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1.
Suspensions of mouse bone marrow cells, spleen cells, and blood leucocytes were cultured in diffusion chambers in dilution series in order to establish the minimum concentrations of haematopoietic stem cells (HSC). The observed frequencies of empty chambers after seven days of culture conformed to the expected frequencies of a null response in a Poisson distribution. The proportions of empty chambers could therefore be used to estimate the concentrations of HSC in the cell suspensions. The following numbers of HSC per 105 cells were found (with 95% confidence limits): Bone marrow: 50 (44–56). Spleen: 3.5 (2.8–4.3). Blood leucocytes: 1.4 (1.2–1.8). The mean (± standard error) HSC-content per femur, spleen, and milliliter blood when pooling cells from three to six donor mice was 8240 ± 600, 7660 ± 490, and 56 ± 6.5 respectively. For comparison, the HSC concentrations were also determined with the spleen colony technique; the ratio between the HSC-concentrations of bone marrow, spleen, and blood determined with the diffusion chamber technique was similar to that determined with the spleen colony technique.  相似文献   

2.
A series of experiments was performed to elucidate the relationship between cells that form granulocytic colonies in fibrin clot diffusion chambers implanted into the peritoneum (i.p.) of irradiated mice (CFU-d) and day 7 and day 14 CFU-c which give rise to colonies after 7 and 14 days in agar cultures in vitro, respectively. Normal human bone marrow cells were cultured in suspension in vitro or in diffusion chambers implanted into irradiated or non-irradiated mice. During these culture conditions there was an initial decrease in the number of CFU-c per culture. This was followed by an increase between day 2 and day 7 of culture. No similar increase of neutrophilic CFU-d was observed. When CFU-d, day 14 and day 7 CFU-c in normal marrow were separated by velocity sedimentation and cultured in suspension culture or in diffusion chambers for 7 days, the maximum increase of day 7 and day 14 CFU-c was observed in slowly sedimenting cell fractions which contained the majority of CFU-d. After 3 days in suspension culture, the maximum increase of day 14 CFU-c was found in fractions which also gave rise to maximum numbers of CFU-c after 7 days. However, day 7 CFU-c were found in fractions which initially contained the majority of day 14 CFU-c. No increase in CFU-d was found in fractions initially containing peak numbers of CFU-c. Between 53 and 71% of CFU-c harvested from diffusion chambers in irradiated mice or from suspension cultures were sensitive to pulse incubation with tritiated thymidine, suggesting that the cells were proliferating during these culture conditions. In diffusion chambers implanted into non-irradiated mice, however, CFU-c were found to be relatively resistant to this treatment (3–11% sensitive to tritiated thymidine). Thus marked increases in CFU-c were also observed during experimental conditions, where no significant DNA synthesis was detected. A reproducible time sequence of increase in CFU-c populations in culture was observed. Day 14 CFU-c and cells that gave rise to clusters on day 7 in agar increased between day 2 and day 4, whereas day 7 CFU-c increased between day 4 and day 7. The results suggested that CFU-d gave rise to CFU-c in culture and that day 14 CFU-c were precursors of day 7 CFU-c.  相似文献   

3.
1. The adelgid Adelges japonicus induces shoot galls in ezo‐spruce Picea jezoensis. The relative importance of shoot characteristics and the number of adelgids that induce a gall in determining resource availability on a shoot was examined. Gall volume, which is highly correlated with the number of larvae that mature in a gall, was used as a measure of resources available for adelgids on a shoot. Gall volume was partitioned into two components: the number of chambers and the chamber volume. 2. Path analysis revealed that the number of chambers in a gall was not affected by the number of leaves (which is an indirect measure of shoot length), the whorl where the gall was attached, or the number of surrounding shoots arising from the 1‐year‐old parent shoot. Each of these shoot characteristics, however, affected chamber volume: chamber volume increased with increasing numbers of leaves and surrounding shoots and with increasing heights of the whorls. 3. Two generations of adelgids contributed to gall enlargement in different ways: the number of gall founders affected the number of chambers, and the number of gall inhabitants affected chamber volume. 4. In a path diagram that incorporated the effects of both shoot characteristics and the numbers of adelgids, the numbers of adelgids in the two generations together accounted for 30% of the variance in gall volume, whereas the three shoot characteristics explained only 3.7% of the variance. This suggests that the amount of resources available in a gall is affected mainly by the numbers of adelgids but only slightly by shoot characteristics.  相似文献   

