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  • 1
    Electronic Resource
    Electronic Resource
    Transformations of osteoclast phenotype in ia rats cured of congenital osteopetrosis (1982)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 174 (1982), S. 141-147 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The reduced bone resorption characteristic of osteopetrosis is accompanied in the incisors-absent (ia) rat mutation by a significant increase in osteoclasts of inactive (mutant) phenotype. Restoration of bone resorption in ia rats by transfer of spleen cells from normal littermates is preceded by a transformation of osteoclasts from mutant to normal phenotype.In this investigation the proportions of osteoclasts of normal phenotype have been determined by light microscopy in untreated ia and normal rats and in ia rats treated with various cell populations from normal rats. Significant increases in numbers of osteoclasts of normal phenotype were seen in the mutant skeleton soon after cell treatments that eventually restored bone resorption and cured the disease. No changes in osteoclast phenotype were seen after cell transfers that did not cure the disease.These data establish transformation of osteoclast phenotype as an early event in the recovery from osteopetrosis and suggest that determination of osteoclast phenotype is a reliable predictor of the success of normal cell populations to restore bone resorption in this mutation.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1051740203
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  • 2
    Electronic Resource
    Electronic Resource
    The effects of lectins on the interaction between macrophages and bone in vitro (1985)
    Springer
    Cell & tissue research 241 (1985), S. 103-109 
    Details
    ISSN: 1432-0878
    Keywords: Macrophages ; Bone ; Lectins ; Tissue culture ; Oligosaccharides ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Recent studies have demonstrated that the attachment of elicited rat macrophages to bone is mediated by specific saccharides located on the cell and/or bone surfaces. We have used a macrophage-bone culture system to study the effects of two lectins, concanavalin A (con A) and soybean agglutinin (SBA), on the morphology of macrophage attachment to a devitalized bone surface and subsequent functional activity. Macrophages were obtained from 3- to 4-week-old rats by peritoneal lavage and the adherent pool was used to prepare cell suspensions. Con A-treated, SBA-treated or control cell suspensions were aliquoted onto the endocranial surface of devitalized rat calvariae. The cells were allowed to attach for 1 h at 37° C, after which, the bone samples were removed from culture and prepared for scanning electron microscopy (SEM). The morphology of con A-treated macrophages attached to bone was markedly different from that of control or SBA-treated cells. Con A altered the attachment and subsequent spreading of macrophages on bone as visualized by SEM. Furthermore, the number of con A-treated cells that attached to bone and the average surface area of cell membrane apposed to the matrix was significantly different from that of control or SBA-treated cells. A 45Ca bone-release assay was performed to evaluate the functional significance of the morphological findings. Lectin-treated or control cell suspensions were allowed to attach to the endocranial surface of 45Ca pre-labeled calvariae for 1 h. Following attachment, the samples were cultured for 72 h. The con A-treated cultures demonstrated a significant decrease in the release of 45Ca after 48 and 72 h in comparison to control cultures, while the 45Ca released from SBA-treated cultures did not differ significantly from controls. These results suggest that certain sugar residues common to membrane-associated glycoconjugates and the organic component of the bone matrix regulate the attachment of macrophages to bone and their subsequent bone-resorbing activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00214631
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  • 3
    Electronic Resource
    Electronic Resource
    Evidence for a relationship between lymphoid cells and osteoclasts: Bone resorption restored in ia (osteopetrotic) rats by lymphocytes, monocytes and macrophages from a normal littermateSupported by Grant DE 03818 from the National Institute for Dental Research, and Grant RR 05712-06 from the National Institutes of Health. (1978)
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 152 (1978), S. 331-341 
    Details
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Osteopetrosis in the ia rat is known to be due to reduced bone resorption and the absence of ruffled borders in osteoclasts. The disease can be cured by the temporary parabiotic union with, or by transfusion of spleen, thymus, or liver cells from, normal littermates. This investigation was pursued in an attempt to determine the cellular basis of this cure by testing the relative effectiveness of populations of mononuclear cells  -  lymphocytes, monocytes, and tissue macrophages, from spleen and thymus of normal rats  -  in curing the disease, compared with cell suspensions of the entire organs. The mononuclear fractions were separated from cell suspensions of spleen or thymus by an adaptation of the Ficoll-Hypaque density-gradient-centrifugation procedure. The mononuclear isolates were evaluated by conventional histological procedures and non-specific esterase histochemistry and found to contain approximately 97% lymphocytes and 3% monocytes