Oscillator control of cell division in Euglena: cyclic AMP oscillations mediate the phasing of the cell division cycle by the circadian clock

1993 ◽  
Vol 104 (4) ◽  
pp. 1163-1173 ◽  
Author(s):  
I.A. Carre ◽  
L.N. Edmunds
Keyword(s):  
Cell Division ◽  
Cyclic Amp ◽  
Cell Division Cycle ◽  
Circadian Oscillator ◽  
Constant Darkness ◽  
Rapid Loss ◽  
Subjective Night ◽  
Circadian Time ◽  
Pharmacological Studies

The achlorophyllous ZC strain of Euglena gracilis exhibits a circadian rhythm of cell division in constant darkness (DD). Mitosis occurs during a restricted part of the circadian cycle, corresponding to the dark intervals in a light-dark cycle comprising 12 h of light and 12 h of darkness. We have demonstrated that division-phased cultures also exhibit bimodal, circadian changes of cyclic AMP level. Maximum cyclic AMP levels occurred at the beginning of the light period (CT (circadian time) 00–02), and at the beginning of darkness (CT 12–14). These variations persisted in cultures that had been transferred into DD and appeared to be under the control of the circadian oscillator rather than to be cell division cycle (CDC)-dependent, since they continued in cultures that had reached the stationary phase of growth. In the experiments reported in this paper, we tested for the possible role of this periodic cyclic AMP signal in the generation of cell division rhythmicity by examining the effects of exogenous cyclic AMP signals and of forskolin, which permanently increased the cyclic AMP level, on the cell division rhythm. Perturbations of the cyclic AMP oscillation by exogenous cyclic AMP resulted in the temporary uncoupling of the CDC from the circadian timer. The addition of cyclic AMP during the subjective day resulted in delays (up to 9 h) of the next synchronous division step. In contrast, mitosis was stimulated when cyclic AMP was administered in the middle of the subjective night. Measurement of the DNA content of cells by flow cytometry indicated that cyclic AMP injected at CT 06–08 delayed progression through S phase, and perhaps also through mitosis. When added at CT 18–20, cyclic AMP accelerated the G2/M transition. The circadian oscillator was not perturbed by the addition of exogenous cyclic AMP: the division rhythm soon returned to its original phase. On the other hand, the permanent elevation of cyclic AMP levels in the presence of forskolin induced a rapid loss of cell division rhythmicity. These findings are consistent with the hypothesis that cyclic AMP acts downstream from the oscillator and that the cyclic AMP oscillation is an essential component of the signaling pathway for the control of the CDC by the circadian oscillator. The receptors for cyclic AMP in Euglena have been shown to be two cyclic AMP-dependent kinases (cPKA and cPKB). Pharmacological studies using cyclic AMP analogs suggested that cPKA mediates cyclic AMP effects during the subjective day, and cPKB during the subjective night.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals The Cell Division Cycle of Euglena gracilis Indicates That the Level of Circadian Plasticity to the External Light Regime Changes in Prolonged-Stationary Cultures

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1475
Author(s):  
Shota Kato ◽  
Hong Gil Nam
Keyword(s):  
Cell Division ◽  
Circadian Rhythms ◽  
Euglena Gracilis ◽  
Cell Cycle Progression ◽  
Light Signal ◽  
Cell Division Cycle ◽  
Regime Changes ◽  
Fitness Advantage ◽  
Subjective Night ◽  
Free Running

In unicellular photosynthetic organisms, circadian rhythm is tightly linked to gating of cell cycle progression, and is entrained by light signal. As several organisms obtain a fitness advantage when the external light/dark cycle matches their endogenous period, and aging alters circadian rhythms, senescence phenotypes of the microalga Euglena gracilis of different culture ages were characterized with respect to the cell division cycle. We report here the effects of prolonged-stationary-phase conditions on the cell division cycles of E. gracilis under non-24-h light/dark cycles (T-cycles). Under T-cycles, cells established from 1-month-old and 2-month-old cultures produced lower cell concentrations after cultivation in the fresh medium than cells from 1-week-old culture. This decrease was not due to higher concentrations of dead cells in the populations, suggesting that cells of different culture ages differ in their capacity for cell division. Cells from 1-week-old cultures had a shorter circadian period of their cell division cycle under shortened T-cycles than aged cells. When algae were transferred to free-running conditions after entrainment to shortened T-cycles, the young cells showed the peak growth rate at a time corresponding to the first subjective night, but the aged cells did not. This suggests that circadian rhythms are more plastic in younger E. gracilis cells.

