Proximal tubular epithelial cells are generated by division of differentiated cells in the healthy kidney

We searched for evidence for a contribution of stem cells in growth of the proximal S3 segments of healthy rats. According to the stem cell model, stem cells are undifferentiated and slow cycling; the bulk of cycling cells are transit amplifying, rapidly cycling cells. We show the following. 1) By continuous application of a thymidine analog (ThA) for 7 days, S3 proximal epithelial cells in healthy kidneys display a high-cycling rate. 2) Slow-cycling cells, identified by lack of ThA uptake during 14 days of continuous ThA application up to death and by expression of the cell cycle protein Ki67 at death, have the same degree of differentiation as quiescent cells. 3) To detect rapidly cycling cells, rats were killed at various time points after injection of a ThA. Double immunofluorescence for ThA and a cell cycle marker was performed, with colocalization indicating successive divisions. During one week after division, daughter cells display a very low proliferation rate, indicating the absence of rapidly cycling cells. 4) Labeling with cyclin D1 showed that this low proliferation rate is due to cycle arrest. 5) More than 50% of the S3 cells entered the cell cycle 36 h after a potent proliferative stimulus (lead acetate injection). We conclude that generation of new cells in the proximal tubule relies on division of differentiated, normally slow-cycling cells. These may rapidly enter the cycle under an adequate stimulus. immunohistochemistry; cell cycle; proliferation; renal stem cells; proximal tubule; renal epithelial cells     

Cell Cycle Changes in the Shoot Apex of Silene coeli-rosa During the Second and Third Days of Floral Induction

The cell cycle in Silene coeli-rosa shoot apices was measuredto test whether or not early components of the floral stimulus,produced during the 2nd and 3rd long days (LD) of an inductiveLD treatment, resulted in an increase in the duration of G₂phase in constant 20–24 h cell cycles. Plants were grownat 20°C in short days (SD) of 8 h light and 16 h darknessfor 28 d (day 0). Starting on day 0, plants were given SD or3 LD each comprising an identical 8 h day and 16 h photo-extension,or 3 dark-interrupted (d.i.) non-inductive LD, interrupted at1700 h of each day with 1 h of darkness. The cell cycle (percentagelabelled mitoses method) and changes in cell number were determinedin the shoot apical meristem. During days 1–2 of the SDtreatment, the cell cycle and mean cell generation time (MCGT)was 18 and 32 h, respectively, giving a growth fraction of 56%.During days 2–3, the cell cycle and MCGT shortened to15 and 23 h, respectively (growth fraction = 65%). During days1–2 of the LD and d.i. LD treatments, cell cycles andMCGTs were 9–10 and 27–29 h, respectively, resultingin smaller growth fractions (about 33%). Thus, shortened cellcycles and altered growth fractions occurred regardless of whetheror not the treatment was inductive. The LD treatment resultedin a marked shortening of G₁ and, to a lesser extent, S-phase,whilst G₂ remained constant. These changes were consistent withincreases in the proportion of cells in G₂ during the photoextensionof each LD which were suppressed during the comparable periodsof the d.i. LD treatment. The latter treatment resulted in eachphase occupying virtually identical proportions of the cellcycle as in the SD treatment. Thus, the unique cell cycle responsesto the initial part of the inductive LD treatment were increasesin the proportion of cells in G₂ coupled with G₁ and G₂ beingof similar duration. Cell cycle, mean cell generation time, shoot apex, Silene coeli-rosa     

Cell Cycle Duration in the Root Meristem of Sonoran Desert Cactaceae as Estimated by Cell-flow and Rate-of-cell-production Methods

Slow rates of cactus growth in the Sonoran Desert and high productivityof some Cactaceae under cultivation suggest that relativelylow growth rates are not the consequence of a long cell divisioncycle but of short optimal periods for growth and adverse environmentalfactors. To verify this hypothesis, the duration of the celldivision cycle (T)in the root apical meristem of seedlings ofthree sympatric species from the Sonoran Desert [Ferocactuspeninsulae(F. A. C. Weber) Britton & Rose ‘Townsendianus’(Britton & Rose) N. P. Taylor, stat. nov.,Stenocereus gummosus(Engelm.)Gibson & Horak andPachycereus pringlei(S. Watson) Britton& Rose] was estimated with the rate-of-cell-production (RCP)and the cell-flow (colchicine) methods. Both methods were appliedduring the steady-state growth phase, which was relatively shortin the first two species because of the determinate patternof root growth. The RCP method permitted estimation ofTin eachroot individually. Durations of the cell division cycle wereinversely proportional to the rate of root growth (r²rangedfrom 0.42 to 0.88,P<0.05).T,determined by the cell-flow method,ranged from 14.4 to 19.3 h in these species and was within thesame range asTdetermined by the RCP method. The averageTdeterminedby the RCP method was 67 to 75% of that determined by the cell-flowmethod. Results obtained with both methods are compared andanalysed. The proposed hypothesis appears to be correct, indicatingthat these species can be more productive under cultivationthan in the wild due to the relatively short duration of thecell division cycle. Adaptive features of these findings arealso considered.Copyright 1998 Annals of Botany Company Cactaceae, cell division cycle, root growth, root meristem, Sonoran Desert     

