Functional morphology of chloride cells in killifish Fundulus heteroclitus, a euryhaline teleost with seawater preference

ABSTRACT:   Recent advances in the functional morphology of chloride cells in killifish Fundulus heteroclitus , a euryhaline teleost with seawater (SW) preference, were reviewed. Immunocytochemical detection of chloride cells with anti-Na⁺/K⁺-ATPase revealed transitional processes of the chloride cell distribution during early life stages. Chloride cells first appear in the yolk-sac membrane at an early embryonic stage, followed by their appearance in the body skin in the later embryonic stages. The principal site for the chloride cell distribution then shifts from the yolk-sac membrane and body skin during embryonic stages to the gills and opercular membrane in larval and later developmental stages. Morphologically distinct SW- and freshwater (FW)-type chloride cells were identified in adult killifish adapted to SW and FW, respectively. Both types of chloride cells are equally active in the two environments, but exert different ion-transporting functions. Following direct transfer of killifish from SW to FW, SW-type chloride cells were transformed into FW-type cells as a short-term response, followed by the promotion of chloride cell replacement as a long-term response. In killifish acclimated to low NaCl (0.1 mM) FW, intense immunoreactivity for vacuolar-type proton pump (V-ATPase) was detected in the basolateral membrane of FW-type chloride cells, whereas the immunoreactivity was much weaker in fish acclimated to FW with higher NaCl concentrations (1.0 and 10.0 mM). These results suggest the occurrence of active ion absorption in FW-type chloride cells and the involvement of V-ATPase in ion-absorbing mechanisms. In view of recent advances in this field, future chloride cell research should be considered in relation to the functional diversity of chloride cells.     

Effects of environmental Ca2+ levels on branchial chloride cell morphology in freshwater-adapted killifish Fundulus heteroclitus

ABSTRACT: To examine the involvement of chloride cells in the uptake of Ca²⁺ in freshwater (FW) killifish, chloride cell morphology was compared in fish acclimated to different defined FW environments with Ca²⁺ concentrations of either 0.1 mM, 0.5 mM, or 2.5 mM. Numerous chloride cells were detected in whole-mount preparations of the gill filaments, which were stained with an antiserum specific for Na⁺, K⁺-ATPase. Chloride cells, located mostly on the afferent–vascular edge of the filaments, were larger at lower Ca²⁺ concentrations. Electron microscopic observations showed that in the 0.1 mM and 0.5 mM Ca²⁺ experimental groups, the apical membrane of chloride cells were flat or slightly projecting and equipped with numerous microvilli. In the 2.5 mM Ca²⁺ group, some chloride cells formed an apical pit, whereas other cells were similar to those observed in the 0.1 mM and 0.5 mM Ca²⁺ groups. Plasma osmolality decreased with decreasing ambient Ca²⁺ concentration, suggesting that environmental Ca²⁺ affects the permeability of the body surfaces. Gill Na⁺, K⁺-ATPase activity in the 0.1 mM and 0.5 mM Ca²⁺ groups were significantly higher than that in the 2.5 mM Ca²⁺ group, implying the involvement of the Na⁺–Ca²⁺ exchanger in Ca²⁺ uptake in the gills. These findings suggest that chloride cells function as the site for Ca²⁺ uptake in killifish acclimated to low Ca²⁺ environments.     

蓝鳃太阳鱼繁殖与苗种培育研究

记述了蓝鳃太阳鱼 (Lepomismacrochirus)的繁殖行为、卵子性状、有效繁殖力和培育鱼苗鱼种的结果。蓝鳃太阳鱼由雄性作巢并护巢。卵粒小 ,沉性、粘性 ,胚胎发育积温为70 0～ 80 0℃·h。生产的鱼苗全年累计每 1 0 0g体重雌鱼可获得鱼苗 640 1尾。鱼苗培育成活率 72 .2 % ,鱼种培育成活率 93.8%。     

