In situ hybridization to lampbrush chromosomes: a potential source of error exposed

1980 ◽  
Vol 41 (1) ◽  
pp. 115-123
Author(s):  
H.G. Callan ◽  
R.W. Old
Keyword(s):  
Lampbrush Chromosome ◽  
Single Pair ◽  
Lampbrush Chromosomes ◽  
Triturus Cristatus ◽  
Rna Transcripts ◽  
Potential Source ◽  
Source Of Error ◽  
Simple Sequence ◽  
Do So

Denatured 3H-labelled DNAs containing Xenopus or human globin sequences hybridize to RNA transcripts on a single pair of lateral loops on lampbrush chromosome IX of Triturus cristatus carnifex, and to no other loops on this chromosome or the rest of the complement. However they do so, not because of the globin sequences in the probes, but rather because the plasmids from which the probes were prepared were constructed with G.C homopolymer tails. Simple sequence poly d(C/G)n probes also hybridize with RNA transcripts on this same pair of loops, and with no others.

scholarly journals Transcription on lampbrush chromosome loops in the absence of U2 snRNA.

1992 ◽  
Vol 3 (3) ◽  
pp. 249-261 ◽  
Author(s):  
A Tsvetkov ◽  
M Jantsch ◽  
Z Wu ◽  
C Murphy ◽  
J G Gall
Keyword(s):  
Physiological Saline ◽  
Lampbrush Chromosome ◽  
Antisense Oligodeoxynucleotides ◽  
Lampbrush Chromosomes ◽  
U2 Snrna ◽  
Complete Destruction ◽  
Small Nuclear Rnas ◽  
Cytological Changes

The five small nuclear RNAs (snRNAs) involved in splicing occur on the loops of amphibian lampbrush chromosomes and in hundreds to thousands of extrachromosomal granules called B snurposomes. To assess the role of these snRNAs during transcription and to explore possible relationships between the loops and B snurposomes, we injected single-stranded antisense oligodeoxynucleotides (oligos) against U1 and U2 snRNA into toad and newt oocytes. As shown before, antisense U1 and U2 oligos caused truncation of U1 and complete destruction of U2 snRNAs, respectively. However, injection of any oligo, regardless of sequence, brought on dramatic cytological changes, including shortening of the chromosomes and retraction of the lateral loops, with concomitant shutdown of polymerase II transcription, as well as disappearance of some or all of the B snurposomes. When injected oocytes were incubated for 12 h or longer in physiological saline, these changes were reversible; that is, the chromosomes lengthened, transcription (detected by 3H-UTP incorporation) resumed on newly extended lateral loops, and B snurposomes reappeared. In situ hybridization showed that loops and B snurposomes had negligible amounts of U2 snRNA after recovery from injection of the anti-U2 oligo, whereas these structures had normal levels of U2 snRNA after recovery from a control oligo. Thus, the morphological integrity of B snurposomes and lampbrush chromosome loops is not dependent on the presence of U2 snRNA. Because transcription occurs in the absence of U2 snRNA, we conclude that splicing is not required for transcription on lampbrush chromosome loops.

scholarly journals Monoclonal antibodies that recognize transcription unit proteins on newt lampbrush chromosomes.

1987 ◽  
Vol 105 (3) ◽  
pp. 1047-1054 ◽  
Author(s):  
M B Roth ◽  
J G Gall
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Lines ◽  
Germinal Vesicle ◽  
Transcription Unit ◽  
Lampbrush Chromosome ◽  
Lampbrush Chromosomes ◽  
Rna Transcripts ◽  
Heterogeneous Nuclear Ribonucleoproteins ◽  
Nascent Rna ◽  
Hybridoma Cell Lines

We prepared hybridoma cell lines from mice injected with newt germinal vesicle proteins. We tested culture supernates from these cell lines for antibodies that bound to specific morphological structures in lampbrush chromosome preparations (nucleoli, loops, chromomeres, etc.). Four mAbs that recognize antigens on the lateral transcription loops are described here. We suggest that these antigens are proteins associated with nascent RNA transcripts, although they are not among the 30-40-kD "core" heterogeneous nuclear ribonucleoproteins.