In situ growth rate and distribution of the ichthyotoxic dinoflagellate Gyrodinium corsicum Paulmier in an estuarine embayment (Alfacs Bay, NW Mediterranean Sea)

Gyrodinium corsicum Paulmier produced massive blooms in Alfacs Bay (Ebro Delta, NW Mediterranean Sea) from December 1994 to April 1995, a period characterized by an initial period of water stability and low outward flux of water during subsequent months. The highest cell densities of G.corsicum were found at the bottom of the bay during the population maintenance period (from January to March). During the day, no differences in the vertical distribution of G.corsicum were observed and no diel in situ vertical migration of the population was recorded. The in situ growth rate of G.corsicum was estimated using the cell cycle method since the cell division cycle was well phased with the daily light cycle. The S phase fraction of G.corsicum reached a maximum near the middle of the dark period. The G2M peak usually occurred in the dark period. The duration of the S phase was 2.1-8.7 h and the duration of the G2M phase was 2.6-7.7 h. The estimated specific in situ growth rate of G.corsicum ranged from 0.94 day^-1 during the initial phases of the bloom to 0.3 day^-1 when the bloom was well developed. Growth rates, measured in different locations at the same time, varied by a factor of 1.5, suggesting that different parts of the same blooming population were at different stages of growth. The persistence of the G.corsicum bloom with high cell densities (10⁵-10⁶ cells l^-1) at the bottom of the bay is discussed in relation to the low cell losses by physical dispersion in this area and the lack of upward vertical migration of G.corsicum.      

First evidence of cell deformation occurrence during a Dinophysis bloom along the shores of the Gulf of Tunis (SW Mediterranean Sea)

Never before observed or cited in Dinophysis studies, deformations in Dinophysis acuminata and Dinophysis sacculus are reported throughout their cellular division phases (cytokinesis, and sulcal list regeneration) in 5 in situ cell cycle studies in the Punic harbors of Carthage (northern Tunisia). Two types of deformation were observed: invaginations in the ventral and dorsal margin and protuberances at the base of the left sulcal list. No virus or bacteria were detected with Syber green stain. In situ division rates (μ) varied among seasons and stations for the same species. D. acuminata exhibited moderate (0.22 day⁻¹) to high (0.68 day⁻¹) μ rates which were however very low (0.02–0.17 day⁻¹) for D. sacculus in autumn and moderate (0.21–0.35 day⁻¹) in late spring. In 2009 the seasonal distribution of Dinophysis indicates maximum Dinophysis cf. ovum abundance in March and a high number of D. acuminata in early June, while in 2010 maximum abundance of the same species was found in mid-June.Molecular and genetic studies and staining with specific fluorescent strains should be addressed to hopefully explain these Dinophysis cell deformations during their in situ division.     

Effects of light and nitrogen limitation on the cell cycle of the dinoflagellate Amphidinium carteri

Cell cycle phase durations of cultures of Amphidinium carteriin light- or nitrogen-limited balanced growth were determinedusing flow cytometry. For both types of growth rate limitation,the increases in generation time caused by increasing degreesof limitation were due solely to expansion of the G₁ phase ofthe cell cycle. The durations of the S and G₂ + M phases wereindependent of growth rate. Furthermore, when cells were deprivedcompletely of light and nitrogen, they arrested in the G₁ phaseof the cell cycle. The results indicate that light- and nitrogen-dependentprocesses are heavily concentrated in the early part of thecell cycle, while DNA replication and cell division, once initiated,are independent of light or nitrogen supply.     

