The factors responsible for producing some degree of order to the arrangement of chromatin in the nucleus are reviewed. They are the following: 1. Chromosomes are attached to the nuclear membrane, nucleolus and intranuclear matrix. As a result they have a relatively fixed position in the nucleus. 2. In some species somatic pairing results in alignment of homologs. This is rare in mammals. 3. The association of ribosomal DNA and 5S DNA with the nucleolous results in the close approximation of the chromosomes carrying these DNA sequences. In man and other animals the most obvious consequence is satellite association. 4. Heterochromatin is condensed onto the inner nuclear membrane and periphery of the nucleolous while genetically active chromatin occupies the more central portion of the nucleus. The results is a peripheral location of late replicating DNA and a central location of early relicating DNA. 5. The DNA replication points tend to be associated with the nuclear matrix. Autoradiography of briefly labelled cells shows a high frequency of grains associated with nuclear matrix material. 6. Heterochromatin association results in chromocenters and ectopic pairing. 7. In addition to all these is the Rabl orientation or alignment of centromeres with centromeres and telomeres with telomeres. This polarization of the chromosomes results from the traction on the centromeres by the spindle fibers. There is no firm evidence for any higher degrees of order that might bring specific functioning genes into close proximity.

Arrangement of chromatin in the nucleus

Using the Counce-Meyer spreading technique, in over 70 spermatocytes it was possible consistently to obtain whole, flattened nuclei containing complete sets of pachytene SCs. The SCs are visible in both the phase and electron microscopes. Each SC is morphologically intact, preferentially stained, and attached to the nuclear envelope by a dense, terminal plaque. It is thus possible to trace each SC for its entire length. Also, a structure representing the kinetochore is clearly visible in each autosomal SC. Karyotypes comparable to the somatic karyotype can be constructed by arranging SCs according to length and kinetochore position. The observed regularity of SC morphology implies structural stability sufficient to withstand the stresses imposed by the procedure.— A coarse network of closely packed nuclear annuli connecting SC attachment plaques often provides end-to-end associations and may tend to immobilize SCs during processing.— Three kinds of perturbation of SC structure are encountered. Twists in the SC frequently occur, but no regular pattern or correspondence with chiasma distribution is observed. SCs occasionally hook around each other without disruption, but in two instances the unpaired axis of the X apparently was interlocked within an autosomal SC. Stretching of the SC is infrequent; it is conspicuous when it occurs and is usually associated with other obvious distortions of the nucleus.— Distinctive morphologies of the X and Y chromosomes facilitate their identification in all preparations. — During zygotene, autosomal synapsis, i.e., the formation of SCs from the pairing of single axial elements, initiates at distal ends and terminates at the kinetochore region; neither initiation nor termination is synchronous among all autosomes.

Synaptonemal complex karyotyping in spermatocytes of the Chinese hamster (Cricetulus griseus). I. Morphology of the autosomal complement in spread preparations.

Biochemical and genetic studies have produced a vast fund of knowledge concerning gene action and regulation in prokaryotes In these organisms the DNA is exposed rather nakedly to the world, protected primarily by the cell membrane In eukaryotes the DNA seems far better shielded, being enmeshed in histone and nonhistone proteins and sequestered behind both the cell and the nuclear membrane These differences have led to a considerable degree of caution in the application of this knowledge of prokaryotes to problems of gene regulation in eukaryotes, and rightly so There are, however, several observations which suggest that higher organisms may have picked up a number of fundamental genetic tricks from their lowly predecessors It has frequently been suggested that eukaryotes must do things differently from prokaryotes, until proven otherwise It may be prudent to reverse this line of thought and suggest that they do things the came until proven different The following similarities suggest this (1) T he basic genetic dogmas concerning DNA replication, transcription, andlation are similar, (2) The genetic code is the same (3) Both systems appear to make use of cyclic AMP as a basic mediator for, humoral or diffusible, signals (4) In both systems DNA synthesis may be controlled at membranes (5) Both make use of different types of RNA polymerase and RNA polymerase cofactors (6) Recent studies of polylysine binding to chromatin suggest the eukaryotic DNA may not be so thoroughly enmeshed in Protein as once thought (7) The visualization of genes in action by electron microscopic techniques intimates that genes are spaced and read in a similar manner And finally, (8) merely because the clustering of related genes is unusual in higher organisms is no reason in itself to totally discard the promoter-operator-repressor concept as a way of regulating single structural genes This system has provided an immense amount of data concerning the manner in which proteins interact with specific DNA sequences to control the attachment and utilization of RNA polymerase It is hard to imagine that eukaryotes, being presented with such a superb mechanism for controlling DNA transcription, would totally discard it and opt for something different It is far more likely that they would build on to this solid foundation

