Biomedical subjects

R W Morton

Publications and source records attributed to R W Morton.

4 recordsLinked to original sources

PubMed1986

Extended sensitivity for the calcium selective electrode.

Sensitivity of calcium-selective electrodes heretofore has been limited to calcium concentrations above 10(-8) M in the absence of competing ions. We describe the use of calcium buffers to stabilize the free calcium in the reference electrode. Electrode calibration is linear to 10(-8) M and is curvilinear to 10(-11) M in the presence of 0.1 M ionic strength. Selectivity with respect to competing cations, magnesium, potassium, sodium, and hydrogen is preserved. Electrode response time is less than 2 s for small changes in calcium activity. Response range is linear over 9 log units of calcium activity. Potential-time stability is less than 10 mV/h at saturation currents. Although the silver-silver chloride terminals are photosensitive throughout the visible and near-ultraviolet regions, housing the reference and indifferent in opaque barrels avoids false photovoltaic response.

Buffers↗

PubMed1985

Simplified concanavalin-A sepharose adsorption method for separation of cone visual pigments from rhodopsin.

Batch adsorption of chicken photoreceptor extract using Concanavalin-A Sepharose enables separation of rod and cone pigments and separation of cone pigments of different color sensitivity. In earlier work, column separations using the same adsorption medium, although effective, with high resolution, were slow, demanding and required many differential bleachings of column fractions for analysis. It is shown here that affinity separations can be performed in the batch adsorption mode to purify cattle, frog and chicken rhodopsin. These procedures are more rapid and much more convenient. An extract from the chicken retina can rapidly be separated into four fractions, including three highly enriched (80% or more) visual pigment fractions: (1) extraneous proteins, carotenoids and phospholipids; (2) short wavelength-sensitive pigments; (3) iodopsin; and (4) rhodopsin. While the resolution is not as great as that of the columns, the selectivity is sufficient to produce cone pigments, which are only slightly contaminated with rhodopsin and free of other proteins, either to experiment with directly or to enable heavier loading on high resolution columns. The method is adaptable both to highly labile pigments and to very small quantities, neither of which perform well in column separations.

Adsorption↗

PubMed1983

Increased uptake of thymidine in the activation of sea urchin eggs. II. Cooperativity with phosphorylation, involvement of the cortex, and partial localization of the kinases.

Uptake and phosphorylation of exogenously supplied thymidine are stimulated in Strongylocentrotus purpuratus eggs after fertilization. Before fertilization, the rate of uptake is low and less than 10% of the thymidine entering the egg is phosphorylated. After fertilization, the rate of uptake increases over 50-fold and greater than 90% of the thymidine is immediately phosphorylated. These results imply that there is close cooperativity between fertilization-induced uptake and phosphorylation of thymidine. To gain insight into the structural basis of this apparent cooperativity and to provide a partial localization of the kinases, uptake and phosphorylation were measured in centrifuged eggs, and in centrifuged nucleate and anucleate merogons. Electron micrographs show that in these cells, the inner cytoplasmic contents are stratified according to density and displaced within the egg, whereas the outer cortical region of the cytoplasm remains intact. Uptake and phosphorylation of thymidine are fully stimulated in these eggs and merogons after fertilization, suggesting that both processes are mediated by an intact egg cortex. In support of this suggestion, we report that controlled disruption of the egg cortex prior to fertilization by treatment with cytochalasin B (CB) significantly reduces the rates of uptake and phosphorylation after fertilization. The full stimulation of phosphorylation in nucleate and anucleate merogons eliminates any localization of the catalyzing enzymes (thymidine kinase and thymidylate kinase) in the maternal nucleus and other inner cytoplasmic contents differentially segregated by centrifugation.

Animals↗

PubMed1983

Effects of cytochalasin B on the cortex of the unfertilized sea urchin egg.

The peripheral cytoplasm of the unfertilized sea urchin egg contains approximately 18,000 cortical granules. These granules remain monolayered within the normal boundaries of the cortex when the egg is centrifuged at forces sufficient to stratify other intracellular inclusions. Exposure of unfertilized eggs to the microfilament disrupting agent, cytochalasin B (CB) causes the granules to rearrange into several layers and occasionally to undergo exocytosis or break down in situ. When these eggs are centrifuged, the cortical granules are dislodged from the cortex and migrate centrifugally among the densest intracellular components. In addition, cytoplasmic inclusions, which normally are excluded from the cortex, impinge directly upon the egg plasma membrane in CB-treated, centrifuged eggs. These results are consistent with the existence of a microfilamentous network which confines the cortical granules within and excludes other intracellular inclusions from the cortex of the unfertilized egg.

Animals↗