scholarly journals The Relation between the Axial Complex of Meiotic Prophase Chromosomes and Chromosome Pairing in a Salamander (Plethodon cinereus)

1958 ◽  
Vol 4 (5) ◽  
pp. 633-638 ◽  
Author(s):  
Montrose J. Moses

An investigation of the structure of meiotic chromosomes from primary spermatocytes of two salamanders, Plethodon cinereus and Desmognathus fusca, has been made using correlated light and electron microscopy. Feulgen squashes were compared with stained sections and these related to adjacent thin sections in the electron microscope. A transition from the familiar cytological preparation to the electron image was thus effected. A linear complex consisting of three parallel strands has been observed with the electron microscope, passing along the central axis of primary spermatocyte chromosomes. The complex is similar to that found in comparable chromosomes from at least a dozen animal species. The structure in Plethodon is described in detail. Synapsis has been positively identified as the stage of meiotic prophase at which the complex occurs. Thus the complex is a part of bivalent chromosomes. It has not been seen in other stages or other divisions and is thus thought to be exclusively of synaptic occurrence. The term synaptinemal complex is suggested for the entire structure. By virtue of the material condensed around it, the complex is also seen in the light microscope where it appears as a fine, densely Feulgen-positive central core along the chromosome. The complex is thus closely associated with DNA, if not at least in part, composed of it. In the stages studied, homologous chromosomes are not always completely paired. The lateral elements of the complex separate and follow the single chromosome axes at these points. The central element disappears and thus may be a phenomenon of pairing. It is concluded that the lateral elements of the synaptinemal complex may more correctly be a "core" of the single meiotic prophase chromosome, possibly being concerned with its linear organization.

1964 ◽  
Vol 23 (1) ◽  
pp. 63-78 ◽  
Author(s):  
James R. Coleman ◽  
Montrose J. Moses

The indium trichloride method of Watson and Aldridge (38) for staining nucleic acids for electron microscopy was employed to study the relationship of DNA to the structure of the synaptinemal complex in meiotic prophase chromosomes of the domestic rooster. The selectivity of the method was demonstrated in untreated and DNase-digested testis material by comparing the distribution of indium staining in the electron microscope to Feulgen staining and ultraviolet absorption in thicker sections seen with the light microscope. Following staining by indium, DNA was found mainly in the microfibril component of the synaptinemal complex. When DNA was known to have been removed from aldehyde-fixed material by digestion with DNase, indium stainability was also lost. However, staining of the digested material with non-selective heavy metal techniques demonstrated the presence of material other than DNA in the microfibrils and showed that little alteration in appearance of the chromosome resulted from DNA removal. The two dense lateral axial elements of the synaptinemal complex, but not the central one to any extent, also contained DNA, together with non-DNA material.


1970 ◽  
Vol 6 (3) ◽  
pp. 655-667
Author(s):  
L. F. LA COUR ◽  
B. WELLS

The 1-4 chromocentres seen in nuclei of Fritillaria lanceolata, which derive from fusion of heterochromatic segments situated proximal to the centromere in all but two of the 24 chromosomes, were studied with the electron microscope in thin sections of pollen mother cells at zygotene and pachytene, in respect of the synaptinemal complex. Prophase stages of meiosis in two plants were also surveyed briefly with the light microscope. The latter observations revealed that the timing of the separation of heterochromatic segments from chromocentres is genetically controlled. In one plant the segments were still contained in chromocentres at pachytene, whereas in the other they were free at zygotene. At this time they could be identified by a near-surface position in the nucleus and an even condensation concomitant with an absence of chromomeres. In thin section, the fine structure of the chromocentres in zygotene nuclei was distinctive in that the chromatin fibrils were less condensed and more widely dispersed than those in euchromatic regions. The fibrillar network was also interspersed with ‘clear areas’ or channels. After further chromosome condensation, the condensation of fibrils in the chromocentres became equivalent at pachytene to those in euchromatic regions. Synaptinemal complexes were seen at zygotene and pachytene both in euchromatic regions and chromocentres. Their presence in the chromocentres signifies that homologous chromosomes must have been closely paired in regions extending from the centromeres to the distal ends of the heterochromatic segments already at telophase of the last pre-meiotic mitosis. Configurations involving entangled pairs of axial cores, peculiar to zygotene and chromocentres and parts of euchromatic regions proximal to them, are interpreted as resulting from restricted movement.


