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Plant regeneration from protoplasts of hydroxyprolineresistant cell line in Onobrychis viciaefolia

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Cell Research 1995年第2期5卷 187-194页

作者： XU ZIQIN JINGFEN JIA(Cell Blology Laboratory, Lanzhou University, Lanzhou 730000, China.) Cell Biology Laboratory Lanzhou University Lanzhou 730000 China.Received 30August1994 revised 27April1995 Accepted 25July1995

An efficient protocol for plant regeneration from protoplasts of hydroxyproline(HYP)resistant cell line of Onoblychis viciaefolia was established. In SH medium supplemented with 1 mg/L 2, 4-dichlorophenoxy-acetic acid (2,4-D), 0.5 mg/L kinetin (KT) and 0.2 mg/L naphthalene acetic acid (NAA), the division frequency of protoplastderived cells reached uP to over 60 %, and microcalli were obtained in 5-6 wk. Upon transferring them on agar solidified MS medium plus 2 mg/L indole-3-acetic acid (IAA), shoots were induced. After cultivating them on MS medium with or without IAA, roots were *** number of all protoplast-regenerated plants examined were normal (2n=28). The protoplast-derived calli and plants grew vigorously on the medium containing 10 mmol/L HYP.

关键词： Sainfoin (Onobrychis Viciaefolia) hydroxyproline resistant cell line protoplast culture plant regeneration

Substance P enhances the proliferation of rat anteriorpituitary cells in vitro

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Cell Research 1995年第2期5卷 197-207页

作者： ZHANG WANHUI YUNLONG ZHU FUZHOU WANG YUZHEN HU QI MEI CHAO ZHAO JIANGKANG CHEN(Laboratory of Neuroendocrinology, Department of Physiology, The 4th Military Medical University, Xi'an 710032,China.) Laboratory of Neuroendocrinology Department of Physiology The 4th Military Medical University Xi'an 710032 China.Received 24October1994 revised 8June1995 Accepted 20October1995

The undecapeptide substance P(SP) was shown to be intimately involved in both the structural and functional aspects of the anterior pituitary. Yet, in addition to its influences on hormonal secretion, SP may well possess more actions in this master gland. The present study was ftherefore aimed to investigate the effect of SP on the proliferation of rat anterior pituitary cells in primary culture. It was found that SP could dose-dependently increase the ineorporation of tritiated thymidine (3H-TdR)into cultured anterior pituitary cells. Other mammalian tachykinins such as neurokinin A and neurokinin B had similar effect but to varying degrees. The equipotent analogue of SP, Norleucine(11) -SP(Nle(11)-SP), also acted likewise, with its action antagonizable by spantide, a SP receptor blocker. To further characterize the nature of cells responsive to the challenge of SP, immunocytochemical staining against S-100 protein and some adenohypophyseal hormones was performed alone or plus autoradiography The results showed that the percentage of S-100 proteinimmunoreactive cells was apparently elevated by the addtion of Nle(11)-SP for 48 h, which indicates a preferential proliferation of folliculo-stellate cells under the regime. This was confirmed by increases in immunocytochemical or autoradiographical labelling indices of anterior pituitary cells treated similarly. Taken togethbr, these results reveal that the trophic action of SP observed previously in other tissues is also present at least in cultured rat anterior pituitary cells, with responding cells being predominantly folliculo-stellate cells as typified by S-100 proteinimmunoreactivity. Therefore, an intra-pituitary trophic action of SP in vivo could be anticipated.

关键词： Substance P anterior pituitary proliferation in vitro rat

A TEM study on pre-excystment cellular structures of Euplotes encysticus

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Cell Research 1995年第1期5卷 125-133页

作者： GU FUKANG AND JUNMEI XU (Department of Biology, East China Normal University, Shanghai 200062, China) Department of Biology East China Normal University Shanghai 200062 China&#x0002A Dedicated to professor Zhen YAO's 80th Birthday.† This work was supported by a grant from the National Natural Science Foundation.Received 14March1995 revised 15June1995 Accepted 20June1995

Right before the excystment of an Euplotes encysti-cus sawtooth-like folds appeared among the pellicle plas- malemma, the inner and outer alveolar membranes were still sticking together, and were not distinguishable. Mi- crotubular layers already formed at the sites beneath the dorsal cortical pellicle corresponding to vegetative cells,but they still proceed to be organized on the ventral struc-tures. Cristae, highly-tangled with tubular-type struc-tures, appeared on the mitochondria, and were morpho-logically similar to that of vegetative cells. In the cor-tical ciliatures, such as ciliary shafts, kinetosomes, sur-rounding fibrillar cirral baskets, and attached structures of ciliatures, etc., they are different from those in rest- ing cysts which are degenerated or lost. All the cilia- ture microtubules of ciliary shafts are of the 9+2 pattern,but the microtubule-like structure aggregates at triplet - microtubule centers of many kinetosmes, are still under various stages of differentiation. Microtubules beneath the kinetosomal rows are of a developmentally elongated stage; crowded chromatins of various shapes and sizes are found in macronucleus, but there are no nuclear pores (formed by nuclear membrane as in resting cysts) on the nuclear membrane where these chromatins attached.

