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H G Callan

Publications and source records attributed to H G Callan.

At least 19 recordsLinked to original sources
PubMed1991

Association of RNA with the B and C snurposomes of Xenopus oocyte nuclei.

We studied the time course of [3H]-uridine incorporation into the B and C snurposomes of Xenopus oocyte nuclei. B snurposomes constitute most of the non-nucleolar granules in the 1-4 micron size range; they contain the five splicing small nuclear RNAs (snRNAs; U1, U2, U4, U5 and U6) plus a variety of associated proteins. The organelles referred to as spheres consist of a C snurposome with one or more B snurposomes on its surface. C snurposomes can exist independently of Bs and many are smaller than the structures usually classified as spheres. C snurposomes contain the trimethylguanosine moiety characteristic of snRNAs, as well as the Sm epitope found on several small nuclear ribonucleoproteins (snRNPs), but it is not known which snRNA(s) they contain. When oocytes are incubated with [3H]uridine, all of the nucleoli and chromosome loops label strongly and rapidly. By contrast, labelled RNA appears slowly in the B snurposomes and then only in a fraction of them. After a 24 h incubation, about half of the Bs are labelled, and half are unlabelled or weakly labelled. This observation suggests that there are "mature" and "immature" B snurposomes, and that only the latter acquire newly synthesized RNA. The nature of this RNA is unknown, but it probably includes the splicing snRNAs. B snurposomes on the surface of Cs also constitute a heterogeneous population, some becoming labelled and some remaining unlabelled during a 24 h incubation. An analysis of the label in "doublets" (one B and one C snurposome) suggests that RNA may pass from the Bs to the Cs.

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PubMed1991

Histone genes are located at the sphere loci of Xenopus lampbrush chromosomes.

In the anuran Xenopus, as has been demonstrated previously in several species of urodele Amphibia, histone genes lie at the sphere organizer loci of the lampbrush chromosomes. They were located by in situ hybridization of a 3H-labelled histone H4 anti-sense cRNA probe applied to lampbrush preparations in which transcript RNA had been retained, and likewise to preparations in which transcripts were absent but whose DNA had been denatured prior to hybridization. In Xenopus the histone genes lie in intimate association with the spheres that are attached to the lampbrush chromosomes, but they are absent from spheres that lie free in the germinal vesicle. The Anura separated from the Urodela several hundred million years ago, so the sphere organizer/histone gene association is of great antiquity. This suggests that the association has a functional significance, though it is one that has yet to be discovered.

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PubMed1991

Small nuclear ribonucleoproteins and heterogeneous nuclear ribonucleoproteins in the amphibian germinal vesicle: loops, spheres, and snurposomes.

We have examined the distribution of snRNPs in the germinal vesicle (GV) of frogs and salamanders by immunofluorescent staining and in situ nucleic acid hybridization. The major snRNAs involved in pre-mRNA splicing (U1, U2, U4, U5, and U6) occur together in nearly all loops of the lampbrush chromosomes, and in hundreds to thousands of small granules (1-4 microns diameter) suspended in the nucleoplasm. The loops and granules also contain several antigens that are regularly associated with snRNAs or spliceosomes (the Sm antigen, U1- and U2-specific antigens, and the splicing factor SC35). A second type of granule, often distinguishable by morphology, contains only U1 snRNA and associated antigens. We propose the term "snurposome" to describe the granules that contain snRNPs ("snurps"). Those that contain only U1 snRNA are A snurposomes, whereas those that contain all the splicing snRNAs are B snurposomes. GVs contain a third type of snRNP granule, which we call the C snurposome. C snurposomes range in size from less than 1 micron to giant structures greater than 20 microns in diameter. Usually, although not invariably, they have B snurposomes on their surface. They may also contain from one to hundreds of inclusions. Because of their remarkably spherical shape, C snurposomes with their associated B snurposomes have long been referred to as spheres or sphere organelles. Most spheres are free in the nucleoplasm, but a few are attached to chromosomes at specific chromosome loci, the sphere organizers (SOs). The relationship of sphere organelles to other snRNP-containing structures in the GV is obscure. We show by immunofluorescent staining that the lampbrush loops and B snurposomes also react with antibodies against heterogeneous nuclear ribonucleoproteins (hnRNPs). Transcription units on the loops are uniformly stained by anti-hnRNP and anti-snRNP antibodies, suggesting that nascent transcripts are associated with hnRNPs and snRNPs along their entire length, perhaps in the form of a unitary hnRNP/snRNP particle. That B snurposomes contain so many components involved in pre-mRNA packaging and processing suggests that they may serve as sites for assembly and storage of hnRNP/snRNP complexes destined for transport to the nascent transcripts on the lampbrush chromosome loops.

