THE INFLUENCE OF INJECTED TRITIATED THYMIDINE ON THE MITOTIC CIRCADIAN RHYTHM IN THE EPITHELIUM OF THE HAMSTER CHEEK POUCH*

The initial effect of an injection of TdR-5-³H (1 μCi/g body weight; 6 Ci/mmol) in the cheek pouch epithelium of the Syrian hamster is an increase in the mitotic index. The increase is observed 1–5 hr after injection, depending upon the time of day when the injection is given, and is followed by compensatory variations in mitotic index. This deviation from the normal circadian rhythm in the mitotic index appears to depend on the fraction of G₂-cells at the time of injection. The main effect is a shortening of t_g2. No effect is observed after injection of non-radioactive TdR or isotonic saline. The results of the present experiment emphasize that unexpected results may be obtained when using mitotic indices from animals labelled with ³H-TdR, as well as the risks of using the PLM-method in a partially synchronized system.     

A QUANTITATIVE STUDY OF THE MITOTIC ACTIVITY IN THE SUBEPENDYMAL PLATE OF ADULT RATS

A quantitative study of the mitotic activity of the cells of the subependymal layer in the rat brain has shown that mitotic activity decreases with the increase in age of the animals. Mitotic activity is still found in rats 16 months old and in the case of animals of two strains of rats, of this age, the mitotic count is significantly higher in males than in females.
The relationship between a reduced rate of brain growth and the decline in mitotic activity and the significance and importance of the higher mitotic count in the subependymal plate of 16-month-old males compared with similar females is discussed.     

EFFECT OF UNDERNUTRITION ON CELL FORMATION IN THE RAT BRAIN

Abstract— Rats were undernourished by approximately halving the normal food given from the 6th day of gestation throughout lactation. Growth of the foetuses was nearly normal, in marked contrast to the severe retardation caused by undernutrition during the suckling period. In comparison with controls the size and the DNA content of the brain were permanently reduced by undernutrition during the suckling period: this effect was relatively small, approx. 15 per cent decrease at 21 and 35 days. The rate of ¹⁴C incorporation into brain DNA at 30 min after administration of [2-¹⁴C] thymidine was taken as an index of mitotic activity; compared with controls there was severe reduction in mitotic activity (maximal decrease by about 80 per cent at 6 days in the cerebrum and by 70 per cent at 10 days in the cerebellum). The rate of acquisition of cells was calculated from the slopes of the logistic curves fitted to the estimated DNA contents. In normal animals the maximal slope was attained at 2·7 days and at 12·8 days after birth in cerebrum and cerebellum respectively; the daily acquisition of cells at these times was 4·8 × 10⁶ and 18 × 10⁶ cells respectively. The fractional increase in cell number at the maximum was 5·4 percent per day in the cerebrum and 15·2 per cent per day in the cerebellum. The rate of acquisition of cells relative to the rate of mitotic activity was higher in the brains of undernourished animals than in controls. One of the compensatory mechanisms for the severe depression of mitotic activity in the brain of undernourished animals Seems to involve a reduction in the normal rate of cell loss.     

III. DECREASE OF LABELED DNA IN CELLS OF THE ADRENAL MEDULLA AFTER INTERMITTENT EXPOSURE TO COLD

Italico rats were injected with thymidine-³H 6 hr after the end of 300 hr of intermittent cold treatment. This plan of experiment ensured replacement in the adrenal medulla of lost DNA which is specifically sensitive to cold treatment and has a labeling index sufficiently high for statistical evaluation. The labeling index in the adrenal medulla decreases to one-half of the initial value within 10 days in animals subjected to further intermittent cold treatment and within 32 days in animals kept at room temperature. The very low mitotic index and the absence of doubling of the labeling index show that the observed labeling cannot be ascribed to pre-mitotic DNA synthesis. The concept of metabolic DNA adequately explains the findings.     