fumble Encodes a Pantothenate Kinase Homolog Required for Proper Mitosis and Meiosis in Drosophila melanogaster

Genetics ◽  
2001 ◽  
Vol 157 (3) ◽  
pp. 1267-1276
Author(s):  
Katayoun Afshar ◽  
Pierre Gönczy ◽  
Stephen DiNardo ◽  
Steven A Wasserman
Keyword(s):  
Cell Division ◽  
Chromosome Segregation ◽  
Cell Division Cycle ◽  
Protein Isoforms ◽  
Pantothenate Kinase ◽  
Male Germline ◽  
Dividing Cells ◽  
Mutant Cells ◽  
Mitosis And Meiosis

Abstract A number of fundamental processes comprise the cell division cycle, including spindle formation, chromosome segregation, and cytokinesis. Our current understanding of these processes has benefited from the isolation and analysis of mutants, with the meiotic divisions in the male germline of Drosophila being particularly well suited to the identification of the required genes. We show here that the fumble (fbl) gene is required for cell division in Drosophila. We find that dividing cells in fbl-deficient testes exhibit abnormalities in bipolar spindle organization, chromosome segregation, and contractile ring formation. Cytological analysis of larval neuroblasts from null mutants reveals a reduced mitotic index and the presence of polyploid cells. Molecular analysis demonstrates that fbl encodes three protein isoforms, all of which contain a domain with high similarity to the pantothenate kinases of A. nidulans and mouse. The largest Fumble isoform is dispersed in the cytoplasm during interphase, concentrates around the spindle at metaphase, and localizes to the spindle midbody at telophase. During early embryonic development, the protein localizes to areas of membrane deposition and/or rearrangement, such as the metaphase and cellularization furrows. Given the role of pantothenate kinase in production of Coenzyme A and in phospholipid biosynthesis, this pattern of localization is suggestive of a role for fbl in membrane synthesis. We propose that abnormalities in synthesis and redistribution of membranous structures during the cell division cycle underlie the cell division defects in fbl mutant cells.

Effect of somatostatin on circadian rhythms of firing and 2-deoxyglucose uptake in rat suprachiasmatic slices

1993 ◽  
Vol 265 (5) ◽  
pp. R1199-R1204 ◽  
Author(s):  
T. Hamada ◽  
S. Shibata ◽  
A. Tsuneyoshi ◽  
K. Tominaga ◽  
S. Watanabe
Keyword(s):  
Circadian Rhythm ◽  
Vasoactive Intestinal Polypeptide ◽  
Phase Changes ◽  
Circadian Clocks ◽  
Constant Darkness ◽  
Phase Resetting ◽  
Firing Activity ◽  
Light Pulses ◽  
Circadian Time

In mammals, the suprachiasmatic nucleus (SCN) of the hypothalamus appears to act as a circadian clock. The SCN vasoactive intestinal polypeptide-like immunoreactive neurons, which may act to mediate photic information in the SCN, receive input from neurons immunoreactive for somatostatin (SST). Therefore we investigated the role of SST as a transmitter for entrainment by analyzing the phase-resetting effect of SST on the circadian rhythm of SCN firing activity. Perfusion of SST increased 2-deoxyglucose uptake at circadian time (CT) 18, but not at CT6. A 1-h or 15-min treatment with SST produced phase delays when it was administered at CT13-14 and phase advances at CT22-23. Thus SST-induced phase changes are similar to those for light pulses to animals under constant darkness. The present findings suggest that SST is a transmitter for mediating information of entrainment to circadian clocks within the SCN.

