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PubMed1996

[The dynamic intracellular distribution of nucleolar protein B23 in the interphase and mitotic cells of mammalian cultures].

Using monoclonal antibodies against B23 (Ochs et al., 1983) and indirect immunofluorescence of cells exposed to different conditions of fixation, distribution of B23 in HeLa (human) PtK1 (rat kangaroo) and PK (pig) cells was studied at interphase and mitosis. In different cells B23 is distributed in a similar manner. At interphase it is located mainly within nucleoli. When cells enter mitosis (at prophase), B23 translocates from nucleoli to the nucleoplasm, and after breakdown of the nuclear envelope at prometaphase-to the cytoplasm to remain there till the end of telophase. As extraction buffers are applied to remove soluble cytoplasmic proteins. B23 is clearly seen on the surface of chromosomes. This unequivocally indicates that B23 is a "chromosomal passenger protein" (Earnshaw, Barnet, 1991). At telophase, the formation of numerous prenucleolar bodies on the surface of chromosomes starts. A long-term incubation of cells with actinomycin D prevents from the appearance of B23 around chromosomes and within prenucleolar bodies.

Animals↗
PubMed1996

Interphase-like chromatin configuration induced by cycloheximide in maturing pig oocytes: effects of protein phosphatase inhibitors.

Embryo cloning methods could greatly benefit from the manipulation of cell cycle in oocytes from large domestic mammals. The present study was undertaken to examine the effects of the protein synthesis inhibitor cycloheximide and the inhibitors of protein phosphatases 1 and 2A okadaic acid and calyculin A on maturing pig oocytes. Cycloheximide treatment (10 micrograms/ml) induced an interphase-like chromatin configuration (ICC) in maturing oocytes. Up to 69% of the oocytes exhibited ICC when treated with cycloheximide after 24 h of in vitro culture. ICC starts to appear after a 4 h exposure to cycloheximide and the ICC percentage reached its plateau after 12 h of cycloheximide treatment. ICC is fully reversible. The addition of okadaic acid (0.5 microM) inhibited the ICC in cycloheximide-treated maturing oocytes and allowed the completion of maturation in 55% of them. In oocytes with ICC, the immunocytochemistry for tubulin revealed the rearrangement of microtubule into an interphase meshwork and these oocytes lost their ability to induce tubulin assembly, as shown after short-time taxol treatment. The addition of okadaic acid prevented this microtubule rearrangement and preserved a certain level of tubulin assembly. Calyculin appeared to be more effective than okadaic acid in the prevention of ICC. It is concluded that de novo protein synthesis is necessary during a certain period of meiotic maturation for the maintenance of metaphase chromatin configuration in pig oocytes. This protein (or proteins) acts through the inhibition of endogenous protein phosphatases, probably protein phosphatase of 2A type.

Animals↗
PubMed1997

Interphase cytogenetic analysis of mucinous ovarian neoplasms.

