Biomedical subjects

E K Wolff

Publications and source records attributed to E K Wolff.

5 recordsLinked to original sources

PubMed1988

A rapid population method for action spectra applied to Halobacterium halobium.

We have developed a simple and rapid technique for measuring the action spectra for phototaxis of populations of microorganisms and applied it to halobacteria. A microscope with a dark-field condenser was used to illuminate the cell suspension in a sealed chamber with light of wavelength greater than 750 nm; in this region of the spectrum, the halobacteria show no phototactic response. A 150-micron spot of light from a xenon arc lamp, whose wavelength and intensity can be varied, was projected through the objective lens into the center of the dark field. The objective lens imaged this measuring spot through a 780-nm cut-off filter on an aperture in front of a photomultiplier. The intensity of the scattered 750-nm light, and therefore the photomultiplier current, is proportional to the number of cells in the measuring spot. A third lamp provided background light of variable wavelength and intensity through the dark-field condenser. To minimize secondary effects due to large changes in cell density, we recorded the initial changes in the photomultiplier current over 1 min after the actinic light had been switched on. By plotting the rate of change against wavelength, we obtained action spectra after the proper corrections for changes in light intensity with wavelength were applied and saturation effects were avoided.

Halobacterium↗

PubMed1986

Color discrimination in halobacteria: spectroscopic characterization of a second sensory receptor covering the blue-green region of the spectrum.

Halobacterium halobium is attracted by green and red light and repelled by blue-green and shorter wavelength light. a photochromic, rhodopsin-like protein in the cell membrane, sensory rhodopsin sR587, has been identified as the receptor for the long-wavelength and near-UV stimuli. Discrepancies between the action spectrum for the repellent effect of blue light and the absorption spectrum of sR587 and its photocycle intermediate S373 strongly suggest the existence of an additional photoreceptor for the blue region of the spectrum. Transient light-induced absorbance changes in intact cells and cell membranes show, in addition to sR587, the presence of a second photoactive pigment with maximal absorption near 480 nm. It undergoes a cyclic photoreaction with a half-time of 150 msec. One intermediate state with maximal absorption near 360 nm has been resolved. The spectral properties of the new pigment are consistent with a function as the postulated photoreceptor for the repellent effect of blue light. The phototactic reactions and both pigments are absent when retinal synthesis is blocked; both can be restored by the addition of retinal. These results confirm and extend similar observations by Takahashi et al. [Takahashi, T., Tomioka, H., Kamo, N. & Kobatake, Y. (1985) FEMS Microbiol. Lett. 28, 161-164]. The archaeobacterium H. halobium thus uses two different mechanisms for color discrimination; it uses two rhodopsin-like receptors with different spectral sensitivities and also the photochromicity of at least one of these receptors to distinguish between three regions covering the visible and near-UV spectrum.

Bacteriorhodopsins↗

PubMed1985

In vivo micro application of adrenaline to capillaries in frog mesentery.

In-vivo studies were carried out on frog mesentery in order to determine the precise and specific actions of micropipette application of adrenaline on arterioles, precapillary sphincters and capillaries. A technique was employed, using liquid paraffin and micropipettes to deliver droplets to a particular microvessel; the droplets varied between 10-20 microns in diameter. All vessel types, including segments of capillaries, demonstrated constrictor responses to the local application of adrenaline. The time course of these constrictor events varied with the vessel type, with the sphincter being the most rapid in response.

Animals↗

PubMed1982

Equilibrium binding of acetylcholine to the membrane-bound acetylcholine receptor.

We have studied the binding of acetylcholine to membrane-bound acetylcholine receptor from Torpedo marmorata employing a highly accurate airfuge assay procedure. At equilibrium the receptor displays two classes of acetylcholine binding sites; these interact with only weak positive cooperativity. As a further difference to binding data deduced from electrophysiological dose/response curves, the equilibrium constants for the two classes of sites (Kd1 = 25 nM, Kd2 = 8 nM) are orders of magnitude lower than the concentration required for half-maximal response. Both the weaker-than-expected cooperativity of sites and the high binding affinities are likely to be due to desensitisation of the receptor during the period of incubation. The positively cooperative interaction of acetylcholine binding sites is only observed with membrane preparations obtained in the presence of appropriate chelating, sulfhydryl-blocking and active-serine-blocking agents. Aged membrane preparations loose the ability of site interactions while only small changes in the total number of binding sites are observed. In the absence of divalent ions, the affinity of binding of acetylcholine to the receptor is reduced. To assess the significance of the binding data obtained, several alternative reaction schemes for non-random binding to two sites at the receptor are considered. In addition, the effects of possible sources of experimental error on the shape of Scatchard plots are analysed.

Acetylcholine↗

PubMedDate not supplied

Imaging cells with the atomic force microscope.

Different types of cells have been imaged with the atomic force microscope. The morphology of the archaebacterium Halobacterium halobium in its dry state was revealed. On a leaf of the small Indian tree Lagerstroemia subcostata a stoma was imaged. The lower side of a water lily leaf was imaged in water showing features down to 12 nm. Finally, fixed red and white blood cells were imaged in buffer showing features down to 8 nm. The images demonstrate that atomic force microscopy can provide high-resolution images of cell surfaces under physiological conditions.

Animals↗