可移植性小鼠白血病(L615)的实验研究——Ⅳ.L615白血细胞增殖动力学

采用体内~3H-TdR标记法测定L615白血细胞的增殖时(Tc)为13.8小时、合成前期(G_1)2.8小时、合成期(S)8.6小时、合成后期(G_2)1.8小时、分裂期(M)为0.65小时。~3H-TdR单次脉冲标记率(LI)为53％,连续标记24小时后标记率>99％,其增殖比例近于1。 用稀释法接种不同浓度L615白血细胞所得的结果表明:接种细胞数与小鼠存活时间呈直线的对数关系。“单个”L615白血细胞接种至小鼠死亡需时13.2天。 由于L615白血细胞的增殖时短而合成期较长,因而使用烷化剂及抑制合成期的抗癌药物可取得较好的疗效。     

甜叶菊的受精作用及胚和胚乳的早期发育

成熟胚囊被珠被绒毡县包围,由卵器、具次生核的中央细胞以及数目为1至6个反足细胞组成。传粉后6小时左右,雌、雄性核融合。配子融合同精核与次生核的融合几乎同时发生;精核与次生核融合速度快于配子融合。甜叶菊受精作用属于有丝分裂前类型。传粉后8小时左右,初生胚乳核分裂,其分裂方向可与胚囊长轴平行或垂直,从而形成最初的两个胚乳细胞。胚乳细胞前5次分裂是同步的。心形胚阶段,胚乳细胞呈现被消化吸收的迹象。胚乳发育属细胞型。传粉后10小时左右,合子第一次分裂,为横分裂。胚胎发育属紫菀型。     

X射线对小白鼠雄性生殖细胞的作用

1.以400伦X射线全身照射昆明种雄性小白鼠,照射后隔1、3、6、12、24小时分别将动物杀死,以Dietrich-Parat法固定其睾丸。制成石蜡切片。观察第Ⅱ、Ⅳ、Ⅵ、Ⅻ期精小管横切面中各类细胞数量的变化。 2.X射线照射后,小白鼠雄性生殖细胞中出现各种坏死及异常细胞,按其性质可大致分为四类:细胞坏死、染色体畸变、异常分裂、巨细胞。类似的坏死及异常细胞在正常睾丸中几乎都能发现,但频率低得多。 3.根据细胞数量、细胞分裂周期各类细胞和细胞坏死频率的变化,可以把X射线对精原细胞的早期(24小时内的)效应大致分为三个阶段:原发效应期、恢复期和转折期。 4.精原细胞数量的减少主要由于细胞死亡。它们的辐射致死和细胞分裂周期有一定的关系。然而,当分裂抑制解除时,精原细胞可以在细胞分裂周期的各个时期中死亡。 5.X射线照射后1小时,三种精原细胞都出现了分裂延滞的现象,但对中-后期细胞分裂有一定的促进作用。实验结果表明X射线对精原细胞有丝分裂的抑制是可逆的。抑制的时期可能是在间-前期之间和分裂之前(前期)两个时期。这两个时期与细胞有丝分裂周期中的“不可逆点”(Point of no return)相符。 6.X射线对减数分裂前期细胞及精细胞的数量和显微结构影响不大,但大大地减少减数分裂中-后期正常细胞的数量     

中华大蟾蜍(Bufo bufo gagarizans)血淋巴细胞的体外培养及其转化

本文报道了中华大蟾蜍血淋巴细胞的培养及其在植物血球凝集素(PHA)的刺激下所引起的转化,并初步查明了转化的淋巴细胞的S期、G_2期所占时间。用氚标记放射自显影手段测定了细胞的转化率。用姐妹染色单体区分着色的方法决定细胞的分裂次数。实验结果表明,在新鲜血液中有0.1—0.2％的白细胞具有合成DNA的能力。培养3天后,淋巴细胞转化率为14.8％,第5天达48％。培养的第6天有6—10％为第2次分裂。在秋水仙素处理3小时的情况下,有丝分裂指数为7％,延长处理时间至10—15小时,则平均达10％。最高有丝分裂指数为29％。转化淋巴细胞的S期为16小时,G_2期为3小时。     