Extending our previous efforts to characterize ovarian neoplasms by interphase cytogenetics, we analyzed a series of 32 mucinous tumors by nonisotopic in situ hybridization with seven different centromere-specific probes as well as by flow and image DNA cytometry; we then compared the data with results of p53 and Ki67 immunohistochemistry and MYC DNA-PCR analysis and of the clinical follow-ups. Of the tumors studied, 11 of 14 (78.6%) mucinous carcinomas, 7 of 7 (100%) mucinous tumors of low malignant potential (LMP), and 7 of 11 (63.6%) mucinous cystadenomas demonstrated chromosomal aberrations. The mean number of chromosomal aberrations (+/- SD) was slightly higher in DNA cytometrically nondiploid cases than in diploid cases (2.0 +/- 1.6 versus 1.6 +/- 1.2, not significant) but did not differ significantly among the study groups (carcinomas: 1.7 +/- 1.4; tumors of LMP; 1.9 +/- 0.7; adenomas: 1.4 +/- 1.4). Aberrations affected chromosomes 1 (14 of 27 cases) and 6 (12 of 31) most frequently, followed by chromosomes 17 (7 of 28), 7 (6 of 29), and X (6 of 28). Signal gain for centromere 1, which was the most prevalent finding (13 of 27), was observed in 3 of 10 mucinous cystadenomas, 2 of 4 mucinous tumors of LMP, and 8 of 13 mucinous carcinomas. All six moderately and poorly differentiated carcinomas demonstrated this aberration. Signal gain of centromere 6 (3 of 13) and centromere 7 (4 of 13) were found only in carcinomas (p < 0.05 and p < 0.025, respectively). The interphase cytogenetic results correlated neither with proliferative activity, immunohistochemical p53 accumulation, MYC DNA amplification, nor postoperative outcome. Compared with serous ovarian neoplasms (Lab Invest 1996, 75:473-485), mucinous tumors demonstrated signal gain for chromosome 1 (p < 0.0001) and signal loss for chromosomes 6 (p < 0.001) and X (p < 0.01) significantly more often. Loss of centromere 17 was more characteristic for serous than for mucinous carcinomas (p < 0.05). Our observations show that chromosomal aberrations in mucinous ovarian neoplasms are apparently not random. These results support the notion that the molecular genetic changes in mucinous neoplasms differ from those in serous tumors.

Adenocarcinoma, Mucinous↗
PubMed1997

[Chromatin structure and chromosomes on preparations of interphase nuclear derivatives after removal of nuclear envelopes. I. Method of preparation and morphology of residual nuclei in human leukocytes and intestinal epitheliocytes of Chironomus].

A method of a stage by stage dismantling of the cell nucleus by a 0.5-5 min resuspending cells in physiological media containing 0.05 and 0.5% of non-ionic detergent Triton X-100 is first described. Depending on the detergent concentration and cell incubation time a suspension was obtained consisting of either practically undamaged nuclei with removed envelopes, cytoplasm-free mitotic figures and isolated envelopes gradually spreading interphase chromatin associated with nucleoplasm proteins. The article describes a technique to make from such a suspension preparations of spread and flattened derivatives of nucleus and mitotic chromosomes to use for optical microscopic research. This method allows to reveal perichromatin components of the nucleus and mitotic chromosome to study structure of chromonemic net and nucleoli of diploid nuclei of human leucocytes and non-polytenic nuclei of chironomus epitheliocytes and the discoidal structure of polynemic (low-polytenic) chromosomes from the midgut epithelium cells of chironomus larvae. An attempt was made to use the obtained preparations of flattened unenveloped nuclei in order to distinguish between human leucocytes according to the structure of their chromonemic net and silver-stained nucleolus, and to find out the manner of impregnation of argyrophilic nucleoplasm proteins. The partially isolated chironomus polynemic chromosomes have morphological manifestations of unproportional endoreduplication, local compactization of chromomers and physical association between a nucleolus and numerous of distant chromosome loci. We failed, however, to find any homology in the band pattern of such polynemic chromosomes and polytenic chromosomes of larval salivary gland cells. Individuality of chromosomes in polynemic nuclei was not so obvious due to a developed network of intra- and interchromosomal ectopic contacts. The discussion touches upon the mechanisms of isolation of nuclear envelopes and ways to restore the original shape of the unenveloped nucleus in the used hydrodynamic system with fading detergent activity. Prospects are mapped out to use this technique in the structural studies of interphase nuclei and mitotic and polynemic chromosomes.

Animals↗
PubMed1997

Interphase cytogenetics of prostatic tumor progression: specific chromosomal abnormalities are involved in metastasis to the bone.