叉尾斗鱼关睾虫(单殖亚纲)病的鳃组织病理学研究

对叉尾斗鱼（Macropodus opercularis）的大钩关睾虫（Heteronchocleidus magnihamatus Zhang et Ji）和小钩关睾虫（H．buschkieli Bychowsky）病进行了病理学研究。结果表明，关睾虫寄生于叉尾斗鱼的初级鳃丝及鳃小片，轻度感染时，病变不明显，导致部分鳃丝组织完整性受到破坏；中度感染时，鳃丝组织增生、融合，有大量黏液细胞浸润，鳃小片与窦状隙分离，形成空隙；重度感染时，鳃丝组织细胞解离、空泡化，呼吸上皮与毛细血管分离严重，形成空腔。     

甲基汞对鲤鱼鳃组织及氯细胞的影响

水体中甲基汞(CH3ClHg)浓度(μg/L)分别为1、5、10、25、50、100、200和1、2．5、5、10、20、40、50染毒鲤鱼种24h和一周。鱼鳃石蜡切片观察结果：50μg/L染毒24h和20μg/L染毒1周鱼，鳃丝组织末见异常变化；100μg/L染毒24h和40μg/L染毒1周鱼，鳃丝出现轻微混乱且向上卷曲；200μg/L染毒24h和80μg/L染毒1周鱼，外表皮层从基膜脱落，鳃丝严重卷曲。鳃氯细胞的结构变化：100μg/L汞染毒24h40μg/L染毒1周鱼，氯细胞的细胞膜质分散，偶尔可见肿胀线粒体和空泡膜管系统；暴露于甲基汞200μg／L中24h和80μg/L1周鱼的氯细胞损伤明显：细胞质内空泡大量出现且无规律，膜管系统空泡增加，线粒体多数变成肿胀或已崩溃解体。     

辛基酚对泥鳅的毒性作用及鳃、肝的组织学影响

【目的】探讨辛基酚(Octylphenol,OP)对泥鳅(Misgurnus anguillicaudatus)的毒性效应及对鳃、肝组织结构的影响,为评价酚类化合物对水生生物的毒害作用提供理论依据。【方法】采用单因子毒性试验法和组织切片法,检测OP对泥鳅的急性毒性效应,统计死亡率,建立回归方程,计算半数致死质量浓度(LC50)和安全质量浓度(SC),并观察OP对鳃、肝组织结构的影响。【结果】OP对泥鳅24,48,72和96hLC50分别为3.27,2.70,2.13和1.84mg/L,SC为0.55mg/L;0.52mg/L OP可导致泥鳅鳃小片顶端膨大、卷曲,细胞坏死脱落,肝细胞肿大、空泡化、核溶解、核变形及细胞溶解。【结论】OP属于高毒等级污染物,对泥鳅存在较强的急性毒性效应,且毒性随着质量浓度的增加及作用时间的延长而增强;当质量浓度小于SC的OP对泥鳅胁迫一段时间后,会导致泥鳅鳃、肝组织结构严重受损。     

Evaluation of fixation methods for demonstration of Neoparamoeba perurans infection in Atlantic salmon,Salmo salar L., gills