温度休克法诱导草履虫的同步分裂

运用细胞周期原理,采用温度休克法,对尾草履虫进行分裂周期同步化的研究,实验中草履虫经过3－5h的处理后,就能观察到大量不同阶段的无性生殖横分裂状态,并获得了大量处于分裂阶段的草履虫。运用这种技术取材容易,获取率稳定,可达61％,可为细胞生理学等领域的研究提供大量的同步分裂个体。     

蚕豆有絲分裂在X射綫照射后初期的变动分析

五、摘要本文对500伦X射綫照射后初期蚕豆根端細胞有絲分裂各期比例的变化进行了分析,所得結果总結如下: 1.照射后30分钟內有絲分裂指数(分裂細胞占全部細胞的9％)与对照组无明显差別。自照射后1小时分裂指数开始表現出下降的趋势,照射后3小时下降才很明显,降为对照組的66％。 2.照射后3分钟看到前期占分裂細胞的比例減少,而中期比例增加。前期比例的减少可能是由于射綫的作用使間期末細胞不能进入前期,而一部分晚前期細胞轉入中期的結果。中期比例的增加是由于一方面有前期细胞进入中期,另方面中期过程受到阻抑。虽然中期过程受阻,但后、末期細胞占分裂細胞的比例并未減少,因此說明几乎沒有末期細胞轉入間期。 3.照射后3分钟看到中期比例增加,30分钟看到早后期比例增加,照射后30分钟至3小时带桥的后、末期細胞数不断增加。这一系列变化是由于輻射对染色体作用的“生理学”效应,即由于射綫作用使染色体表面的粘着性升高,从而中期和早后期过程变慢,并产生染色体桥。 4.照射后30分钟看到中期比例比照射后3分钟減少,而后期比例增加。这說明前期細胞未进入中期,而一部分中期細胞已轉入后期,另外后期过程遭到了阻抑。这时看到末期比例比照射后3分钟減少,可能有一小部分末期細胞完成分裂过程轉入了間期。 5.照射后1小时后期比例比照射后30分钟减少很多,說明在照射后30分钟至1小时期間已有一部分后期細胞进入末期。虽然如此,末期比例并未增加。因此,說明末期細胞也有一部分进入了間期。 6.照射后3小时前期占全部細胞的％与照射后1小时并无差別,但占分裂細胞的％大大增加。中期比例略有減少,而后期和末期減少甚多,并且这时看到的后、末期細胞大多具染色体桥。說明这时后、末期細胞大多数已进入間期,而那些不正常的剩下未动。前期过程遭受阻抑而未进入中期,中期細胞只有一部分进入了后期。过去有些作者对照射后2、3小时前期比例增加現象的原因提出过各种推测意見,本文对这些意見进行了分析討論。     

PHA活体注射诱发黑斑蛙淋巴细胞转化的观察

本文应用PHA一次性注入黑斑蛙腹腔的方法,系统地研究了不同剂量的PHA对黑斑蛙的细胞遗传学效应。结果表明,PHA剂量为0.3mg/g(体重)时,骨髓和血淋巴细胞转化率分别为77.49％和79.17％;分裂指数为9.40％和10.87％;此外,统计结果说明,PHA处理24小时和处理48、72小时相比,骨髓和血淋巴细胞转化率均有显著性差异(P<0.05)。     

大蒜根尖细胞有丝分裂同步化诱导与中期染色体分离

本研究以大蒜（Ａｌｌｉｕｍｓａｔｉｖｕｍ）根尖为材料,用ＨＵ－ＡＰＭ双阻断法对细胞有丝分裂中期同步化和前,后,末期部分同步化作了探讨。表明在１．２５ｍｍｏｌ／ＬＨＵ,４μｍｏｌ／ＡＰＭ处理可使细胞有丝分裂中期指数（Ｍｅｔ．Ｉ）达３５％,１８ｈｒ的ＨＵ单独处理可检测到２７％的前期分裂细胞,ＨＵ对后,末期的部分同步化也有明显作用,约有１７％的后,末期细胞被检测,用Ｓｃｈｕｂｅｒｔ介绍的方法,分离出中期     