Only limited data are available on chromosomes specifically involved in the multistep tumorigenesis of prostate cancer. To investigate the cytogenetic status at different stages of prostatic tumor development, we have applied interphase in situ hybridization (ISH) with a set of (peri) centromeric DNA probes--specific for chromosomes 1, 7, 8, and Y--to routinely processed tissue sections of prostatic specimens from 75 different individuals. Our panel consisted of: 16 normal/benign prostatic hyperplasia specimens; 23 primary, localized, prostatic tumors (N0M0 stage); 20 regional lymph node metastases (M0 stage); and 16 distant metastases. Numerical aberrations of at least one chromosome were not observed in normal/benign prostatic hyperplasia cases, but were present in localized tumors (39%), regional lymph node metastases (40%), and distant metastases (69%). Within the different pTNM groups, we observed the following aberrations (listed, within each series, in decreasing order of frequency): -Y, +8, -8, +7 in primary tumors; +8, +7, -Y, +Y, -8 in regional lymph node metastases; and +8, +7, +1, -Y, -8 in distant metastases. In primary tumors, the number of aberrant cases increased significantly with local tumor stage (p < 0.05). A significant increase in gain of chromosome 8 was also observed (p < 0.02). Gain of chromosome 7 and/or 8 showed a significant increase with progression of local tumor stage (p < 0.02). Specific involvement of chromosome 8 was seen in bone metastases, but not in hematogenous metastases to other sites (p = 0.02). Comparative genomic hybridization analysis of these bone metastases disclosed centromere 8 gains as amplifications of the (whole) 8q arm, whereas centromeric loss appeared to be due to loss of 8p sequences. With progression toward metastatic disease, an accumulation of genetic changes was seen as exemplified by gain of chromosome 1, which was solely observed in distant metastases. With tumor progression, gain of chromosomes 7 and/or 8 significantly increased (p = 0.03), whereas the number of cases with aberrations of the Y chromosome did not change. Furthermore, ploidy status determined by ISH revealed a significant increase in the number of aneuploid cases along with advancement of pTNM stage (p = 0.04). Collectively, the data strongly suggest that: (a) gain of chromosome 7 and/or 8 sequences is implicated in prostatic tumor progression; (b) gain of chromosome 8 sequences is related to local tumor growth; (c) overrepresentation of 8q sequences, most likely by isochromosome 8q formation, is involved in metastatic spread to the bone; and (d) changes in the centromeric copy number, as detected by interphase ISH, might in some cases represent structural alterations, such as an isochromosome.

Aged↗
PubMed1997

Imbalance of morphologically addressed telophases reflects interphase DNA aneuploidy in tumorigenesis.

Chromosome division figures (CDFs) are quantitatively different from normal mitoses and represent a novel cytogenetic phenomenon. This investigation was focused on morphologically addressed bipolar telophases in histologically defined human biopsies and in the tumour breast cell-line MDA231. Single cell nuclei were recorded by image microphotometry on inflamed and premalignant lesions of skin (49 cases), oral mucosa (43) and colon mucosa (46). DNA content and replication status were analysed in interphase nuclei as well as in mitoses and in CDFs. In contrast to inflamed lesions, premalignancies were characterised by pronounced endoreplication, when the rate exceeding 5 c was > or = 10% in interphase nuclei. CDFs from the corresponding lesions showed an aberrant DNA content beyond 5 c even more frequently. DNA profiles of metaphases and telophases resembled those of prophases. Therefore, the DNA content of corresponding telophase hemispheres was measured. Severe differences averaged 0.3 c in MDA231 and up to 0.5 c in premalignant lesions. The mean difference between two corresponding hemispheres was 0.39 +/- 0.09 c in Bowenoid keratosis (n = 31), 0.40 +/- 0.08 c in high-grade dysplasia of oral mucosa (n = 16) and 0.21 +/- 0.03 c in high-grade dysplasia of colon adenoma (n = 65 telophases). As a control, the telophase difference was only 0.07 +/- 0.02 c (n = 23) in foetal liver and 0.06 +/- 0.01 c in 24 amnion cells. Thus, genomic instability and, in consequence, genomic imbalance can best be quantified from the DNA profiles of telophase CDFs and from the various DNA amounts in their hemispheres. A strong selection against telophases was observed in neoplasias developing DNA aneuploidy. Those aberrant telophases which escape selection are thought to enhance tumour progression.

Aneuploidy↗
PubMed1998

Detection of deletions in the short arm of chromosome 3 in uncultured renal cell carcinomas by interphase cytogenetics.

