Biomedical subjects

Z J Chen

Publications and source records attributed to Z J Chen.

At least 37 records · Page 2Linked to original sources

PubMed2001

[Comparative effects of carvedilol and metoprolol in preventing from left ventricular remodeling after acute myocardial infarction in rats].

OBJECTIVE: To compare the effects of carvedilol and metoprolol in preventing from left ventricular remodeling (LVRM) after acute myocardial infarction (AMI) in rats. METHODS: Twenty-four hours after ligating left coronary artery, 105 surviving female SD rats were randomly assigned to AMI control, carvedilol 1 mg/(kg.d) and metoprolol 2 mg/(kg.d) groups. Sham-operated rats (n = 16) were selected randomly as non-infarction control. After four weeks of drugs therapy, hemodynamic studies and pathologic analysis were performed. Exclusive of MI size < 35% or > 55%, complete experimental variables were obtained in 46 rats, which were comprised of AMI (n = 11), carvedilol (n = 12), metoprolol (n = 11), and Sham-operated (n = 12) groups. RESULTS: Compared with sham-operated group, left ventricular (LV) end diastolic pressure (LVEDP), volume (LVV) and weight (LVW), were all significantly increased (P < 0.05-0.001), while maximal rate of rise and fall (+/- dp/dt) of LV pressure as well as their corrected values (+/- dp/dt/LVSP) were all significantly decreased (P < 0.01-0.001) in AMI group. In comparison with AMI group, the LVEDP and LVV were all significantly decreased (all P < 0.001), while +/- dp/dt and +/- dp/dt/LVSP were significantly increased (P < 0.05-0.001) in both carvedilol and metoprolol group, with LVW and RVW only decreased in carvedilol group (P < 0.05-0.01). CONCLUSIONS: 1. Carvedilol can effectively attenuate LVRM, and improve hemodynamics and LV function after AMI in rats, 2. Metoprolol has equivalent beneficial effects as carvedilol on hemodynamics, LV dilatation and function, but not LV hypertrophy.

Adrenergic beta-Antagonists↗

PubMed2000

Molecular cloning of a regulatory protein for membrane-bound guanylate cyclase GC-A.

Activation of membrane-bound guanylate cyclase GC-A by atrial natriuretic factor (ANF) may require the involvement of accessory proteins. To identify these postulated proteins, we isolated a 1. 0-kb cDNA clone from a rat brain expression library using a polyclonal antiserum against mastoparan. The 1.0-kb cDNA encodes a protein of 111 amino acids. Expression of this cDNA in COS-7 cells potentiated ANF-stimulated GC-A activity. Therefore, the 1.0-kb gene encodes a guanylate cyclase regulatory protein (GCRP). Fluorescence microscopy studies using the fusion protein of GCRP with green fluorescence protein (GFP) indicated that GCRP was present in the cytosol in PC12 cells, but translocated toward the plasma membrane in the presence of ANF. Coimmunoprecipitation experiments indicate that GCRP associates with GC-A in the presence of ANF. These results suggest that ANF induces the association of GCRP with GC-A and this association contributes to the activation of GC-A.

Amino Acid Sequence↗

PubMed2000

Activation of the IkappaB kinase complex by TRAF6 requires a dimeric ubiquitin-conjugating enzyme complex and a unique polyubiquitin chain.

TRAF6 is a signal transducer in the NF-kappaB pathway that activates IkappaB kinase (IKK) in response to proinflammatory cytokines. We have purified a heterodimeric protein complex that links TRAF6 to IKK activation. Peptide mass fingerprinting analysis reveals that this complex is composed of the ubiquitin conjugating enzyme Ubc13 and the Ubc-like protein Uev1A. We find that TRAF6, a RING domain protein, functions together with Ubc13/Uev1A to catalyze the synthesis of unique polyubiquitin chains linked through lysine-63 (K63) of ubiquitin. Blockade of this polyubiquitin chain synthesis, but not inhibition of the proteasome, prevents the activation of IKK by TRAF6. These results unveil a new regulatory function for ubiquitin, in which IKK is activated through the assembly of K63-linked polyubiquitin chains.