关键词： Euplotes encysticus excystment pellicle kinetosome macronucleus

Characterization of Giardia cell nucleus:Its implication on the nature and origin of the primitive cell nucleus

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Cell Research 1995年第1期5卷 115-124页

作者： LI JINGYAN (Department of Evolutionary Cell Biology, Laboratory of Cellular and Molecular Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China.) Department of Evolutionary Cell Biology Laboratory of Cellular and Molecular Evolution Kunming Institute of Zoology Chinese Academy of Sciences Kunming 650223 China.Received 20October1994 revised 8June1995 Accepted 18June1995

Inhibition of invasiveness and expression of epidermal growth factor receptor in human colorectal carcinoma cells induced by retinoic acid

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Cell Research 1995年第1期5卷 135-142页

作者： SUN BAODONG JINDAN SONG (Key Laboratory of Cell Biology, Ministry of Public Health of China, China Medical University, Shenyang 110001,China.) Key Laboratory of Cell Biology Ministry of Public Health of China China Medical University Shenyang 110001 China.&#x0002A Dedicated to 80th anniversary of Professor Zhen YAO.† This work was supported by National Natural Science Foundation.Received 6July1995 revised 12July1995 Accepted 14July1995

Human amniotic basement membrane (HABM) model and agarose drop explant method were used to in-vestigate the effects of retinoic acid(RA) on the invasive-ness alld adhesiveness to the basement membrane, and the migration of a highly invassive human colorectal cancer cell line CCL229. Results showed that 5 ×106 MRA markedly reduced the in vitro invasiveness and adhesiveness to the HABM, and the migration of the CCL229 cells. In addi-tion, to elucidate the relation between expression of epider-mal growth factor receptor (EGFR) and the invasiveness of the colorectal carcinoma cells, two well-differentiated, but with different invasiveness colorectal cancer cell lines were compared at mRNA level for expressioll of EGFR by using EGFR cDNA probe labeled with digoxigenin (DIG). Expression of EGFR was showll to be markedly higher in the highly invassive CCL229 cells than that in the low in- vasive CX-1 cells. Furthermore, expression of EGFR in RA treated CCL229 cells gradually decreased with time,the level being the lowest on day 6 of the RA treatment.

关键词： Retinoic acid colorectal carcinoma cell invasiveness epidermal growth factor receptor inhibition

Apoptosis of human leukemic HL-60 cells induced to differentiate by treatment with RA or DMSO

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Cell Research 1995年第2期5卷 181-186页

作者： SUN HONG YONGCHAO WANG(Key Laborutory of Cell Proliferation and Regulation Biology, Beijing Normal University, Beijing 100875, China.) Key Laboratory of Cell Proliferation and Regulation Biology Beijing Normal University Beijing 100875 China.&#x0002A This work was supported by the National Medical and Scientific Key Item Grant of “ 8.5 Plan” .Received 9November1994 revised 13June1995 Accepted 17September1995

In present study we studied the effect of all-trans retinoic acid(ATRA) and dimethylsulfoxide (DMSO) on the induction of apoptosis in HL-60 cell line. Based on morphological changes by Hochest 33342 staining and identification of internucleosomal DNA cleavage by gel electrophoresis, we observed aberrant nuclear chromatin eondensation and ladder-like pattern of DNA degradation. Using Flow Cytometric method, we found sub-G1 peak in RA-treated HL-60 cells starting 5 to 6 d after the initiation of the treatment. However, such an obvious apoptotic peak was not identified in DMSO-differentiated cells. Combining the research accomplished before, our study approves further that apoptosis could be a common mode of death of terminally differentiated HL-60 cells.

关键词： Apoptosis HL-60 cell RA DMSO

Changes of macromolecular organizations in nonjunctional sarcolemmas after cross-innervation──a study offast-and slow-twitch muscle fibres in rats

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Cell Research 1995年第2期5卷 143-154页

作者： LU JIANLI TIEFENG ZHANG FUJIN QU BEIHUA QIAN SHIKAI HUANG(Shanghai Institute of Physiology, Chinesc Academy of Scicnces, 320 Yue-Yang Road, Shanghai 200031, China) Shanghai Institute of Physiology Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China.Received 11January1994 revised 12December1994 Accepted 16February1995

