Proteins of the newt oocyte nucleus: analysis of the nonhistone proteins from lampbrush chromosomes, nucleoli and nuclear sap

1975 ◽  
Vol 17 (3) ◽  
pp. 579-588
Author(s):  
K. Maundrell
Keyword(s):  
Total Protein ◽  
Electrophoretic Analysis ◽  
Lampbrush Chromosome ◽  
Chromosome Preparation ◽  
Oocyte Nucleus ◽  
Protein Species ◽  
Lampbrush Chromosomes ◽  
Banding Patterns ◽  
Nonhistone Proteins

An electrophoretic analysis has been carried out on the total protein of lampbrush chromosomes, nucleoli and nuclear sap, obtained from newt oocyte nuclei. In each case, distinctive and heterogeneous banding patterns are observed. The absence of detectable quantities of histones in occyte chromatin is noted. In the case of the lampbrush chromosome preparation, it is concluded that all protein species are derived from the ribonucleoprotein matrix of the lateral loops.

Nonhistone Proteins of the Oocyte Nucleus of the Newt

1974 ◽  
Vol 15 (1) ◽  
pp. 145-161
Author(s):  
R. J. HILL ◽  
K. MAUNDRELL ◽  
H. G. CALLAN
Keyword(s):  
Nuclear Membrane ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Germinal Vesicle ◽  
Oocyte Nucleus ◽  
Lampbrush Chromosomes ◽  
Nonhistone Proteins ◽  
Aerial Oxidation ◽  
A Minor

Evidence has been obtained which indicates that disulphide bond crosslinks contribute to the morphological integrity of isolated lampbrush chromosomes (both chromomeres and lateral loops) and nucleoli. It is suggested that the progressive formation of these bonds in vitro by aerial oxidation may provide the basis for the previously recognized time-dependent hardening or ‘denaturing’ of these structures. Manually isolated germinal vesicle nuclei have been massed and fractionated by low-speed centrifugation into nucleoplasm and chromatin. Phase-contrast microscopy demonstrates the chromatin to consist of nucleoli, lampbrush chromosomes and nuclear membranes. Urea gel electrophoresis has been employed to resolve the reduced and S-carboxymethylated proteins of whole nuclei into some 12 components, negatively charged at pH 8. The nucleoplasm alone gives an essentially similar pattern, but with the distinct depletion of one component and slight depletion of another. Both of these components are much enriched in the chromatin pellet where they predominate over all other proteins. The total chromatin has been subfractionated by microdissection, taking advantage of the differential attachment of nucleoli to the nuclear membrane at different stages of oogenesis. It is concluded that the nuclear membrane per se does not contribute to the major chromatin proteins. The two major polypeptides are components of the nucleoli. Preparations of isolated lampbrush chromosomes have not, to date, provided sufficient material to give a distinctive electropherogram; only one faint band, a major component of whole nuclei, was apparent. Sodium dodecyl sulphate gel electrophoresis has resolved some 25 components in whole nuclei, and again demonstrates the enrichment of the two major species in the total chromatin fraction. The apparent molecular weights of these two species are 43 kilodaltons and 110 kilodaltons. Approximately 20 minor species are also present in the chromatin and are obviously good candidates as components of the nucleolar and chromosomal structures. Histones, at most, make only a minor contribution to the overall chromatin protein population.

scholarly journals Chromosomes and Nucleoli of the Axolotl, Ambystoma Mexicanum

1966 ◽  
Vol 1 (1) ◽  
pp. 85-108
Author(s):  
H. G. CALLAN
Keyword(s):  
Low Temperature ◽  
Nuclear Periphery ◽  
Great Majority ◽  
Lampbrush Chromosome ◽  
Ambystoma Mexicanum ◽  
Oocyte Nucleus ◽  
Lampbrush Chromosomes ◽  
Mitotic Chromosomes ◽  
Nucleolar Material ◽  
Secondary Constrictions