Diel Division Cycle and Growth Rates of Synechococcus in Lakes Huron and Michigan1

A clone of Synechococcus isolated from Lake Huron and natural populations of Synechococcus from lakes Huron and Michigan were studied in 1989 to examine the diel division cycle and to provide estimates of the in situ growth rate based on the frequency of dividing cells (FDC) method. Cultured populations of Synechococcus exhibited a consistent diel division pattern with a midday/afternoon (1100–1800 h) peak in the percent of dividing cells. The maximum percent of dividing cells varied among cultures (8-27%) and was related to the growth rate. A small fraction of dividing cells (3-5%) remained throughout the dark period, suggesting that some cells were arrested in the doublet stage prior to division. The duration of division (t_d) ranged from 2.6-4.9 h, with a 3.7 h mean for cultures with growth rates ≥0.34 d⁻¹ but increased to 8 h at a lower growth rate of 0.20 d⁻¹. The diel division pattern for natural populations was very similar to the laboratory clone; an afternoon peak (1400-2100 h) in dividing cells and a small fraction of dividing cells (2-5%) remained during the dark period. The maximum percent of dividing cells for natural populations ranged from 6-10%. In situ growth rates, determined from the FDC and assuming a constant t_d of 3.7 h, ranged from 0.30-0.42 d⁻¹. The FDC method may provide accurate estimates of in situ growth, particularly in environments where the growth rate is >0.34 d⁻¹, but in lakes Huron and Michigan where growth rates can be lower and t_d values may increase, FDC-growth rates must be viewed with caution.     

The Effects of Cell Cycle Parameters on Cell Wall Regeneration and Cell Division of Cotton Protoplasts (Gossypium hirsutum L.)

Protoplasts of cotton cotyledons were isolated and culturedto undergo cell wall regeneration and cell division. DNA contentand cell cycle parameters of nuclei from cotyledons and/or protoplastswere determined by flow cytometry. The DNA content of cotton,Gossypium hirsutum L., was estimated to be 4·34±0·12pg DNA per nucleus. There was a strong positive correlation between G₂ or Sand G₂,and cell wall regeneration and cell division and a strong negativecorrelation between G₁, and cell wall regeneration and celldivision of cotton cotyledon protoplasts. The cell cycle statusof cotyledons changes during their development; as the cotyledonsenlarge, the proportion of cells in G₀ and G₁ phases of thecell cycle increases. The implication of these results in relationto protoplast growth and development is discussed. Key words: Cell cycle parameters, cell wall regeneration, cell division, flow cytometry, Gossypium     

Control of cell division in the yeast Saccharomyces cerevisiae cultured at different growth rates

Saccharomyces cerevisiae has been grown with different generation times by alterations in media richness and by altering the flow rate of the limiting nutrient, glucose in a chemostat. Within the generation time range 2.89-approx. 8.0 h the time from the initiation of DNA synthesis to cell division was independent of generation time and was approx. 2 h. Thus the cell cycle of yeast can be divided into an expandable phase from cell division to the initiation of DNA synthesis, the length of which is dependent on growth rate and a constant phase from the initiation of DNA synthesis to cell division which takes a constant time independent of generation time. In cells growing with generation times longer than 8.6 h this constant phase expands somewhat in time. These results are reminiscent of the observation that in the bacterium Escherichia coliB/R the time from initiation of DNA synthesis to cell division is constant except at very slow growth rates.     

Photoperiodic Regulation of Cell Division and Chloroplast Replication in Heterosigma akashiwo

Cell division and chloroplast replication in Heterosigma akashiwo(Hada) Hada occurred as separate synchronous events during thecell cycle when cells were subjected to light-dark regimes.Under three different photoperiodic cycles of 10L/14D (10 hlight/14 h dark), 12L/12D or 16L/8D, cell division began athour 19–20 and finished at hour 23–26 after theonset of the light period, while chloroplast replication beganat hour 20–22 after the onset of the dark period. Almostall the cells divided only once in the 12L/12D cycle. The rateof increase in chloroplast number during one light-anddark cyclewas always equal to that in cell number in every photoperiodexamined. Light was essential for both cell division and chloroplast replication,but the minimum light period necessary for each event differed.When the light period was shorter than 6 h, no cell divisionoccurred; when it was shorter than 3 h, no chloroplast replicationoccurred. (Received February 26, 1987; Accepted June 17, 1987)     

Inductive and Inhibitory Effects of Light on Cell Division in Chattonella antiqua