Amino Acid Sequence↗

PubMed2000

Cytochrome c deficiency causes embryonic lethality and attenuates stress-induced apoptosis.

Cytochrome c released from mitochondria has been proposed to be an essential component of an apoptotic pathway responsive to DNA damage and other forms of cell stress. Murine embryos devoid of cytochrome c die in utero by midgestation, but cell lines established from early cytochrome c null embryos are viable under conditions that compensate for defective oxidative phosphorylation. As compared to cell lines established from wild-type embryos, cells lacking cytochrome c show reduced caspase-3 activation and are resistant to the proapoptotic effects of UV irradiation, serum withdrawal, or staurosporine. In contrast, cells lacking cytochrome c demonstrate increased sensitivity to cell death signals triggered by TNFalpha. These results define the role of cytochrome c in different apoptotic signaling cascades.

Animals↗

PubMed2000

[Pharmacokinetics of topically applied in-situ-forming gels of fluconazole in rabbit eyes].

AIM: To study the pharmcokinetics of fluconazole (FCZ) in the aqueous humors and tears of New Zealand white rabbits following topically applied in-situ-forming gels of 0.5% FCZ (ISG-FCZ) and 0.5% FCZ eye drops to rabbit eyes. METHODS: The rabbit tears and aqueous humors were obtained and quantified at different times after topically applying single dose of ISG-FCZ and FCZ eye drops to the rabbit eyes. The drug levels were assayed by megabore capillary gas-liquid chromatography with nitrogen-selective detector. The pharmacokinetic parameters were calculated with nonlinear least square method with computer. RESULTS: FCZ concentrations in rabbit tears within 180 min after applying ISG-FCZ were found to be significantly higher compared with those of FCZ eye drops. The FCZ levels in the aqueous at 10, 30, 60, 90 and 120 min after applying ISG-FCZ are significantly higher compared with FCZ eye drops. The time for arriving peak concentration (Tmax) and the area under concentration (AUC)-time curve as well as the half-life (T1/2) of FCZ in rabbit aqueous humors were markedly higher after application of ISG-FCZ. The peak levels in the aqueous humors showed no difference between the two groups. CONCLUSION: The ISG-FCZ, which upon exposure to physiological conditions will shift the gel phase, can significantly extend FCZ release and increase the precorneal residence time of the drug, thus enhance ocular bioavailability compared with FCZ eye drops.

Acrylic Resins↗

PubMed2000

[Application of autotransfusion hemodilution in laryngectomy].

OBJECTIVE: To study the application and clinical significance of autotransfusion hemodilution in laryngectomy. METHOD: Autotransfusion hemodilution was adopted in 30 patients with III or IV stage laryngeal carcinoma during operation as the experimental group, and the blood transfusion only was adopted in the patients with excessively blood loss. The other 30 patients with the same stages of disease and the same kinds of operation were performed as the control group, without autotransfusion hemodilution and the blood transfusion was adopted if necessary. RESULT: Blood transfusion was necessary to be performed only in 8 patients of the experimental group, on the contrary, it was necessary in 19 of 30 patients of the control group (P < 0.01). There were no complications during the operations, and the postoperative complications were no significance (P > 0.05) between two groups. CONCLUSION: The application of autotransfusion hemodilution may be of help to a plenty of patients with laryngeal carcinoma in advanced stage to keep off the blood transfusion during laryngectomy.

Adult↗

PubMed1999

What is the significance of nasal involvement in nasopharyngeal carcinoma?