Amongst the axolotl's haploid complement of fourteen mitotic chromosomes, one of the four largest, with a greater arm asymmetry than the other three, shows a nucleolar constriction subterminally in its shorter arm. Low-temperature treatment causes further secondary constrictions to appear; these constrictions enable most of the mitotic chromosomes to be identified; the constrictions occur at similar sites in the chromosomes of tail-fin epithelial cells, hepatocytes, and brain cells. Homology between the mitotic and oocyte (lampbrush) nucleolar organizers has been established, and thus the several hundred free nucleoli in oocytes are genetically related to the two nucleoli of diploid somatic interphases. During oocyte development the free nucleoli transform from solid structures to rings and back to solid structures again without detectable increase in number. During the contraction and aggregation of the lampbrush chromosomes within the oocyte nucleus as maturity approaches, in most axolotls the free ring-shaped nucleoli become stretched between the nuclear periphery and central chromosome group, and take on a characteristic beaded appearance. These transformations of the free nucleoli are largely paralleled by forms which nucleoli attached subterminally to the shorter arm of lampbrush chromosome III concurrently assume. The question as to whether fully developed nucleoli detach from the organizer loci and add to the population of free nucleoli in oocytes remains undecided. It may well be that virtually all the DNA-generators of free nucleoli detach from the organizer loci before starting to carry out nucleolar functions, and before there is any significant accumulation of protein and RNA around them. If so, the variability in quantity of attached nucleolar material may not reflect different states in a nucleolar synthesis and detachment cycle, but rather variation in the number of nucleolar DNA Anlagen which happen to remain attached to the organizer loci after the synthesis and detachment of the great majority of the Anlagen has ceased. In occasional oocytes the only chromosomal continuity maintained across the organizer locus consists of a nucleolar ‘double bridge’; this indicates that the genetically persistent (i.e. chromosomal) organizer DNA bears the same structural relationship to neighbouring parts of a lampbrush chromosome as any other chromomere with its attendant pair of lateral loops. The lampbrush chromosomes of the axolotl have been provisionally mapped. The centromeres are represented by short portions of chromosome axis without lateral loops, and there are two spheres close to the centromeres of both chromosome VI and chromosome XIII. Other recognition characters are inconspicuous or not very reliable, and features of the lampbrush chromosomes related to the low-temperature induced secondary constrictions of mitotic chromosomes have not been identified.

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 Isoflavone and protein content in soybeans grains submitted to flooding at different stages of development

2011 ◽  
Vol 41 (12) ◽  
pp. 2224-2229 ◽  
Author(s):  
Camila Argenta Fante ◽  
Patrícia de Fátima Pereira Goulart ◽  
José Donizeti Alves ◽  
Paôla de Castro Henrique ◽  
Daniela Deitos Fries
Keyword(s):  
Protein Content ◽  
Chronic Diseases ◽  
Total Protein ◽  
Electrophoretic Analysis ◽  
Stages Of Development ◽  
Major Barrier ◽  
Human Food ◽  
Soil Flooding ◽  
Sds Page ◽  
Page Electrophoresis

The stress imposed on plants by soil flooding constitutes a major barrier to growth and productivity. The identification of soybean varieties that produce higher levels of isoflavones, is necessary as soybeans have been used as human food to reduce risks of chronic diseases. Thus, this study was conducted with the objective of quantifying proteins and isoflavones in soybean cultivars subjected to flooding at various stages of development. The cultivars 'BRS267', 'BRS257' and 'BRS213' were subjected to 15 days of flooding, starting at the stages V6 and V8 and 11 days under stress starting at the stage R4. The proteins in the grain were extracted and quantified and analyzed by SDS-PAGE electrophoresis. Isoflavones were extracted, separated and quantified on HPLC. The electrophoretic analysis of the three cultivars under study revealed the same pattern of banding relative to the total protein regardless of the treatment. However, it was noted that flooding led to an increase in the total contents of isoflavones in the BRS 267 plants flooded in stage R4, remaining constant in other cultivars.

scholarly journals Production of Somatic Hybrid and Autotetraploid Breeding Parents for Seedless Citrus Development

HortScience ◽  
1992 ◽  
Vol 27 (10) ◽  
pp. 1125-1127 ◽  
Author(s):  
Jude W. Grosser ◽  
Frederick G. Gmitter ◽  
E.S. Louzada ◽  
J.L. Chandler
Keyword(s):  
Somatic Hybrid ◽  
Somatic Hybrids ◽  
Sweet Orange ◽  
Electrophoretic Analysis ◽  
Root Tip ◽  
Banding Patterns ◽  
Hybrid Plants ◽  
Sexual Hybrid ◽  
Seedling Leaf ◽  
Cell Chromosome

