Intraspecific phylogeography of the gopher tortoise, Gopherus polyphemus: RFLP analysis of amplified mtDNA segments.

The slow rate of mtDNA evolution in turtles poses a limitation on the levels of intraspecific variation detectable by conventional restriction fragment surveys. We examined mtDNA variation in the gopher tortoise (Gopherus polyphemus) using an alternative restriction assay, one in which PCR-amplified segments of the mitochondrial genome were digested with tetranucleotide-site endonucleases. Restriction fragment polymorphisms representing four amplified regions were analysed to evaluate population genetic structure among 112 tortoises throughout the species' range.

Fibroblast growth factor is a direct neural inducer, which combined with noggin generates anterior-posterior neural pattern.

Neural tissue in developing Xenopus embryos is induced by signals from the dorsal mesoderm. Induction of anterior neural tissue could be mediated by noggin, a secreted polypeptide found in dorsal mesoderm. We show that bFGF, a known mesoderm inducer of blastula staged ectoderm, induces neural tissue from gastrula stage ectoderm.

Photoreceptor spectral sensitivities: common shape in the long-wavelength region.

Previous measurements of mammalian photoreceptor spectral sensitivity have been analysed, with particular attention to the long-wavelength region. The measurements selected for study come from rod and cone systems, and from human, monkey, bovine and squirrel sources. For the spectra from photoreceptor electrophysiology and from psychophysical sensitivity, the frequency scaling applied by Mansfield (1985, The visual system, pp.

Identification of mammalian noggin and its expression in the adult nervous system.

The multiple roles of noggin during dorsal fate specification in Xenopus embryos, together with noggin's ability to directly induce neural tissue, inspired an effort to determine whether a similar molecule exists in mammals. Here we describe the identification of human and rat noggin and explore their expression patterns; we also localize the human NOGGIN gene to chromosome 17q22, and the mouse gene to a syntenic region of chromosome 11. Mammalian noggin is remarkably similar in its sequence to Xenopus noggin, and is similarly active in induction assays performed on Xenopus embryo tissues.

Phylogeny of six Sciurus aberti subspecies based on nucleotide sequences of cytochrome b.

The tassel-eared squirrel, Sciurus aberti, is separated into six subspecies which occupy restricted and comparable habitats in ponderosa pine forests in the south-western United States and Mexico. These forests and squirrel populations are currently isolated by large arid areas and, as such, S. aberti appears to offer an example of incipient speciation.

The slider turtle as an environmental sentinel: multiple tissue assays using flow cytometric analysis.

We used flow cytometry (FCM) to conduct a multiple-tissue assay on slider turtles (Trachemys scripta) inhabiting radioactive seepage basins. Duplicate samples of blood, heart, spleen and kidney were analysed on two different cytometers (Leitz MPV and Coulter Profile II), each employing distinct staining protocols (DAPI and PI, respectively). Both DAPI and PI assays of spleen cells demonstrated significantly greater variation in DNA content for the basin turtles than for 'control' animals from nearby, uncontaminated sites.

Phylogeny of mitochondrial DNA clones in tassel-eared squirrels Sciurus aberti.

The tassel-eared squirrel, Sciurus aberti, includes six subspecies which occupy restrictive and apparently identical habitats in Ponderosa pine forests in the south-western United States and Mexico; the strict habitat requirement of this species is based on dietary requirements which are only fulfilled in these forests. To examine evolutionary relationships among certain subspecies of S. aberti, we obtained estimates of nucleotide diversity within subspecies as well as nucleotide divergence between subspecies using mitochondrial DNA (mtDNA) analysis.

A molecular phylogeny of the gopher tortoises, with comments on familial relationships within the Testudinoidea.

Sequences from the mitochondrial cytochrome b gene were obtained to examine molecular phylogenetic relationships among the North American gopher tortoises. Data from 352 aligned positions generated a single most-parsimonious tree for each of three analytical approaches: (1) equal weighting, all substitutions; (2) equal weighting, third position changes limited to transversions; and (3) transversions weighted 10 times transitions. Identical topologies for the resulting trees depict the gopher tortoises as a monophyletic group comprising two well-defined clades.

Dark adaptation of toad rod photoreceptors following small bleaches.

The recovery of toad rod photoreceptors, following exposure to intense lights that bleached 0.02-3% of the rhodopsin, has been investigated using the suction pipette technique. The post-bleach period was accompanied by reduced flash sensitivity, accelerated kinetics, and spontaneous fluctuations (noise).

Stochastic simulation of activation in the G-protein cascade of phototransduction.

Activation of the G-protein cascade underlying phototransduction has been modeled by simulating the two-dimensional diffusional interactions that occur at the rod disc membrane between the three reacting protein species, which are the activated rhodopsin (R*), the G-protein (G), and the effector protein (E, the phosphodiesterase, PDE). The stochastic simulations confirm the main predictions of a simplified analytical model (Lamb, T. D.

Gating, selectivity and blockage of single channels activated by cyclic GMP in retinal rods of the tiger salamander.

1. Patches in the inside-out configuration were excised from the membrane of outer and inner segments of the larval tiger salamander, Ambystoma tigrinum. The current flowing through single channels opened by cyclic GMP was studied with the voltage clamp technique.

Analysis of ERG a-wave amplification and kinetics in terms of the G-protein cascade of phototransduction.

Purpose: To test rigorously the hypothesis that the a-wave of the electroretinogram (ERG) is proportional to the rod photocurrent by examining the applicability to a-waves of a recent model of the activation steps in the G-protein cascade of phototransduction.