1977 ◽  
Vol 74 (3) ◽  
pp. 794-815 ◽  
Author(s):  
J A Schloss ◽  
A Milsted ◽  
R D Goldman

Fluorescein-labeled heavy meromyosin subfragment-1 (F-S-1) has been purified by ion exchange chromatography and characterized in terms of its ability to bind specifically to actin. F-S-1 activates the Mg++-adenosine triphosphatase activity of rabbit skeletal muscle actin and decorates actin as shown by negative stains and thin sections of rabbit actin and rat embryo cell microfilament bundles, respectively. Binding of F-S-1 to cellular structures is prevented by pyrophosphate and by competition with excess unlabeled S-1. The F-S-1 is used in light microscope studies to determine the distribution of actin-containing structures in wnterphase and mitotic rat embryo and rat kangaroo cells. Interphase cells display the familiar pattern of fluorescent stress fibers. Chromosome-to-pole fibers are fluorescent in mitotic cells. The glycerol extraction procedures employed provide an opportunity to examine cells prepared in an identical manner by light and electron microscopy. The latter technique reveals that actin-like microfilaments are identifiable in spindles of glycerinated cells before and after addition of S-1 or HMM. In some cases, microfilaments appear to be closely associated with spindle microtubles. Comparison of the light and electron microscope results aids in the evaluation of the fluorescent myosin fragment technique and provides further evidence for possible structural and functional roles of actin in the mitotic apparatus.


Author(s):  
Robert L. Corbett

The ammoniacal silver stain for light microscopy of plastic embedded sections has been adapted for use in electron microscopy. Since the silver will stain even very thin sections, i.e., silver and gold for light microscopy, and silver deposits are sufficiently electron dense to be seen in the electron microscope, the results are very useful for correlating light and electron microscopy. Compared to the conventional stains for electron microscopy which usually take over one-half hour, the silver procedure can be done in five minutes or less and thus provides a quick look at sections This stain has more contrast, so it is especially good for low power electron microscopy. The ability of the silver to stain very thin sections enables a correlation between light and electron microscopy in three ways. First, thin sections can be stained with silver on a glass slide and compared with immediately adjacent thin sections on grids stained the usual way for electron microscopy.


1974 ◽  
Vol 16 (2) ◽  
pp. 261-299 ◽  
Author(s):  
H. G. DAVIES ◽  
A. B. MURRAY ◽  
M. E. WALMSLEY

Previously it was shown that when condensed chromatin from several different types of cell is stained with uranyl-lead and examined in thin sections in the electron microscope, the stain is distributed into a dot-dash pattern arising from threads, with lesser-staining intermediate areas. We now show that when a section through chicken erythrocyte chromatin is stained with ethanolic phosphotungstic acid (PTA) the stain distribution is homogeneous. This shows that the lesser-staining regions after uranyl-lead, cannot be an overlap artifact. We conclude that the stains and hence the molecules in chromatin are distributed between 2 phases, an o- and an e-phase, so called because the structural units in chromatin are arranged in an orderly way at the surface of the nucleus and give rise to oddly (o) and evenly (e) numbered bands. Measurements of electron density per unit thickness, proportional to the number of stain molecules per unit volume, are made in thin sections through erythrocytes and reticulocytes from adult hen, 4-day-old chicks and 17-day embryos. The results indicate differences in the packing of the molecules in chromatin and further show that the e-phase is quite likely to have a higher DNA to protein ratio than the o-phase. After uranyl-lead stain the visibility of the dot-dash pattern in cells from adult hen is relatively low due, we propose, to closer packing. In micrographs through condensed chromatin treated with uranyl-lead the eye selects out only the densely stained dots and dashes, width 17 nm. When erythrocyte chromatin is partially or completely disrupted in various ways, threads 25-30 nm then become visible. We propose that condensed chromatin in intact cells contains structural units which consist of a central element, width 17 nm previously referred to as the unit thread, forming the e-phase, surrounded by a cylindrical shell forming the o-phase. This socalled superunit thread is similar in width, about 25-30 nm, to that reported by other workers in preparations of chromosomes spread on water surfaces. The hypothesis therefore helps explain what appeared to be discrepancies in thread dimensions. Certain other ultrastructural features of erythrocyte nuclei are also reported which are either pertinent to the general aim of this study, namely the way in which nucleoproteins fold up in chromosomes, or to biochemical studies, to be reported shortly, in which attempts are made to locate the proteins removed from isolated erythrocyte nuclei during subsequent washing in salt solutions.