Faunal transport within event horizons in the British Upper Silurian

1994 ◽  
Vol 131 (4) ◽  
pp. 485-498 ◽  
Author(s):  
Charlie J. Underwood
Keyword(s):  
Shallow Water ◽  
Outer Shelf ◽  
Event Horizons ◽  
Potential Source ◽  
Upper Silurian ◽  
Source Of Error

AbstractMany marine fossil concentrations are considered the result of episodic sedimentological events, and in particular those due to storms. Most storm or tempestite concentrations are identified as autochthonous or parautochthonous assemblages created by a variety of winnowing processes within shallow water environments. In contrast, samples described here from both a ‘shelf’ and a ‘basinal’ setting within the Ludlow (Upper Silurian) succession of the Welsh Basin reveal the presence of a biota transported by tempestite activity into a setting dominated by a more offshore biota. Tempestite horizons from within an ‘outer shelf’ mud dominated setting include shelly lenses with a transported fauna abounding in gastropods, tentaculitids and atrypid brachiopods, the background sediment being rich in graptolites, cephalopods and small strophomenid brachiopods. Within the ‘basinal’ area, distal tempestites range from minor siltstone layers to thicker bioclastic limestone lenses. The siltstones are largely graptolitic (dominated byBohemograptus), with some small brachiopods, whilstSaetograptus colonusis the only common graptolite in the limestones, which also contain a fauna of broken brachiopods and bryozoa. The transport of assemblages distally into a variety of settings represents a potential source of error in palaeoecological analysis. Transported assemblages may, however, provide evidence of the composition of both benthic and pelagic shallower water faunas no longer knownin situ.

Some Palaeoliths from South Arabia

1954 ◽  
Vol 19 (2) ◽  
pp. 189-218 ◽  
Author(s):  
G. Caton-Thompson
Keyword(s):  
Geographical Society ◽  
South Arabia ◽  
First Time ◽  
Do So

The material here described was found in the Hadhramaut by Elinor Gardner and myself between November 1937 and March 1938. My personal investigation of the Palaeolithic Age was limited by pre-Islamic excavations, and I am therefore indebted to her for the gathering of most of the specimens in situ in terrace gravels, and to her detailed study of their positions.The collection consists mainly of groups from four fairly widely separated localities; the physiography of these has already been outlined in a comprehensive paper published in the Geographical Journal. Whenever appropriate to the purpose of this account, which is to place for the first time on illustrated record all we observed about the palaeoliths, I have reused in this different context illustrations of Quaternary environment which appeared in that Journal. With thanks I acknowledge the permission of the Royal Geographical Society to do so.

scholarly journals Histone H4 acetylation and transcription in amphibian chromatin.

1993 ◽  
Vol 120 (2) ◽  
pp. 277-290 ◽  
Author(s):  
J Sommerville ◽  
J Baird ◽  
B M Turner
Keyword(s):  
Transcriptional Activation ◽  
Histone Deacetylation ◽  
Dense Matrix ◽  
Lampbrush Chromosomes ◽  
Loop Formation ◽  
Triturus Cristatus ◽  
Immature Oocytes ◽  
Intense Fluorescence ◽  
Histone H4 Acetylation ◽  
Lysine Residues