scholarly journals Bright light during lactation alters the functioning of the circadian system of adult rats

2000 ◽  
Vol 278 (1) ◽  
pp. R201-R208 ◽  
Author(s):  
M. M. Canal-Corretger ◽  
T. Cambras ◽  
J. Vilaplana ◽  
A. Díez-Noguera
Keyword(s):  
Motor Activity ◽  
Activity Rhythm ◽  
Bright Light ◽  
Circadian System ◽  
Constant Darkness ◽  
Adult Rats ◽  
Light Pulses ◽  
Circadian Time ◽  
Role Of Light

To examine the role of light in the maturation of the circadian pacemaker, twelve groups of rats were raised in different conditions of exposure to constant bright light (LL) during lactation: both duration and timing of LL were varied. We studied the motor activity rhythm of the rats after weaning, first under LL and then under constant darkness (DD). In DD, two light pulses [at circadian time 15 (CT15) and CT22] were applied to test the response of the pacemaker. Greater exposure to LL days during lactation increased the number of rhythmic animals and the amplitude of their motor activity rhythm in the LL stage and decreased the phase delay due to the light pulse at CT15. The timing of LL during lactation affected these variables too. Because the response of the adult to light depended on both the number and timing of LL days during lactation, the exposure to light at early stages may influence the development of the circadian system by modifying it structurally or functionally.

245 EXPRESSION AND FUNCTIONAL ROLE OF CELL DIVISION CYCLE 42 IN MOUSE OOCYTES MATURATION AND PRE-IMPLANTATION DEVELOPMENT

2006 ◽  
Vol 18 (2) ◽  
pp. 230
Author(s):  
X.-S. Cui ◽  
X.-Y. Li ◽  
N.-H. Kim
Keyword(s):  
Protein Synthesis ◽  
Cell Division ◽  
Mrna Expression ◽  
Oocyte Maturation ◽  
Germinal Vesicle ◽  
Cell Division Cycle ◽  
Mrna Levels ◽  
Mouse Oocytes

Cell division cycle 42 (Cdc42), a member of the Rho family of small guanosine triphosphatase (GTPase) proteins, regulates multiple cell functions, including motility, proliferation, apoptosis, and cell morphology. In order to gain insight into the role of Cdc42 in embryo development, we first characterized mRNA and protein levels of Cdc42 in mouse oocytes and early embryogenesis. We then examined the possible role of the gene in oocyte maturation and pre-implantation development using RNA interference analysis. The relative abundance of Cdc42 transcripts were measured by real time RT-PCR. After normalization with histone H2a mRNA levels, the mRNA expression of Cdc42 was abundant in immature oocytes and reduced slightly in zygotes and 2- to 8-cell stage embryos. The expression levels were significantly increased during the morula and blastocyst stages. Indirect immunocytochemistry showed protein synthesis of Cdc42 in oocytes and embryos of all stages. Introducing small interference RNA (siRNA) of Cdc42 into germinal vesicle stage oocytes or zygotes specifically reduce both mRNA expression and protein synthesis of Cdc42 in metaphase II stage oocytes and early embryos developing in vitro. Meiotic maturation was significantly reduced following siRNA injection into germinal vesicle stage oocytes. It is evident that actin distribution in siRNA treated blastocysts is morphologically abnormal following injection of siRNA for Cdc42. Injection of siRNA into zygotes did not influence cleavage, but significantly decreased in vitro development to morulae and blastocysts. While housekeeping genes such as tissue plasminogen activator were not altered by siRNA, wiskott-aldrich syndrome protein family 1 (WASP1) mRNA was down-regulated in the morula. Interestingly, mRNA of WASP1, tubulin alpha 1 (Tuba1), and actin-related protein 2/3 complex subunit V (Arpc5) increased at the blastocyst stage following siRNA injection. These results suggest that Cdc42 plays an important role during oocyte maturation and early pre-implantation development, likely through linkage with several other genes. This work was funded by a grant from National Research Laboratory Program in Korea.

