Stereological estimation of surface area and barrier thickness of fish gills in vertical sections

Previous morphometric methods for estimation of the volume of components, surface area and thickness of the diffusion barrier in fish gills have taken advantage of the highly ordered structure of these organs for sampling and surface area estimations, whereas the thickness of the diffusion barrier has been measured orthogonally on perpendicularly sectioned material at subjectively selected sites. Although intuitively logical, these procedures do not have a demonstrated mathematical basis, do not involve random sampling and measurement techniques, and are not applicable to the gills of all fish. The present stereological methods apply the principles of surface area estimation in vertical uniform random sections to the gills of the Brazilian teleost Arapaima gigas. The tissue was taken from the entire gill apparatus of the right‐hand or left‐hand side (selected at random) of the fish by systematic random sampling and embedded in glycol methacrylate for light microscopy. Arches from the other side were embedded in Epoxy resin. Reference volume was estimated by the Cavalieri method in the same vertical sections that were used for surface density and volume density measurements. The harmonic mean barrier thickness of the water‐blood diffusion barrier was calculated from measurements taken along randomly selected orientation lines that were sine‐weighted relative to the vertical axis. The values thus obtained for the anatomical diffusion factor (surface area divided by barrier thickness) compare favourably with those obtained for other sluggish fish using existing methods.     

福建牡蛎新基因SMRP1的功能分析

通过对福建牡蛎幼体不同发育时期的转录组文库比对分析,结果显示牡蛎附着变态相关蛋白1(settlement and metamorphosis related protein1,SMRP1)基因在幼体附着变态时期高表达,利用定量即时聚合酶连锁反应(quantitative real time polymerase chain reaction,qRT-PCR)技术确认了SMRP1基因在幼体附着变态时期高表达,且进一步发现SMRP1集中表达在牡蛎鳃和外套膜组织中,具有明显的组织特异性。利用组织和整体原位杂交技术检测SMRP1基因在不同时期幼体以及成体的外套膜和鳃组织中的表达定位情况,发现SMRP1基因集中表达在幼体壳顶时期到眼点幼虫的时期,在成体鳃和外套膜组织集中表达在组织的外层上皮细胞中。结合牡蛎幼体在附着变态时期鳃丝的快速生长发育及其在成体外套膜中的功能作用分析表明SMRP1基因对牡蛎幼体鳃和外套膜的形成和生长发育具有重要的作用。该研究为解析胰岛素相关多肽受体信号通路的功能提供数据支持。     

The Living: Surface Tensions

‘Our work begins with the premise of a dynamic world. Political and cultural conditions change: what if the walls and windows morphed in response?’ New York-based team The Living, led by David Benjamin and Soo-in Yang, have developed a practice of deep, open-source interfaces of participation. Jordan Geiger describes how for The Living their home city and its environs is a site of evolving forms of public space whether on land, air or water. Copyright © 2010 John Wiley & Sons, Ltd.     

In Vivo Functional Assay in Fish Gills: Exploring Branchial Acid-Excreting Mechanisms in Zebrafish

Molecular and physiological analyses in ionoregulatory organs (e.g., adult gills and embryonic skin) are essential for studying fish ion regulation. Recent progress in the molecular physiology of fish ion regulation was mostly obtained in embryonic skin; however, studies of ion regulation in adult gills are still elusive and limited because there are no direct methods for in vivo functional assays in the gills. The present study applied the scanning ion-selective electrode technique (SIET) in adult gills to investigate branchial H⁺-excreting functions in vivo. We removed the opercula from zebrafish and then performed long-term acid acclimation experiments. The results of Western blot and immunofluorescence showed that the protein expression of H⁺-ATPase (HA) and the number of H⁺-ATPase-rich ionocytes were increased under acidic situations. The SIET results proved that the H⁺ excretion capacity is indeed enhanced in the gills acclimated to acidic water. In addition, both HA and Na⁺/H⁺ exchanger (Nhe) inhibitors suppressed the branchial H⁺ excretion capacity, suggesting that H⁺ is excreted in association with HA and Nhe in zebrafish gills. These results demonstrate that SIET is effective for in vivo detection in fish gills, representing a breakthrough approach for studying the molecular physiology of fish ion regulation.     

Palp to gill area ratio of bivalves: A sensitive indicator of elevated suspended solids

Area measurements were made of the gills and labial palps of several freshwater bivalve species from sites with distinctly different suspended solids concentrations. Without exception, the palp to gill area ratio (PA:GA) was markedly higher for populations from sites with high versus low suspended solids concentrations. The average PA:GA (expressed as a percentage) ranged from 9·3 to 11·5 for bivalves from high suspended solids concentration sites and from 2.5 to 4.8 for bivalves from low suspended solids concentration sites. Such interpopulation differences were observed for two introduced species, Dreissena polymorpha and Corbicula fluminea, despite extremely brief residence times in an evolutionary sense. At any particular location, different species of native unionids had similar PA:GA. These results suggest that PA:GA is a sensitive biological indicator of suspended solids concentration, with interpopulation differences probably reflecting ecophenotypic rather than genetic variation.     