赤麂和小麂♀×赤麂♂的杂种外周淋巴细胞动力学和姐妹染色单体交换率的研究

赤麂及其杂种(赤麂×小麂)外周淋巴细胞在离体培养条件下,培养了24小时均未出现分裂相。到36小时,赤麂只有1.4%而杂种麂已有10.5%的分裂细胞进入第2次分裂。在48小时,赤麂细胞尚未进入第3次分裂,而杂种麂却已有4.7%和0.2%的分裂细胞分别进入第3次和第4次分裂。在72小时,赤麂及其杂种的大多数分裂细胞仍处于第2次分裂阶段,进入第3次和第4次分裂的细胞数量也随之增多。赤麂的姐妹染色单体交换率在48和72小时分别为5.08±0.33/细胞和5.33±0.30/细胞;向杂种麂的姐妹染色单体交换率分别为4.76±0.21/细胞和5.40±0.74/细胞。     

家兔供体细胞的发育周期与重构胚发育的关系

采用血清饥饿法处理体外培养的兔子胎儿成纤维细胞,并将其作为供体细胞移入去核卵母细胞内构建重构胚胎。检查供体细胞的细胞周期对重构胚的融合率、分裂率和着床率的影响。实验结果表明：培养基中血清含量在0．5％的情况下,G0／G1期的细胞比例由正常培养条件下(培乔液中含有10％FCS)的73．2％明显地增加到86％以上。饥饿1～3天的细胞作为供体细胞构建重构胚时,可明显提高重构胚的融合率,但是不同的饥饿时间其融合率并无显著的差异。饥饿处理可明显增加重构胚的分裂率,以饥饿处理3天为最佳。     

Photosynthetic Activity during the Life Cycle of Synchronous Skeletonema Cells

A culture of Skeletonema costatum grown at a light intensity of 3 klux and at 20°C was synchronized in diurnally intermittent illumination of 12 hour light and 12 hour dark. The culture was hardly fully synchronous as the cell division period lasted about 9 hours. The cell division started in the middle of the light period. The concentration of the pigments: chlorophyll a, chlorophyll 6 and fucoxanthin and the rate of light-saturated photosynthesis were followed every hour during the 24 hour period. Both the concentration of pigments and the photosynthetic activity showed a rhythmical variation. The concentration per cell of all three pigments examined increased during the development of the cells and decreased automatically during the period of cell division. An increase in the pigment concentration was found only in the light period. The rate of light-saturated photosynthesis calculated per unit of cell number increased during the cell development and decreased during the division period. The increase in the photosynthetic activity at light-saturation started about 4 hours after the end of cell division, which was 4 hours before the light was turned on while the increase in the concentration of chlorophyll a first started 1–2 hours after this moment. The variation in photosynthetic activity was compared with that found by other workers. The results found with Chlorella ellipsoidea by Japanese scientists (Nihci et al.) was explained as an inhibition phenomenon because the cells were not adapted to the experimental conditions.     

Synchronous Growth and Plastid Replication in the Naturally Wall-less Alga Olisthodiscus luteus

Olisthodiscus luteus is a unicellular biflagellate alga which contains many small discoidal chloroplasts. This naturally wall-less organism can be axenically maintained on a defined nonprecipitating artificial seawater medium. Sufficient light, the presence of bicarbonate, minimum mechanical turbulence, and the addition of vitamin B₁₂ to the culture medium are important factors in the maintenance of a good growth response. Cells can be induced to divide synchronously when subject to a 12-hour light/12-hour dark cycle. The chronology of cell division, DNA synthesis, and plastid replication has been studied during this synchronous growth cycle. Cell division begins at hour 4 in the dark and terminates at hour 3 in the light, whereas DNA synthesis initiates 3 hours prior to cell division and terminates at hour 10 in the dark. Synchronous replication of the cell's numerous chloroplasts begins at hour 10 in the light and terminates almost 8 hours before cell division is completed. The average number of chloroplasts found in an exponentially growing synchronous culture is rather stringently maintained at 20 to 21 plastids per cell, although a large variability in plastid complement (4-50) is observed within individual cells of the population. A change in the physiological condition of an Olisthodiscus cell may cause an alteration of this chloroplast complement. For example, during the linear growth period, chloroplast number is reduced to 14 plastids per cell. In addition, when Olisthodiscus cells are grown in medium lacking vitamin B₁₂, plastid replication continues in the absence of cell division thereby increasing the cell's plastid complement significantly.     