4.
1. Habitat loss and fragmentation potentially affect the performance of bees that forage nectar and pollen of plants in their habitats. In forest landscapes, silvicultural conifer plantations often have reduced and fragmented natural broadleaf forests, which seem to provide more floral resources for bees than do the plantations. 2. This study evaluated the effects of forest characteristics (i.e. elevation, area, edge length, and tree size of natural forests) on pollen diets (plant taxa assemblages of pollen grains in provisions) and total provision mass in oviposited chambers in nests made by a standardised number of Osmia cornifrons bees at 14 sites in a forestry area in central Japan. 3. From April to May, the numbers of nests and chambers per nest increased, and the provision mass per chamber decreased. Main pollen sources were Prunus at higher elevations in April and Wisteria at lower elevations in May, foraging on which increased the numbers of nests and chambers per nest. The provision mass per chamber was smaller at higher elevations in more fragmented natural forests. Decreases in the area of natural forests within the foraging range (400‐m radii) of O. cornifrons increased the utilisation of Rubus pollen and decreased the total provision mass. 4. These findings suggest that the loss and fragmentation of natural broadleaf forests change pollen diets and reduce the provision mass of mason bees, which may reduce the number and size of their offspring.  相似文献   

5.
Summary Specimens of Haliclona elegans (Bowerbank, 1866) are covered by a thin, double layered dermal membrane extending over large subdermal spaces. The pores in the dermal membrane are formed by single porocytes with one or sometimes several pores in the center of the cell. The subjacent tissue shows a faintly developed mesenchyme and numerous big choanocyte chambers projecting into lacunar spaces of the incurrent canal system. The outer surface of the chambers is directly covered by the pinacocyte epithelium of the incurrent canal wall, which also separates them completely from the mesenchyme. Water influx into the chambers is guaranteed by prosopylar openings in the pinacocyte cover at the outer chamber surface. The chambers are connected to the excurrent canal system in the eurypylous way by wide apopyles, each of which is surrounded by a small ring of flagellated cone cells. About 15% of the choanocyte chambers in H. elegans contain central cells, which are thought to derive from migrating pinacocytes of the canal systems.  相似文献   

6.
Both murine and human bone marrow cells were cultured in plasma clots which were formed inside diffusion chambers implanted into cyclophosphamide- and saline-treated mice. After an initial fall, the number of mouse bone marrow cells and numbers of mouse myeloid stem cells (CFU-C) and agar cluster-forming units rose faster in the cyclophosphamide-treated animals. These hosts also favored formation of myeloid (CFU-D-G) and erythroid (CFU-D-E) colonies and myeloid clusters in the plasma clot. The number and growth rate of mouse CFU-D-G were higher than those of CFU-C from the same marrow population. These observations suggest the existence of humoral factors stimulating granulocyte progenitor cell replication and differentiation. At its best the increment of CFU-D-E number was equivalent to that caused by a single 0·1 unit erythropoietin dose. Culture of normal human marrow cells resulted in colonies in the plasma clot containing only granulocytes and macrophages. Cyclophosphamide-treated host animals were essential for human CFU-D-G development. Plating efficiency for human marrow myeloid colonies was better in the conventional in vitro agar cultures than in diffusion chambers.  相似文献   

7.
Dispersed primary tissue culture cells from adrenal, aorta, heart and cornea of adult rabbits were placed into diffusion chambers. The chambers were implanted into serologically compatible litter mates. Chimaeric aggregates resembling embryonal organs formed in the chambers within 3 weeks. The ‘adrenal’had two to three cortical zones to both sides of a medulla. The ‘aorta’had a central fibromuscular layer and a surface layer of endothelial cells. The ‘heart’consisted of a muscular layer coated with endothelial cells; the aggregate assumed a chamber-like shape. The ‘cornea’had a loose matrix covered on the convexity by epithelial cells. Upon transfer of the aggregates from in vivo to in vitro culture, cells dispersed within 1 week and propagated as a mixed population. Analysis of the phenomenon has been attempted and the course of future studies suggested. The study contributes to the discussion of cell automatism.  相似文献   

