盐度胁迫对尼罗罗非鱼鳃氯细胞调节变化的影响

采用扫描电镜和免疫组化技术,研究了尼罗罗非鱼(Oreochromis niloticus)鳃中氯细胞的分布,及其不同盐度(0、10、20、30)胁迫对氯细胞数目和形态变化的影响.扫描电镜结果表明:氯细胞分布在鳃丝的鳃小片基部,根据其表面开口长度,可分为Ⅰ型(＞6.5μm)、Ⅱ型(3.2～6.5μm)和Ⅲ型(＜3.2 μm)3种亚型;不同盐度下氯细胞总数目变化趋势为盐度10＜盐度20＜盐度0＜盐度30;从盐度0转移到盐度10后,氯细胞总数目减少,主要是由于Ⅰ型氯细胞数目显著下降;盐度20中的氯细胞数量高于盐度10,但不显著;盐度30中的氯细胞数量随Ⅲ型氯细胞数量的提高而显著增加.免疫组织化学进一步证实了不同盐度条件下Na+-K+-ATPase免疫反应性细胞均分布在鳃丝的鳃小片基部.本研究结果表明,尼罗罗非鱼可通过改变鳃氯细胞数量和形态结构来适应环境中的盐度变化,推测Ⅰ型氯细胞和Ⅲ型氯细胞分别在低盐、高盐适应中起着重要作用.     

斑马鱼鳃的光镜和透射电镜观察

应用光学显微镜和透射电镜对斑马鱼(Danio rerio)鳃的组织结构及鳃丝、鳃小片超微结构进行了观察。结果表明,斑马鱼有4对全鳃,鳃耙呈长锥状,鳃丝呈梳状排列在鳃弓上,鳃小片均匀排列在鳃丝两侧。鳃小片由上皮细胞、柱细胞、内皮细胞和毛细血管网组成,鳃小片基部和血管周围分布有泌氯细胞,胞内有丰富的线粒体和排泄小泡,根据线粒体形态特征和细胞质电子密度可将其分为两个亚型。黏液细胞通常与泌氯细胞对生存在,并且有通外的开口。斑马鱼鳃组织结构与其他硬骨鱼鳃结构相似,其结构和功能有密切的关系。     

低盐胁迫对大底鳉(Fundulus grandis)血浆渗透压、鳃上皮细胞形态和通道蛋白mRNA 表达的影响

为了研究水环境中的低盐度对大底鳉(Fundulus grandis)的适应性影响, 采用生理和荧光定量PCR 方法探讨了盐度为5、2、1、0.5 和0.1 的情况下, 不同饲养时间大底鳉的血浆渗透压, 鳃上皮细胞超微结构及通道蛋白mRNA 表达的变化。饲养时间小于1 d、盐度小于0.5 的胁迫可以导致血浆渗透压明显降低(P<0.001); 鳃上皮表面泌氯细胞的体积增大、顶隐窝开口扩大或其细胞的形状变为三角形或不规则形。当饲养时间超过3 d 时, 血浆渗透压以及鳃上皮表面泌氯细胞的形态都恢复到对照组水平 (盐度为5); 低盐胁迫上调了六种鳃通道蛋白mRNA 的表达, 并下调了两种鳃通道蛋白mRNA 的表达。结果显示: 广盐性的大底鳉通过调整鳃上皮的形态及通道蛋白mRNA 的表达来维持机体渗透压的平衡状态。     

