The Atlantic Salmon Mhc Class II α and β Promoters are Active in Mammalian Cell Lines

The major histocompatibility complex class II (MHCII) genes are only constitutively expressed in certain immune response cells such as B cells, macrophages, dendritic cells and other antigen presenting cells. This cell specific expression pattern and the presence of conserved regions such as the X-, X2-, Y-, and W-boxes make the MHCII promoters especially interesting as vector constructs. We tested whether the Atlantic salmon (Salmo salar L.) MHCII promoters can function in cell lines from other organisms. We found that the salmon MHCII α and MHCII β promoters could drive expression of a LacZ reporter gene in adherent lymphoblast cell lines from dog (DH82) and rabbit (HybL-L). This paper shows that the promoters of Atlantic salmon MHCII α and β genes can function in mammalian cell lines.     

Sound localization by the Hawaiian squirrelfishes, Myripristis berndti and M. argyromus

Sound localization was investigated in a large pond open to a bay and similar to the normal environment of the animals. Observations were made of fish movements towards one of two underwater loud-speakers emitting squirrelfish alarm calls normally produced in response to predators. When the sound source was within 2·0 m of the test cage housing the fish, the subjects faced and moved toward the speaker. The animals responded some of the time when the source was within 3·0 m but generally did not orient to the sound source when the speaker was beyond 3·0 m. Response loss was correlated with the fish being in the acoustic far-field. Possible cues which release and direct localization remain unknown, but include particle velocity information alone, or some change in particle velocity: pressure relationships.     

Mitigating the effects of high-head dams on the Columbia River, USA: experience from the trenches

Worldwide, humans have tremendously altered freshwater ecosystems and arguably, construction of dams has had the greatest effect. Maintaining natural ecological processes and developing mitigation strategies that will maintain species while retaining dam benefits is challenging. In the Columbia River, USA, over the last 30 years more than US$7 billion has been spent on efforts to save historically large runs of salmon. These efforts have included improving passage conditions at dams through construction of efficient fish ladders for adult salmon, effective fish passage facilities for downstream migrating juvenile salmon, voluntarily spilling water to decrease the number of downstream migrants that pass through turbines, modifying dam operations to provide more constant flow and providing additional flow from storage reservoirs to create more natural flow through areas inundated by dams. Construction of hatcheries to offset losses in habitat for wild fish has also occurred. Further, for salmon from the Snake River, the largest tributary to the Columbia River, a large percent of juvenile salmon smolts are collected at upstream dams and transported in barges to the lower river to avoid passage through dams, turbines, and reservoirs. Experiences in the Columbia River suggest that the sum of all of these actions may keep salmon stocks from going extinct, but the technological fixes will not likely provide complete mitigation for altered freshwater ecosystems. Guest editors: R. L. Welcomme & G. Marmulla Hydropower, Flood Control and Water Abstraction: Implications for Fish and Fisheries     

Oxygen consumption and blood flow distribution in perfused skeletal muscle of chinook salmon

An isolated, perfused salmon tail preparation showed oxyconformance at low oxygen delivery rates. Addition of pig red blood cells to the perfusing solution at a haematocrit of 5 or 10% allowed the tail tissues to oxyregulate. Below ca. 60 ml O₂ kg⁻¹ h⁻¹ of oxygen delivery (DO₂), VO₂ was delivery dependent. Above this value additional oxygen delivery did not increase VO₂ of resting muscle above ca. 35 ml O₂ kg⁻¹ h⁻¹. Following electrical stimulation, VO₂ increased to ca. 65 ml O₂ kg⁻¹ h⁻¹, with a critical DO₂ of ca. 150 ml O₂ kg⁻¹ h⁻¹. Dorsal aortic pressure fell to 69% of the pre-stimulation value after 5 min of stimulation and to 54% after 10 min. Microspheres were used to determine blood flow distribution (BFD) to red (RM) and white muscle (WM) within the perfused myotome. Mass specific BFD ratio at rest was found to be 4.03 ± 0.49 (RM:WM). After 5 min of electrical stimulation the ratio did not change. Perfusion with saline containing the tetrazolium salt 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) revealed significantly more mitochondrial activity in RM. Formazan production from MTT was directly proportional to time of perfusion in both red and WM. The mitochondrial activity ratio (RM:WM) did not change over 90 min of perfusion.     

Regional permeability differences between the proximal and distal portions of the isolated salmonid posterior intestine

The objective of this study was to assess regional variations in the permeability of the salmon posterior intestine and to evaluate the effect of permeability enhancers as a basis for oral delivery of biologically active peptides. Proximal and distal portions of the posterior intestine of the chinook salmon (Oncorhynchus tshawytscha) were removed, mounted as flat sheets in Ussing chambers and superfused with trout Ringer's. Intestinal permeability was assessed under short-circuit conditions by measurement of ¹⁴C-mannitol (mucosal to serosal) flux. Tissues were treated either with the mucolytic agent dithiothreitol (10 mmol · l⁻¹), the permeability enhancer sodium deoxycholate (5.0 mmol · l⁻¹) or both and compared to untreated controls. Both proximal and distal control tissues had low permeabilities, but the distal region had a lower transepithelial electrical resistance and produced significantly less mucus. Treatment with either dithiothreitol or sodium deoxycholate alone reduced mucus adhering to tissue in both regions but did not increase permeability or change transepithelial electrical resistance. In the distal region, sequential treatment with both agents significantly reduced adhering mucus, decreased transepithelial electrical resistance, and increased tissue permeability. The salmon posterior intestine can be divided into proximal and distal regions. The distal region is more likely to have the necessary permeability and responsiveness to enhancement for the successful delivery of peptides or polar drugs. Accepted: 14 January 1997     

Effect of addition of ‘carrier’ DNA during transient protein expression in suspension CHO culture

Transient protein expression using polyethyleneimine as a transfection agent is useful for the rapid production of small amounts of recombinant proteins. It is known that an increase in extracellular DNA concentration during transfection can lead to a nonlinear increase in intracellular DNA concentration. We present an approach that hypothesizes that this nonlinearity can be used to decrease the amount of plasmid required for productive transfections. Through addition of non coding ‘carrier’ DNA to increase total DNA concentration during transfection, we report a statistically significant increase in protein (IgG) expression per unit plasmid used for transfection. This approach could be useful to increase protein yields for large scale transfections under conditions where plasmid availability is limited.     

Ionic movements across the chorion in newly shed salmon eggs (Salmo salar L.)

Summary Sodium and chloride exchange and trans-chorion potentials were investigated in newly shed eggs from Atlantic salmon. Exposure of eggs to pH 3.5 caused the non-labile sodium fraction at pH 7.0 to become labile and lost from the egg. Chloride fluxes appear unaffected by the pH of the external medium. Trans-chorion potential was inside negative (about –100 mV) in dilute media (10^–5 mol·l^–1 NaCl or KCl) and immediately decreased as external cation concentration was increased, by about 46 mV/decade change in cation concentration, reaching about 0 mV in 10^–2 mol·l^–1 cation. Return to dilute cation solutions resulted in a slow increase in potential (repolarisation) and the time course of these potential changes was paralleled by the rate of sodium efflux, although chloride efflux was very rapid. After exposure to acid conditions repolarisation of the egg on return to dilute cation concentrations was preceeded by a phase when the pvp became inside positive. The results are discussed in terms of chorion structure, anionic charge on the perivitelline molecules and unstirred layers within the chorion.Abbreviations pvp perivitelline potential