THE RELATIONSHIP BETWEEN DIURNAL VARIATION OF THE NUMBER OF CELLS IN MITOSIS AND OF THE NUMBER OF CELLS SYNTHESIZING DNA IN THE EPITHELIUM OF THE HAMSTER CHEEK POUCH

1. The numbers of cells in mitosis and in DNA synthesis in the epithelium of the hamster cheek pouch have been studied at different times of the day and night. 2. By accumulation of mitotic cells using colcemid, both the rate of entry of cells into mitosis and the duration of mitosis have been estimated at two different times of day. 3. A diurnal variation has been demonstrated in both the mitotic index and in the tritiated thymidine labelling index. Although these variations are of different amplitude and timing, the experimental data fit closely to the hypothesis that the diurnal mitotic variation is the result of a partially synchronous population moving through the DNA synthetic period. No direct action on the mitotic process need be postulated. 4. From the results of mitotic accumulation, it is clear that the rate of entry of cells into mitosis depends on the time of day at which this is studied. There is also the possibility that the duration of mitosis is slightly longer when the mitotic index is high. 5. It is concluded that, at least in the epithelium of the hamster cheek pouch, the diurnal rhythm in the number of mitoses present is a reflection of the diurnal variation in the number of cells synthesizing DNA at some time earlier. Small fluctuations in the mitotic pattern imposed by this partially synchronous population moving from S into mitosis, could be caused by slight variations in the duration of mitosis.     

REGULATION AND KINETICS OF SPERMATOGONIAL PROLIFERATION IN ARENICOLA MARINA (ANNELIDA, POLYCHAETA) II. KINETICS

The rate of accumulation of mitoses in the testis of Arenicola marina has been estimated by the colchicine method. The rate is low in February but increases during spring to a maximum in June. The rate remains high during July and August but decreases in September. A second temporary and minor rise was observed in October or November prior to spawning.
The rate of accumulation of mitoses (with respect to the duration of metaphase arrest) is logarithmic from February to June, but is arithmetic during the latter part of the reproductive cycle (July-October). The transition from one phase to the next coincides with the initiation of spermatocyte release from the testis into the coelom.
The rate of accumulation of mitoses has also been recorded in testes cultured in vitro. The rate in vitro does not differ from that in vivo in May when the rate is high, but in September when the rate in vivo is depressed, culture in vitro significantly increases the mitotic rate.
The cell cycle time of the proliferating spermatogonia has been estimated by a modified per cent labelled mitosis technique. The cell cycle time is unusually long (in excess of 20 days) and the G₂ period is at least 5 days. It is suggested that the testis of A. marina is an exponentially growing cell population from February until early June when coelomic spermatocytes first appear. The rate of cell release then temporarily exceeds the rate of renewal: eventually they become equal and the testis remains in steady state until September when the mitotic rate is depressed due to the presence of maturing spermatocytes. The depression of the mitotic rate in September is achieved by the arrest of spermatogonia in G₂. Explanting the gonads in organ culture removes the inhibition and causes an increased flow of cells from G₂ into mitosis.     

CHANGES IN THE DNA SYNTHETIC PERIOD DURING THE PHASE OF RAPID GROWTH IN CULTURED DIPLOID FIBROBLASTIC CELLS

The DNA synthetic period in diploid fibroblastic cell populations was studied during the phase of rapid growth in vitro , using the labelled mitotic wave method and quantitative autoradiography.
It was found that the length of the DNA synthetic period increased progressively, from early passage levels, with the number of generations in vitro. the increase was mainly in the time taken to synthesize the first 50%, and especially the first 10%, of DNA.
The change was detectable despite the complication due to a low level of tritium incorporation after the removal of tritiated thymidine from the cultures.     

THE ROLE OF GLUCOCORTICOID HORMONES IN THE CONTROL OF EPIDERMAL MITOSIS

It has previously been established that the epidermal chalone inhibits epidermal mitotic activity more powerfully in the presence of adrenalin, although adrenalin itself is not a mitotic inhibitor. It is now shown that in low concentrations hydrocortisone has little if any antimitotic activity, that when it is present together with chalone and adrenalin it does not markedly increase their antimitotic activity, but that it does act to prolong the mitotic depression which they induce. It is known that, without hydrocortisone, adrenalin rapidly escapes from epidermal cells so that the chalone action is weakened. It appears that the role of hydrocortisone may be to reduce the rate of adrenalin loss and thus to prolong the chalone-adrenalin activity. It is also shown that the rate of loss of adrenalin from epidermal cells is inhibited, though to a much lesser extent, in the presence of excess chalone.     

