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Biomedical subjects

M J Moses

Publications and source records attributed to M J Moses.

43 records · Page 3Linked to original sources

Assembly of microtubules onto kinetochores of isolated mitotic chromosomes of HeLa cells.

The kinetochores of isolated HeLa cell chromosomes attached to an electron microscope specimen grid, fixed in formaldehyde, and stained with alcoholic phosphotungstic acid are visible as dark, preferentially stained structures distinct from the chromatin with which they are associated. When unfixed chromosomes are immobilized by attachment to grids and incubated with chick brain tubulin, microtubules are observed to assemble onto the kinetochores. This demonstrates the competence of kinetochores in isolated chromosomes to act in vitro as microtubule assembly sites and suggests that they also possess this capacity in vivo. In addition, the results provide a possible means for isolating and characterizing kinetochores.

Chromosomes↗

The structure of the central region in the synaptonemal complexes of hamster and cricket spermatocytes.

The fine structure of bivalents from golden hamster and house cricket spermatocytes has been studied with a whole mount surface-spreading method combined with negative staining. The elements of the synaptonemal complex show detail of structure which is absent in other preparative procedures. The transverse filaments found in the central region of the synaptonemal complex from both species are straight and have a similar width, 1 6-1 8 nm These filaments occur mainly in bundles The central element differs in architecture in the two species In hamster bivalents it is formed of longitudinal stretches of filaments 1.6-1 8 nm wide and a small amount of an amorphous material similar to that of the lateral elements In the cricket, the central element contains transverse fibrils which are continuous with the transverse filaments of the central region, and an amorphous material lying mainly along the sides of the central element All of the components of the central region of the synaptonemal complex are resistant to pancreatic DNase. The overlapping ends of the transverse filaments, together with additional protein material, make up the central element The widespread occurrence and close morphological and histochemical interspecies similarities of the transverse filaments indicate that they serve an essential role, probably one concerned with holding synapsed bivalents together via the lateral elements. Restrictions placed by the observations reported here on current models of the synaptonemal complex are discussed.

Animals↗

Protein synthesis in pancreatic beta cells of the normal and diabetic Egyptian sand rat (Psammomys obesus). An autoradiographic study with the electron microscope.

The pattern of protein synthesis was studied in the pancreatic beta cells of the Egyptian sand rat (Psammomys obesus). When fed a standard Purina Laboratory Chow diet instead of a leafy vegetable diet, these animals develop the characteristic signs of diabetes mellitus. Tritiated leucine was injected intravenously into pairs of sand rats (one on a vegetable diet and one on a Purina Laboratory Chow diet). Two pairs of animals were sacrificed at 5-, 20- and 60-minute intervals, and pancreatic tissue was studied by electron microscopic autoradiography. At 5 minutes, the relative grain density was greatest over the rough endoplasmic reticulum; at 20 minutes it was greatest over the Golgi complex and at 60 minutes, over the granules. There were no statistically significant differences in the relative grain densities over the rough endoplasmic reticulum, over the Golgi complex or over the secretion granules between the sand rats on the vegetable diet and Chow diet. These results show that in the early phase of the development of diabetes mellitus, the pattern of protein synthesis in the beta cells of the normal and diabetic sand rat compares with that of other endocrine glands. The tritiated leucine was apparently incorporated into the newly synthesized secretory product in the rough endoplasmic reticulum during the first 5 minutes. The formed product migrated to the Golgi complex at 20 minutes, and at 1 hour was seen mainly over the light granules. In addition, there was no obvious difference in this pattern of protein synthesis between the normal and diabetic sand rats. This suggests that the secretory product, considered to be mainly insulin, is produced in the usual or in increased amounts, but it is not fully utilized by the diabetic animal and remains in circulation, thus increasing the plasma insulin level.

Animal Feed↗

Ultrastructure and cytochemistry of metabolic DNA in Tipula.

A DNA body is present in the females of the fly Tipula oleracea and is formed in contact with the sex chromosomes in the oogonial interphases. At each oogonial mitosis, the DNA body follows the chromosomes to one anaphase group and is included in one of the telophase nuclei. The body increases appreciably in size during the interphase of meiosis. All oocytes have the body, but only a few nurse cells possess it. The DNA body synthesizes its DNA at a different time than the chromosomes, as is shown by incorporation of tritiated thymidine, and contains 59% of the DNA of the nucleus, as is disclosed by spectrophotometric measurements. At late diplotene the DNA body disintegrates, releasing its DNA into either the nucleus or the cytoplasm. When studied in the electron microscope, the DNA body appears composed of a tight mass of intertwined fibrils. Demonstration that the main mass of the body is composed of DNA is obtained from cytochemical tests which reveal that the DNA body is Feulgen positive, stains green with azure B, incorporates H(3)-thymidine, and after digestion with DNase is Feulgen negative. The DNA of the body is complexed with histone, like the DNA of the chromosomes, as is revealed by an intense alkaline fast green staining. Electron microscope examination of oocytes reveals that one side of the DNA body is in close contact with the nuclear envelope and that the other side possesses an outer shell composed mainly of particles 150 to 250 A in diameter. Between the outer shell and the chromosomes there is a band of low electron opacity, 4000 to 7000 A thick. In the light microscope, this light band together with the outer shell is Feulgen negative and stains violet with azure B; this is confirmation of the presence of RNA. In the oocytes the nucleoli are found inside the DNA body. These nucleoli have a nucleolonema composed mainly of particles 150 to 250 A. The nucleoli are Feulgen negative, alkaline fast green negative, stain violet with azure B, and do not stain with azure B after RNase digestion, thus confirming their RNA content. The presence of the nucleoli inside the DNA body and of a band of RNA between the body and the chromosomes is indicative of a high RNA synthetic activity. Since the DNA of the body is complexed with histone, as in the chromosomes, and the nucleoli are located inside the body, the simplest interpretation of the DNA body is that it represents hundreds of copies of the operons of the nucleolar organizing region or neighboring regions. The situation found in Tipula has several basic features in common with the polytene chromosomes of other Diptera and with the hundreds of nucleoli present in Triturus oocytes. In all three cases, genes seem to be copied hundreds of times but are kept in different types of packages. A DNA body like the one in Tipula oleracea is found in other species of Diptera and in the Coleoptera. There is no indication, from the present investigation, that the DNA body is in any way associated with a virus.

Cell Division↗