scholarly journals COMPARTMENTALIZATION OF THE DEVELOPING MACRONUCLEUS FOLLOWING CONJUGATION IN STYLONYCHIA AND EUPLOTES

1970 ◽  
Vol 47 (2) ◽  
pp. 395-407 ◽  
Author(s):  
John A. Kloetzel

The development of the macronucleus following conjugation in the hypotrichous ciliates Euplotes and Stylonychia has been examined with the electron microscope. Banded polytene chromosomes can be seen in thin sections of the macronuclear anlagen during the early periods of exconjugant development. As the chromosomes reach their maximum state of polyteny, sheets of fibrous material appear between the chromosomes and transect the chromosomes in the interband regions. Individual bands of the polytene chromosomes thus appear to be isolated in separate compartments. Subsequently, during the stage when the bulk of the polytenic DNA is degraded (1), these compartments swell, resulting in a nucleus packed with thousands of separate spherical chambers. Individual chromosomes are no longer discernible. The anlagen retain this compartmentalized condition for several hours, at the end of which time aggregates of dense material form within many of the compartments. The partitioning layers disperse shortly before replication bands appear within the elongating anlagen, initiating the second period of DNA synthesis characteristic of macronuclear development in these hypotrichs. The evidence presented here suggests that the "chromatin granules" seen in the mature vegetative macronucleus represent the material of single bands of the polytene chromosomes seen during the earlier stages of macronuclear development. The possibility is also discussed that the degradation of DNA in the polytene chromosomes may be genetically selective, which would result in a somatic macronucleus with a different genetic constitution than that of the micronucleus from which it was derived.

1973 ◽  
Vol 13 (2) ◽  
pp. 479-509 ◽  
Author(s):  
K. G. MURTI

This report describes an ultrastructural investigation of macronuclear development following conjugation in Stylonychia mytilus (a spirotrichous ciliate) and Tetrahymena pyriformis (a holotrichous ciliate). In S. mytilus, polytene chromosomes are formed in the young macronucleus (macronuclear Anlage). They are subsequently broken between the bands by ‘membranous’ partitions; the assembly of the membranes appears to be concomitant with the formation of the polytene chromosomes. The membranes in the Anlage appear to originate from fibrous material seen in the early Anlage. This fibrous material in the earlier stages is seen concentrated at several points along the border of the inner nuclear membrane. In the later stages it is seen in the interior of the Anlage, outlining the developing polytene chromosomes. As the chromosomes reach the maximum degree of polyteny, the fibrous material condenses to acquire a membranous appearance and extends into the interband regions. The Anlage throughout this period shows a progressive increase in size. Subsequently, the membranes enclose individually each band plus portions of the 2 adjacent interband regions of the polytene chromosomes to form a large number of vesicles. After vesicle formation the Anlage shrinks, and the chromatin inside the vesicles shows degradative changes. Finally, the vesicles disappear, the membrane degradation products appear at the nuclear membrane, and the Anlage now contains nucleoli. The Anlage increases its DNA content by multiple rounds of replication to become a mature macronucleus. The ultra-structural changes observed in the Anlage support the idea of genetic diminution (i.e. extensive DNA synthesis, elimination of many DNA nucleotide sequences, and amplification of the remaining DNA nucleotide sequences in a second period of DNA synthesis) proposed earlier on the basis of cytochemical, biochemical, and limited electron-microscope studies. In T. pyriformis, the macronuclear development differs substantially from that of Stylonychia. Features such as the formation and degradation of polytene chromosomes are absent in the macronuclear development of Tetrahymena; the young macronucleus in this cell becomes a mature macronucleus by progressive increment in size and chromatin content with no apparent genetic diminution. These observations agree with cytochemical studies on the macronuclear development of Tetrahymena.


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.