Methods: ERGs were recorded in response to flashes of graded intensity, from six dark-adapted normal subjects and from two patients, one with retinitis pigmentosa (RP) and one with cone retinal dystrophy with rod involvement (CRD). The a-wave portions of the responses were analyzed with a model of the activation steps of the G-protein cascade.

Neural induction by the secreted polypeptide noggin.

The Spermann organizer induces neural tissue from dorsal ectoderm and dorsalizes lateral and ventral mesoderm in Xenopus. The secreted factor noggin, which is expressed in the organizer, can mimic the dorsalizing signal of the organizer. Data are presented showing that noggin directly induces neural tissue, that it induces neural tissue in the absence of dorsal mesoderm, and that it acts at the appropriate stage to be an endogenous neural inducing signal.

CT of Clonorchis Sinensis pancreatitis.

Clonorchis Sinensis infestation can involve the pancreatic duct and induce attacks of acute pancreatitis. Ct findings in a case of Clonorchis Sinensis pancreatitis revealed significant enlargement of the pancreas with compression and obstruction of the common duct. Medical treatment leads to resolution of the clinical and CT findings.

Amplification and kinetics of the activation steps in phototransduction.

We can summarize our investigation of amplification in the activation steps of vertebrate phototransduction as follows. (1) A theoretical analysis of the activation steps of the cGMP cascade shows that after a brief flash of phi photoisomerizations the number of activated PDE molecules should rise as a delayed ramp with slope proportional to phi, and that, as a consequence, the cGMP-activated current should decay as a delayed Gaussian function of time (Eqn. 20).

Different channel-gating properties of two classes of cyclic GMP-activated channel in vertebrate photoreceptors.

We report that two types of cGMP-activated channel coexist in the photoreceptor plasma membrane, with the most commonly encountered class appearing broadly similar to the channel reported in previous patch-pipette experiments. However, we find that flickering of this channel between the open and closed states is so rapid that a discrete single-channel conductance cannot unequivocally be resolved; the occurrence of flickering is largely independent of membrane voltage and of the presence of cytoplasmic Ca2+ or Mg2+. In recordings from the inner segment we occasionally find a second class of cGMP-gated channel, with activity resembling that reported for cloned channels.

The application of bioassays in risk assessment of environmental pollution.

Increased contamination of the environment by toxic chemicals has resulted in the need for sensitive assays to be used in risk assessment of polluted sites. Traditional tests are useful to detect and measure concentrations of chemicals in the environment and in tissues. However, physicochemical assays possess deficiencies that impair their use in evaluating complex environmental contamination.

G-protein cascades: gain and kinetics.

G-protein cascades provide amplification in a wide variety of biological signal transducers--from hormonal and synaptic systems to the receptor cells of vision and olfaction. Through recent understanding of the molecular mechanisms involved, it is possible to construct a quantitative description of the amplification and speed of response of the cascade. The gain and kinetics can now be described in terms of physical parameters, such as enzyme activities and the densities and lateral diffusion coefficients of the proteins involved.

Mitochondrial DNA evolution at a turtle's pace: evidence for low genetic variability and reduced microevolutionary rate in the Testudines.

Evidence is compiled suggesting a slowdown in mean microevolutionary rate for turtle mitochondrial DNA (mtDNA). Within each of six species or species complexes of Testudines, representing six genera and three taxonomic families, sequence divergence estimates derived from restriction assays are consistently lower than expectations based on either (a) the dates of particular geographic barriers with which significant mtDNA genetic clades appear associated or (b) the magnitudes of sequence divergence between mtDNA clades in nonturtle species that otherwise exhibit striking phylogeographic concordance with the genetic partitions in turtles. Magnitudes of the inferred rate slowdowns average eightfold relative to the "conventional" mtDNA clock calibration of 2%/Myr sequence divergence between higher animal lineages.

A quantitative account of the activation steps involved in phototransduction in amphibian photoreceptors.

1. We have undertaken a theoretical analysis of the steps contributing to the phototransduction cascade in vertebrate photoreceptors. We have explicitly considered only the activation steps, i.

Genetic damage in a population of slider turtles (Trachemys scripta) inhabiting a radioactive reservoir.

Turtles inhabiting a radioactive reservoir appear to experience genetic damage due to environmental exposure to low concentrations of long-lived radionuclides. Total body burdens for the 50 reservoir turtles examined in the survey ranged from 164.7-4679.

The role of steroids in reproduction in female elasmobranchs and reptiles.

Adequate evidence exists to suggest the importance of temporal changes in steroid hormone ratios in the normal reproductive/vitellogenin cycle in oviparous and viviparous elasmobranchs and reptiles. In oviparous species, where the cycle is relatively short, secretion of gonadal hormones is synchronous; thus inhibitory actions of progesterone (P) on hepatic or reproductive tract functions would be offset by stimulatory actions of estradiol (E), resulting in appropriate vitellogenin secretion and reproductive tract development. In viviparous species, temporal asynchrony of E and P secretion occurs, and the actions of the individual hormones can be more easily dissected out.

Cyclic GMP and calcium: the internal messengers of excitation and adaptation in vertebrate photoreceptors.

The roles of cyclic GMP (cGMP) and calcium (Ca2+) in vertebrate rod phototransduction are reviewed, with the emphasis on developments since the discovery of the cGMP-activated conductance of the rod outer segment. The first hypothesis subjected to critical examination is that cGMP acts as the sole internal messenger of excitation. This hypothesis is evaluated with a formal, quantitative model of the biochemical actions of cGMP.

Light adaptation in cone photoreceptors of the salamander: a role for cytoplasmic calcium.

1. Light adaptation has been studied in isolated red-sensitive cone photoreceptors of the salamander, using suction pipette recordings of circulating current. 2.