from the spleen and 99% lymphocytes and less than 1% monocytes from the thymus.The results of these transplantations were evaluated radiographically and histologically three weeks after treatment. No difference in the rate of skeletal cure was observed in the ia rats that received cells from either whole spleen or thymus, or the mononuclear isolate from these organs. Osteoclasts from the ia animals cured by the transplantation of the mononuclear isolate were examined by electron microscopy and found to have ruffled borders.We conclude that the mononuclear cells isolated from either the spleen or thymus of normal littermates have positive effects on osteoclasts and bone resorption in the ia rat, either indirectly, by elaboration of some local factor that affects the mutant osteoclasts, or directly by transformation into osteoclasts.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aja.1001520306
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  • 4
    Electronic Resource
    Electronic Resource
    Binding of concanavalin-A to critical-point-dried and freeze-dried human lymphocytesSupported i n part by NIH Grants DE-05291 and NS-11425. (1979)
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 156 (1979), S. 121-129 
    Details
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: When human lymphocytes are treated with concanavalin-A (con A) and hemocyanin, the hemocyanin marker, which demonstrates con A binding sites, can be visualized by scanning (SEM) and transmission electron microscopy (TEM) on both critical-point-dried and freeze-dried cells. The ability to visualize the hemocyanin marker by SEM, its quantity and distribution, were all similar in lymphocytes prepared by both drying procedures. By TEM, hemocyanin was seen adjacent to the plasma membrane on critical-point-dried lymphocytes. In contrast, freeze-dried cells showed hemocyanin labeling at some distance from the plasma membrane (40-70 nm) as well as adjacent to it. The distribution of hemocyanin corresponded to the thickness of the amorphous coat seen on fixed, freeze-dried cells. Therefore, the extracellular coat on freeze-dried lymphocytes is a carbohydrate-containing glycocalyx.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aja.1001560113
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  • 5
    Electronic Resource
    Electronic Resource
    Role of the adrenal cortex in maturation of the lymphoid system and immunological competence; the effects of aminoglutethimide in suckling miceSupported in part by grants from the National Institutes of Health (AI-08791) and the National Science Foundation (GB 7574). (1973)
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 137 (1973), S. 231-255 
    Details
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Newborn rats and mice have rudimentary peripheral lymphoid tissues and are immunologically incompetent. The lymphoid system matures late in the third week after birth, shortly before weaning. The adrenal cortex also is relatively inactive neonatally and begins to secrete glucocorticoids in a mature fashion two weeks after birth. These experiments were designed to test the hypothesis that adrenal glucocorticoids induce maturation of the lymphoid system and immunological competence during the third postnatal week. Mice 10-12 days old were injected daily for 4-11 days with aminoglutethimide, an inhibitor of steroid biosynthesis. The adrenal glands and lymphoid tissues were prepared for histological examination 24 hours after the last injection and evaluated without knowledge of the treatment received. Aminoglutethimide caused adrenal dysplasia with a frequency proportional to dose. Effective doses also inhibited growth, and slightly larger doses were fatal. Therefore it was concluded that large doses of aminoglutethimide caused adrenal insufficiency, but the completeness of this insufficiency in surviving animals was not ascertained. Three phases in postnatal maturation of the lymphoid system were identified by examination of untreated littermate controls and reference to previous work. During the first week after birth, the thymus-dependent lymphoid tissues grow by immigration of thymus-derived cells that soon have the capacity for cell-mediated immunity. During the second postnatal week, a new population of wandering lymphoid cells, presumptively derived from bone marrow, settles in lymphoid organs in response to antigenic stimulation, to form primary lymphoid nodules and a few plasma cells. Late in the third week after birth the machinery for humoral antibody synthesis matures with the appearance of germinal centers and numerous plasma cells, coincident with a great increase in production of immunoglobulin. This third phase of maturation was retarded in mice injected with near-fatal doses of aminoglutethimide. Because these mice suffered neither involution of lymphoid tissue nor suppression of proliferation in thymus or thymus-dependent lymphoid-tissue, it was concluded that the effects of aminoglutethimide upon the development of germinal centers and plasma cells were selective and specific. Therefore these experiments support the hypothesis that glucocorticoid secretion plays a decisive role in maturation of immunological competence during the third week after birth in mice. This role appears to be the potentiation of cellular proliferation and differentiation of B cells in response to antigens, culminating in antibody synthesis. Maturation of adrenal glucocorticoid synthesis has also been implicated in the initiation of physiological involution of the thymus and cessation of intestinal absorption of antibodies during the third postnatal week in rats and mice.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aja.1001370302