PURPOSE: Analysis of genetic alterations may facilitate the differential diagnosis of renal cell carcinoma (RCC) subtypes. For genetic classification, deletion of the short arm of chromosome 3 (3p), the hallmark of nonpapillary/clear cell RCC, is a major diagnostic criterion. Because of the limited routine applicability of cytogenetics and molecular genetic techniques we investigated interphase fluorescence in situ hybridization (FISH) for the detection of this aberration in RCC. MATERIALS AND METHODS: Using seven chromosome 3 specific probes FISH was performed on isolated nuclei from 26 uncultured sporadic RCC. RESULTS: Alterations of chromosome 3 were identified in 19 RCC (73%). Monosomy and/or 3p-deletions were observed in 15 of 19 (79%) non-papillary/clear cell RCC but not in other morphologic subgroups. The median percentage of cells in a specimen containing loss of 3p was 45%. Deletion mapping indicated that large deletions affecting different regions in 3p are predominant. Chromosomal region 3p24 was recurrently involved in all RCC with a deletion in 3p. CONCLUSION: Interphase FISH for the detection of loss in 3p provides a sensitive and feasible method for the genetic classification of kidney tumors and the delineation of recurrently deleted regions in 3p.

Adult↗
PubMed1996

Cell cycle-dependent regulation of cellular ATP concentration, and depolymerization of the interphase microtubular network induced by elevated cellular ATP concentration in whole fibroblasts.

In the present work, evidence is presented indicating that an increased cellular ATP concentration during mitosis may, in conjunction with other factors [Verde et al., 1990: Nature 343:233-238; Andersen et al., 1994: J Cell Biol. 127:1289-1299], induce depolymerization of the interphase microtubular network in cultured fibroblasts. It is shown here that the cellular ATP concentration varies through the cell cycle, reaching a peak at G2M- and minimum at late G1/early S-phase. Furthermore, we have found, using indirect immunofluorescent staining with an antitubulin antibody, that depolymerization of the interphase microtubular network may be induced by increasing the intracellular ATP concentration in cultured fibroblasts from 2.2 mM to 4.1 mM. This may be obtained through addition of adenosine and P1 to the growth medium. Our results indicate that this effect of adenosine and Pi is not mediated via adenosine receptors, but through an elevated cellular ATP concentration. ATP is suggested to act through a concentration-dependent effect on the exchangeable GTP site on tubulin, and not through the action of protein kinases or microtubule-associated proteins.

3T3 Cells↗
PubMed2000

Non-random trisomies of chromosomes 5, 8 and 12 in the prolactinoma sub-type of pituitary adenomas: conventional cytogenetics and interphase FISH study.

Specimens from 53 pituitary adenomas (PAs), including 17 NFPA, 16 PRL-, 9 ACTH-, 9 GH- and 2 TSH-secreting tumors, underwent cytogenetic analysis by the direct and short-term culture methods. Only 8 tumors (15%) appeared to have an abnormal karyotype. To increase the resolution of cytogenetic analysis, direct preparations from 31 PAs were investigated by interphase FISH with probes specific for chromosomes 5, 8, 12 and X, for which gain in pituitary tumors has been reported. Of these 31 PAs, 17 (54.8%) had an abnormal dosage of one or more of the 4 chromosomes tested. Separate or combined trisomies of chromosomes 5, 8 and 12 were found in 10/10 prolactinomas and in 4/9 NFPA, whereas the combined loss of chromosomes 5 and 8 was observed in 1/6 ACTH- and 1/6 GH-secreting PAs. Present and earlier data on 23 PAs showed that tumors with the highest frequency of abnormal karyotypes revealed by cytogenetics and/or interphase FISH were PRL (78%), followed by NFPA (26%) and GH (18%). Recurrent structural rearrangements affecting chromosomes 1, 3 and 12 were also identified in prolactinomas, which therefore appear to be the only pituitary adenoma sub-type with a defined trend of tumor-specific chromosomal changes. Cytogenetic and FISH analyses of different pituitary tumor sub-types indicate that they may harbour genetically distinct lesions.