POPULATION DYNAMICS OF MICROALGAE IN THE UPPER LAND-FAST SEA ICE AT A SNOW-FREE LOCATION

The population dynamics of interior ice microalgae were investigated at a snow-free site on annual land-fast sea ice in McMurdo Sound, Antarctica, during the austral spring and summer of 1995 – 96. A dynamic successional sequence was observed with life history transformations playing an important role. During late November and early December (austral spring), cryo- and halotolerant dinoflagellates and chrysophytes bloomed in brine channels within the upper ice. At this time, competition and grazing pressure are low because of the inability of most marine species to grow under the extreme environmental conditions found in the upper ice during the austral spring. In November and December, dinoflagellates, chrysophytes, and prasinophytes contributed an average of 66%, 44%, and < 1% of the phytoflagellate biomass, respectively. Both the dinoflagellates and the chrysophytes encysted in December, with cyst formation most intense just prior to surface melt and flushing of the ice. The cysts appear to be an adaptation for survival and dispersal in the plankton during ice decay and/or overwintering in the sea ice. In January (austral summer), when ice temperatures were similar to those in the water column, pennate diatoms replaced flagellates as the photosynthetic dominants in the upper sea ice. The upper land-fast sea ice undergoes dramatic seasonal changes in light availability, temperature, brine salinity, and inorganic nutrient availability. Ephemeral blooms of cyst-forming phytoflagellates exploit this habitat in the austral spring, when both inorganic nutrients and light are available but temperatures <− 2° C and brine salinities elevated.     

MITOTIC ACTIVITY OF THE PARS INTERMEDIA IN THE FEMALE MOUSE: AGE-ASSOCIATED VARIATIONS IN PROLIFERATION RATE AND CIRCADIAN PERIODICITY

We previously reported daily variations in the mitotic activity of the endocrine cells in the pars intermedia of 21- and 28-day-old male mice. Since cellular proliferation might be affected by factors such as sex and age, we undertook the present experiments to study the mitotic activity of the pars intermedia from 14-, 28-, and 150-day-old female mice. Inbred C3H/S mice, grouped according to age, were housed under standard conditions (12h each of light and dark [LD 12:12]) for periodicity analysis and were killed in lots of 5–11 animals every 4h over a single 24h cycle, with each mouse receiving 2 μg/g of colchicine 4h before decapitation. Pituitaries were excised, extracted, fixed in buffered formaldehyde, embedded in celloidin-paraffin, sectioned at 5 μm, and stained with hematoxylin and eosin. We counted the total number of nuclei to estimate the total number of cells monitored and then calculated the mitotic index (metaphases/1000 nuclei). Differences were analyzed for statistical significance by the Student t test. While the 14-day-old animals manifested no significant changes in mitotic activity during the 24h cycle, the 28- and 150-day-old mice showed higher mitotic indices during the period of darkness. The average mitotic activity over the entire cycle, however, was higher in the two groups of younger animals than in the 150-day-old mice. Moreover, the averages for the 28-day-old females were higher than the corresponding values previously reported by us for male mice of the same age. (Chronobiology International, 17(6), 751–756, 2000)     

THE EARLY ACTION OF THE FLORAL STIMULUS ON MITOTIC ACTIVITY AND DNA SYNTHESIS IN THE APICAL MERISTEM OF XANTHIUM STRUMARIUM

Vegetative plants of Xanthium strumarium (a short-day species) were induced to flower by exposure to a single 16-hr long night. By cutting off the induced leaf (half-expanded leaf) at various times, it was established that, by 8 hr after the end of the long night, a sufficient amount of floral stimulus had reached the meristem to induce a flowering response. The following sequence of events occurred in both the peripheral and central zones of the apical meristem of induced plants: 1) a rise in the mitotic index beginning at 28 hr after the end of the long night and culminating at 36 and 56 hr; 2) a stimulation of DNA synthesis starting at 32–36 hr and reaching a maximum at 60 hr; 3) an increase in nucleolus diameter starting at 32 hr. The cell population in the meristems of both vegetative and induced plants displayed a similar distribution, with about 80 % of the nuclei with the 2C amount of DNA. The comparison of the kinetic data concerning the mitotic index and DNA synthesis indicated that one of the early effects of the floral stimulus in the peripheral and central zones was the release in mitosis of cells whose nuclei were in the postsynthetic (G₂) phase of the mitotic cycle. In the pith-rib meristem, the following events were recorded: 1) a stimulation of DNA synthesis starting at 20 hr; 2) a rise of the mitotic index beginning at 28 hr; 3) the vacuolation and elongation of cells starting at 48 hr. All these events occurred well before the initiation of bract and flower primordia, which began at 96 and 136 hr, respectively. Neither stimulation of mitotic activity nor flowering occurred in the meristems of plants subjected to a long night interrupted at its midpoint by a 5-min light break. The results are discussed in relation to the early events which are known to occur in the meristems of other photoperiodic species in transition to flowering.     