scholarly journals Regulation of light's action in the mammalian circadian clock: role of the extrasynaptic GABAA receptor

2009 ◽  
Vol 296 (5) ◽  
pp. R1606-R1612 ◽  
Author(s):  
J. Christopher Ehlen ◽  
Ketema N. Paul
Keyword(s):  
Wheel Running ◽  
Phase Shifts ◽  
Constant Darkness ◽  
Receptor Subtypes ◽  
Mrna Levels ◽  
Subjective Night ◽  
Behavioral Phase ◽  
Free Running ◽  
Light Input

GABAA receptor agonists act in the suprachiasmatic nucleus (SCN) to reset circadian rhythms during the day but inhibit the ability of light to reset rhythms during the night. In the present study, we examined whether these paradoxical differences in the effect of GABAA receptor stimulation on the circadian system are mediated by separate GABAA receptor subtypes. 4,5,6,7-Tetrahydroisoxazolo[5,4- c]pyridin-3-ol (THIP), a GABAA receptor agonist, preferentially activates GABAA receptors in extrasynaptic locations. THIP, muscimol (a GABAA agonist), or vehicle were microinjected into the SCN region of Syrian hamsters free-running in constant darkness during the mid-subjective day, early subjective night, or late subjective night. The subjective night injections were followed by a light pulse or sham control. Behavioral phase shifts of wheel running rhythms and both Period1 ( Per1) and Per2 mRNA levels in the SCN were assessed. Animals that received THIP during the subjective day did not exhibit significant phase alterations. During the early and late subjective night, however, THIP abolished the phase-shifting effects of light and the ability of light to increase Per1 and Per2 mRNA levels. The ability of N-methyl-d-aspartic acid to phase-shift wheel running rhythms was also attenuated by THIP. Together these data demonstrate that THIP does not produce phase shifts during the subjective day, but does inhibit the ability of light to produce phase shifts. Thus, extrasynaptic GABAA receptors appear to play a role in regulating light input to the SCN, while a different population of GABAA receptors appears to be responsible for daytime effects of GABA.

Circadian changes in cyclic AMP levels in synchronously dividing and stationary-phase cultures of the achlorophyllous ZC mutant of Euglena gracilis

1989 ◽  
Vol 94 (2) ◽  
pp. 267-272
Author(s):  
ISABELLE A. CARRÉ ◽  
DANIELLE L. LAVAL-MARTIN ◽  
LELAND N. EDMUNDS
Keyword(s):  
Cell Division ◽  
Cyclic Amp ◽  
Euglena Gracilis ◽  
Phase Response Curve ◽  
Circadian Variation ◽  
Light Signal ◽  
Circadian Oscillator ◽  
Circadian Oscillation ◽  
Continuous Darkness ◽  
Free Running

Oscillations in adenosine 3′,5′-cyclic monophosphate (cyclic AMP) level have been proposed to be part of the biochemical feed-back loop(s), or ‘clock(8)’, believed to underlie circadian rhythmicity. This possibility has been examined for a cellular circadian oscillator in synchronously dividing (or nondividing) cultures of the photosynthesis- deficient ZC mutant of the alga Euglena gracilis Klebs (Z). We have demonstrated a bimodal, autonomously oscillating, circadian variation of cyclic AMP content in this unicell. Rhythmic changes of the cyclic AMP level, which may reflect the transition of the cell population through the different phases of the cell division cycle (CDC) in division-phased cultures, also persisted after the culture medium had become limiting and the cells had stopped dividing. We have also shown that the free-running, circadian oscillation of cyclic AMP content displayed by nondividing cells in continuous darkness could be phase-shifted by a light signal (a property inherent to most circadian systems), in a manner that could be predicted from the phase-response curve previously obtained for the cell division rhythm in the ZC mutant. These results suggest a possible role for cyclic AMP, either as an element of the coupling pathway for the control of the CDC by the circadian oscillator, or as a ‘gear’ of the clock itself.

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