Formaldehyde‐based fixatives are generally employed in histopathology despite some significant disadvantages associated with their usage. Formaldehyde fixes tissue by covalently cross‐linking proteins, a process known to mask epitopes which in turn can reduce the intensity of immunohistochemical stains widely used in disease diagnostics. Additionally, formaldehyde fixation greatly limits the ability to recover DNA and mRNA from fixed specimens to the detriment of further downstream molecular analyses. Amoebic gill disease (AGD) has been reliably diagnosed from histological examination of gills although complementary methods such as in situ hybridization (ISH) and polymerase chain reaction (PCR) are required to confirm the presence of Neoparamoeba perurans, the causative agent of AGD. As molecular techniques are becoming more prevalent for pathogen identification, there is a need to adapt specimen collection and preservation so that both histology and molecular biology can be used to diagnose the same sample. This study used a general approach to evaluate five different fixatives for Atlantic salmon, Salmo salar L., gills. Neutral‐buffered formalin and seawater Davidson's, formaldehyde‐based fixatives commonly used in fish histopathology, were compared to formalin‐free commercial fixatives PAXgene^®, HistoChoice?MB* and RNAlater?. Each fixative was assessed by a suite of analyses used to demonstrate AGD including routine histochemical stains, immunohistochemical stains, ISH and DNA extraction followed by PCR. All five fixatives were suitable for histological examination of Atlantic salmon gills, with seawater Davidson's providing the best quality histopathology results. Of the fixatives evaluated seawater Davidson's and PAXgene^® were shown to be the most compatible with molecular biology techniques. They both provided good DNA recovery, quantity and integrity, from fixed and embedded specimens. The capacity to preserve tissue and cellular morphology in addition to allowing molecular analyses of the same specimens makes seawater Davidson's and PAXgene^® appear to be the best fixation methods for diagnosis and research on AGD in Atlantic salmon gills.     

Acute ammonia toxicity and gill morphological changes of Japanese flounder Paralichthys olivaceus in normal versus supersaturated oxygen

Ammonia toxicity and morphological changes in gills of juvenile Japanese flounder Paralichthys olivaceus (5.76 ± 0.12 g) were investigated when fish were separately exposed to normal dissolved oxygen (DO) at 6.5 ± 0.5 mg L^?1 and supersaturated oxygen at 16.0 ± 2.0 mg L^?1 at different ammonia concentrations. Under normal oxygen, ammonia concentrations were tested from 0.04 (control) to 93.3 mg L^?1 total ammonia nitrogen (TAN), whereas under oxygen supersaturation, ammonia concentrations ranged from 0.04 (control) to 226.7 mg L^?1 TAN in the trial. After exposure to ammonia for 96 h, the ammonia LC₅₀ for fish was 62.48 mg L^?1 TAN (0.50 mg L^?1 NH₃–N) at normal oxygen and 160.71 mg L^?1 TAN (0.65 mg L^?1 NH₃–N) at oxygen supersaturation. Light microscopic observations confirmed that gill damage in normal oxygen was more profound than in oxygen supersaturation when fish were exposed to the same level of TAN (93.3 mg L^?1). Furthermore, electron microscopic scanning also showed more crimple, retraction and fibrosis on the secondary lamella surface in fish exposed to normal oxygen than those in fish exposed to supersaturated oxygen at the same TAN (93.3 mg L^?1). This study suggests that supersaturated oxygen can increase ammonia tolerance in Japanese flounder through reducing gill damage by ammonia, which partially explains the merit of using pure oxygen injection in intensive fish farming.     

大口黑鲈和蓝鳃太阳鱼鳃结构的扫描电镜观察

应用扫描电镜对大口黑鲈Micropterus salmoides和蓝鳃太阳鱼Lepomis macrochirus鳃的表面微细结构进行了观察。结果表明：大口黑鲈和蓝鳃太阳鱼鳃的表面结构和微细结构与其它硬骨鱼类基本相似,鳃弓和鳃耙表面被具环形微嵴的上皮细胞所覆盖,表面观凹凸不平;大口黑鲈鳃耙的尖齿直径和长度分别为蓝鳃太阳鱼鳃耙的3.4-3.8倍和2.1-3.8倍;两种鱼的鳃丝末端略为膨大呈勺状,鳃丝表面都具有规则或不规则分布的环形微嵴、沟、坑、孔等结构;大口黑鲈鳃丝之间的距离相对较大,蓝鳃太阳鱼的鳃丝间距离仅为大口黑鲈的83%-88%;大口黑鲈鳃小片的高度、厚度和片间距都大于蓝鳃太阳鱼。文中还比较探讨了大口黑鲈和蓝鳃太阳鱼的形态结构与其生理功能之间的关系。