Garofalo F, Amelio D, Icardo J M, Chew S F, Tota B, Cerra M C, Ip Y K
Department of Biology, Ecology and Earth Science, University of Calabria, 87030 Arcavacata di Rende, CS, Italy.
Department of Anatomy and Cell Biology, University of Cantabria, Santander, Spain.
Nitric Oxide. 2015 Jan 30;44:71-80. doi: 10.1016/j.niox.2014.11.017. Epub 2014 Dec 9.
African lungfishes are obligate air breathers, with reduced gills and pulmonary breathing throughout their life. During the dry season they aestivate on land, with the collapse of secondary lamellae of their gills and the establishment of an exclusive aerial ventilation through the vascularization and expansion of their lungs. To date, the mechanisms underlining the respiratory organ remodeling in aestivating lungfishes are unknown. This study aimed to identify key switch components of the stress-induced signal transduction networks implicated in both rapid and medium-long term remodeling of the gills and lungs of the African lungfish Protopterus annectens during aestivation. Through immunofluorescence microscopy and Western blotting, the localization and the expression of nitric oxide synthase (NOS), Akt, Hsp-90 and HIF-1α were evaluated in both gills and lungs exposed to three experimental conditions: freshwater (FW), 6 months of experimentally induced aestivation (6mAe), and 6 days after arousal from 6 months of aestivation (6mAe6d). After 6mAe, the expression of NOS (p-eNOS antibody), Akt (p-Akt antibody), and Hsp-90 decreased in the gills, while NOS and Hsp-90 expression increased with Akt remained unchanged in the lungs. Upon 6mAe6d, NOS, Akt and Hsp-90 expression in the gills returned to the respective FW values. In the lungs of the aroused fish, NOS and Akt decreased to their respective FW levels, while Hsp-90 expression was enhanced with respect to aestivation. In both respiratory organs, the qualitative and quantitative patterns of HIF-1α expression correlated inversely to those of NOS. Overall, our findings suggest that the molecular components of the NOS/NO system changed in a tissue-specific manner in parallel with organ readjustment in the gills and lungs of P. annectens during aestivation and arousal.
非洲肺鱼是专性空气呼吸者,终生鳃退化且通过肺呼吸。在旱季,它们在陆地上夏眠,鳃的次生薄片塌陷,通过肺的血管化和扩张建立起唯一的空气通气方式。迄今为止,夏眠肺鱼呼吸器官重塑的潜在机制尚不清楚。本研究旨在确定在非洲肺鱼原鳍鱼夏眠期间,参与鳃和肺快速及中长期重塑的应激诱导信号转导网络的关键开关组件。通过免疫荧光显微镜和蛋白质印迹法,评估一氧化氮合酶(NOS)、Akt、热休克蛋白90(Hsp-90)和缺氧诱导因子1α(HIF-1α)在暴露于三种实验条件下的鳃和肺中的定位及表达:淡水(FW)、6个月实验诱导夏眠(6mAe)以及夏眠6个月苏醒后6天(6mAe6d)。6mAe后,鳃中NOS(p-eNOS抗体)、Akt(p-Akt抗体)和Hsp-90的表达下降,而肺中NOS和Hsp-90表达增加,Akt保持不变。在6mAe6d时,鳃中NOS、Akt和Hsp-90的表达恢复到各自的FW值水平。在苏醒鱼的肺中,NOS和Akt降至各自的FW水平,而Hsp-90表达相对于夏眠时增强。在两个呼吸器官中,HIF-1α表达的定性和定量模式与NOS的模式呈负相关。总体而言,我们的研究结果表明,在原鳍鱼夏眠和苏醒期间,鳃和肺的器官重新调整过程中,NOS/NO系统的分子组件以组织特异性方式发生变化。