A ‘TEA+-insensitive’ mutant with increased potassium conductance in Paramecium aurelia

1976 ◽  
Vol 65 (1) ◽  
pp. 51-63
Author(s):  
Y. Satow ◽  
C. Kung

A single-gene mutant of Paramecium aurelia is analysed electrophysiologically. (a) The regenerative Ca-response, triggered by small or moderate current, was smaller and slower in the mutant than in wild type. (b) Input resistance of the mutant membrane is about half of that of wild type bathed in various solutions. This is true for the zero-current input resistance and the chord resistance measured with high depolarizing current. (c) Membrane resistance of the mutant measure with hyperpolarizing currents is smaller than that of wild type only when K+ is the major external cation. (d) Internally applied TEA+ or externally applied Ba+ increases the membrane resistance of the mutant to that of wild type similarly treated. We conclude that the mutant has an increased K conductance.

1970 ◽  
Vol 12 (1) ◽  
pp. 70-79
Author(s):  
Philip G. Miles

The puff mutant of Schizophyllum commune is characterized by bead-like puffs of hyphae formed by extensive dichotomous branching of lateral branches from main hyphae. Puff segregates as a single gene mutant when mated with wild-type although the expression of the puff phenotype is variable. A modifier gene of puff, p-mod-1, is described. Strains bearing the gene for puff (p) and the p-mod-1 gene are of wild-type appearance macroscopically, but can generally be distinguished microscopically by the dichotomous branching of the hyphal tips. The p-mod-1 gene segregates independently of p. The puff mutant is elasticotropic (i.e. grows along lines of stress), but the presence of the p-mod-1 gene removes this effect. The p-mod-1 gene arose spontaneously in a wild-type strain, but modifiers of mutant morphology have also been obtained by mutagenic treatment. Such modifiers have great utility in morphogenetic studies of hyphal development.


2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Godai Suzuki ◽  
Yutaka Saito ◽  
Motoaki Seki ◽  
Daniel Evans-Yamamoto ◽  
Mikiko Negishi ◽  
...  

AbstractMorphological profiling is a combination of established optical microscopes and cutting-edge machine vision technologies, which stacks up successful applications in high-throughput phenotyping. One major question is how much information can be extracted from an image to identify genetic differences between cells. While fluorescent microscopy images of specific organelles have been broadly used for single-cell profiling, the potential ability of bright-field (BF) microscopy images of label-free cells remains to be tested. Here, we examine whether single-gene perturbation can be discriminated based on BF images of label-free cells using a machine learning approach. We acquired hundreds of BF images of single-gene mutant cells, quantified single-cell profiles consisting of texture features of cellular regions, and constructed a machine learning model to discriminate mutant cells from wild-type cells. Interestingly, the mutants were successfully discriminated from the wild type (area under the receiver operating characteristic curve = 0.773). The features that contributed to the discrimination were identified, and they included those related to the morphology of structures that appeared within cellular regions. Furthermore, functionally close gene pairs showed similar feature profiles of the mutant cells. Our study reveals that single-gene mutant cells can be discriminated from wild-type cells based on BF images, suggesting the potential as a useful tool for mutant cell profiling.


Genetics ◽  
1972 ◽  
Vol 72 (3) ◽  
pp. 411-417
Author(s):  
C W H Partridge ◽  
Mary E Case ◽  
Norman H Giles

ABSTRACT A color test has been developed for the selection and identification of mutants in Neurospora crassa, constitutive for the three normally inducible enzymes which convert quinate to protocatechuate. By this means seven such mutants have been recovered after ultra violet irradiation of wild type and have been shown to be allelic (or very closely linked) to the qa-1C mutants previously obtained by other means. Thus, the regulation of the synthesis of these three catabolic enzymes is indicated to be under the control of a single gene, qa-1+.


1995 ◽  
Vol 182 (5) ◽  
pp. 1415-1421 ◽  
Author(s):  
T C Wu ◽  
A Y Huang ◽  
E M Jaffee ◽  
H I Levitsky ◽  
D M Pardoll

Introduction of the B7-1 gene into murine tumor cells can result in rejection of the B7-1 transductants and, in some cases, systemic immunity to subsequent challenge with the nontransduced tumor cells. These effects have been largely attributed to the function of B7-1 as a costimulator in directly activating tumor specific, major histocompatibility class I-restricted CD8+ T cells. We examined the role of B7-1 expression in the direct rejection as well as in the induction of systemic immunity to a nonimmunogenic murine tumor. B-16 melanoma cells with high levels of B7-1 expression did not grow in C57BL/6 recipient mice, while wild-type B-16 cells and cells with low B7-1 expression grew progressively within 21 d. In mixing experiments with B7-1hi and wild-type B-16 cells, tumors grew out in vivo even when a minority of cells were B7-1-. Furthermore, the occasional tumors that grew out after injection of 100% B-16 B7-1hi cells showed markedly decreased B7-1 expression. In vivo antibody depletions showed that NK1.1 and CD8+ T cells, but not CD4+ T cells, were essential for the in vivo rejection of tumors. Animals that rejected B-16 B7-1hi tumors did not develop enhanced systemic immunity against challenge with wild-type B-16 cells. These results suggest that a major role of B7-1 expression by tumors is to mediate direct recognition and killing by natural killer cells. With an intrinsically nonimmunogenic tumor, this direct killing does not lead to enhanced systemic immunity.