盐度对大麻哈鱼幼鱼存活率、鳃ATP酶活力及其组织结构的影响

为研究不同盐度对大麻哈鱼幼鱼存活率、鳃ATP酶活力及其组织结构的影响, 试验共设置4个盐度组(S0、S8、S16、S24), 试验周期42d, 解剖取鱼鳃测定ATP酶活力, 并运用组织切片及扫描电镜技术观察其鳃组织结构的变化。结果显示: S8和S16组大麻哈鱼幼鱼存活率最高, 均达到98.89%, S0组存活率为94.45%, 而S24组存活率最低, 为83.34%。随着盐度的升高, 大麻哈鱼幼鱼鳃组织Na+/K+-ATP和Ca2+/Mg2+-ATP酶活力均呈现出先升高后下降的趋势且酶活力最高的均为S8组、最低的为S24组。鳃丝宽度随盐度升高逐渐增大且各组之间呈现出显著性差异(P<0.05), 而鳃小片长度和宽度均随盐度升高逐渐减小。扫描电镜结果表明随着盐度的升高线粒体丰富细胞数量逐渐增多、顶膜变小且微绒毛消失; 同S0组相比, S8组和S16组鳃丝表面扁平上皮细胞之间的轮廓更加清楚且环形微脊条纹清晰, 而S24组鳃丝表面扁平上皮细胞之间界限模糊, 环形微脊间有融合或间断情况; 鳃小片底部扁平上皮细胞表层结构由清晰逐渐变得混乱、气孔数量逐渐减少且孔径变小。因此推测大麻哈鱼幼鱼在降海阶段适宜的盐度生存范围可能介于8‰—16‰, 具体有待进一步研究。     

遮目鱼幼鱼鳃线粒体丰富细胞的形态结构及其在不同盐度下的变化

该实验通过普通光学显微镜、透射电子显微镜和扫描电子显微镜的方法,研究不同盐度条件下(盐度0、10、20、27、35)广盐性海水鱼类遮目鱼(Chanos chanos)幼鱼鳃器官结构和鳃上线粒体丰富细胞分布及结构的变化。鳃线粒体丰富细胞呈椭圆形或卵圆形,内含有大量线粒体,细胞核较大。在不同盐度条件下,遮目鱼幼鱼出现两种鳃线粒体丰富细胞:一种是具有顶端小窝、线粒体体积较大的A型线粒体丰富细胞;另一种是单独存在、线粒体体积较小的B型线粒体丰富细胞。随着盐度降低,A型线粒体丰富细胞及其线粒体数量减少、体积减小,电子密度降低,顶端开口变小甚至关闭。盐度降至淡水条件下,鳃小片肿胀、脱落,鳃小片上增生出具有大面积平滑或波状的顶端开口的B型线粒体丰富细胞。结果表明,在高渗环境下,A型线粒体丰富细胞较为丰富和发达,其结构特征适应了离子分泌的功能,为海水型线粒体丰富细胞;在低渗环境下,B型线粒体丰富细胞较为丰富,其结构特征适应了离子吸收的功能,为淡水型线粒体丰富细胞。不同结构类型鳃线粒体丰富细胞的存在使得广盐性海水鱼类可以适应较广的盐度范围变化。     

不同盐度下人工选育卵形鲳鲹(Trachinotus ovatus)子代鳃线粒体丰富细胞结构变化

该文采用光学显微镜和透射电镜技术观察不同盐度下(5、20、30)人工选育卵形鲳鲹(Trachinotus ovatus)鳃线粒体丰富细胞的分布和超微结构变化。结果表明,线粒体丰富细胞主要分布于鳃丝和鳃小片基部,且随盐度升高而体积增大,数量增多;三个盐度组均存在由线粒体丰富细胞、扁平细胞和附细胞构成的顶端小窝,盐度5组线粒体丰富细胞顶膜面积较大,微脊发达,顶端小窝内凹,盐度20和30组线粒体丰富细胞顶膜面积相对较小,微脊不发达,顶端小窝明显内陷;盐度5和30组线粒体丰富细胞胞质内存在发达的微细小管系统,线粒体内脊丰富,盐度20组胞质内微细小管系统分布不均匀,结构松散,部分收缩成珠泡状结构,与粗面内质网相混杂。线粒体丰富细胞的结构变化与其所处的渗透压环境相适应。     

华鲮鳃表面形态结构扫描电镜观察

应用扫描电镜对华鲮Sinilabeor rendahli鳃耙、鳃弓、鳃丝和鳃小片的形态结构进行了观察.鳃耙和鳃弓表面凹凸不平,分布着大量丘突.鳃丝和鳃耙表面有大量粘液细胞分布,鳃丝上皮细胞表面密布微嵴,氯细胞附着在鳃丝表面和鳃小片侧表面.鳃小片薄、表面凹凸不平,垂直排列在鳃丝上.鳃丝和鳃小片的表面形态结构特征有助于提高鱼鳃的气体交换效率.     