The structure and function of chromatin.

Chromosomal arrangement in the interphase nucleus has two main aspects: (1) arrangement of chromosomes with respect to nuclear polarity and to other nuclear components, and (2) arrangement of chromosomes with respect to one another The latter aspect consists of two main types of spatial relationships; (a) relationships between different members of one chromosomal set, (b) relationships between different chromosomal sets Data concerning various aspects of chromosomal arrangement in the interphase nucleus are presented and discussed and the genetic control as well as subcellular mechanisms which are involvled in nuclear organization, are elucidated Evidence is presented indicating that, in common wheat, the gene system that determines the specific pattern of chromosomal arrangement in the nucleus is operating via the microtubular elements of the spindle system The significance of ordered arrangement of chromosomes in the nucleus for the regularity of genetic activity and chromosomal behavior, is pointed out

Arrangement of chromosomes in the interphase nucleus of plants

SYNOPSIS

Micronuclear changes of variety 1 of Tetrahymena pyriformis during meiotic prophase have been observed by the light microscope. Morphologic changes in the micronucleus are divided into 6 stages. In stage I, chromatin begins to polarize; in stage II, the micronucleus becomes spindle shaped; and in stage III, one end of the micronucleus protrudes to form a “neck.” In stage IV, where the micronucleus elongates to maximal length, the whole micronucleus consists of 2 chromatin threads pairing longitudinally. One thread probably contains one genome. In stage V, the elongated thread becomes shorter and thicker. Finally, in stage VI, separate chromosomes appear and enter into metaphase.



To discover the role of the elongation of the micronucleus, called crescent formation, autoradiographic analysis of RNA and DNA synthesis were undertaken using [3H]uridine and [3H]thymidine. Pulse label and chase experiments show that the crescent in stages II and III is actively synthesizing RNA. Though no remarkable DNA synthesis was observed during meiosis, a small amount of DNA synthesis occurred during the 1st and 2nd prezygotic divisions.

Cytologic and autoradiographic studies of the micronucleus at meiotic prophase in Tetrahymena pyriformis.

In an exceptionally well-defined first cleavage metaphase in a sectioned egg of Polychoerus carmelensis all 34 (2n) chromosomes were present in a single 8-micron section, and it was possible to identify the homologous chromosomes derived from the two parent gametes. Three groups of chromosomes from one parent, containing 7, 7, and 3 chromosomes, respectively, could be exactly matched by corresponding groups of homologues derived from the other gamete. The probability of such an ordered pattern occurring by chance is somewhat less than 1 in 248. The simplest explanation of this arrangement is that the 17 chromosomes contributed by the egg were in precisely the same linear order as those contributed by the spermatozoon. This suggests that during certain stages preceding first cleavage metaphase, the 17 members of each haploid set of chromosomes may have been attached, end-to-end, in a linear fashion, by highly specific bonds.

http://www.pnas.org/content/67/4/1951.full.pdf

Donald P. Costello

Papers

Identical Linear Order of Chromosomes in Both Gametes of the Acoel Turbellarian Polychoerus carmelensis: A Preliminary Note