THE EFFECT OF X-RAYS ON CHROMOSOMES IN DIFFERENT STAGES OF MEIOSIS

1. Pollen mother cells exposed to low dosages of x-rays at various stages show different frequencies of chromosome abnormalities in the first meiotic anaphase. 2. Maximum frequencies of abnormalities were obtained in buds irradiated in the pachytene stage of the meiotic prophase and in the preceding mitosis. 3. These results are taken to indicate that the x-ray-sensitive portions of the chromonemata are closely approximated in pairs in pachytene and in the early mitotic prophase. The significance of this in relation to non-homologous pairing is indicated. 4. From the nature of the chromosome configurations observed it is concluded that chromonemata are two-parted when they synapse and that a chromonematic division occurs between pachytene and anaphase and during the mitotic prophase. 5. The frequencies of abnormalities show a linear relationship to dosage. 6. The diameter of the sensitive volume of the chromonema is calculated and found to approximate the diameter of some known protein molecules. 7. The linkage mechanism is found to make up about 90 per cent of the total sensitive volume which corresponds with the approximate reduction in length of the chromonema from pachytene to anaphase. 8. The relation of these sensitive volumes to the gene is discussed.     

THE EFFECT OF VESSEL NOISE ON THE VOCAL BEHAVIOR OF BELUGAS IN THE ST. LAWRENCE RIVER ESTUARY,CANADA

During June-July 1991, we monitored the vocal behavior of belugas before, during, and after exposure to noise from a small motorboat and a ferry to determine if there were any consistent patterns in their vocal behavior when exposed to these two familiar, but different sources of potential disturbance. Vocal responses were observed in all trials and were more persistent when whales were exposed to the ferry than to the small boat. These included (1) a progressive reduction in calling rate from 3.4–10.5 calls/whale/min to 0.0 or <1.0 calls/whale/min while vessels were approaching; (2) brief increases in the emission of falling tonal calls and the theree pulsed-tone call types; (3) at distances <1 km, an increase in the repetition of specific calls, and (4) a shift in frequency bands used by vocalizing animals from a mean frequency of 3.6 kHz prior to exposure to noise to frequencies of 5.2-8.8 kHz when vessels were close to the whales.     

蓄水对三峡库区重庆段长江干流浮游植物群落结构的影响

为了分析三峡库区长江干流重庆段因蓄水产生的三类不同水体(自然河流型、过渡型、类湖泊型)对浮游植物的影响,分别于不同季节和蓄水期对水体浮游植物进行了群落调查,结果显示各群落指标仍呈现亚热带河流型水库的季节变化趋势:春季绿藻、隐藻占优势,夏季蓝藻所占比例增加,冬季硅藻占绝对优势,但不同类型水体对季节的响应程度不同:(1)2010年3月春季调查期间绿藻的优势种的优势度以及所占比例均呈现随水体类型改变逐渐增加趋势,并在类湖泊型水体中达到最大(优势度0.62、所占比例达67.8%),而夏秋季(9月蓄水前)蓝藻的优势种数、优势度、藻密度以及所占比例也存在同样的变化,2009年9月蓝藻所占比例由河流型水体的8.5%增加到类湖泊型水体的52.6%,到2010年9月类湖泊型水体蓝藻比例甚至达到63.8%。冬季蓄水期间(2010年1月),硅藻占绝对优势,但绿藻在类湖泊型水体中的比例要大于其他两类型水体。(2)Shannon-Wiener指数平均值为2.43,总体呈现出河流型水体>过渡型水体>类湖泊型水体,类湖泊型水体多样性指数夏秋季>冬季>春季,而自然河流型和过渡型水体冬季多样性指数低于其他季节。     

DEVELOPMENT OF A CIRCADIAN RHYTHM IN THE ACTIVITY OF PINEAL SEROTONIN N-ACETYLTRANSFERASE

Abstract— Pineal serotonin N -acetyltransferase (EC 2.3.1.5) is a neurally regulated enzyme. It is detectable in the rat as early as 4 days prior to birth. A circadian rhythm in enzyme activity appears on the fourth day after birth. It develops most rapidly during the second week and achieves an adult magnitude by the end of the third week at which time nocturnal values are more than 30-fold greater than daytime values. Norepinephrine, which appears to be the neurotransmitter regulating this enzyme, can cause a 2- to 3-fold stimulation of N -acetyltransferase in organ cultures of pineal glands from 4-day-old animals and a 17-fold increase in the activity of glands from 15-day-old animals. Apparently the norepinephrinesensitive system controlling pineal N -acetyltransferase activity also develops most rapidly during the first few weeks of life. The circadian rhythm in the activity of serotonin N -acetyltransferase develops in the pineal glands of both male and female rats at the same rate. A similar rhythm for the enzyme was not observed in twelve other tissues of the rat.     