Author(s):  
S. Q. Xiao ◽  
S. Baden ◽  
A. H. Heuer

The avian eggshell is one of the most rapidly mineralizing biological systems known. In situ, 5g of calcium carbonate are crystallized in less than 20 hrs to fabricate the shell. Although there have been much work about the formation of eggshells, controversy about the nucleation and growth mechanisms of the calcite crystals, and their texture in the eggshell, still remain unclear. In this report the microstructure and microchemistry of avian eggshells have been analyzed using transmission electron microscope (TEM) and energy dispersive spectroscopy (EDS).Fresh white and dry brown eggshells were broken and fixed in Karnosky's fixative (kaltitanden) for 2 hrs, then rinsed in distilled H2O. Small speckles of the eggshells were embedded in Spurr medium and thin sections were made ultramicrotome.The crystalline part of eggshells are composed of many small plate-like calcite grains, whose plate normals are approximately parallel to the shell surface. The sizes of the grains are about 0.3×0.3×1 μm3 (Fig.l). These grains are not as closely packed as man-made polycrystalline metals and ceramics, and small gaps between adjacent grains are visible indicating the absence of conventional grain boundaries.


Author(s):  
Julio Sepúlveda-Saavedra ◽  
Beatriz González-Corona ◽  
Víctor A. Tamez Rodríguez ◽  
Ma. Victoria Bermúdez de Rocha ◽  
Alfredo Piñeyro López

It has been shown in previous studies that the toxin T-514 isolated from K. humboldtiana induces severe damage to the lung in treated rodents. Histopathological findings include edema, and alveolar hemorrage. However, the ultraestructure of the lesion has not been investigated. In this study we used two species of rodents: Hamster and guinea pig, and a primate: Macaca fascicularis. Animals received different single dosis of the toxin via intraperitoneal. Control animals received only the vehicle (propylen glycol). Inmediately after spontaneous death, lung samples were fixed in Karnovsky-Ito fixative, post fixed in osmium tetroxide and embedded in epon. Thin sections were prepared with an Ultratome V LKB, stained with uranly acetate and lead citrate, and studied in an electron microscope Zeiss-EM109.


Author(s):  
W. P. Wergin ◽  
S. Roy ◽  
E. F. Erbe ◽  
C. A. Murphy ◽  
C. D. Pooley

Larvae of the nematode, Steinernema carpocapsae Weiser strain All, were cryofixed and freezesubstituted for 3 days in acetone containing 2% osmium tetroxide according to established procedures. Following chemical fixation, the nematodes were brought to room temperature, embedded in Spurr's medium and sectioned for observation with a Hitachi S-4100 field emission scanning electron microscope that was equipped with an Oxford CT 1500 Cryotrans System. Thin sections, about 80 nm thick, similar to those generally used in conventional transmission electron microscope (TEM) studies were mounted on copper grids and stained with uranyl acetate for 30 min and lead citrate for 5 min. Sections about 2 μm thick were also mounted and stained in a similar fashion. The grids were mounted on an Oxford grid holder, inserted into the microscope and onto a cryostage that was operated at ambient temperature. Thick and thin sections of the larvae were evaluated and photographed in the SEM at different accelerating voltages. Figs. 4 and 5 have undergone contrast conversion so that the images would resemble transmitted electron micrographs obtained with a TEM.


1953 ◽  
Vol 98 (2) ◽  
pp. 157-172 ◽  
Author(s):  
William H. Gaylord ◽  
Joseph L. Melnick

The intracellular development of three pox viruses has been studied with the electron microscope using thin sections of infected tissue. Cells infected with vaccinia, ectromelia, and molluscum contagiosum viruses all form developmental bodies preliminary to the production of mature virus. Developmental bodies, believed to be virus precursors, are round to oval, slightly larger than mature virus particles, less dense to electrons, and have a more varied morphology. It is suggested as a working hypothesis that the process of maturation of a virus particle takes place as follows. In the earliest form the developmental bodies appear as hollow spheres, imbedded in a very dense cytoplasmic mass constituting an inclusion body, or in a less dense matrix near the nucleus in cells without typical inclusion bodies. The spheres become filled with a homogeneous material of low electron density. A small, dense granule appears in each developmental body and grows in size at the expense of the low density material. Following growth of the granule, particles are found with the dimensions of mature virus and having complex internal structure resembling bars or dumbells. Mature virus is ovoid and very dense to electrons. An "empty" interior may be found within its thick walls.


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