The cell division of a red tide flagellate, Chattonella antiqua,was synchronously induced under light and dark regimes of 10L14D(a light period, L, for 10 h followed by a dark period, D, for14 h), 12L12D and l4L10D. In all regimes cell number began toincrease ca. 14 h after the onset of L and almost doubled duringone LD cycle. When the light-off timing of the last L was changedor the whole L was shifted, cells that had been synchronizedunder 12L12D invariably began to divide ca. 14 h after the onsetof L. This shows that the timing of cell division was determinedby the time of the onset of L. When cells were continuously exposed to light after a cell division,the subsequent cell division was inhibited. This effect waslimited to cells that had been synchronized under short-dayconditions. Thus it can be concluded that light has both inductive and inhibitoryeffects on cell division in this alga, the latter effect dependingupon the previously given light and dark regimes. (Received December 21, 1984; Accepted February 28, 1985)     

Simultaneous production of buds on mother and daughter cells of Saccharomyces cerevisiae in the presence of hydroxyurea

Individual budding yeast cells, Saccharomyces cerevisiae, enclosedin small culture chambers were observed through two buddingcycles to examine their behavior during growth and division.In the nutrient medium (YHG medium), the duration of the buddingcycles was 77 min for mother cells and 90 min for daughter cells;a 13-min time lag between the two durations. Continuous exposureof cells to 16 or 32 mM hydroxyurea extended the duration ofthe cycles and increased the volume of the cells, resultingin the formation of abnormally large and equal-sized mother-daughterpairs. Each cell of these pairs subsequently produced buds simultaneously.Stained cell nuclei showed simultaneous nuclear division. Thissynchronous budding on mother-daughter pairs was repeated inthe next budding cycle. The coordination of growth with divisionis discussed in relation to these results. (Received August 11, 1979; )     

Variation in the Rate of Cell Division in the Apical Meristem of Sinapis alba During Transition to Flowering

Rates of cell division in the central and the peripheral zonesof vegetative and evoked meristems of Sinapis alba have beenmeasured by accumulation of metaphases after colchicine treatment.The cells of the central zone had a longer cycle than the cellson the flanks of both kinds of meristems. The duration of thecell cycle was shortened in both zones of the meristem duringtransition to flowering. It was shown that the mitotic indicesof the two regions of the meristem were closely comparable totheir rates of cell division and therefore could be consideredrepresentative of the rates of cell division.     

Application of the frequency of dividing cells technique to estimate the in situ growth rate of Microcystis (Cyanobacteria)

1. Diel patterns of the frequency of dividing cells (FDC) of the bloom‐forming cyanobacteria Microcystis were investigated using both culture strains and natural populations. 2. In laboratory experiments, diel division cycles were examined twice in a 24‐h light/dark cycle during time‐course batch incubations of six culture conditions using two strains (morphospecies) of Microcystis (M. aeruginosa and M. wesenbergii). While both strains clearly showed phased cell division in the light period during the logarithmic growth phase, the peaks of FDC became unclear towards the stationary phases. Some dividing cells were always found in the dark period regardless of whether or not division had paused at the same time. 3. This result implied the inadequacy of applying the model of McDuff & Chisholm [Limnology and Oceanography (1982) vol. 27 , pp. 783–787] directly to calculate the duration of cell division. Modified equations are proposed to calculate the duration of cytokinesis as a terminal event, in which the FDC values at night are regarded as 0% and all FDC values are subtracted by the minimum FDC value. 4. The diel FDC in natural populations of M. aeruginosa and M. wesenbergii were examined at five sites from a harbour to several distances offshore in Lake Biwa. While both species showed phased cell division patterns in the daytime at the harbour, no peaks in FDC were discernible in the samples taken from the offshore sites. These results strongly suggested that Microcystis growth was higher inshore than offshore. The in situ growth rates were estimated using the new equations.     

Timing and Regulation of Nuclear and Cortical Events in the Cell Cycle of Tetrahymena pyriformis*

SYNOPSIS. Using continuous flow cultures based on the chemostat principle, we varied the cell generation times of the ciliate Tetrahymena pyriformis strain GL, from 4.9 to 22.2 hr and studied various parameters of the cell cycle at 28 C. These included: the duration of the periods required for oral morphogenesis, macronuclear division, cell division, G₁ S, and G₂. The size of individual cells was also measured. Independent of the growth rate, the period of oral morphogenesis occurred during the last 90 min of the cell cycle. In all cases macronuclear and cell divisions took place during the last part of these 90 min, and the final macronuclear separation occurred just before final cell separation. The S-period increased slightly, while the G₁ and G₂ both increased in roughly the same relative proportion to the increasing generation times. Slowly growing cells (generation time 20.5 hr) were shorter but broader and somewhat larger in volume than quickly growing cells (generation time 4.9 hr).     