珠江口池养梭鱼鳃的光镜、扫描和透射电镜观察

应用光学显微镜、扫描电镜和透射电镜对珠江口池塘养殖梭鱼Liza haematocheila鳃的组织结构、表面形态特征及鳃小片超微结构进行了观察。结果表明梭鱼具有4对鳃,每个鳃由鳃弓、鳃丝、鳃小片和鳃耙组成。梭鱼鳃丝和鳃小片的表面结构和超微结构与其他硬骨鱼类的基本相似,鳃丝表面分布有众多规则或不规则的环形微嵴、沟、坑、孔等结构。鳃丝分为呼吸区和非呼吸区,呼吸区较为平滑,上皮细胞表面无微嵴,呈皱褶状;非呼吸区分布有沟、坑、孔等结构,上皮细胞有较规则的指纹状微嵴。鳃小片是最主要的呼吸场所,由基膜、上皮细胞、内皮细胞、柱细胞和毛细血管网组成。泌氯细胞主要分布在鳃小片基部,并有开口通往外界。本文还探讨了梭鱼鳃的结构与其功能的密切关系。     

不同NaHCO3碱度对瓦氏雅罗鱼鳃组织结构的影响

研究采用组织学方法观察和比较了达里诺尔湖瓦氏雅罗鱼(Leuciscus waleckii, 碱水种)和松花江瓦氏雅罗鱼(淡水种)在相同碳酸盐碱度胁迫下(C_A0、C_A30和C_A50)鳃组织结构的差异, 探究瓦氏雅罗鱼碱水种耐高碱特性与鳃组织结构微观调整的适应性关系。结果显示, 随着碱度增加, 碱水种鳃丝变宽、鳃小片变长、鳃小片间距变大(P<0.05); 淡水种鳃丝变宽、鳃小片间距变大(P<0.05), 鳃小片长度在C_A30时显著变长(P<0.05), 而在C_A50时与对照组无明显差异(P>0.05)。碱水种在C_A30和C_A50的氯细胞数量与对照组相比明显增加, C_A50的氯细胞排列更加紧密并且有叠加现象, 扁平上皮细胞变大, 细胞表面增厚; 淡水种在C_A30时的氯细胞数量明显多于C_A50, 但在碱度胁迫下, 鳃小片出现破损, 扁平上皮细胞、柱细胞和血细胞融合、脱落现象严重。另外, 在碱水种和淡水种鳃耙上皮细胞中发现了大量黏液细胞分泌, 随着碱度增加, 黏液细胞由大而稀疏变为小而密集, 其中碱水种的黏液细胞数量较淡水种多, 而且排列更为整齐、密集。鳃组织学研究结果表明, 瓦氏雅罗鱼碱水种通过保持鳃组织结构和生理功能的完整性达到对高碱环境的长期适应, 而淡水种则因鳃细胞融合、脱落造成生理功能丧失, 不能长期适应高碱环境。研究结果可为淡水鱼类在盐碱水的移植驯化提供依据和指导。     

急性低温胁迫对四指马鲅幼鱼肝脏、肌肉以及鳃组织结构的影响

为探究急性低温胁迫对四指马鲅Eleutheronema tetradactylum 幼鱼肝脏、肌肉以及鳃组织结构的影响。该研 究设置了常温(27±0.5 ℃)组、20 ℃组以及15 ℃组共三个温度梯度进行试验, 以常温组作为对照,在试验进行的2 h、6 h 以及12 h 分别采集肝脏、肌肉以及鳃样品进行观察和测定。结果显示: 随着低温处理时间的延长, 20℃组的肝脏细胞空泡结构呈增多, 细胞核偏离, 染色变浅趋势; 肌纤维出现弯曲现象, 肌纤维之间与内部均出现较大间隙; 线粒体丰富细胞数目增多, 鳃小片末端轻微的膨大到鳃小片整体水肿且严重弯曲, 血管以及血窦内出现大量的红细胞, 部分鳃小片因红细胞过多而涨破; 同时, 15 ℃组肝脏细胞排列混乱, 肝板结构不清晰, 细胞核溶解, 肝脏整体失去固有形态。肌纤维间隙增大, 部分断裂直至肌纤维之间与内部均严重开裂, 部分肌纤维溶解并暴露出细胞核。鳃小片表皮出现轻微的脱落, 少部分鳃小片胀大到其吸水涨破, 鳃小片基本形态不可见。该研究结果为优化四指马鲅养殖生产温控管理措施, 提高越冬成活率提供参考资料。     