Adult↗
PubMed1998

Detection of t(11;14) using interphase molecular cytogenetics in mantle cell lymphoma and atypical chronic lymphocytic leukemia.

The chromosomal translocation t(11;14)(q13;q32) fuses the IGH and CCND1 genes and leads to cyclin D1 overexpression. This genetic abnormality is the hallmark of mantle cell lymphoma (MCL), but is also found in some cases of atypical chronic lymphocytic leukemia (CLL), characterized by a poor outcome. For an unequivocal assessment of this specific chromosomal rearrangement on interphase cells, we developed a set of probes for fluorescence in situ hybridization (FISH). Northern blotting was performed for analysis of the cyclin D1 expression in 18 patients. Thirty-eight patients, with either a typical MCL leukemic phase (17 patients) or atypical CLL with an MCL-type immunophenotype, i.e., CD19-, CD5+, CD23-/low, CD79b/sIgM(D)++, and FMC7+ (21 patients), were analyzed by dual-color interphase FISH. We selected an IGH-specific BAC probe (covering the JH and first constant regions) and a commercially available CCND1 probe. An IGH-CCND1 fusion was detected in 28 of the 38 patients (17 typical MCL and 11 cases with CLL). Cyclin D1 was not overexpressed in two patients with typical MCL and an IGH-CCND1 fusion. In view of the poor prognosis associated with MCL and t(11;14)-positive CLL, we conclude that this set of probes is a valuable and reliable tool for a rapid diagnosis of these entities.

Adult↗
PubMed2004

Rapid detection of the t(11;14) translocation in mantle cell lymphoma by interphase fluorescence in situ hybridization on archival cytopathologic material.

BACKGROUND: The cytomorphologic diagnosis of mantle cell lymphoma (MCL) can be difficult and requires ancillary studies for accurate subclassification. More than 95% of MCLs are known to carry the t(11;14) chromosomal translocation. However, traditional cytogenetic studies on cytologic material can be both difficult technically and time consuming. Interphase fluorescence in situ hybridization (FISH) can be a powerful tool for detecting chromosomal changes in individual tumor cells. The authors evaluated the utility of interphase FISH for the rapid detection of t(11;14) in archival cytologic material. METHODS: The cytopathology data bases at two institutions were searched for patients with well characterized MCL (biopsy, immunophenotyping). Ten patients with MCL (8 fine-needle aspiration samples and 2 body cavity fluid samples) were identified. The area of interest on the cytology slides was marked and hybridized with two-color, locus-specific identifier DNA probes. A dual-fusion probe signal was used to detect the juxtaposition of the immunoglobulin heavy-chain (IgH) (14q32) locus with cyclin D1 (CCND1) gene sequences (11q13). Samples with tumor cell nuclei that showed at least one yellow fusion signal in addition one green signal (IgH) and one orange signal (CCND1) were interpreted as positive. Positive and negative controls were used. RESULTS: The t(11;14) translocation was detected by FISH in 10 of 10 patients (100%) with MCL. CONCLUSIONS: The cytomorphology of small-to-intermediate cell lymphomas, including MCL, follicular lymphoma, and marginal zone/mucosa-associated lymphoid tissue lymphoma, can show overlapping cytomorphologic features with one another as well as with reactive lymphoid proliferations. In selected samples in which specific classification is not possible or when confirmation is required on a small sample size, molecular analysis and cytogenetics may be helpful in arriving at an unambiguous cytodiagnosis and subclassification. Distinction of MCL from other lymphomas is important, because the clinical course is aggressive, and response to conventional chemotherapy is poor. This study showed that the detection of t(11;14) by FISH can be performed rapidly and easily on archival cytologic material for the molecular diagnosis of MCL.

Adult↗
PubMed1995

Interphase cytogenetic analysis of erbB2 and topoII alpha co-amplification in invasive breast cancer and polysomy of chromosome 17 in ductal carcinoma in situ.