PURPOSE: The purpose of this study was to differentiate the patterns of nasal fossa involvement in nasopharyngeal carcinoma (NPC) and to clarify its prognostic influence on local control and survival after radiation therapy. METHODS AND MATERIALS: Between November 1989 and July 1991, 218 patients with histologically proven local-regional NPC were treated with radiotherapy following the protocol at the Department of Radiation Oncology, Cancer Hospital, Shantou University School of Medicine. All patients had pretreatment CT scans. Fiberoptic endoscopic examination was performed every week during treatment and at the time of every follow-up visit to define the initial extent of disease and to evaluate treatment response. No chemotherapy or brachytherapy was given. RESULTS: Of the 218 patients, 87 had nasal involvement. Sixty of them had a pattern of mucosal infiltration (MI), another 27 had an exophytic protruding (EP) component. The likelihood of residual disease after irradiation, the local relapse rate, 5-year freedom from progression rate (FFP), and death rate associated with local relapse (DRALR) of MI and EP were 36.7% vs. 3.7%, 30.0% vs. 7.4%, 26.7% vs. 51.8%, and 25.0% vs. 3.7% with p<0.004, p<0.005, p<0.02, and p<0.03, respectively. Multivariate analysis in this selected group demonstrated that infiltration of nasal fossa mucosa was an independent prognostic factor on primary control and freedom from progression. CONCLUSION: Differentiation of nasal fossa involvement according to MI or EP is of value in predicting the outcome of treatment. We suggest that only the MI group should be considered as nasal involvement in the staging of NPC.

Carcinoma↗

PubMed1999

Signal-induced ubiquitination of IkappaBalpha by the F-box protein Slimb/beta-TrCP.

Signal-induced phosphorylation of IkappaBalpha targets this inhibitor of NF-kappaB for ubiquitination and subsequent degradation, thus allowing NF-kappaB to enter the nucleus to turn on its target genes. We report here the identification of an IkappaB-ubiquitin (Ub) ligase complex containing the F-box/WD40-repeat protein, beta-TrCP, a vertebrate homolog of Drosophila Slimb. beta-TrCP binds to IkappaBalpha only when the latter is specifically phosphorylated by an IkappaB kinase complex. Moreover, immunopurified beta-TrCP ubiquitinates phosphorylated IkappaBalpha at specific lysines in the presence of Ub-activating (E1) and -conjugating (Ubch5) enzymes. A beta-TrCP mutant lacking the F-box inhibits the signal-induced degradation of IkappaBalpha and subsequent activation of NF-kappaB-dependent transcription. Furthermore, Drosophila embryos deficient in slimb fail to activate twist and snail, two genes known to be regulated by the NF-kappaB homolog, Dorsal. These biochemical and genetic data strongly suggest that Slimb/beta-TrCP is the specificity determinant for the signal-induced ubiquitination of IkappaBalpha.

Animals↗

PubMed1999

RNA polymerase I transcription in a Brassica interspecific hybrid and its progenitors: Tests of transcription factor involvement in nucleolar dominance.

In interspecific hybrids or allopolyploids, often one parental set of ribosomal RNA genes is transcribed and the other is silent, an epigenetic phenomenon known as nucleolar dominance. Silencing is enforced by cytosine methylation and histone deacetylation, but the initial discrimination mechanism is unknown. One hypothesis is that a species-specific transcription factor is inactivated, thereby silencing one set of rRNA genes. Another is that dominant rRNA genes have higher binding affinities for limiting transcription factors. A third suggests that selective methylation of underdominant rRNA genes blocks transcription factor binding. We tested these hypotheses using Brassica napus (canola), an allotetraploid derived from B. rapa and B. oleracea in which only B. rapa rRNA genes are transcribed. B. oleracea and B. rapa rRNA genes were active when transfected into protoplasts of the other species, which argues against the species-specific transcription factor model. B. oleracea and B. rapa rRNA genes also competed equally for the pol I transcription machinery in vitro and in vivo. Cytosine methylation had no effect on rRNA gene transcription in vitro, which suggests that transcription factor binding was unimpaired. These data are inconsistent with the prevailing models and point to discrimination mechanisms that are likely to act at a chromosomal level.