THE REPLICATION TIME AND PATTERN OF THE LIVER CELL IN THE GROWING RAT

Three-week-old male rats of the Wistar strain were given tritiated thymidine, 1 µc/gm body weight, intraperitoneally and were killed at intervals from 0.25 to 72 hours later. Autoradiographs were made from 5 µ sections, stained by the Feulgen method. The replication time and its component intervals were determined from the scoring of the labeling of interphase nuclei as well as of prophase, metaphase, anaphase, and telophase nuclei. Absorption of the intraperitoneally injected label is rapid and is attended by "flash" labeling during interphase. The results show that at any one time about 4 per cent of the liver cells are synthesizing DNA preliminary to cell division. These cells alternate with waves of other cells and it is estimated that about 10 per cent of the liver cell population is engaged in cell duplication. The replication time is about 21.5 hours, and its component intervals occupy the following times: DNA synthesis, 9 hours; post-DNA synthesis gap, 0.50 hour; prophase, 1.3 hours; metaphase, 1.0 hour; anaphase, 0.4 hour; telophase, 0.3 hour; postmitosis gap, 9.0 hours. A group of liver cells has been recorded in at least 3 successive replication cycles.     

Shifts in nuclear and cytoplasmic nucleic acid content in temperature-synchronized Tetrahymena pyriformis (HSM)

Nuclei were isolated by exposing temperature synchronized Tetrahymena pyriformis (HSM) to Triton-X-100. Cell division synchrony was induced with a repetitive 12-hour temperature cycle (9.5 hours at 13°, 2.5 hours at 29°). Increase in nucleic acid content was biphasic: primarily during the last two hours of the cold period well in advance of the synchronous burst of division and secondarily in the last hour of the warm period. Nuclear RNA content rises almost two hours ahead of cytoplasmic RNA which shows a maximum 0.5 hour before the onset of the warm period. The DNA content reaches a peak 30 minutes later. On the basis of these shifts there appears to be not net synthesis of nucleic acids during cell division. The changes in RNA/DNA of the isolated macronuclei and micronuclei suggest enhanced RNA turnover, loss to the cytoplasm and enhanced ribonuclease activity prior to cell division. Cytoplasmic RNA also appears to be subject to enzymic degradation.     

Uterine epithelial cells in primary culture: a model system to study cell proliferation

Guinea-pig uterine glandular epithelial cells were grown in primary culture. During the 4-day initial culture period, a 6.7 fold increase in DNA synthesis and a doubling time of approximately 30 hours were observed. Then the cells were submitted to serum depletion (60 hours) and the quiescent cells were stimulated with 15% fetal calf serum (FCS). The control cells were submitted to 1% heated and dextran-coated charcoal stripped FCS. In stimulated cells, the DNA synthesis increased and peaked between the 12th and 24th hour. In these cells, c-fos mRNAs increased rapidly, within 30 min., peaked at 75 min. (ratio to the control = 2.5), and returned to basal level within 90 min. These results prove that uterine epithelial cells in primary culture are able to respond to unspecific mitogen by both rapid expression of c-fos gene and DNA synthesis, suggesting that this cell culture system will be useful in studying the growth regulation in endometrium.     