Branchial osmoregulatory response to salinity in the gilthead sea bream, Sparus auratus

The branchial osmoregulatory response of gilthead sea bream (Sparus auratus L.) to short-term (2-192 hr) and long-term (2 weeks) exposure to different environmental salinities (5 per thousand, 15 per thousand, 25 per thousand, 38 per thousand and 60 per thousand) was investigated. A "U-shaped" relationship was observed between environmental salinity and gill Na+,K+ -ATPase activity in both long- and short-term exposure to altered salinity, with the increase in activity occurring between 24 and 96 hr after the onset of exposure. Plasma osmolality and plasma ions (sodium, chloride, calcium and potassium) showed a tendency to increase in parallel with salinity. These variables only differed significantly (P<0.05) in fish adapted to 60 per thousand salinity with respect to fish adapted to full-strength sea-water (SW). Plasma glucose remained unchanged whereas plasma lactate was elevated at 5 per thousand and 60 per thousand. Muscle water content (MWC) was significantly lower in fish adapted to 60 per thousand. Chloride cells (CC) were only present on the surface of the gill filaments and absent from the secondary lamellae. CC distribution was not altered by external salinity. However, the number and size of CC were significantly increased at salinity extremes (5 per thousand and 60 per thousand), whereas fish exposed to intermediate salinities (15 per thousand and 25 per thousand) had fewer and smaller cells. Furthermore, the CC of fish exposed to diluted SW became rounder whereas they were more elongated in fish in full-strength and hypersaline SW. This is consistent with previous reports indicating the existence of two CC types in euryhaline fish. At likely environmental salinities, gilthead sea bream show minor changes in plasma variables and the effective regulation of gill Na+,K+ -ATPase. However, at very low salinities both haemodilution and up-regulation of gill Na+,K+ -ATPase predict a poor adaptation most likely related to deficiency or absence of specific components of the CC important for ion xuptake.     

Developmental and environmental regulation of chloride cells in young American shad, Alosa sapidissima

Location, abundance, and morphology of gill chloride cells were quantified during changes in osmoregulatory physiology accompanying early development in American shad, Alosa sapidissima. During the larval-juvenile transition of shad, gill chloride cells increased 3.5-fold in abundance coincident with gill formation, increased seawater tolerance, and increased Na(+),K(+)-ATPase activity. Chloride cells were found on both the primary filament and secondary lamellae in pre-migratory juveniles. Chloride cells on both the primary filament and secondary lamellae increased in abundance (1.5- to 2-fold) and size (2- to 2.5-fold) in juveniles held in fresh water from August 31 to December 1 (the period of downstream migration) under declining temperature. This proliferation of chloride cells was correlated with physiological changes associated with migration (decreased hyperosmoregulatory ability and increased gill Na(+),K(+)-ATPase activity). Increases in chloride cell size and number of fish in fresh water were delayed and of a lower magnitude when shad were maintained at constant temperature (24 degrees C). When juveniles were acclimated to seawater, chloride cell abundance on the primary filament did not (though size increased 1.5- to 2-fold), but cells on the secondary lamellae disappeared. Na(+),K(+)-ATPase was immunolocalized to chloride cells in both fresh water and seawater acclimated fish. The disappearance of chloride cells on the secondary lamellae upon seawater acclimation is evidence that their role is confined to fresh water. The proliferation of chloride cells in fresh water during the migratory-associated loss of hyperosmoregulatory ability is likely to be a compensatory mechanism for increasing ion uptake. J. Exp. Zool. 290:73-87, 2001.     

