Intragenic duplication and divergence in the spectrin superfamily of proteins

Many structural, signaling, and adhesion molecules contain tandemly repeated amino acid motifs. The alpha-actinin/spectrin/dystrophin superfamily of F-actin-crosslinking proteins contains an array of triple alpha-helical motifs (spectrin repeats). We present here the complete sequence of the novel beta-spectrin isoform beta(Heavy)- spectrin (beta H). The sequence of beta H supports the origin of alpha- and beta-spectrins from a common ancestor, and we present a novel model for the origin of the spectrins from a homodimeric actin-crosslinking precursor. The pattern of similarity between the spectrin repeat units indicates that they have evolved by a series of nested, nonuniform duplications. Furthermore, the spectrins and dystrophins clearly have common ancestry, yet the repeat unit is of a different length in each family. Together, these observations suggest a dynamic period of increase in repeat number accompanied by homogenization within each array by concerted evolution. However, today, there is greater similarity of homologous repeats between species than there is across repeats within species, suggesting that concerted evolution ceased some time before the arthropod/vertebrate split. We propose a two-phase model for the evolution of the spectrin repeat arrays in which an initial phase of concerted evolution is subsequently retarded as each new protein becomes constrained to a specific length and the repeats diverge at the DNA level. This evolutionary model has general applicability to the origins of the many other proteins that have tandemly repeated motifs.      

Evolutionary implications of mitochondrial genetic variation: mitochondrial genetic effects on OXPHOS respiration and mitochondrial quantity change with age and sex in fruit flies

The ancient acquisition of the mitochondrion into the ancestor of modern‐day eukaryotes is thought to have been pivotal in facilitating the evolution of complex life. Mitochondria retain their own diminutive genome, with mitochondrial genes encoding core subunits involved in oxidative phosphorylation. Traditionally, it was assumed that there was little scope for genetic variation to accumulate and be maintained within the mitochondrial genome. However, in the past decade, mitochondrial genetic variation has been routinely tied to the expression of life‐history traits such as fertility, development and longevity. To examine whether these broad‐scale effects on life‐history trait expression might ultimately find their root in mitochondrially mediated effects on core bioenergetic function, we measured the effects of genetic variation across twelve different mitochondrial haplotypes on respiratory capacity and mitochondrial quantity in the fruit fly, Drosophila melanogaster. We used strains of flies that differed only in their mitochondrial haplotype, and tested each sex separately at two different adult ages. Mitochondrial haplotypes affected both respiratory capacity and mitochondrial quantity. However, these effects were highly context‐dependent, with the genetic effects contingent on both the sex and the age of the flies. These sex‐ and age‐specific genetic effects are likely to resonate across the entire organismal life‐history, providing insights into how mitochondrial genetic variation may contribute to sex‐specific trajectories of life‐history evolution.     

Retinal mechanisms of visual adaptation in the skate

Electrical potentials were recorded from different levels within the skate retina. Comparing the adaptive properties of the various responses revealed that the isolated receptor potential and the S-potential always exhibited similar changes in sensitivity, and that the b-wave and ganglion-cell thresholds acted in concert. However, the two sets of responses behaved differently under certain conditions. For example, a dimly iluminated background that had no measurable effect on the senitivities of either of the distal responses, raised significantly the thresholds of both the b-wave and the ganglion cell responses. In addition, the rate of recovery during the early, "neural" phase of dark adaptation was significantly faster for the receptor and S-potentials than for the b-wave or ganglion cell discharge. These results indicate that there is an adaptive ("network") mechanism in the retina which can influence significantly b-wave and gaglion cell activity and which behaves independently of the receptors and horizontal cells. We conclude that visual adaptation in the skate retina is regulated by a combination of receptoral and network mechanisms.     

Regulation of Dopamine Release from Interplexiform Cell Processes in the Outer Plexiform Layer of the Carp Retina

The gamma-aminobutyric acid (GABA) antagonists bicuculline and picrotoxin stimulate a four- to fivefold increase in endogenous dopamine release from isolated intact carp retina. The release evoked by these agents is Ca2+ dependent, a finding suggesting a vesicular release. Using light microscopic autoradiography, we have localized the sites of dopamine release to the dopaminergic interplexiform cell processes of the outer plexiform layer, which synapse onto horizontal cells. Our findings support previous suggestions that the dopaminergic interplexiform cells receive GABAergic inhibitory input and that the effects of GABA antagonists on horizontal cells are mediated by dopamine release from the interplexiform cells.     

The interplexiform-horizontal cell system of the fish retina: effects of dopamine, light stimulation and time in the dark

Interplexiform cells contact cone horizontal cells in the fish retina and probably release dopamine at synaptic sites. The effects of dopamine, certain related compounds, and light and dark régimes were tested on the intracellularly recorded activity of horizontal cells in the superfused carp retina to elucidate the functional role of the interplexiform cell. Dopamine application onto retinae kept in the dark for 30-40 min increased the size of the responses of cone horizontal cells to small-spot stimuli but decreased response size to large- and full-field stimuli. Dopamine also altered the response waveform of these cells; the transient at response onset increased in size and the depolarizing afterpotential decreased in size. Haloperidol, a dopamine antagonist, blocked these effects of dopamine application. Forskolin, an adenylate cyclase activator, increased the size of the responses of the cells to small-spot stimuli. Superfusion of vasoactive intestinal peptide did not produce any effects on horizontal cells. The results indicate that dopamine produces multiple physiological effects on cone horizontal cells by activation of an intracellular enzyme system. We propose that some of these effects are probably related to an uncoupling of the gap junctions between horizontal cells, but that other effects are most likely not explained on this basis and reflect additional changes induced in the cells by dopamine. After prolonged periods of darkness (100-110 min), compared with short periods (30-40 min), L-type cone horizontal cells exhibited responses similar to those obtained during dopamine application. Dim flickering or continuous light backgrounds did not mimic the effects of dopamine. Although dopamine application onto retinae after short-term darkness produced dramatic effects on L-type cone horizontal cells, little or no effect was observed when dopamine was applied while the effects of a previous dopamine application were still present or after prolonged darkness. These results suggest that interplexiform cells may release dopamine after prolonged darkness and that interplexiform cells may regulate lateral inhibitory effects mediated by L-type cone horizontal cells as a function of time in the dark.     

Measurement of faecal bile acid sulphates

A method is described for the measurement, by difference, of the sulphate fractions of faecal bile acids. A solvolysis step (for the deliberate hydrolysis of the bile acid sulphates) was added to the procedure of sample homogenisation, extraction, enzymatic hydrolysis and thin-layer chromatography. The bile acids were quantitated by gas—liquid chromatography of their methyl ester and trifluoroacetate methyl ester derivatives on 3% QF-1 columns. The total bile acid excretion in 15 control subjects was 603 ± 71 mg/24 h (x̄ ± S.E.M.). The major bile acid peaks (mg/24 h) were: lithocholic acid, without solvolysis 118 ± 26 and including solvolysis 175 ± 30; deoxycholic acid 60 ± 8 and 90 ± 18 and chenodeoxycholic acid 13 ± 7 and 15 ± 7. It was concluded that bile acid sulphates may form a considerable proportion of the total bile acids excreted in man.