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Potentiometric measurement of cell volume changes and intracellular ion concentrations under voltage-clamp conditions in invertebrate nerve cells.

Because changes in cell volume might disturb the normal function of animal cells, most cells are endowed with volume-regulating mechanisms. Experimentally induced changes in cell volume are often paralleled by changes in the membrane potential, which might affect a variety of transport processes across the cell membrane and, in turn, volume regulation [1, 2]. We have shown previously that multi-barrelled ion-selective microelectrodes are useful tools for measurement of cell volume and the intracellular concentrations of ions that might be relevant to volume regulation [1]. To investigate whether voltage-dependent transport processes are involved in cell-volume regulation we combined the potentiometric technique of ion-selective microelectrodes with the voltage-clamp technique. This combination enables simultaneous recording of cell volume, relevant intracellular ion concentrations, and ion currents across the cell membrane at a fixed membrane potential.

Animals↗

Invertebrate species with nonpelagic larvae have elevated levels of nonsynonymous substitutions and reduced nucleotide diversities.

Under a nearly neutral model in which most amino acid substitutions are slightly deleterious, variation in demography, population structure, and other ecological factors among closely related species can potentially modify the effective population size or the selective regime, leading to differences in the rate of nonsynonymous substitution. Ratios of nonsynonymous to synonymous substitutions (d(N)/d(S)) between species were analyzed in a sea star genus (Patiriella) and a molluscan genus (Littorina), each with diverse modes of reproduction, including multiple lineages with pelagic and nonpelagic larvae. In both genera, lineages with nonpelagic larvae had significantly higher d(N)/d(S) ratios than lineages with pelagic larvae. The hypothesis that the elevated d(N)/d(S) ratios in species with nonpelagic larvae was due to reduced effective population size was tested by comparing nucleotide diversities in three genera of gastropod mollusks (Littorina, Crepidula, and Hydrobia), each with several modes of reproduction. Overall, there was a significant (p < 0.05) reduction in nucleotide diversity in species with nonpelagic larvae compared to species with pelagic larvae.

Animals↗

Fate and effects of 2,4,6-trinitrotoluene (TNT) from dumped ammunition in a field study with fish and invertebrates.

2,4,6-Trinitrotoluene (TNT) is the major explosive ingredient in ammunition dumped into lakes and sea after World War II. The aim of the present field study was to study the fate and effect of TNT and its degradation products from dumped ammunition. Artillery shells were cleaved longitudinally to expose TNT and placed in open boxes filled with sediment, and then placed at the sea bottom. Sediment samples were taken in each box at the start and after 3, 9, 13, 20, 24, 33, and 36 months, and the sediments were tested for toxicity with bioassays using Nitocra spinipes (96 h), Hyalella azteca (96 h), and Daphnia magna (24 and 48 h). The result from the bioassays showed no impact of dumped ammunition on the survival of H. azteca and mobility of D. magna. Bioassays with N. spinipes showed significant differences in toxicity between control boxes and boxes with shells after 9 months and thereafter. The mean mortality (+/- SD) of N. spinipes in boxes with shells was 63 +/- 22%, and the mortality in control boxes was 23 +/- 17%. No continuous increase in sediment toxicity over time was found. After 3 years, cages with European flounder (Platichtys flesus) and blue mussels (Mytilus edulis) were attached to the boxes. The fish were examined for biochemical and physiological effects 8 weeks later. Exposure to ammunition, which had rested on the sea bottom 3 years, caused no significant effects on body indices, hematological variables, and detoxification and antioxidant enzymes activities in the flounder. The sediment, bile, and blood plasma of exposed fish, and hepatopancreas of exposed mussels, contained no detectable levels of TNT and its metabolites. Only minor disappearance of TNT from the shells could be detected by visual inspection on site (by scuba divers). This study suggests that the survival of sensitive benthic organisms, e.g., N. spinipes, might be negatively affected at an ammunition dumping site.

Animals↗

[Fundamental mechanisms of apoptosis in the simplest invertebrates, the Porifera].

Sponges (phylum Porifera) represent the oldest metazoan taxon. Through a variety of molecular biological methods the existence of elaborated mechanisms such as programmed cell death was established in those organisms that are regarded today as living fossils. Among the apoptotic proteins identified in sponges are the key molecules of apoptotic pathways, effectors (caspases, death domain proteins) and regulators (Bcl-2 homologues) of programmed cell death. Furthermore, when introduced in vertebrate cells one sponge anti-apoptotic Bcl-2 homologue displayed its pro-survival function in the heterologous system, confirming the conserved character of apoptotic cell death. Comparing apoptotic elements of both the most ancient and the most recent of all metazoans might elucidate the evolution of programmed cell death and processes involved, such as aging.

Aging↗

Muscarinic acetylcholine receptor compounds alter net Ca2+ flux and contractility in an invertebrate smooth muscle.

Responses of a holothurian smooth muscle to a range of muscarinic (M(1) to M(5)) acetylcholine receptor (mAChR) agonists and antagonists were surveyed using calcium (Ca(2+))-selective electrodes and a mechanical recording technique. Most of the mAChR agonists and antagonists tested increased both contractility and net Ca(2+) efflux, with M(1)-specific agents like oxotremorine M being the most potent in their action. To investigate the possible sources of Ca(2+) used during mAChR activation, agents that disrupt intracellular Ca(2+) ion sequestration [cyclopiazonic acid (CPA), caffeine, ryanodine], the phosphoinositide signaling pathway [lithium chloride (LiCl)], and L-type Ca(2+) channels (diltiazem and verapamil) were used to challenge contractions induced by oxotremorine M. These contractions were blocked by treatment with CPA, caffeine, LiCl, and by channel blockers, diltiazem and verapamil, but were unaltered by ryanodine. Our data suggest that this smooth muscle had an M(1,3,5)-like receptor that was associated with the phosphoinositide signaling pathway that relied on intracellular Ca(2+) stores, but secondarily used extracellular Ca(2+) via the opening of L-type channels.

Animals↗

Locomotor responses and neuron excitability in conditions of haloperidol blockade of dopamine in invertebrates and vertebrates.

Levels of movement activity were used to identify two groups of rats: those with high- and low-activity levels. Blockade of dopamine receptors with haloperidol led to suppression of locomotor activity in both groups of rats; in common snails, haloperidol decreased the rate of locomotion. The excitability of spinal centers in rats decreased 5 min after single i.v. injections, with gradual recovery seen by 30 min. Chronic administration of haloperidol suppressed post-tetanic potentiation of the H response in the gastrocnemius muscle of spinal rats. Prolonged use of haloperidol induced significant hyperpolarization of the membrane potential of command neurons in common snails and increased the action potential generation threshold. Selective pharmacological exclusion of the brain dopamine system was found to lead to decreases in the excitability of defined neurons in snails and the spinal motor centers in rats, also producing impairments in locomotor responses in these animals.

Animals↗