Ultrastructural alterations in branchial chloride cells of Atlantic salmon, Salmo salar, during parr-smolt transformation and early development in sea water

The ultrastructure of the gill primary lamellae of juvenile Atlantic salmon was examined during the parr-smolt transformation and for 42 days after smolts were exposed to sea water. Scanning electron microscopy indicated that primary lamellae were characterized by rough convoluted surfaces that became rougher throughout the experimental period and that crypts did begin to form in freshwater fish. Crypt formation increased in sea water.
Transmission electron microscopy indicated that parr preadapt for life in sea water in part by changes in chloride cells. Chloride cells show an elaboration of rough endoplasmic reticulum in fresh water and a decline of rough endoplasmic reticulum after 42 days of sea water exposure. The tubular membrane system becomes well developed in fresh water, and apical vesicles become abundant only after seawater exposure. Mitochondria are both spherical and elongate through the period and contain well developed cristae. No evidence of mitochondrial rupture was observed. The junctions between chloride cells and adjacent cells were characterized in fresh water by long tight junctions with desmosomes. This type of junction continued in sea water and was the norm between chloride cells and accessory cells after 42 days of seawater exposure. While leaky junctions appeared to be forming, no evidence was found of membrane interdigitation between accessory cells and chloride cells after 42 days of seawater exposure. It also appeared that seawater exposure influenced the number of chloride cells exposed to the external milieu.
Pavement cells showed an elaboration in fresh water of free ribosomes and rough endoplasmic reticulum and these elements became less prominent after seawater exposure.     

Late migration and seawater entry is physiologically disadvantageous for American shad juveniles

Juvenile American shad Alosa sapidissima were subjected to isothermal transfers into sea water (salinity 24)‘early’(1 September; 24° C) and ‘late’(10 November; 10° C) in the autumn migratory season. Early acclimation resulted in a modest osmotic perturbation that recovered rapidly. Haematocrit declined by 14% at 24 h, recovering within 48 h. Plasma osmolality increased by 6% at 4 h, recovering within 8 h. Early acclimation caused a two‐fold increase in gill Na⁺, K⁺‐ATPase activity by 24 h and a four‐fold increase by 4 days. The number of chloride cells on the primary gill filament increased two‐fold by 4 days. Chloride cells on the secondary lamellae rapidly decreased from 22 cells mm^?1 to <2 cells mm^?1 within 4 days. Late acclimation resulted in a severe and protracted osmotic perturbation. Haematocrit levels declined by 23% at 4 days, recovering by 14 days. Plasma osmolality increased by 36% by 48 h, recovering by 4 days. Initial gill Na⁺, K⁺‐ATPase activity was two‐fold greater than in ‘early’ fish and did not change during acclimation. Initial numbers of chloride cells on the primary filament were two‐fold greater than ‘early’ fish and did not increase during acclimation. Initial number of chloride cells on the secondary lamellae was five‐fold greater than ‘early’ fish (116 v. 22 cells mm^?1) and declined to negligible numbers over 14 days. Differences between initial measures for ‘early’ and ‘late’ fish reflect previously described physiological changes associated with migration. These data indicate that late migrants face a greater physiological challenge during seawater acclimation than early migrants. Physiological performance apparently limits the observed duration of autumnal migration.     

Thyroid status alters gill ionic metabolism and chloride cell morphology as evidenced by scanning electron microscopy in a teleost Anabas testudineus (Bloch): short and long term in vivo study

Gill is the main organ of osmotic regulation in teleosts and chloride cells are the sites of ion transport across gill epithelium. Thyroid hormones are implicated in the regulation of osmotic balance in teleosts also. Treatment with 6-propyl thiouracil (6-PTU) inhibited the membrane bound enzyme Na+K+ ATPase in the gill while triiodothyronine (T3) injection stimulated it in a short-term in vivo study in the teleost Anabas testudineus. Na+, K+ and Ca2+ ions were also decreased in the 6-PTU treated fish and the T3 treatment increased their concentrations in the gill lamellae. The gill morphology also changed according to the thyroid status in the long term study. 6-PTU treatment altered the typical serrated morphology of the gill lamellae, while the T3 treatment reversed it. T3 injection increased the density of pavement and chloride cells as evidenced by scanning electron microscopy. The results demonstrate that physiological status of the thyroid influences gill Na+ pump activity and chloride cell morphological changes. Further, the study suggests a regulatory role of T3 on gill ions (Na+, K+ and Ca2+), Na+K+ and Ca2+ ATPase activity and the different gill cell types in A. testudineus.     

