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Biomedical subjects

H Schraer

Publications and source records attributed to H Schraer.

At least 37 records · Page 2Linked to original sources

Effect of estrogen on rate of 59Fe uptake by hemopoietic tissue in Japanese quail.

Injection of 2 mg estradiol valerate into male Japanese quail caused the rate of 59Fe uptake by the skeleton to drop 70% within 48 h. This phenomenon was due to an increase in the iron-binding capacity of the plasma induced by estrogen that caused a substantial decline in the availability of the label for hematopoiesis and to partial inhibition of hematopoiesis by estrogen directly. The changes in 59Fe uptake induced by estrogen were accompanied by a significant drop in circulating red blood cell concentration and a substantial rise in plasma volume. These altered physiological conditions were similar to those normally found in the laying hen.

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Estrogen-induced sequential changes in avian bone metabolism.

Medullary bone was induced in male Japanese quail by administration of estradiol valerate. An increase in the organic weight of the femur was observed by 36 h after estrogen and an increase in ash weight was observed by 96 h. A complex sequence of metabolic changes in the femur occurred after estrogen treatment. A large increase in uptake of 3H-proline associated with enhanced collagen formation began 36 h after estrogen and reached a peak 3.5 times the control rate at 60 h. The onset of mineralization of the newly formed bone matrix, as determined by 45Ca uptake, began about 24 h after onset of collagen synthesis indicating that synthesis of the bone matrix and its mineralization occur sequentially. Large increases in the rates of uptake of 3H-uridine and 3H-thymidine occurred prior to the onset of medullary bone formation and appear to reflect the differentiation of osteogenic precursors to bone-forming cells.

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Matrix vesicles in newly synthesizing bone observed after ultracryotomy and ultramicroincineration.

Matrix vesicles were observed in femurs of 8-day-old chick embryos prepared by ultracryotomy. Some of the sections were subjected to ultramicroincineration. The unfixed tissues never came into contact with solutions, and thereby artifacts due to dissolution, redistribution, or rearrangement of the mineral constituents were avoided. In the osteoid, electron dense objects with the size and appearance of matrix vesicles were seen, although limiting membranes were not visible. After ultramicroincineration the vesicles were observed to contain small crystals and a less dense amorphous mineral material which may te the precursor of bone mineral. In addition, a ring of ash enclosed the crystalline and amorphous mineral and appeared to occupy the position of the vesicle membrane as seen in conventionally prepared material.

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Frozen thin-sections of rapidly forming bone: bone cell ultrastructure.

Frozen thin-sections of fresh medullary bone, which had been calcifying for 2 to 4 days, were prepared and examined in the electron microscope. The bone was obtained from male Japanese quail treated with estradiol valerate. Large numbers of 200-800 A electron dense granules, which consisted of 50-75 A subparticles, were seen within mitochondria. A population of electron-dense particles slightly smaller than ribosomes was also observed. Most granules were not present after conventional fixation, dehydration, embedding, sectioning and staining. Oother structures which were visible, also on the basis of their intrinsic electron density, were nuclei with regions which resembled condensed chromatin, ribosomes and rough endoplasmic reticulum. By the use of osmium tetroxide vapor staining, inner and outer membranes and cristae of mitochondria were delineated. Other membranous components were not detected. Since very little loss or dislocation of cell components can occur during the preparation of frozen thin-sections, the micrographs obtained may be a more accurate representation of cell ultrastructure.

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Ultrastructural observations of amorphous bone mineral in avian bone.

Mineral from medullary bone of three avian species was examined with the electron microscope in order to clarify the ultrastructure of amorphous bone mineral (ABM) in a mineralized tissue. Powders from freeze-dried bone revealed bone mineral with morphology and behavior identical to synthetic amorphous calcium phosphate (ACP). These powders exhibited spherically shaped particles 180--400 A in diameter with uniform electron density when viewed at low beam intensity. Thin sections of embedded freeze-dried bone also revealed spherically shaped particles 100--350 A in diameter with electron beam sensitivity characteristic of ACP. Regions of bone mineral with irregular outline and morphology were observed which closely resemble the discoidal form of synthetic ACP. More electron-dense spherical particles (150 A in diameter) could be seen budding from these regions. Some of these buds exhibited electronlucent centers characteristic of AMB. The inorganic nature of these features of bone mineral was confirmed using ultramicroincineration. Preliminary exploration of a freeze-substitution technique showed spherical bone mineral particles which were similar in morphology to those observed in freeze-dried samples. A limited degree of preservation of cellular material was observed using this freeze-substitution technique.

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Low temperature ultramicroincineration of thin-sectioned tissue.

Low temperature ultramicroincineration was employed to determine the morphological localization of "structure-bound" mineral and/or metallic elements within biological cells at the electron microscope level. This technique chemically removes organic material from thin sections of tissues with reactive, excited oxygen instead of heat as used in a furnace. The remaining ash representing the mineral/metallic ultrastructure of cells is advantageous for ultrastructural studies because incineration without applying heat is less destructive than the burning associated with high temperatures. This low temperature incineration method was applied to thin-sectioned avian shell gland mucosa, a calcium transporting system, as a sample tissue. The results include: recognition of many subcellular organelles in the ash patterns, identification of dense, ash-containing granules (possibly organic-metallic complexes) in epithelial cells which may be involved in calcium transport, description of ashed erythrocytes and collagen, comparison of ashed glutaraldehyde fixed tissue with and without osmium postfixation, description of lead-stained cells after ashing, demonstration that ash preservation is dependent upon section thickness, illustration of the fine resolution obtainable because the ash residues remain relatively near their in situ origins, discussion of technical problems in this relatively new field, and demonstration of the presence of Ca and P in the ash with electron microprobe X-ray analysis.

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The intracellular distribution of calcium in the mucosa of the avian shell gland.

The intracellular distribution of calcium has been studied in the mucosa of the avian shell gland, a tissue which transports large quantities of calcium during discrete time intervals. Ca(45) was administered to hens either in a single dose followed by sacrifice 5 min later or in repeated doses over an extended period followed by sacrifice 2 hr or 24 hr after the last injection. Subcellular fractions were isolated by differential centrifugation and analyzed for Ca(45). The Ca(45) was located principally in the particulate fractions; the concentration (CPM Ca(45)/mg N) was highest in the mitochondrial fraction. Comparisons of (1) the Ca(45) distribution in shell gland cells with that of liver cells, (2) the alterations which occur due to the phase of the egg laying cycle, (3) the effects due to the time elapsed since the last injection of Ca(45), and (4) the Ca(45) distribution of the short term experiments with that of the long term experiments revealed that the mitochondrial fraction of the shell gland appeared to be active in the movement of calcium. The microsomal fraction showed increased values in CPM Ca(45)/mg N when calcification was occurring, which may indicate that the subcellular components of this fraction have a role in calcium transport. The nuclear and supernatant fractions did not seem to be involved in the transport process. The implications of these results concerning the manner by which calcium may be controlled on a cellular level in this system are discussed.

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