Lampbrush chromosomes from oocytes of the amphibian Triturus cristatus have been used to examine the role of histone acetylation in transcription by indirect immunofluorescence with antisera to H4 acetylated at specific lysine residues. Electrophoresis on acid-urea-Triton gels and Western blotting have confirmed the specificity of these antisera and defined the order in which particular lysine residues are acetylated in amphibian cells. As in mammals, lysine 16 is acetylated first, followed by 8 and/or 12 and then 5. With lampbrush chromosomes from immature (previtellogenic) oocytes, antisera to H4 acetylated at lysines 8, 12, and 16 labeled fluorescent foci at the bases of transcription loops. Antisera to H4 acetylated at lysine 5 labeled weakly (i.e., the tri- and tetraacetylated isoforms must be rare). Loops showed weak labeling of the chromatin axis but intense fluorescence at particular points, which probably represent incompletely decondensed chromatin. The RNP matrix of loops, including the RNP-rich sphere bodies and the dense matrix of "marker" loops, was not labeled. Treatment of immature oocytes with butyrate for 12 h to inhibit histone deacetylation did not affect immunolabeling, suggesting that turnover of H4 acetates is slow. In contrast, in chromosomes from mature oocytes, in which loops have retracted and transcription is low, butyrate caused an increase in labeling with all antisera, followed by the appearance of vestigial loops, weakly labeled, but with regions of intense fluorescence. These loops contain RNP and are presumably transcriptionally active. We conclude that H4 acetates turn over more rapidly in mature than immature oocytes and that histone hyperacetylation precedes, and possibly induces, loop formation and transcriptional activation.

Echocardiographic Study of Premature Infants

PEDIATRICS ◽  
1984 ◽  
Vol 73 (6) ◽  
pp. 883-883
Author(s):  
GREGORY L. JOHNSON ◽  
Albert B. Chandler
Keyword(s):  
Premature Infants ◽  
Multicenter Study ◽  
Aortic Root ◽  
Leading Edge ◽  
Potential Source ◽  
Echocardiographic Study ◽  
Edge Method ◽  
American Society ◽  
Source Of Error

In Reply.— Levine has correctly pointed out an important potential source of error in performing echocardiographic measurements in small infants. Measurements of aortic root dimension in the multicenter study we reported were, indeed, performed utilizing the outer-to-inner (ie, leading edge) method as recommended by the American Society of Echocardiography and noted in the methods portion of our manuscript. Due to the reduction in figure size associated with publication, our figure 1 (Pediatrics 1984;72:865) appears, at first glance, to demonstrate inner-to-inner measurement as Levine points out.

Sequential silver staining and in situ hybridization reveal a direct association between rDNA levels and the expression of homologous nucleolar organizing regions: a hypothesis for NOR structure and function

1998 ◽  
Vol 111 (10) ◽  
pp. 1433-1439
Author(s):  
F. Zurita ◽  
R. Jimenez ◽  
M. Burgos ◽  
R.D. de la Guardia
Keyword(s):  
Transcription Factors ◽  
In Situ Hybridization ◽  
Silver Staining ◽  
Organizer Region ◽  
Nucleolar Organizer ◽  
Interindividual Variation ◽  
Nucleolar Organizer Regions ◽  
Single Pair ◽  
Ribosomal Cistrons

We have developed a procedure for sequential silver staining and in situ hybridization to analyze the relationship between the amount of rDNA present in nucleolar organizer regions, as estimated by in situ hybridization, and their level of expression, as estimated by the silver signal. For simplicity we used cells from the insectivorous mole Talpa occidentalis, which have a single pair of nucleolar organizer regions in chromosome pair 3. The relative content of ribosomal cistrons was also related to the hierarchy of activation of the nucleolar organizer regions present in this chromosomal pair. Statistical analyses demonstrated that both the relative level of expression and the activation hierarchy depended mainly on the number of ribosomal cistrons in nucleolar organizer regions. We propose a functional two-step hypothesis, which is consistent with most known data concerning interchromosomal, intercellular and interindividual variation in a number of plant and animal species, including Talpa occidentalis. In step one, the first available transcription factors bind randomly to the ribosomal promoters, such that larger nucleolar organizer regions are more likely to recruit them. In the second step the remaining transcription factors are recruited in a cooperative way, thus completing activation of one nucleolar organizer region, before the next one becomes active.

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