Breast cancer is a genetically complex disease. Fluorescence in situ hybridisation can be used to analyse the genetics of breast-cancer progression in interphase cytogenetics. We have analysed the histological distribution of erbB2 and topoll alpha co-amplification in paraffin sections of invasive breast cancer and show that the co-amplified loci share the same histological distribution in the tumour and have a similar nuclear distribution within individual nuclei. Regions of the tumours without amplification are easily recognized and tumours with erbB2 and topoll alpha co-amplification can be distinguished from those with erbB2 amplification alone. In addition, FISH was used to show polysomy of chromosome 17 in non-invasive ductal carcinoma in situ of the breast and erbB2 amplification in both the invasive and non-invasive components of a breast cancer biopsy. This report of an interphase cytogenetic analysis of non-invasive breast carcinoma in situ demonstrates the usefulness of FISH for the genetic study of breast cancer progression.

Aneuploidy↗
PubMed2004

Local properties of a functionally graded interphase between cementum and dentin.

The study of natural interfaces may provide information necessary to engineer functionally graded biomaterials for bioengineering applications. In this study, the mechanical, structural, and chemical composition variations associated with a region between cementum and dentin were studied with the use of nanoindentation, microindentation, optical microscopy, and Raman microspectroscopy techniques. Three-millimeter-thick transverse sections (N = 5) were obtained from the apical one-third of the roots of sterilized human molars. The samples were ultrasectioned at room temperature with the use of a diamond knife and an ultramicrotome. Longitudinal ground sections of 100 microm thickness were prepared and stained with von Kossa stain to determine the mineralized regions within the molar roots. Raman microspectroscopy was used to determine the relative inorganic content, mainly apatite (PO4(3-)nu1 mode at 960 cm(-1)) and organic content, mainly collagen (C--H stretch at 2940 cm(-1)) between cementum and dentin bulk tissues. The microindentation and nanoindentation results indicated a gradual transition in hardness from cementum to dentin over a width ranging from 100 to 200 microm. However, the variation in hardness data for cementum and dentin by nanoindentation was larger (0.62 +/- 0.21, 0.77 +/- 0.14 GPa) than from microindentation (0.49 +/- 0.03, 0.69 +/- 0.07 GPa). Within the 100 to 200 microm region there was a 10 to 50 microm fibrillar hydrophilic cementum-dentin junction (CDJ) with mechanical properties significantly lower than either the cementum or the dentin side of CDJ. Light microscopy revealed a 100 to 200 microm translucent region between cementum and dentin. Raman microspectroscopy results showed a variation in organic and inorganic composition 80 to 140 microm wide. It was concluded that a morphologically and biomechanically different CDJ lies within a wider cementum-dentin interphase. Hence, cementum, dentin, and the interphase can be classified as a functionally graded dental tissue within the root of a tooth.

Biomechanical Phenomena↗
PubMed1975

Effect of polysorbate 60 on interphase transport of cholesterol.

Interphase cholesterol transport was investigated at 24 +/- 1 degrees in a stirred diffusion cell and in various oil-in-water emulsions. Cholesterol uptake by vegetable oil from a cholesterol-surfactant-rich aqueous phase was extremely slow in the stirred cell; no measurable transport had occurred after 500 hr. Cholesterol transport in oil-in-water emulsions following dilution with a cholesterol-surfactant-rich aqueous phase was much faster due to the greatly increased interfacial area available for mass transfer. Equilibration half-lives, t(50), varied from 2.02 to 28.1 hr. Variations in the t(50) were due to: (a) differences in the mean oil droplet diameter among various emulsions, and (b) differences in cholesterol-polysorbate 60 micelle sizes among various dilution media. When polysorbate 60 was omitted from the dilution medium, transport occurred in a two-stage process. In the first stage, transport was extremely rapid, with the t(50) less than 30 sec; in the second stage, transport was comparable to previous emulsion rates, with the t(50) varying from 7.9 to 8.1 hr. The significance of this two-stage transport to mechanisms of interphase cholesterol transport is briefly discussed.