1985 ◽  
Vol 54 (6) ◽  
pp. 1375-1382 ◽  
Author(s):  
C. W. Bourque ◽  
J. C. Randle ◽  
L. P. Renaud

Intracellular recordings of rat supraoptic nucleus neurons were obtained from perfused hypothalamic explants. Individual action potentials were followed by hyperpolarizing afterpotentials (HAPs) having a mean amplitude of -7.4 +/- 0.8 mV (SD). The decay of the HAP was approximated by a single exponential function having a mean time constant of 17.5 +/- 6.1 ms. This considerably exceeded the cell time constant of the same neurons (9.5 +/- 0.8 ms), thus indicating that the ionic conductance underlying the HAP persisted briefly after each spike. The HAP had a reversal potential of -85 mV and was unaffected by intracellular Cl- ionophoresis of during exposure to elevated extracellular concentrations of Mg2+. In contrast, the peak amplitude of the HAP was proportional to the extracellular Ca2+ concentration and could be reversibly eliminated by replacing Ca2+ with Co2+, Mn2+, or EGTA in the perfusion fluid. During depolarizing current pulses, evoked action potential trains demonstrated a progressive increase in interspike intervals associated with a potentiation of successive HAPs. This spike frequency adaptation was reversibly abolished by replacing Ca2+ with Co2+, Mn2+, or EGTA. Bursts of action potentials were followed by a more prolonged afterhyperpolarization (AHP) whose magnitude was proportional to the number of impulses elicited (greater than 20 Hz) during a burst. Current injection revealed that the AHP was associated with a 20-60% decrease in input resistance and showed little voltage dependence in the range of -70 to -120 mV. The reversal potential of the AHP shifted with the extracellular concentration of K+ [( K+]o) with a mean slope of -50 mV/log[K+]o.(ABSTRACT TRUNCATED AT 250 WORDS)


1980 ◽  
Vol 84 (1) ◽  
pp. 187-199
Author(s):  
D. Mellon ◽  
G. Lnenicka

The morphologies and passive electrical parameters of fibres in two eye muscles of a surface- and a cave-dwelling crayfish were compared. In the cave-dwelling form the muscles contained fewer fibres, of less diameter, and hence had a smaller cross-sectional area. Current-voltage relationships were similar in both species. Input resistance was higher in the cave-dweller, but the difference was not as great as would be expected on the basis of geometry alone. Accordingly, the specific membrane resistance of muscle fibres in the cave-dweller is 50–60% smaller than that in the surface-dweller. This may account partially for the observation that identified excitatory junctional potentials in muscles of cave- and surface dwellers have similar amplitudes. We conclude that a functional oculomotor system is maintained in cave-dwelling crayfish, and that this system confers some positive selective advantage.


1984 ◽  
Vol 246 (5) ◽  
pp. G574-G579
Author(s):  
G. W. Kidder ◽  
M. G. Elrod

Changing the potential across the isolated frog gastric mucosa by voltage clamping changes the measured resistance of the tissue in two ways. An immediate change in resistance results from changing the measuring position on the nonlinear current-voltage (I-V) plot. Subsequent to this, the resistance changes slowly with a half-time of about 3 min, a change that is not predicted by a previous model for voltage transients and that implies slow changes in membrane resistance following changes in intracellular ion content. The I-V plot over the range examined shows three breakpoints; changing clamp voltage alters the position of two of these breakpoints as well as the slope of the connecting resistances. The central breakpoint agrees with the potential at zero current and varies with it as the clamp potential is changed, as predicted from a diode model for breakpoint generation.


1990 ◽  
Vol 258 (1) ◽  
pp. H145-H152 ◽  
Author(s):  
O. F. Schanne ◽  
M. Lefloch ◽  
B. Fermini ◽  
E. Ruiz-Petrich

We compared the passive electrical properties of isolated ventricular myocytes (resting potential -65 mV, fast action potentials, and no spontaneous activity) with those of 2- to 7-day-old cultured ventricle cells from neonatal rats (resting potential -50 mV, slow action potentials, and presence of spontaneous activity). In myocytes the specific membrane capacity was 0.99 microF/cm2, and the specific membrane resistance increased from 2.46 k omega.cm2 at -65 mV to 7.30 k omega.cm2 at -30 mV. In clusters, the current-voltage relationships measured under current-clamp conditions showed anomalous rectification and the input resistance decreased from 1.05 to 0.48 M omega when external K+ concentration was increased from 6 to 100 mM. Using the model of a finite disk we determined the specific membrane resistance (12.9 k omega.cm2), the effective membrane capacity (17.8 microF/cm2), and the lumped resistivity of the disk interior (1,964 omega.cm). We conclude that 1) the voltage dependence of the specific membrane resistance cannot completely explain the membrane resistance increase that accompanies the appearance of spontaneous activity; 2) a decrease of the inwardly rectifying conductance (gk1) is mainly responsible for the increase in the specific membrane resistance and depolarization; and 3) approximately 41% of the inward-rectifying channels are electrically silent when spontaneous activity develops in explanted ventricle cells.


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