Presence of chloride cells in gill filaments and lamellae of the skate Torpedo marmorata

The morphology of chloride cells in the gill of the skate Torpedo marmorata is described from a light and an electron microscopic study. Chloride cells have been regularly observed in both the gill filament (or primary epithelium) and the lamellae (or secondary epithelium). Chloride cells located in the filament usually display convex apical regions with microvilli protruding amongst microridges of neighbouring pavement cells, whereas chloride cells in the lamellae are located between the two epithelial layers, and contact with the external milieu is via a narrow apical opening. Present observations are discussed in relation to data on the presence of chloride cells in the lamellae of marine teleost fish, and it is suggested that the occurrence of chloride cells all along the lamellae might be in some way inversely related to fish activity.     

Effect of salinity on osmoregulatory patch epithelia in gills of the blue crab Callinectes sapidus

In euryhaline crabs, ion-transporting cells are clustered into osmoregulatory patches on the lamellae of the posterior gills. To examine changes in the branchial osmoregulatory patch in the blue crab Callinectes sapidus in response to change in salinity and to correlate these changes with other osmoregulatory responses, crabs were acclimated to a range of salinities between 10 and 35 ppt. When crabs that had been acclimated to 35 ppt were subsequently transferred to 10 ppt, both the size of the osmoregulatory patch on individual gill lamellae and the specific activity of Na+, K+-ATPase in whole-gill homogenates increased only after the first 24 h of exposure to dilute seawater. Enzyme activity and size of patch area increased gradually and reached their maxima (increasing by 200% and 60%, respectively) 6 days following transfer to 10 ppt seawater and then remained at these levels. Patch size at acclimation varied inversely with the salinity for seawater dilutions below 26 ppt (the isosmotic point of the crab), although it did not vary in salinities at or above 26 ppt. Thus, the size of the patch clearly is modulated with acclimation salinity, but it increases only in those salinities in which the crab hyperosmoregulates. An increase in the total RNA/DNA ratio in gill homogenates, the lack of mitotic figures in the lamellae, and the lack of incorporation of bromodeoxyuridine into nuclei of lamellar epithelial cells during acclimation to dilute seawater were interpreted as evidence that no cell proliferation had occurred and that increases in the size of the osmoregulatory patch occurred through differentiation of existing gas exchange cells or of undifferentiated epithelial cells into ion-transporting cells.     

久效磷对美国红鱼鳃Na^＋／K^＋-ATP酶活性和超显微结构的影响

经不同试验浓度的久效磷(0．25、0．5、1．0和2．0mg·L^-1)处理美国红鱼4d后,分别对鱼鳃Na^+／K^+—ATP酶活性和氯细胞密度进行了测定和计数,并观察了鱼鳃组织显微结构和超微结构的变化。结果表明,低浓度久效磷(0．25mg·L^-1)处理可以诱导鱼鳃氯细胞大量增生,Na^+／K^+—ATP酶活性增强,随着试验浓度的增加,久效磷对鳃组织的损伤越来越重,Na^+／K^+—ATP酶活性逐渐降低;久效磷对鱼鳃显微结构的损伤表现为鳃小片上皮细胞水肿、脱离。鳃小片基部粘连。鳃小片上皮细胞角质化;超微结构变化主要为内质网、线粒体、微小管和核膜的水肿及部分溶解,这种损伤表现为由细胞膜到细胞核的动态过程。     

Cellular and epithelial adjustments to altered salinity in the gill and opercular epithelium of a cichlid fish (Oreochromis mossambicus)

Morphological features of the gill and opercular epithelia of tilapia (Oreochromis mossambicus) have been compared in fish acclimated to either fresh water (FW) or hypersaline water (60 S) by scanning electron and fluorescence microscopy. In hyperosmoregulating, i.e., FW-acclimated, tilapia only those mitochondria-rich (MR) cells present on the filament epithelium of the gill were exposed to the external medium. After acclimation of fish to hypersaline water these cells become more numerous, hypertrophy extensively, and form apical crypts not only in the gill filament but also in the opercular epithelium. Regardless of salinity, MR cells were never found to be exposed to the external medium on the secondary lamellae. In addition, two types of pavement cells were identified having distinct morphologies, which were unaffected by salinity. The gill filaments and the inner operculum were generally found to be covered by pavement cells with microridges, whereas the secondary lamellae were covered exclusively by smooth pavement cells.