Role of histone deacetylase activity in the developing lateral line neuromast of zebrafish larvae

Histone deacetylases are involved in many biological processes and have roles in regulating cell behaviors such as cell cycle entry, cell proliferation and apoptosis. However, the effect of histone deacetylases on the development of hair cells (HCs) has not been fully elucidated. In this study, we examined the influence of histone deacetylases on the early development of neuromasts in the lateral line of zebrafish. Hair cell development was evaluated by fluorescent immunostaining in the absence or presence of histone deacetylase inhibitors. Our results suggested that pharmacological inhibition of histone deacetylases with inhibitors, including trichostatin A, valproic acid and MS-275, reduced the numbers of both HCs and supporting cells in neuromasts. We also found that the treatment of zebrafish larvae with inhibitors caused accumulation of histone acetylation and suppressed proliferation of neuromast cells. Real-time PCR results showed that the expression of both p21 and p27 mRNA was increased following trichostatin A treatment and the increase in p53 mRNA was modest under the same conditions. However, the expression of p53 mRNA was significantly increased by treatment with a high concentration of trichostatin A. A high concentration of trichostatin A also led to increased cell death in neuromasts as detected in a TUNEL assay. Moreover, the nuclei of most of these pyknotic cells were immunohistochemically positive for cleaved caspase-3. These results suggest that histone deacetylase activity is involved in lateral line development in the zebrafish and might have a role in neuromast formation by altering cell proliferation through the expression of cell cycle regulatory proteins.

The Zebrafish Model for the Study on Hair Cell Development / 听力学及言语疾病杂志

Objective This study aims to examine the development of the posterior lateral line of the zebrafish and establish ant model to study the process of hair cell differentiation and regeneration .Methods We observed the posterior lateral line system formation by DAPI immunohistochemistry and whole mount in situ hybridization .We further evaluated hair cells differentiation within neuromast by using Transgenic Tg (Brn3c:mGFP) zebrafish and stained the functional hair cells by the mechanotransduction marker FM 1 -43FX .We labelled proliferating cells in primordium and neuromast by addition of BrdU to the system water .Results The posterior lateral line primordium originated from a sensory placode and started its journey at around 20 hours post fertilization to migrate along the horizontal myoseptum to the tail -tip with a constant speed (1 .7somite/hour) .The primordium depositd five or six neuromasts spaced along the body ,and two or three terminal neuromasts at the tail -tip at 48 hours post fertiliza-tion .At 3 ,5 and 7 days post fertilization ,zebrafish contained 5 .68 ± 1 .46 ,10 .1 ± 0 .99 ,and 12 .45 ± 1 .32 hair cells per neuromast ,respectively .Furthermore ,the average number of FM1-43FX stained hair cells within each neuro-mast were 3 .68 ± 1 .11 ,8 .18 ± 1 .86 ,and 10 .22 ± 1 .24 ,respectively .Conclusion We establish the development model of hair cells in zebrafish lateral line neuromast and suggest that 3 to 7 days post fertilization is an important period for lateral line neuromast differentiation .This study would be useful for underlying the mechanisms of hair cell differentiation and regeneration .

Morphology of lateral line canals in Neotropical freshwater stingrays (Chondrichthyes: Potamotrygonidae) from Negro River, Brazilian Amazon

The relationship between the distribution of the lateral line canals and their functionality has not been well examined in elasmobranchs, especially among Neotropical freshwater stingrays of the family Potamotrygonidae. The spatial distribution of the canals and their tubules and the quantification of the neuromasts were analyzed in preserved specimens of Potamotrygon motoro, P. orbignyi, Potamotrygon sp. "cururu", and Paratrygon aiereba from the middle Negro River, Amazonas, Brazil. The hyomandibular, infraorbital, posterior lateral line, mandibular, nasal and supraorbital canals were characterized and their pores and neuromasts quantified. The ventral canals are known to facilitate the accurate localization of prey items under the body, and our results indicate that the dorsal canals may be employed in identifying the presence of predators or potential prey positioned above the stingray's body. The presence of non-pored canals in the ventral region may be compensated by the high concentration of neuromasts found in the same area, which possibly allow the accurate detection of mechanical stimuli. The concentration of non-pored canals near the mouth indicates their importance in locating and capturing prey buried in the bottom substrate, possibly aided by the presence of vesicles of Savi.

A relação entre a distribuição dos canais da linha lateral e a sua funcionalidade é pouco conhecida para os elasmobrânquios, especialmente para as raias neotropicais da família Potamotrygonidae. A distribuição espacial dos canais e seus túbulos e a quantificação dos neuromastos foram analisadas em exemplares preservados das raias Potamotrygon motoro, P. orbignyi, Potamotrygon sp. "cururu" e Paratrygon aiereba, provenientes do médio rio Negro, Amazonas, Brasil. Foram identificados os canais hiomandibular, infra-orbital, linha lateral posterior, mandibular, nasal e supra-orbital, e os poros e neuromastos de cada um foram quantificados. Nossos resultados mostram que os canais dorsais podem ter a função de identificar a presença de predadores ou presas posicionadas acima do corpo da raia, enquanto os canais ventrais provavelmente permitem uma localização precisa da posição de presas sob o corpo. A existência de canais sem poros na região ventral pode estar relacionada com a alta concentração de neuromastos, que compensaria a falta de contato direto dos neuromastos com o meio externo na detecção de estímulos mecânicos. A concentração dos canais sem poros na proximidade da boca indica sua importância para a localização e captura de presas enterradas no substrato, possivelmente auxiliadas pela presença das vesículas de Savi.