Assay and Characteristics of Circadian Rhythmicity in Liquid Cultures of <i>Neurospora crassa</i>

Perlman, J.; Nakashima, Hiroaki; Feldman, Jerry F.

doi:10.1104/pp.67.3.404

Cited by 46 publications

(20 citation statements)

References 9 publications

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“…This time was calculated using the method described by Dharmananda and Feldman (4). The phase in the liquid culture was calculated from the first band in the race tubes on the assumption that the clock runs in liquid medium with the same period length as on agar medium and that it is not affected by transfer from liquid to solid medium (12,15). All data are the average of 6 race tubes.…”

Section: Methodsmentioning

confidence: 99%

Effects of Respiratory Inhibitors on Respiration, ATP Contents, and the Circadian Conidiation Rhythm of Neurospora crassa

Nakashima¹

1984

Plant Physiol.

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Effects of respiratory inhibitors on the circadian clock, respiratory activity, and ATP content were examined in Neurospora crassa. AU inhibitors, potassium cyanide, sodium azide, antimycin A, and carbonyl cyanide m-chlorophenyl hydrazone (CCCP), shifted the phase of the conidiation rhythm. All the phase response curves were similar and resembled that for cycloheximide, but were different from the phase response curve for light. Phase shifting by azide and CCCP was proportional to the lowering of respiratory activity and ATP content, but such a correlation was not observed for cyanide and antimycin A. In particular, cyanide at a concentration of 0.5 millimolar completely depleted ATP of the cultures but did not significantly shift their phase. Their results suggest that large shifts caused by these inhibitors are not due to a decrease in energy from respiratory activity.Circadian rhythms are observed in many organisms from lower eukaryotic plants to higher animals. However, the molecular mechanism ofthe clock which regulates these rhythms is still not known. Bunning (2) proposed that the clock is composed of two different phases: a physiological night phase which requires energy and a physiological day phase in which energy is not required. In fact, deprivation of oxygen for short times (1, 18) or treatment with respiratory inhibitors (3, 6, 9-11) resulted in phase shifting at specific phases in several organisms. These results suggest that certain phases of the clock, in particular the subjective night, may be energy dependent. Koyama and Feldman treated Neurospora with several respiratory inhibitors to examine the energy dependence of clock function (cited in Feldman and Dunlap [7]). They obtained phase-shifting data but did not measure inhibition ofrespiration or ATP levels, nor have such measurements been made in other organisms. The correlations such measurements might provide are needed before it can be concluded that energy depletion caused by these inhibitors results in phase shifting of the clock. In this report, both phase shifting by respiratory inhibitors and levels of the respiratory activity and ATP were measured. MATERIALS AND METHODSCulture Conditions and Analysis of the Conidiation Rhythm. The bd (band) strain of Neurospora crassa was used. Procedures for maintaining stock cultures and for liquid culture were the same as reported previously (12). Conidia (13 x 1O') suspended in distilled H20 were added to 25 ml ofliquid medium containing Fries' salts (8), 0.3% glucose, and 0.5% arginine. Conidial con-'Supported in part by the Japan-United States Cooperative Program. centration were determined by A at 480 nm. After culturing for 36 h in continuous light at 26C, discs were cut from the mycelial mats with a cork borer 11 mm in diameter. Six discs from each mat were transferred to a 125-ml Erlenmeyer flask with 25 ml of liquid medium containing Fries' salts, 0.03% glucose and 0.05% arginine and cultured in continuous darkness with reciprocal shaking (100 cycles per min) at 26C. The mediu...

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Section: Methodsmentioning

confidence: 99%

Effects of Respiratory Inhibitors on Respiration, ATP Contents, and the Circadian Conidiation Rhythm of Neurospora crassa

Nakashima¹

1984

Plant Physiol.

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“…The typical light single pulse PRC in Neurospora ( frq>) shows a slight phase advance at the beginning of the period (ct 0}ct 4), while phase delays are observed from ct 4 to ct 16 up to ct 22 with the breakpoint occurring between ct 16 and ct 22 (Sargent & Briggs, 1967;Dharmananda, 1980;Perlman et al, 1981;Nakashima, 1985;Schulz et al, 1985;Rensing & Schill, 1987). Figure 3(a) shows the experimental frq> light PRC by Perlman et al (1981) together with the calculated PRC using the Goodwin model.…”

Section: Phase Resetting With Single Light Pulsesmentioning

confidence: 99%

“…Figure 3(a) shows the experimental frq> light PRC by Perlman et al (1981) together with the calculated PRC using the Goodwin model. The "gure also shows experimentally determined frq>-mRNA levels and the calculated X levels.…”

Section: Phase Resetting With Single Light Pulsesmentioning

confidence: 99%

The Goodwin Model: Simulating the Effect of Light Pulses on the Circadian Sporulation Rhythm of Neurospora Crassa

Ruoff

Vinsjevik

Monnerjahn

et al. 2001

Journal of Theoretical Biology

129

111

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“…The phase of the liquid cultures was calculated from the first band in the race tube with the assumptions that periodicity in liquid medium is the same as that in the race tubes and that the rhythm is unaffected by transfer from liquid to solid medium (9,13). Divalent cation-free medium contained (g/l): NH4Cl, 7.2; NH4NO3, 5; KH2PO4, 5; NaCl, 0.5.…”

mentioning

confidence: 99%

Assay and Characteristics of Circadian Rhythmicity in Liquid Cultures of Neurospora crassa

Cited by 46 publications

References 9 publications

Effects of Respiratory Inhibitors on Respiration, ATP Contents, and the Circadian Conidiation Rhythm of Neurospora crassa

Effects of Respiratory Inhibitors on Respiration, ATP Contents, and the Circadian Conidiation Rhythm of Neurospora crassa

The Goodwin Model: Simulating the Effect of Light Pulses on the Circadian Sporulation Rhythm of Neurospora Crassa

Calcium Inhibits Phase Shifting of the Circadian Conidiation Rhythm of Neurospora crassa by the Calcium Ionophore A23187

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