Allotetraploid somatic hybrid plants of `Nova' tangelo [a sexual hybrid of `Clementine mandarin (C. reticulata Blanco) × `Orlando' tangelo (C. reticulata × C. paradisi Macf.)] + `Succari' sweet orange (C. sinensis L. Osbeck), and `Hamlin' sweet orange (C. sinensis L. Osbeck) + `Dancy' tangerine (C. reticulata) were regenerated following protoplast fusion. `Nova' and `Hamlin' protoplasts were isolated from ovule-derived embryogenic callus and suspension cultures, respectively, and fused using a polyethylene glycol method with seedling leaf-derived protoplasts of `Succari' and `Dancy', respectively. Plants were regenerated via somatic embryogenesis, and somatic hybrids were identified on the basis of leaf morphology, root-tip cell chromosome number, and electrophoretic analysis of peroxidase and phosphoglucose mutase isozyme banding patterns. Diploid plants were regenerated from unfused protoplasts of `Hamlin', `Nova', and `Succari'. Tetraploid plants of `Hamlin' and `Succari' were also recovered, apparently resulting from homokaryotic fusions. No `Dancy' plants were recovered. The somatic hybrid and autotetraploid plants can be used for interploid hybridization with selected monoembryonic scions to generate improved seedless triploid tangor/tangelo cultivars. The lack of suitable tetraploid breeding parents has previously inhibited the development of quality seedless cultivars by this method.

scholarly journals Protein fractions of oats and possibilities of oat utilisation for patients with coeliac disease

2011 ◽  
Vol 22 (No. 4) ◽  
pp. 151-162 ◽  
Author(s):  
I. Capouchová ◽  
J. Petr ◽  
H. Tlaskalová-Hogenová ◽  
I. Michalík ◽  
O. Faměra ◽  
...  
Keyword(s):  
Coeliac Disease ◽  
Nitrogen Content ◽  
Protein Complex ◽  
Total Protein ◽  
Electrophoretic Analysis ◽  
Gluten Free Diet ◽  
Protein Fractions ◽  
Gluten Free ◽  
Protein Nitrogen ◽  
Average Percentage

The applicability was evaluated of 16 different oats species and varieties of different provenance in the coeliac diet in view of the composition of the protein complex and immunological testing during two-year experiments (2001 and 2002). Determination was carried out of total nitrogen content (average of evaluated oats collection in 2001 was 2.21%, in 2002 2.78%), protein nitrogen content (average 2001 1.94%, 2002 2.28%), and crude protein (N × 6.25) content (average 2001 13.80%, 2002 17.37%). The proportions of different protein fractions play a decisive role for the aims of this study because, based on the existing knowledge, coeliacally active protein components are present particularly in the prolamin fraction. The percentage of prolamins (determined by discontinual fractionation after Osborne) in the author’s evaluated collection of oats species and varieties under the conditions of Central Bohemia reached on average 17.68% of the total protein in 2001, and 15.36% in 2002. The average percentage of albumins and globulins of the total protein reached 36.97% in 2001 and 41.04% in 2002, the average percentage of glutelins of the total proteins was 37.61% in 2001 and 34.10% in 2002, and residual was on average 7.55% in 2001 and 8.70% in 2002, respectively, of the total protein. Electrophoretic analysis of reserve (gluten) proteins (SDS-PAGE ISTA) showed in the oats collection evaluated the percentage of LMW + prolamins in the range 56–77% of the total reserve proteins in 2001, and 52–73% in 2002. The results of A-PAGE electrophoretic analysis of prolamin proteins confirmed the presence of α-prolamins, that ranged in the total content of prolamins from 50 to 88% in 2001, and from 77 to 100% in 2002, while β- + γ-prolamins ranged in 2001 from 11 to 49%, and in 2002 from 0 to 22%. These values do not give serious guarantees for the possible utilisation of oats in the gluten-free diet. The results of the immunological evaluation of the amount of prolamins in oats grains using ELISA showed great differences between different varieties and the experimental years. In 2001, 7 oats samples out of 13 evaluated, and in 2002 10 samples out of 12 evaluated were below the limit for the gluten-free diet (10 mg prolamins (gliadins)/100 g of sample dry matter), but the other varieties exceeded the limit, particularly in 2001, very significantly. The results obtained in the evaluated collection of species and varieties of oats revealed a great variability in the structure of the protein complex and in the immunological testing. In addition a significant effect of the year on the results of all analyses was evident. Based on our results, the use of oats in the diet for coeliac disease can be very risky for these reasons.  