Development of Lateral Root Primordia in Vicia faba, Pisum sativum, Zea mays andPhaseolus vulgaris: Rates of Primordium Formation and Cell Doubling Times

Lateral root primordium development has been examined in primaryroots of Vicia faba L., Pisum sativum L., Zea mays L. and Phaseolusvulgaris L. Following their initiation from an estimated minimumnumber of 77–162, 20–57, 17 and 12 cells respectivelyin Vicia, Phaseolus, Pisum and Zea, the primordia rapidly increasedin cell number to emerge as secondary roots about 2.8–3.6days later depending on the species being examined. Cell doublingtimes were estimated directly from cell numbers at differenttimes following primordium inception and were found to increasewith increase in primordium size in each of the species investigated. The number of primordia formed per cm of root growth per daywas greatest in Zea and least in Pisum. A comparison of thedata obtained for Vicia with that in the literature led to theconclusion that although the number of primordia produced percm of root growth was independent of the rate of primary elongation,the number produced per day increased in a linear fashion withincrease in the rate at which the primary lengthened. Vicia faba L, Pisum sativum L, Zea mays L, Phaseolus vulgaris L, broad bean, garden pea, maize, dwarf bean, root primordia, cell division, cell doubling time     

The Nuclear Endoreduplication Cycle in Metaxylem Cells of Primary Roots of Zea mays L.

The nuclear DNA content of metaxylem cells in roots of Zea mayscv. Golden Bantam reaches 16C or 32C by successive rounds ofDNA endoreduplication. Each phase of endoreduplication (endo-S)is separated by a non-DNA synthetic phase (endo-G). These phasesseem to occur in zones at fixed distances from the root tip.The duration of the phases in two of the endoreduplication cycles(4C–8C, 8C–16C) has been estimated in two ways.The first makes use of the rate of movement of cells throughthe positions along the root where the different phases of thecycle are occurring, the second uses labelling with methyl-[³H]thymidineand autoradiography. Both methods indicate that the endo-S phaseswhich cause the nuclear DNA content to rise from 4C to 8C andfrom 8C to 16C last 8–10 h, and that the intervening endo-Gphase lasts 8–12 h. DNA endoreduplication keeps pace withthe increase of nuclear volume; cell volume increases at a morerapid rate, however. Comparison of the endoreduplication cyclein the metaxylem with the mitotic cycle in the adjoining filesof parenchyma cells shows that the mitotic cells complete theircycle more slowly. DNA synthesis, endoreduplication cycle, mitotic cycle, root apex, Zea mays     

Gibberellin does not accelerate rates of cell division in the dwarf pea shoot apical meristem

Although GA₃ doubled the numbers of cells in dwarf pea internodes,it caused no significant acceleration of cell division ratesin the apical meristem, estimated using cell doubling times,mitotic indices, or percentage labelled mitoses data. Increasedcell numbers in GA₃-treated pea stems must be generated withinthe extending internodes. Key words: Cell division cycle, gibberellin, pea, Pisum, shoot apical meristem     

Interrelation Between Growth Rate of Individual Potato (Solanum tuberosum L.) Tubers and Their Cell Number

In potato plants fast and slow growing tubers develop on thesame plant. A hypothetical causality between tuber growth rateand tuber cell number was investigated by determining the tubercell number with the aid of an automatic counting procedure.Our data show a close correlation between tuber size and cellnumber over the whole range of tuber volumes considered (3–28cm³). If the influence of tuber size on cell number is eliminatedby means of a partial correlation analysis, the cell numberof the entire tuber is not significantly correlated with itsgrowth rate. An exclusive consideration of the smaller cells(10–30 µm) in the apical tuber region, where thecell division rate in potato tubers is highest, reveals a loosebut significant partial correlation to tuber growth rate (r= 0.383, P < 0.05). The growth rate of the slow growing tubers of any potato plantmay be enhanced by removing the fast growing tubers. In thefirst few days this enhanced growth rate is not due to a stimulationof cell division rate, but rather due to cell expansion. Potato, Solanum tuberosum L., tuber growth rate, tuber cell number     

Interpretation of in vivo fluorescence and cell division rates of natural phytoplankton using a cell cycle model

A natural population of phytoplankton was collected from theMenai Straits and incubated in a continuous culture system undernatural illumination. In vivo fluorescence data were used toderive cell cycle parameters, and the division rates of cellsin the population were analysed using a transition-point modelof the cell cycle. The results indicated that for modellingpurposes the mixed population could be regarded as being composedof a single species having properties equivalent to the averagefor the population. Fluorescence measurements were subsequcntlyrepeated on samples of the in situ population at the collectionsite, and the cell cycle model used to interpret the growthrate of the phytoplankton in the natural environment.