Binding, Competitive↗

PubMed1998

Gene dosage and stochastic effects determine the severity and direction of uniparental ribosomal RNA gene silencing (nucleolar dominance) in Arabidopsis allopolyploids.

Nucleolar dominance is an epigenetic phenomenon in which one parental set of ribosomal RNA (rRNA) genes is silenced in an interspecific hybrid. In natural Arabidopsis suecica, an allotetraploid (amphidiploid) hybrid of Arabidopsis thaliana and Cardaminopsis arenosa, the A. thaliana rRNA genes are repressed. Interestingly, A. thaliana rRNA gene silencing is variable in synthetic Arabidopsis suecica F1 hybrids. Two generations are needed for A. thaliana rRNA genes to be silenced in all lines, revealing a species-biased direction but stochastic onset to nucleolar dominance. Backcrossing synthetic A. suecica to tetraploid A. thaliana yielded progeny with active A. thaliana rRNA genes and, in some cases, silenced C. arenosa rRNA genes, showing that the direction of dominance can be switched. The hypothesis that naturally dominant rRNA genes have a superior binding affinity for a limiting transcription factor is inconsistent with dominance switching. Inactivation of a species-specific transcription factor is argued against by showing that A. thaliana and C. arenosa rRNA genes can be expressed transiently in the other species. Transfected A. thaliana genes are also active in A. suecica protoplasts in which chromosomal A. thaliana genes are repressed. Collectively, these data suggest that nucleolar dominance is a chromosomal phenomenon that results in coordinate or cooperative silencing of rRNA genes.

Arabidopsis↗

PubMed1998

Stimulation of membrane-bound guanylate cyclase activity by 17-beta estradiol.

Estrogen is known to exert both long-term genomic and short-term nongenomic actions. The long-term genomic actions of estrogen are mediated by the binding of estrogen to its cytoplasmic/nuclear receptor that subsequently triggers the transcription of target genes. In contrast, little is known about the molecular basis of the nongenomic actions of estrogen. We have found that 17-beta estradiol activates membrane-bound guanylate cyclase and enhances atrial natriuretic factor (ANF)-stimulated guanylate cyclase activity in PC12 cell membranes. Membrane-bound guanylate cyclase, GC-A, contains an extracellular ANF-binding domain and an intracellular kinase-like domain plus catalytic domain. The kinase-like domain plus the catalytic domain of guanylate cyclase (GC-kc) can be expressed in E. Coli and functionally renatured. 17-beta estradiol also activates the bacteria expressed GC-kc, indicating that 17-beta estradiol can directly interact with membrane-bound guanylate cyclase. These results demonstrate that 17-beta estradiol exerts a non-genomic action on membrane-bound guanylate cyclase.

Animals↗

PubMed1998

Polyreactive antigen-binding B cells are the predominant cell type in the newborn B cell repertoire.

Polyreactive antibodies bind to a variety of different self and non-self antigens. The B cells that make these antibodies express the polyreactive lg receptor on their surface. To determine the frequency of polyreactive antigen-binding B cells in peripheral blood, we incubated two different antigens, one (insulin) labeled with fluorescein isothiocyanate and the other (beta-galactosidase) with phycoerythrin, with peripheral B cells. The percentage of cells that bound these antigens was determined with the fluorescence-activated cells sorter. Approximately 21% of adult B cells bound insulin, 28% bound beta-galactosidase, and 11% bound both antigens. In contrast to B cells in the adult repertoire, 49% of B cells in cord blood bound insulin, 54% bound beta-galactosidase, and 33% bound both antigens. The properties of polyreactive antigen-binding B cells in adult and cord blood were similar, except for the fact that almost all the polyreactive antigen-binding B cells in cord blood were CD5 positive (93%), whereas only 40% of the polyreactive antigen-binding B cells in adult peripheral blood were CD5 positive, indicating that the CD5 marker is not directly linked to polyreactivity. The percentage of polyreactive antigen-binding B cells in patients with Sjögren's syndrome, systemic lupus erythematosus and rheumatoid arthritis was equal to or slightly below that found in the normal adult B cell repertoire. It is concluded that polyreactive antigen-binding B cells are a major constituent of the normal adult B cell repertoire and are the predominant cell type in the newborn B cell repertoire.