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  • 6
    Electronic Resource
    Electronic Resource
    The ontogeny of cell-mediated immunity: The response of the mouse to dinitrofluorobenzene (DNFB)Supported in part by grants GB-7547 from the National Science Foundation and AI-08791 from the National Institutes of Health. (1973)
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 137 (1973), S. 257-279 
    Details
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Cellular immunocompetence begins to develop in mice during the first week after birth, as indicated by skin allografting and graft-versus-host reactions, but delayed hypersensitivity has not yet been detected at this early age. The purpose of this study was to investigate the ontogeny of delayed hypersensitivity in suckling mice.Dinitrofluorobenzene (DNFB) is an antigenic hapten which, when applied to the skin, can elicit both delayed hypersensitivity and a humoral immune response. In adult mice the induction period of delayed hypersensitivity to DNFB is a blastogenic reaction (accumulation of lymphoblasts) in the diffuse, thymusdependent cortex of the lymph node draining the site of application. This blastogenic reaction reaches a peak three days after initial application of antigen and then subsides, to be followed by the development of germinal centers and numberous plasma cells  -  indicative of a humoral immune response  -  a week after initial sensitization.Suckling mice were sensitized by application of DNFB to the ears, and the auricular lymph nodes wer prepared for light and electron microscopy three and eight days after sensitization. The development of delayed hypersensitivity was assayed by applying DNFB to an ear, 5 to 12 days after initial sensitization, and measuring the increase in thickness of the ear 24 hours later.The results of these studies indicate that by two days of age, mice have the capacity to develop delayed hypersensitivity as shown by both a blastogenic reaction in the draining lymph node and a positive skin test. Mice sensitized at birth or on the next day did not respond in either way. Mice sensitized prior to two weeks of age did not develop the late phase of the response (the formation of germinal centers and plasma cells), which matured in parallel with the normal maturation of the machinery for antibody production during the third postnatal week.These observations strengthen the conclusion of previous investigations that the blastogenic response in the regional lymph node is the inductive phase of delayed hypersensitivity. Because the late phase of the response does not appear in mice sensitized before two weeks of age, these animals offer an unusual opportunity to study delayed hypersensitivity relatively uncomplicated by humoral antibody production.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aja.1001370303
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  • 7
    Electronic Resource
    Electronic Resource
    Surface coats on human lymphocytes: Freeze-drying and staining with cationsSupported in part by NIH Grants NS-11425 and RR-05712. (1979)
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 154 (1979), S. 267-276 
    Details
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Surface coats can be demonstrated on human peripheral blood lymphocytes by staining with ruthenium red, alcian blue, Thorotrast, and cationized ferritin, which are similar in distribution to a 40- to 65-nm layer of amorphous extracellular material recently reported on fixed, freeze-dried lymphocytes. Several additional lines of evidence, including X-ray micro-analysis, suggest that the latter is not a contaminant added by freeze-drying. Freeze-drying may provide the means for a morphological assessment of the lymphocyte surface, including the extracellular coat, which may give additional insight into the immune response.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aja.1001540210
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  • 8
    Electronic Resource
    Electronic Resource
    Pluripotent hemopoietic stem cells give rise to osteoclasts (1986)
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 177 (1986), S. 505-511 
    Details
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Osteopetrosis in the ia (incisors absent) rat is the result of reduced bone resorption due to abnormal osteoclasts. The mutant osteoclasts lack a ruffled border - the membrane specialization involved in osteolysis. Studies in the ia mutant have shown that when pluripotent hemopoietic stem cells from normal littermates are transplanted into ia recipients, normal osteoclasts are formed and the skeletal sclerosis is eventually cured. The present study was conducted to provide evidence for the mechanism of the cure. Do the transplanted stem cells provide a helper function, i.e. secrete soluble factor(s) which transform pre-existing osteoclasts, or do they fuse with each other or pre-existing osteoclasts, to form functional osteoclasts? Using the procedures described by Goldschneider and co-workers, and fluorescence-activated cell sorting (FACS), pluripotent hemopoietic stem cells were isolated from normal rat bone marrow, labeled with saturated FITC, and injected intravenously into irradiated ia rats. After 48 hr, the recipients' long bones were removed and split longitudinally, and the endosteal surface was scraped. The resulting cellular suspension containing osteoclasts was examined by phase contrast and fluorescence microscopy. Fluorescing mononuclear cells of donor origin that had homed to the bone marrow demonstrated moderate cytoplasmic fluorescence. Approximately 30% of the osteoclasts observed demonstrated light cytoplasmic fluorescence. When cellular pools incapable of curing osteopetrosis (thymocytes) were labeled and injected into ia recipients, no labeled osteoclasts were observed. These studies indicated that pluripotent hemopoietic stem cells, when transplanted into ia hosts, fuse with each other and differentiate into osteoclasts or fuse with pre-existing osteoclasts.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aja.1001770408