Formation of flagella during interphase in secondary spermatocytes from Xenopus laevis in vitro

In cell culture, single motile flagella, 1 micron in length, were observed to grow from secondary spermatocytes of Xenopus laevis within 2-3 hours after telophase I, at 22 degrees C. About 90% of the secondary spermatocytes formed flagella as observed by phase-contrast microscopy. The flagella grew up to 2-6 microns in length during interphase II, which lasted about 18 hours. The presence of the "9 + 2" microtubular structure of the flagellar axonemes of secondary spermatocytes was confirmed by electron microscopy. When chromosomal condensation began (prophase II), the flagella were resorbed into the cells and, after the second meiotic division, a flagellum was formed again by each of the round spermatids. Thus, there appears to be a close relationship between the meiotic division cycle and the formation of flagella. The possible contribution of Sertoli cells to the formation of flagella in secondary spermatocytes was examined by reducing the number of Sertoli cells to less than ten per culture. Under these conditions, flagella formed in secondary spermatocytes with very high efficiency. It is very likely that secondary spermatocytes form flagella in vivo, since the secondary spermatocytes were observed to have flagella immediately after dissociation of the testes.     

Effect of concanavalin a on ganglioside metabolism of human lymphocytes

The effect of Con-A on the incorporation of radioactivity from [¹⁴C]-glucosamine into gangliosides of human lymphocytes was investigated. Compared with non-stimulated lymphocytes there was increased incorporation into gangliosides and total lipids within the first 24 hours of exposure to Con-A. Ganglioside synthesis also occurred in later time intervals within the 96 hour incubation period. GM3 accounted for 80% of the labeled ganglioside in Con-A stimulated cells at all times studied. Thus ganglioside synthesis is not only associated with cellular division, but also occurs within a few hours of lymphocyte activation representing an extremely early prereplicative event.     

Changes in mitotic indices in roots of Vicia faba

Roots of Vicia faba were treated with solutions of colchicine or IAA or both. Mitotic indices and the frequencies of the different stages of mitosis were determined immediately after a three hour treatment or following a 24 hour period of recovery. Roots scored after treatment with colchicine for three hours showed several effects, none of which were reversed by simultaneous treatment with IAA. Treatment with IAA for three hours had little detectable effect on mitotic index (MI) on the frequencies of the various stages of mitosis. After a recovery period, following a three hour treatment, of 24 hours, colchicine treated roots showed a significant increase in their MI; this was due largely to an increase in the number of metaphases but it was also due in part to the presence of tetraploid cells in division. IAA treated roots revealed an inhibition of mitotic activity, which was most marked at 3.13–6.26×10^–4 M IAA. The results from roots treated with mixtures of colchicine and IAA for three hours and fixed 24 hours later showed: 1) the increase in MI induced by colchicine is reversed by IAA, the intensity of the reversal increasing with increasing concentrations of IAA; 2) reductions in the total numbers of cells in prophase or in metaphase occur after treatment with different concentrations of IAA; 3) IAA leads to a reduction in the number of tetraploid cells seen in division.It appears that colchicine induces a change in the pattern of mitotic activity 24 hours after the end of treatment and its effects are reversed by IAA. At 4.2×10^–4 M IAA a balance occurs between the opposing effects of colchicine and IAA and the MI is not significantly different from that of the controls. It is suggested that one result of a treatment with colchicine is a change in the level of growth factors in root meristems. This change, which appears to result in a temporary increase in MI is reversed by the addition of IAA. Thus one of the growth factors, the level of which has been affected, is replaceable by exogenous IAA.     

The effect of chloramphenicol and cycloheximide on meiotic zoosporogenesis and mitotic gametogenesis in Ulva mutabilis.

Ulva mutabilis cells induced to form zoospores or gametes were exposed to chloramphenicol or cycloheximide during the 48 hours preparatory period. Chloramphenicol inhibited protein and RNA accumulation to the same degree at all concentrations, and appeared to have a general toxic effect. Cycloheximide at low concentrations inhibited primarily protein accumulation, while at higher concentrations it had also an effect on the net synthesis of RNA. Most of the inhibited cells were found in interphase, but a small percent of vegetative mitoses were observed and may indicate a return to this type of division. The gametophyte appeared to be more sensitive to chloramphenicol than the sporophyte. The effect of cycloheximide on the two processes seems to be more or less the same. A transition period, after which the sporulation process could no longer be inhibited, was found about 10 hours before the cells entered meiotic prophase in zoosporogenesis.