Adolescent↗

PubMed1998

Distribution of macromolecular dyes in brain using positive pressure infusion: a model for direct controlled delivery of therapeutic agents.

BACKGROUND: Direct infusion of therapeutic agents into the brain is a novel technique that has the potential for bypassing the blood-brain barrier and delivering high concentrations of therapeutic agents into the brain parenchyma. We have developed a model to characterize the distribution of Evans Blue (MW 960) and Blue Dextran (MW 2 x 10(6)) in rat brain using a positive pressure infusion system. METHODS: Evans Blue and Blue Dextran were infused in volumes of 20, 40, 60, 100, 140, and 180 microL into the caudate putamen of female Fischer rats over a period of 2 h with rates of infusion varying between 0.167 microL and 1.5 microL/min. During the infusions, the pressure generated in the infusion system and intracranial pressure were measured using a fiberoptic pressure monitoring system. After infusions, the volumes of distribution of the dye molecules were measured from 3-mm thick sections using video microscopy and computer image analysis. Histologic changes during the infusion were studied using snap freezing and hematoxylin/eosin staining of cryosections. RESULTS: Volumes of distribution for Evans Blue were greater than those for Blue Dextran. There was extensive spread of each dye in the ipsilateral hemisphere and also across the corpus callosum to the opposite hemisphere. Infusion/interstitial pressures peaked during the first 5 min of the infusion period, after which pressures dropped to a plateau value that remained relatively constant during the remainder of the infusion. Histologic findings suggest that this phenomenon is an important transition process that is likely to play a role in the pattern of distribution of macromolecules infused by this technique. No marked changes in intracranial pressure were noted during the infusion procedure. CONCLUSIONS: Direct positive pressure infusion into the brain has great potential in the treatment of brain tumors and other central nervous system disorders using both high and low molecular weight compounds (immunotoxins, protein conjugates, pharmacologic agents, oligonucleotides, and viral vectors).

Animals↗

PubMed1998

Distribution and stability of antisense phosphorothioate oligonucleotides in rodent brain following direct intraparenchymal controlled-rate infusion.

OBJECT: High-flow microinfusion is a novel technique for delivery of compounds directly into brain parenchyma, bypassing the blood-brain barrier. The feasibility of this technique has been demonstrated with low-molecular-weight compounds, macromolecular dyes, and proteins. Delivery of antisense oligonucleotides into brain parenchyma represents an additional potential application of this technique not previously described. In this report the authors sought to examine the distribution and disposition of phosphorothioate oligodeoxynucleotide (PS-ODN) for this reason. METHODS: An 18-mer 35S-PS-ODN (Mr approximately 6000) was infused over 1 hour into the caudate putamen of Fischer 344 rats. At 1, 6, 12, 24, and 48 hours after beginning the infusion, the brains were extracted and analyzed using quantitative autoradiographic techniques. Cerebrospinal fluid (CSF) was also aspirated from the cisterna magna and was analyzed to determine the radioactivity and stability of the 35S-PS-ODN. At 1 hour, the infused ODN was uniformly distributed in brain tissue, with a maximum average concentration of 4806.5 +/- 210.5 nCi/g. This represents a tissue concentration of 19.2 +/- 0.84 microM. Extensive spread into surrounding parenchyma was observed over the ensuing 47 hours. The 35S-PS-ODN radioactivity peaked in the CSF at the end of the 1-hour infusion, containing 1% (50 +/- 20 nCi) of the infused radioactivity. Activity then decayed exponentially over 11 hours, but stabilized at a lower CSF content of 0.2% (1 +/- 0.1 nCi) thereafter. The volume of distribution was 105 +/- 7.9 mm3 at 1 hour, representing a volume of distribution/volume of infusion ratio of 5.2. The volume of distribution increased to 443 +/- 62.3 mm3 at the end of 48 hours, whereas the average minimum tissue concentration decreased from 15.2 microM to 3.2 microM. Undegraded 18-mer was observed throughout the 48-hour period by means of 20% polyacrylamide/7 M urea gel electrophoresis. The animals tolerated the infusion without evidence of toxicity and minimal structural changes in tissue were observed on histological investigation. CONCLUSIONS: The authors found that PS-ODNs can be safely delivered in high concentrations to wide areas of rat brain by using high-flow microinfusion and are stable even after 48 hours in situ.