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PubMed1989

The sphere organelle contains small nuclear ribonucleoproteins.

We show by immunofluorescence microscopy of amphibian oocyte nuclei that small nuclear ribonucleoproteins (snRNPs) occur in lampbrush chromosome loops, in a few dozen extrachromosomal organelles previously described as "spheres," and in thousands of smaller granules. Spheres are variable in size (up to approximately 20 microns in diameter in the newt Notophthalmus and approximately 10 microns in the frog Xenopus) and are easily distinguishable from nucleoli by morphology and composition. Spheres occur both free in the nucleoplasm and attached to specific chromosome loci, the sphere organizers. Oocyte nuclei of a cricket and a spider contain essentially similar organelles, suggesting that spheres may be common throughout the animal kingdom. We suggest that spheres play a role in the assembly of snRNP complexes for the nucleus comparable to the way that nucleoli assemble ribosomal RNP complexes for the cytoplasm.

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PubMed1987

The lampbrush chromosomes of Xenopus laevis: preparation, identification, and distribution of 5S DNA sequences.

Details are given of a technique for making permanent preparations of the lampbrush chromosomes of Xenopus laevis. Stained preparations allow all 18 bivalent chromosomes to be identified, and a working map showing the major features has been constructed. Fifteen of the Xenopus chromosomes have one telomere conspicuously larger than the other; the two smallest chromosomes, and one other, lack large telomeres. Similar preparations, extracted with RNase and denatured, have been hybridized in situ with a 3H-labelled 5S cRNA probe. Chromosomes can be identified in the resulting autoradiographs. 5S DNA sequences are present at all the larger telomeres and at three of the smaller ones, but are absent from the telomeres at both ends of the two smallest chromosomes. There are also five interstitial sites of hybridization. At one of these, label is on the chromosome axis; at the other four, label extends well away from the axis.

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PubMed1982

The Croonian Lecture, 1981. Lampbrush chromosomes.

Lampbrush chromosomes were first observed nearly 100 years ago, and this lecture attempts a historical survey of what has been learnt from their study, particularly that over the past 30 years. There have been many controversies concerning the structure and functional significance of lampbrush chromosomes, and although their general structural layout has now, after several misconceptions, been firmly established their functional significance remains controversial. Research on lampbrush chromosomes played a significant part in establishing that chromatids in the germ lines of eukaryotic organisms are unineme in regard to DNA, and thereby exposed the C-value paradox. It also helped to establish that a DNA duplex is continuous throughout the length of a chromatid, but that the DNA/histone complex is at intervals reflected back on itself to form lateral loops. This organization, at one time thought to be a special feature of lampbrush chromosomes, now appears to be widespread in chromosomes undergoing compaction. However, despite attempts to determine the sequence organization of those portions of the DNA that are transcribed by lampbrush chromosomes, the function of these transcripts remains an open question, and the C-value paradox is still unresolved.

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PubMed1980

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

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.

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PubMed1979

An experimental analysis of bivalent interlocking in spermatocytes of the new Triturus vulgaris.