The availability of cadmium to perfused rainbow trout gills in different water qualities

The uptake of cadmium in perfused gills from rainbow trout (Salmo gairdneri, Rich.) in different water qualities has been studied.In the naturally hard alkaline Uppsala tap water, the calcium concentration was the dominating factor controlling the cadmium transfer through the gills. A strong inverse relationship was measured between the external Ca²⁺ concentration up to 3 mmol l^?1 (< 120 mg Ca²⁺ l^?1), and the Cd transfer. A higher Ca²⁺ concentration (10 mmol l^?1) did not further affect the transfer. Magnesium decreased the transfer, but only at concentrations 4 to 5 times as high as calcium. At a constant Ca²⁺/Mg²⁺ hardness, the transfer was a function of the free cadmium ion activity (Cd²⁺). Ca²⁺ and Mg²⁺ selectively reduced the transfer by a biological mechanism, probably by changing the permeability of the gill epithelium.Cd transfer was not dependent on pH in the range pH 5–7. A lower transfer at pH 7.6 was related to a lower Cd²⁺ activity, probably due to the formation of non-available bicarbonate/carbonate complexes.Cd transfer decreased in the presence of 121 mmol l^?1 NaCl (0.7% salinity). This decrease was not caused by the altered osmotic gradient over the gills. The transfer was proportional to the Cd²⁺ activity. CdCl⁺ and CdCl₂ were not available for the gills.The retention of Cd in perfused gill tissue was proportional to the Cd²⁺ activity in all water qualities tested. Tissue accumulation of Cd was not sensitive to external Ca and Mg and in this respect different from the transfer.     

Morphological investigation of the gills of the dusky grouper Epinephelus marginatus (Lowe 1834) using gross anatomy and scanning electron microscopy

The current study has designed to investigate the morphological characteristics of the gills of the dusky grouper (Epinephelus marginatus) by gross morphology, scanning electron microscopy, and morphometric analysis. The study was carried out on 10 fresh fish. The fish has four-gill arches on each side. The lengths of the first-, second-, third-, and fourth-gill arches were 5.27, 4.2, 3.2, and 2.8 cm. The gill arches carried a longitudinal band, the bases of the gill filaments, and gill rakers that varied from rectangular to circular shapes. Each gill arch had two main lateral and medial rows and two accessory rows of gill rakers in an alternative manner with each other. The dusky grouper fish had long rakers, whereas the longest one was the lateral rakers of the first arch, which were 467 and 1271.9 μm in width and length. Three types of spines appeared on the gill arches and rakers. The long spine had detected on the apex of the short rakers. Each gill arch carried on its ventral side two rows of gill filaments. The long filaments were at the middle of the gill arch, while the short ones were at the rostral end of the gill arch. The study has demonstrated the taste buds, mucous, chloride cells, and significant features of the epithelial covering of the gill arches and rakers. The morphology of the gills of the dusky grouper indicates the adaptation to their marine environment.     

Expression of Antimicrobic Peptide Piscidin1 in Gills Mast Cells of Giant Mudskipper Periophthalmodon schlosseri (Pallas, 1770)

The amphibious teleost Giant mudskipper (Periophthalmodon schlosseri, Pallas 1770) inhabit muddy plains and Asian mangrove forests. It spends more than 90% of its life outside of the water, using its skin, gills, and buccal-pharyngeal cavity mucosa to breathe in oxygen from the surrounding air. All vertebrates have been found to have mast cells (MCs), which are part of the innate immune system. These cells are mostly found in the mucous membranes of the organs that come in contact with the outside environment. According to their morphology, MCs have distinctive cytoplasmic granules that are released during the degranulation process. Additionally, these cells have antimicrobial peptides (AMPs) that fight a variety of infections. Piscidins, hepcidins, defensins, cathelicidins, and histonic peptides are examples of fish AMPs. Confocal microscopy was used in this study to assess Piscidin1 expression in Giant Mudskipper branchial MCs. Our results demonstrated the presence of MCs in the gills is highly positive for Piscidin1. Additionally, colocalized MCs labeled with TLR2/5-HT and Piscidin1/5-HT supported our data. The expression of Piscidin1 in giant mudskipper MCs highlights the involvement of this peptide in the orchestration of teleost immunity, advancing the knowledge of the defense system of this fish.     

Gross morphological and surface ultrastructural investigation on the gills of the European barracuda Sphyraena sphyraena

The present work examined 10 gill systems of European barracuda grossly and by scanning electron microscopy (SEM). Grossly, there were four pairs of the gill arches. The convex border of the gill arch carried the gills filaments, but there were abundance of spines near to its concave border and the gill rakers were absent. Laterally near to the convex border of the gill arch, SEM observations revealed that the first gill arch carried small elliptical, oval, cuboidal, and triangular groups of two shapes of spines; spearhead-like spines and canine-like spines, but the other three gill arches carried larger groups of the same shaped spines with the appearance of waterfalls. Medially near to the convex border of the first gill arch, the canine-like spines were observed only in the form of vertical rectangular groups that adhered in some areas. Laterally near to the concave border, the two shapes of spines were present in the form of longitudinal groups separated by spaces at the first gill arch, but these spaces were absent in the other three gill arches. Medially near to the concave border of the first gill arch, the two shapes of spines were presented in oval groups, while the other three gill arches were covered entirely by cuboidal groups of the two shapes of spines. The absence of the gill rakers in conjunction with an abundance of spines helps the European barracuda to control the food particles from escaping to the gill filaments to prevent its suffocation.