Animals↗

PubMed1998

Effect of catecholamic acid on detoxication and distribution of NiCl2 in mice and rats.

AIM: To study the effect of catecholamic acid (CBMIDA) on detoxication of NiCl2. METHODS: Mice and rats were injected s.c. or i.m. CBMIDA immediately after i.p. NiCl2. Each mouse was injected i.p. CBMIDA after i.v. 63NiCl2 185 kBq, and radioactivities of various tissues were measured with liquid scintillation counter at 24 h. The localization of 63Ni was shown by the whole-body autoradiography. RESULTS: CBMIDA s.c. 0.5-1.5 g.kg-1 markedly reduced the mortality from acute poisoning of i.p. NiCl2 500 mg.kg-1. After i.p. NiCl2 in mice, the LD50 was 82.7 mg.kg-1. Mice were injected s.c. CBMIDA 1.5 or 2.5 g.kg-1 after Ni poisoning, the LD50 of NiCl2 were raised to 789 or 820 mg.kg-1, respectively. The LD50 of NiCl2 was 39 mg.kg-1 in rat. If CBMIDA was injected i.m. 0.5 g.kg-1 after i.p. NiCl2, the LD50 was 332 mg.kg-1. CBMIDA 1.5 g.kg-1 i.m. after i.v. 63NiCl2, decreased the contents of 63Ni in blood and lung of mice vs control mice at 24 h. The contents of 63Ni in brain, heart, spleen, and kidney were similar to those of the control mice. The content of 63Ni in bone was more than the control. The excretions of 63Ni through urine and feces were not increased by CBMIDA at 24 h. The whole-body autoradiography showed that the radioactivity was highly localized in the kidney, lung, and Harder's gland. There was a moderate level of 63Ni in the liver, bone, skin, and blood. A pronounced accumulation occurred in the bone. There was a marked reduction of 63Ni in the lung, skin, liver, and blood after i.p. CBMIDA. CONCLUSION: The CBMIDA markedly raised the survival rate of nickel-poisoned mice and rats, and decreased 63Ni levels in lung and blood.

Animals↗

PubMed1997

Distribution and stability of antisense phosphorothioate oligonucleotides in rodent brain following direct intraparenchymal controlled-rate infusion.