MITOTIC ACTIVITY IN THE ROOT APEX OF THE WATER FERN MARSILEA VESTITA HOOK. AND GREV.

Roots of Marsilea vestita ranging from 1–120 mm in length, as well as root primordia, were analyzed to determine mitotic activity and ploidy levels in the apical cell, five well-defined regions of the root proper, and two regions in the root cap. The mitotic index of the apical cell tended to be above the overall mean mitotic index for the entire apical meristem. No diurnal rhythm in mitotic index was apparent. The cell-cycle duration of the apical cell ranged from 12.1–25.2 hr, that of other regions of the root from 16.1–41.5 hr. There was no indication of polyploidy in any part of the apical meristem except in a few procambial cells. Thus, the results support the classical concept that the apical cell is the ultimate source of cells in the root.     

Annual mitotic activity in the intestinal epithelium of the musselCrenomytilus grayanus

The seasonal dynamics of cell reproduction in the intestinal epithelium of the musselCrenomytilus grayanus are described in detail. Mitotic indices in the intestinal epithelium varied throughout the year from 0.005 to 0.26% (averaged data) and from 0.003 to 0.37% (individual data). Cyclic seasonal changes were found in the mussel’s intestinal epithelium. In general, the average values of mitotic activity in the intestinal epithelium were low (the mitotic index was 0.13%); there was a rise in activity in late April–June and September and a decline in July–August and especially in January–March. The winter-early spring period was characterized by a profound inhibition of cell reproduction and the transition of cells to the resting state. An outburst of proliferation occurred in the spring, due to a manifold increase in the number of cells in the mitotic cycle. The musselC. grayanus may be a good model for the study of the two extreme states of proliferation and their alternation in marine animals in nature. The diel dynamics of mitotic activity in the intestinal epithelium were followed during the most active growth period (May). The mitotic index (MI) varied during the day within a narrow range, deviating from the daily average value by no more than one third; no pronounced diel rhythm was found. Optimum water temperatures for cell reproduction ranged from 5 to 18°C.     

MITOTIC ACTIVITY IN THE CAMBIAL ZONE OF PINUS STROBUS

Mitotic activity in the cambial zone of 20-year-old Pinus strobus L. trees was determined quantitatively, using mitotic counts from serial tangential sections of sample pieces. Counts from nine cores 1.6 mm in diameter from each sample piece were averaged and expressed as the number of mitoses per core with the sampling error. Estimation of the number of cambial zone cells per core permitted calculation of mitotic indices. In the fourth internode from the ground the first mitoses were observed on April 15 and the last on September 15. The number of mitoses per sample piece was comparable throughout each internode at any one sampling, but the number was higher in internodes within the crown than in those below it. Mitotic index was not appreciably higher within the crown, but it decreased from 3.7 in spring to 2.0 in fall. An internode in the crown sampled May 11–13 showed afternoon peaks in mitotic activity, but an internode below the crown showed no peaks, nor did other internodes sampled later in the season.     

WEIGHTS OF SOME PALAEARCTIC WADERS WINTERING IN KENYA

Palaearctic waders were caught and weighed at Lake Nakuru in the Kenyan rift valley on 22 occasions in the non-breeding season between March 1967 and May 1969. The weights of four species are presented:Curlew Sandpiper, Little Stint, Ruff and Marsh Sandpiper.
Winter weights were relatively low and varied little. It is thought that waders had very little fat reserve at this time.
The mean weights of early autumn samples tended to be higher than corresponding mean winter weights, and autumn populations included some markedly heavy birds.
An increase in the mean weight, and a dramatic rise in the maximum weights of each species occurred shortly before its main spring departure time. The great majority of waders apparently gained considerable weight before migration from the area in April and May.
The spring migration of waders from Kenya is briefly discussed.