An introduction to Programmes for Development

Development ◽  
1984 ◽  
Vol 83 (Supplement) ◽  
pp. 1-6
Author(s):  
Herbert C. Macgregor ◽  
Alma P. Swan
Keyword(s):  
Oocyte Nucleus ◽  
Nuclear Chromatin ◽  
Lampbrush Chromosomes ◽  
Unicellular Alga ◽  
Complex Molecules ◽  
Animal Development ◽  
Initial Tendency ◽  
Dna Strand ◽  
Acetabularia Mediterranea ◽  
The University

The symposium of which this book is a record was first suggested in the autumn of 1982. At that time, all members of the BSDB committee were persons who were primarily concerned with studies of animal development, and the initial tendency was to think along these lines. The cytology laboratory in the University of Leicester has a long tradition in studies of the lampbrush chromosomes that are found in the growing oocytes of most animals as well as in certain stages of the life cycle of at least one simple plant, the giant unicellular alga Acetabularia mediterranea (Callan, 1982). At some stage in very early diplotene of oogenesis of an amphibian, the oocyte nucleus begins to enlarge and many regions of the nuclear chromatin begin to transcribe RNA. RNA polymerase molecules attach to hundreds of sites along the chromosomes and move progressively along the DNA strand, synthesizing long and complex molecules of RNA as they go.

The role of Lampbrush Chromosomes in the formation of Nucleoli in Amphibian Oocytes

1965 ◽  
Vol s3-106 (75) ◽  
pp. 215-228
Author(s):  
H. C. MACGREGOR
Keyword(s):  
Phase Contrast ◽  
Nucleolar Organizer ◽  
Plethodon Cinereus ◽  
Oocyte Nucleus ◽  
Lampbrush Chromosomes ◽  
Triturus Cristatus ◽  
Amphibian Oocyte ◽  
Ambystoma Jeffersonianum ◽  
And Function

Theories concerning the mode of origin of peripheral nucleoli in amphibian oocytes have been examined and tested. In Triturus cristatus the giant fusing loops of the 3 shortest lampbrush bivalents resemble nucleoli when viewed in phase contrast and may be considered as possible sites of production of nucleoli. Giant fusing loops, however, differ from peripheral nucleoli in certain important respects, and animals lacking giant fusing loops on their lampbrush chromosomes nevertheless have normal peripheral nucleoli. Therefore, similarity in appearance between objects attached to lampbrush chromosomes and free peripheral nucleoli may not be significant. In oocytes of T. c. carnifex, T. c. karelinii, and T. c. danubialis, peripheral nucleoli do not increase in number during the lampbrush phase of oogenesis, except by division of pre-existing nucleoli towards the end of oogenesis. There are about 1,000 nucleoli per oocyte nucleus in each of these sub-species. In T. c. cristatus there are more nucleoli in large oocytes than in small ones, and it seems likely that in this sub-species the giant fusing loops add to the existing population of nucleoli in an oocyte by successively growing and shedding new nucleoli. A similar situation probably holds in Plethodon cinereus. Hexaploid oocytes from triploid females of Ambystoma jeffersonianum have 3 times as many nucleoli as diploid oocytes from diploid females of the same species. The number of nucleoli in an amphibian oocyte nucleus is therefore related to the number of sets of chromosomes in the cell. In yolky oocytes from hypophysectomized newts most peripheral nucleoli are firmly attached to the inner surface of the nuclear membrane; whereas in similar oocytes from unoperated or gonadotrophin-treated animals none of the nucleoli is so attached. On the basis of these observations 2 mechanisms are suggested for the formation of amphibian oocyte nucleoli. The first of these mechanisms probably operates in T. c. carnifex, where all peripheral nucleoli are formed before or soon after the chromosomes assume the lampbrush form, and no part of a lampbrush chromosome is involved in a process which adds to the existing population of nucleoli. The second mechanism probably operates in T. c. cristatus, where most of the peripheral nucleoli are formed before the lampbrush phase of oogenesis but a nucleolar organizer on the lampbrush chromosomes continues to grow and detach nucleoli throughout oogenesis. Both these mechanisms are discussed in terms of what is known of the chemical composition and function of peripheral nucleoli.

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.

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