High-flow microinfusion is a novel technique for delivery of compounds directly into the brain parenchyma, bypassing the blood-brain barrier. The feasibility of this technique has been demonstrated with low-molecular-weight compounds, macromolecular dyes, and proteins. Delivery of antisense oligonucleotides into the brain parenchyma represents an additional potential application of this technique not previously described. In this report, the authors examined the distribution and disposition of phosphorothioate oligodeoxynucleotide (PS-ODN) infused for this reason. An 18-mer (35)S-PS-ODN (molecular weight approximately 6000) was infused over 1 hour into the caudate putamen of Fischer 344 rats. At 1, 6, 12, 24, and 48 hours after beginning the infusion, the brains were extracted and analyzed using quantitative autoradiographic techniques. Cerebrospinal fluid (CSF) was also aspirated from the cisterna magna and analyzed for radioactivity and stability of the (35)S-PS-ODN. At 1 hour, the infused ODN was uniformly distributed in brain tissue, with a maximum average concentration of 4806.5 +/- 210.5 nCi/g. This represents a tissue concentration of 19.2 +/- 0.84 microM. Extensive spread into surrounding parenchyma was observed over the ensuing 47 hours. The (35)S-PS-ODN radioactivity peaked in the CSF at the end of the 1-hour infusion, containing 10% (50 +/- 20 nCi) of the infused radioactivity. Activity then decayed exponentially over 11 hours, stabilizing at a lower CSF content of 0.2% (1 +/- 0.1 nCi). The volume of distribution (V(d)) was 105 +/- 7.9 mm3 at 1 hour, representing a ratio of V(d)/V(i) (volume of infusion) of 5.2. The V(d) increased to 443.4 +/- 62.3 mm(3) at the end of 48 hours, whereas the average minimum tissue concentration decreased from 15.2 to 3.2 microM. Undegraded 18-mer was seen throughout the 48-hour period using 20% polyacrylamide/7M urea gel electrophoresis. The animals tolerated the infusion without evidence of toxicity, and minimal structural changes in tissue were observed on histological examination. Thus, PS-ODN can be safely delivered in high concentrations to wide areas of the rat brain by using high-flow microinfusion, and the concentrations remain stable even after 48 hours in situ.

Journal Article↗

PubMed1997

Epigenetic silencing of RNA polymerase I transcription: a role for DNA methylation and histone modification in nucleolar dominance.

Nucleolar dominance is an epigenetic phenomenon that describes nucleolus formation around rRNA genes inherited from only one progenitor of an interspecific hybrid or allopolyploid. The phenomenon is widespread, occurring in plants, insects, amphibians, and mammals, yet its molecular basis remains unclear. We have demonstrated nucleolar dominance in three allotetraploids of the plant genus Brassica. In Brassica napus, accurately initiated pre-rRNA transcripts from one progenitor, Brassica rapa are detected readily, whereas transcripts from the approximately 3000 rRNA genes inherited from the other progenitor, Brassica oleracea, are undetectable. Nuclear run-on confirmed that dominance is controlled at the level of transcription. Growth of B. napus seedlings on 5-aza-2'-deoxycytidine to inhibit cytosine methylation caused the normally silent, under-dominant B. oleracea rRNA genes to become expressed to high levels. The histone deacetylase inhibitors sodium butyrate and trichostatin A also derepressed silent rRNA genes. These results reveal an enforcement mechanism for nucleolar dominance in which DNA methylation and histone modifications combine to regulate rRNA gene loci spanning tens of megabase pairs of DNA.

Azacitidine↗

PubMed1997

Transcriptional analysis of nucleolar dominance in polyploid plants: biased expression/silencing of progenitor rRNA genes is developmentally regulated in Brassica.

Nucleolar dominance is an epigenetic phenomenon that describes the formation of nucleoli around rRNA genes inherited from only one parent in the progeny of an interspecific hybrid. Despite numerous cytogenetic studies, little is known about nucleolar dominance at the level of rRNA gene expression in plants. We used S1 nuclease protection and primer extension assays to define nucleolar dominance at a molecular level in the plant genus Brassica. rRNA transcription start sites were mapped in three diploids and in three allotetraploids (amphidiploids) and one allohexaploid species derived from these diploid progenitors. rRNA transcripts of only one progenitor were detected in vegetative tissues of each polyploid. Dominance was independent of maternal effect, ploidy, or rRNA gene dosage. Natural and newly synthesized amphidiploids yielded the same results, arguing against substantial evolutionary effects. The hypothesis that nucleolar dominance in plants is correlated with physical characteristics of rRNA gene intergenic spacers is not supported in Brassica. Furthermore, in Brassica napus, rRNA genes silenced in vegetative tissues were found to be expressed in all floral organs, including sepals and petals, arguing against the hypothesis that passage through meiosis is needed to reactivate suppressed genes. Instead, the transition of inflorescence to floral meristem appears to be a developmental stage when silenced genes can be derepressed.

Base Sequence↗