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  • 9
    Electronic Resource
    Electronic Resource
    Morphological and histochemical comparison of the cells elicited by ectopic bone implants and tibial osteoclasts (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 192 (1991), S. 45-54 
    Details
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Pellets of mineralized and demineralized bone and a composite mixture of mineralized and demineralized, devitalized bone particles were implanted subcutaneously on the dorsal body wall of young adult rats. Two weeks post-implantation, the pellets were removed and processed for histochemical and morphological analyses. Rat proximal tibia was also processed for evaluation. The levels of tartrate-resistant acid phosphatase (TRAP) activity in the multinucleated giant cells (MNGCs) from each of the three implants and from osteoclasts were assessed using an image analyzer. The osteoclasts from the proximal tibia and the majority of MNGCs from the demineralized implants demonstrated high levels of TRAP activity. MNGCs from the mineralized implants showed either a low level or absence of TRAP activity. Most MNGCs from the composite implants exhibited a low level of TRAP activity; however, there was a population of cells that demonstrated a high level of reaction product, similar to that seen in the tibia and demineralized implant. Morphologically, osteoclasts from the proximal tibia and from the osteogenic demineralized implant exhibited ruffled borders. A small population of MNGCs from the composite implant also revealed osteoclastic features. In summary, MNGCs from the mineralized implant did not exhibit a level of TRAP reaction product or morphology similar to osteoclasts, while the majority of cells from the demineralized implant and a subpopulation of the MNGCs elicited by the composite implant did demonstrate TRAP expression and morphology similar to osteoclasts. The expression of osteoclastic characteristics in cells at an ectopic site may be dependent on accessory signals from the skeletal microenvironment; such signals appear to be absent from or incomplete in the mineralized implants but appear to be present when demineralized bone particles are implanted.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aja.1001920106
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  • 10
    Electronic Resource
    Electronic Resource
    Proliferative response to the skin-sensitizing contact agent dinitrofluorobenzene in lymph nodes in miceSupported in part by grants GB-38360 and GB-7547 from the National Science Foundation and AI-08791 from the National Institutes of Health. (1974)
    New York, NY [u.a.] : Wiley-Blackwell
    American Journal of Anatomy 141 (1974), S. 219-233 
    Details
    ISSN: 0002-9106
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: When dinitrofluorobenzene (DNFB) is applied to the skin in mice, it induces a state of delayed hypersensitivity. During the three days following the application of this contact agent, lymphoblasts accumulate in the diffuse cortex of the lymph node draining the area of sensitization. This accumulation of blast cells - referred to as a blastogenic response - appears to be part of the induction phase of delayed hypersensitivity.This is an investigation into the origin of the cells of the blastogenic response. There is evidence from other studies that lymphocytes immigrate to the lymph nodes draining a site of sensitization. This paper provides complementary evidence that cellular proliferation in the diffuse cortex of the sensitized lymph node is another source for the accumulation of blast cells. This proliferation has been studied by counting mitotic figures, and by autoradiography after giving 3H-thymidine in order to determine the number of blast cells undergoing DNA synthesis and the duration of the various phases of the cellular reproductive cycle.The results show a high degree of correlation between mitotic figures and blast cells in the diffuse cortex of the sensitized lymph nodes, consistent with a population of dividing blast cells. Approximately 80 percent of the blast cells were in the DNA-synthetic phase of the reproductive cycle at the peak of the blastogenic response, and the cells were dividing with approximately an eight hour generation time.These results can be interpreted to indicate that the cells of the blastogenic response, although accumulating initially by immigration to the draining lymph node, become a homogeneous, nondifferentiating population of blast cells, all proliferating at near maximal rate to provide the great numbers of blast cells seen three days after application of the sensitizing agent.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/aja.1001410205
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