period following gentamicin-induced necrosis of hair cells in a defined region of the lateral line. The assay was also designed to allow extension of the analysis to the individual hair cell level when a higher level of resolution is required. Video LinkThe video component of this article can be found at

Inhibition of H3K27me3 Histone Demethylase Activity Prevents the Proliferative Regeneration of Zebrafish Lateral Line Neuromasts

“…There is also no separation into inner and outer hair cells within a neuromast. Furthermore, unlike their mammalian counterparts, zebrafish have the remarkable ability to regenerate hair cells after hair cell death (López-Schier and Hudspeth, 2006 ; Ma et al, 2008 ; Pisano et al, 2014 ). Follow-up mammalian studies are required to elucidate whether a similar role of H3K27me3 histone demethylase activity is observed in the mammalian inner ear and what the underlying mechanisms are.…”

“…In the mature mammalian inner ear, the majority of damaged hair cells do not regenerate, and this leads to irreversible and permanent hearing loss (Forge et al, 1993 ; Warchol et al, 1993 ; Brigande and Heller, 2009 ). In contrast, non-mammalian vertebrates are capable of regenerating lost sensory hair cells after damage (Balak et al, 1990 ; Lombarte et al, 1993 ; Harris et al, 2003 ; Pisano et al, 2014 ). New hair cells are frequently produced by proliferation of non-sensory supporting cells, which are the source of hair cell precursors that subsequently differentiate into new hair cells and supporting cells (Raphael, 1992 ; Stone and Cotanche, 1994 ; Jones and Corwin, 1996 ).…”

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

“…2010 ; Mekdara et al. 2018 ), while the time course of neuromast recovery using this chemical is less established (but see Pisano et al. 2014 ).…”

The early development and physiology of Xenopus laevis tadpole lateral line system

“…To visualize the LL neuromasts in tadpole skin, an assay by Pisano et al ( 44 ) was adapted for use in the present study. Tadpoles were placed in 0.2 mg/mL solution of vital dye 2-[4-(dimethylamino)styryl]- N -ethylpyridinium iodide (DASPEI, Sigma-Aldrich, UK) in saline for 30 min.…”