Author(s):  
E. B. Masurovsky ◽  
H. H. Benitez ◽  
M. R. Murray

Recent light- and electron microscope studies concerned with the effects of D2O on the development of chick sympathetic ganglia in long-term, organized culture revealed the presence of rod-like fibrillar formations, and associated granulofibrillar bodies, in the nuclei of control and deuterated neurons. Similar fibrillar formations have been reported in the nuclei of certain mammalian CNS neurons; however, related granulofibrillar bodies have not been previously described. Both kinds of intranuclear structures are observed in cultures fixed either in veronal acetate-buffered 2%OsO4 (pH 7. 4), or in 3.5% glutaraldehyde followed by post-osmication. Thin sections from such Epon-embedded cultures were stained with ethanolic uranyl acetate and basic lead citrate for viewing in the electron microscope.


Author(s):  
Roberta M. Bruck

An unusual structure in the cochlea is the spiral limbus; this periosteal tissue consists of stellate fibroblasts and collagenous fibers embedded in a translucent ground substance. The collagenous fibers are arranged in vertical columns (the auditory teeth of Haschke). Between the auditory teeth are interdental furrows in which the interdental cells are situated. These epithelial cells supposedly secrete the tectorial membrane.The fine structure of interdental cells in the rat was reported by Iurato (1962). Since the mouse appears to be different, a description of the fine structure of mouse interdental cells' is presented. Young adult C57BL/6J mice were perfused intervascularly with 1% paraformaldehyde/ 1.25% glutaraldehyde in .1M phosphate buffer (pH7.2-7.4). Intact cochlea were decalcified in .1M EDTA by the method of Baird (1967), postosmicated, dehydrated, and embedded in Araldite. Thin sections stained with uranyl acetate and lead citrate were examined in a Phillips EM-200 electron microscope.


Author(s):  
J. Temple Black

Since its introduction by Fernandez-Moran, the diamond knife has gained wide spread usage as a common material for cutting of thin sections of biological and metallic materials into thin films for examination in the transmission electron microscope. With the development of high voltage E.M. and scanning transmission E.M., microtomy applications will become increasingly important in the preparation of specimens. For those who can afford it, the diamond knife will thus continue to be an important tool to accomplish this effort until a cheaper but equally strong and sharp tool is found to replace the diamond, glass not withstanding.In Figs. 1 thru 3, a first attempt was made to examine the edge of a used (β=45°) diamond knife by means of the scanning electron microscope. Because diamond is conductive, first examination was tried without any coating of the diamond. However, the contamination at the edge caused severe charging during imaging. Next, a thin layer of carbon was deposited but charging was still extensive at high magnification - high voltage settings. Finally, the knife was given a light coating of gold-palladium which eliminated the charging and allowed high magnification micrographs to be made with reasonable resolution.


Author(s):  
T. Guha ◽  
A. Q. Siddiqui ◽  
P. F. Prentis

Tilapia, Oreochromis niloticus, is an economically important fish in Saudi Arabia. Elucidation of reproductive biology of this species is necessary for successful breeding program. In this paper we describe fine structure of testicular sperm cells in O, niloticus.Testes from young adult fish were fixed in gluteraldehyde (2%) and osmium tetroxide (1%), both in cacodyl ate buffer. Specimens were processed in the conventional way for electron microscopy and thin sections of tissues (obtained by cutting the blocks with a diamond knife) were stained by ura- nyl acetate and lead citrate. These were examined in a Carl Zeiss electron microscope operated at 40 kV to 60 kV. Sperm cells were obtained from testes by squeezing them in cacodyl ate buffer. They were fixed in gluteraldehyde (2%) in the same buffer, air dried, gold coated and then examined in a Philips scanning electron microscope (SEM) operated at 25kV.The spermatozoon of O. niloticus is consisting of head, midpiece and tail (Fig. 1).


Author(s):  
J. Edie

In TEM image formation, the observed contrast variations within thin sections result from differential electron scattering within microregions of varying mass thickness. It is possible to utilize these electron scattering properties to obtain objective information regarding various specimen parameters (1, 2, 3).A pragmatic, empirical approach is described which enables a microscopist to perform physical measurements of thickness of thin sections and estimates of local mass, volume, density and, possibly, molecular configurations within thin sections directly in the microscope. A Faraday cage monitors the transmitted electron beam and permits measurements of electron beam intensities.


Sign in / Sign up

Export Citation Format

Share Document