Cholesterol↗
PubMed1978

Kinetic and thermodynamic aspects of in vitro interphase transfer of tetracyclines I: influence of hydroxyl group substitution.

The influence of hydroxyl group substitution on the kinetic and thermodynamic aspects of the interphase transfer of three tetracycline derivatives was studied in a two-phase in vitro system composed of an aqueous pH 5.5 buffer and 1-octanol. Data are reported on the kinetic and thermodynamic parameters of activation, the net thermodynamic parameters for interphase transfer, and the contribution of hydroxyl group substituents to the energy changes associated with partitioning. For each derivative, deltaH, deltaS, T deltaS, and deltaF were positive with enthalpy dominating the entropic energy contribution. Introduction of a hydroxyl group at C-5 on the tetracycline nucleus promoted partitioning through an entropy-dominated decrease in the "apparent" free energy of partitioning, whereas shifting the hydroxyl group to C-6beta retarded partitioning due to an enthalpy-dominated gain in the apparent free energy.

Diffusion↗
PubMed2001

Identification of a novel plant-specific kinesin-like protein that is highly expressed in interphase tobacco BY-2 cells.

Through reverse transcription-polymerase chain reaction and Northern blot analysis, we identified TBK5, a novel plant-specific kinesin-like protein (KLP) that is highly expressed in interphase tobacco BY-2 cells. TBK5 mRNA was present at a high level throughout the growth cycle, even in cells that had entered the stationary phase, where cell proliferation had ceased. However, transcripts for five other tobacco KLPs that we have identified were preferentially expressed in mitotic cells, and either not or only slightly accumulated in cells that had entered the stationary phase. Thus, TBK5 appears to be a KLP whose cellular function most closely relates to the cortical array of microtubules that plays a key role in plant cell morphogenesis. The predicted structure of TBK5 is characterized by a central motor domain that is phylogenetically distant from those of other reported KLPs, coiled-coil domains located on both sides of the motor domain, and a basic C-terminal domain. In addition, TBK5 has a putative neck domain which is closely related to the neck domain of KLPs with C-terminal motor domains, previously shown to control the direction of KLP movement towards the minus ends. Antibodies against truncated TBK5 recognized a polypeptide with a molecular mass of 74 kDa in cytoplasmic extracts of interphase cells, and this polypeptide cosedimented with microtubules assembled in the cytoplasmic extracts. The 74 kDa polypeptide corresponding to TBK5 dissociated from microtubules with high concentrations of NaCl but was not dissociated by MgATP. We hypothesize that TBK5 functions in the regulation of the arrangement of cortical microtubules.

Algal Proteins↗
PubMed1977

Orientation of Giemsa C-bands in interphase cells of Allium cepa L.

Orientation of Giemsa C-bands in Allium cepa was studied in both mitotic and interphase cells. It has been shown that telophase orientation of the chromosome is maintained throughout the interphase and early phophase. It has been assumed that this non-random orientation is due to anchorage of the telomeres with the nuclear membrane. Contrary to earlier observations, 2 by 2 pairing of the telomers could not be traced to this species.

Chromosomes↗
PubMed1996

Rapid fluorescence in situ hybridization on interphasic nuclei to discriminate between homozygous and heterozygous transgenic mice.

Homozygous and heterozygous transgenic mice of the Tg152 line overexpressing the human copper/zinc superoxide dismutase (hSOD-1) were rapidly differentiated by fluorescence in situ hybridization (FISH) using interphase lymphocyte nuclei. We have devised a simple and fast method for preparing interphase nuclei with very small quantities of whole mouse blood, avoiding several steps of the classical FISH technique. Lymphocyte separation and cell culture were not required. This technique provides an excellent tool for the unambiguous detection of homozygous and heterozygous transgenic mice in a litter. It can be used to check young animals since 2 microliters of whole blood is sufficient. We also show that in this transgenic line numerous copies of the hSOD-1 transgene are integrated at a single autosomal locus, in tandem head-to-tail organization.

Animals↗