The purpose of the present study was to analyse the changes of macromolecular organizations in nonjunctional sarcolemmas of different types of skeletal muscle fibres after cross-innervation. In normal rats the mean density of square arrays (6 nm particles organized in orthogonal arrays) was 9.02/μm2 for the nonjunctional sarcolemmas of fast-twitch extensor digitorum longus (control EDL,CE) nuscle fibres and 0.34/μm2 for the nonjunctional sarcolemmas of slow-twitch soleus (control SOL, CS) muscle fibres. After cross-innervation between the fast-twitch EDL and slow-twitch SOL muscle fibres by slow and fast muscle merves respectively for three months, the mean density was 0.45/μm2 for the nonjunetional sarcolemmas of the operated-EDL (OE) and 8.3/μm2 for the nonjunctional sarcolemmas of the operated SOL (OS). This indicates that the cross-innervation causes a reciprocal transformation of the number and distribution of such macromolecular organizations in the electrically excitable nonjunctional sarcolemmas.

关键词： Sarcolemma square array cross-innervation freeze-fracturing rat(Wistar)

Laser scanning fluorescence microscopic measurement of the movement of cleaving egg surface of Rana Amurensis

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Cell Research 1995年第1期5卷 9-24页

作者： GU GUOYAN (FORMERLY KU KUOYEN) CHENGTANG XU KONGHUA ZHANG QIRONG GAO.(Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China) Shanghai Institute of Cell Biology Chinese Academy of Sciences Shanghai 200031 China.&#x0002A Dedicated to Prefessor Zhen YAO on the occasion of his 80th birthday.† This work was supported by the director's foundation of Shanghai Institute of Cell BiologyReceived 27October1994 revised 6April1995 Accepted 25April1995

By laser scanning fluorescence microscopy for quan-titative measurement of fluorescence intensity changes on egg surface stained with fluorescein isothiocyanate duxing cleavage furrow extending forward, it was found that in area of presumptive cleavage furrow the scanning curve became ∨ shape, indicating dark stripe appeared in that place. Then the fluorescence intensity increased at the place where the botton of ∨ shape had located, and the scanning curve tuxned to ∧ shape, indicating single stripe was formed. While enhanced fluorescence appeared on the borders of ∧ shape, an M shape curve was found, show-ing double stripe occurred. During the distance between two borders of M shape incresing from 50 μm to 100μm,a fluorescence peak came to sight in the middle of the M shape, which being the cleavge furrow bottom. The two lateral sides of furrow bottom with decreasing fluorescence were nascent membrane. At that time the curve became W shape. By the sides of cleavage furrow the the stress folds became conspicous after double stripe stage, showing the stretching of the egg surface being increased. With our[31, 33] and others[32] reports that polylysine could induce the appearance of nascent membrane and phyto-hemagglutinins could decrease or prevent the appearance of nascent membrane, we believed the idea of Schroeder[25] that increasing mechanical stress could initiate nascent membrane formation and thought that the stress lay to the outsides of cleavage furrow.

关键词： cleavage egg surface movement initiating nascent membrane formation laser scanning fluorescence microscopy Rana Amurensis

Affinity chromatography-dependent selection (ACDS)of genomic DNA fragments bound specifically to bacterial synthesized Myc/Myn proteins

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Cell Research 1995年第1期5卷 25-34页

作者： SHI CAN PEI WANG YONGJUN HU LIAN XU. (Oncogene Group, Laboratory of Molecular and Cellular Oncology, Shanghai Institute of Cell Biology, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai 200031, China) Oncogene Group Laboratory of Molecular and Cellular Oncology Shanghai Institute of Cell BiologyChinese Academy of Sciences 320 Yue-Yang RoadShanghai 200031 China&#x00023 ]Both made equal contribution to the work.* The work is specifically dedicated to Prof. Zhen YAO for his 80-years birthday.† Supported by grant No. 39270742 of the National Natural Science Foundation.Received 20December1994 revised 21April1995 Accepted 26April1995

This paper describes an approach to seek for mouse c-Myc/Myn proteins-bound specific sequences among ge-nomic DNA. cDNA fragment of myn gene was obtained through RT-PCR technique from RNA of NIH3T3 cells. DNA fragments encoding BR/HLH/LZ structure of Myc and Myn proteins were cloned in frame into pGEX-2T vec-tor respectively Fusion GST-Myc and GST-Myn synthe-sized in *** hosts showed affinity to CACGTG E-boxDNA and subsequently interacted with genomic fragments prepared through whole-genome-PCR. A PCR-assisted procedure which combines protein-DNA interaction and affinity chromatography was designed to enrich Myc/Myn bound DNA. At least two genomic DNA fragments ob- tained exhibit specifical binding capacity to Myc/Myn complex but not to GST alone. Significance of the work and of the technique itself as well as identification of the DNAs are discussed.

关键词： Myc/Myn proteins ACDS genomic DNA binding