Interlocked bivalents at 1st meiotic metaphase are relatively uncommon in spermatocytes of the newt Triturus vulgaris, but their frequency of occurrence can be significantly increased by subjecting newts to a 24-h heat shock. Newt spermatocytes are sensitive to a heart shock at any stage between the end of premeiotic S and mid to late pachytene. The heat shock does not cause evidence desynapsis, nor does it significantly affect chiasma frequency; therefore the interlocked condition induced in spermatocytes which were subjected to a heat shock when they were in zygotene or pachytene is unlikely to be a consequence of synaptic trapping. By way of explanation it is suggested that a heat shock may cause telomers to detach from the nuclear membrane, or from the synaptonemal complex where the latter is attached to the membrane, thus allowing non-homologous chromonemata to become interwined before chiasmata have formed. If this explanation is valid, it is then further suggested that the recombination process which results in chiasma formation probably takes place in chromosomal regions lying outside the synaptonemal complex, rather than inside, between its 2 lateral elements.

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PubMed1978

The organization of transcription on lampbrush chromosomes.

The meiotic lampbrush chromosomes of amphibian oocytes display readily distinguishable regions of transcription (lateral loops) which extend from axial condensates of chromatin (chromomeres). The chromomeres contain most of the chromosomal ENA which, along with histone, is tightly compacted as regular arrays of DNP. Many RNA transcripts are generated on the lateral loops, and heterogeneous non-histone proteins associate with these transcripts, forming periodic condensates of 20--30 nm ribonucleoprotein (RNP) particles. These unit particles aggregate in various ways and to varying degrees and thereby confer distinctive gross morphologies to particular loops. There are about 10(4) lateral loops per haploid complement of newt chromosomes and this figure is similar to the experimentally derived number of different messenger RNA sequences found in oocytes. From cytological and biochemical studies it is now possible to consider individual lateral loops from various aspects: as morphologically distinct units; as units of inheritance; as units of functional activity; as units of transcription; as units of transcribed repetitive sequences; and as units containing one coding sequence. The difficulties in arriving at a simple explanation of the organization of transcription in lampbrush chromosomes are discussed.

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PubMed1978

RNA transcription on the giant lateral loops of the lampbrush chromosomes of the American newt Notophthalmus viridescens.

By counting silver grains in autoradiographs of lampbush chromosomes the rates of incorporation of [3H]adenine, [3H]cytidine, [3H]guanosine and [3H]uridine, administered separately, into RNA on the giant loops of chromosome II of Notophthalmus viridescens, were compared with the rates of incorporation of these same precursors into RNA on other, unidentified loops. The overall rate of RNA transcription on the giant loops is only about half that on the generality of other loops, and the RNA transcribed on the giant loops is computed to have a base ratio of approximately 25 A:39 C:9 G:27 U, implying that there must be about 4 times as many guanine residues on the transcribed as on the non-transcribed strand of the giant loops' DNA.

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PubMed1977

Recombination in male and female meiocytes contrasted.

For technical reasons studies of chiasma frequency and distribution, and hence of intrachromosomal recombination, have mostly been confined to male meiosis. However, there is now sufficient comparative data on male and female meiosis, in both plants and animals, to show that the extent of intra-chromosomal recombination in some organisms may be much the same on the female as on the male side, whereas other organisms show extreme sexual divergence in this regard. The evolutionary significance of such diversity remains enigmatic.

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PubMed1977

Inversion heterozygosity in females of the newt Notophthalmus viridescens and its influence on chiasma distribution.

In a wild population of the American newt Notophthalmus viridescens 15 females out of a total of 94 were found to be heterozygous for a paracentric inversion which includes almost the whole of the longer arm of the smallest chromosome (XI). The inversion was recognized in preparations of lampbrush chromosomes because it transfers the sequential loops, which normally lie close to the telomere, to a position neighbouring the centromere. Because of inversion the transcriptional polarity of the sequential loops is reversed vis-à--vis the chromosome as a whole. In normal bivalents XI (both in male and female meiosis) each arm pair generally forms a single chiasma close to the telomeres (proterminal localization). In bivalents XI heterozygous for the inversion no chiasmata are formed between the mutually inverted longer arm pairs, presumably because they fail to synapse, but chiasma frequency in the non-inverted shorter arm pairs is increased, and the normal restraint on chiasma distribution in this arm pair is lifted. An explanation is offered in terms of the availability of recombination nodules, and the time of their association with the synaptonemal complex.

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