8.
Shoot apices of a short-day sensitive line of Nicotiana tabacum L. cv. NCTG-22 have been examined by electron microscopy for ultrastructural changes which occurred in the central zone over a 17-day period during the transition from vegetative to reproductive growth. Plants were grown in controlled-environment chambers of the NCSU Phytotron and exposed to an inductive photoperiod after a 6-wk juvenile phase. Ultrastructural changes were investigated from photomicrographs using a semi-automatic stereological procedure and a microcomputer. After exposure to only one inductive cycle cell and nuclear cross sectional areas in short-day plants were significantly larger than in long-day controls. Subsequently, under short days cross sectional areas of cells, nuclei, vacuoles and proplastids decreased, while mitochondrial cross sectional area and relative volume increased during the course of the induction period. In induced apices as cross sectional areas and relative volumes of vacuoles and proplastids decreased, their profile numbers increased. The reduction in cross sectional areas of cells and most organelles was associated with an increase in rate of leaf initiation and size of the apical dome. The demand for sufficient energy input to maintain the surge in growth and activity preceding floral initiation was reflected by the significant increases in cross sectional area, profile numbers and density of the mitochondria population. Even though the transition period is quite long for Nicotiana, cells and organelles in the central zone were observed to progress through similar changes in morphology that are known to occur in Sinapis and Xanthium which exhibit a more rapid and absolute response to photo-induction.  相似文献   

9.
We describe the features of programmed cell death occurring in the egg chambers of Drosophila virilis during mid-oogenesis and late oogenesis. During mid-oogenesis, the spontaneously degenerating egg chambers exhibit typical characteristics of apoptotic cell death. As revealed by propidium iodide, rhodamine-conjugated phalloidin staining, and the TUNEL assay, respectively, the nurse cells contain condensed chromatin, altered actin cytoskeleton, and fragmented DNA. In vitro caspase activity assays and immunostaining procedures demonstrate that the atretic egg chambers possess high levels of caspase activity. Features of autophagic cell death are also observed during D. virilis mid-oogenesis, as shown by monodansylcadaverine staining, together with an ultrastructural examination by transmission electron microscopy. During the late stages of oogenesis in D. virilis, once again, the two mechanisms, viz., nurse cell cluster apoptosis and autophagy, operate together, manifesting features of cell death similar to those detailed above. Moreover, an altered form of cytochrome c seems to be released from the mitochondria in the nurse cells proximal to the oocyte. We propose that apoptosis and autophagy function synergistically during oogenesis in D. virilis in order to achieve a more efficient elimination of the degenerated nurse cells and abnormal egg chambers. The present study was co-financed within Op. Education by the European Social Fund and by National Resources via a grant (HRAKLEITOS 70/3/7164) to Professor L.H. Margaritis.  相似文献   

10.
The effect of an oil spill on estuarine meiofauna was examined in a controlled colonization experiment. Forty-five replicate azoic sediment chambers treated with 0,133 or 381 mg hydrocarbons: 100 g dry sediment of South Louisiana crude oil were each quantitatively sampled with three replicate cores for colonizing meiofauna. Chambers were sampled on days 2, 5, 10, 30 and 60 postplacement in a Louisiana Spartina alterniflora (Loisel) salt marsh. Polychaetes showed a delayed colonization and reduced densities in oiled relative to non-oiled sediments. Nematode numbers were significantly depressed in the high oil treatment but no delay in colonization was identified. Only one species of meiobenthic copepods, Enhydrosoma woodini, displayed a reaction to the presence of the oil but only in the top centimeter of sediment. This species showed significantly decreased densities due to the heavily oiled treatment throughout the study until day 60 when numbers in the heavily oiled chambers were significantly higher than those in the non-oiled chambers. Species diversity (H') was calculated on the meiobenthic copepod assemblage and showed that diversity in the high oil treatment was generally lower than that in other treatments through day 30. Fewer species colonized the heavily oiled chambers before day 30. Principle Components Ordination conducted on the copepod assemblage could not identify an oil effect separate from a chamber effect for copepod community structure.  相似文献   

11.
The choanocyte chambers of the marine spongeReniera sp. protrude with their curved outer surface free into the incurrent canals. The water is sucked into the chambers by cavities between the choanocytes. Particles up to 1 µm in diameter may enter the chambers with the water current. These particles are trapped on the outer surface of the choanocyte collars and are ingested by the choanocytes and processes of the pinacocyte epithelium of the incurrent canal system, which project into the chambers. Bigger particles are retained in the incurrent canals mainly on the outer surface of the choanocyte chambers. They are ingested by pinacocytes of the canal wall and transported to cells of the mesenchyme. The present investigation shows the great importance of the pinacocyte epithelium of the incurrent canal system for suspension feeding inReniera sp.  相似文献   

12.
 In Drosophila a remarkable feature of oogenesis is the regression of the nurse cells after dumping their cytoplasmic contents into the oocyte. We have studied the nature of this process at the late stages of egg chamber development. In egg chambers DAPI staining shows highly condensed chromatin from stage 12 and TUNEL labelling shows DNA fragmentation up to stage 14. Gel electrophoresis of the end-labelled DNA, extracted from isolated egg chambers at the same stages of development, shows a ladder typical of apoptotic nuclei. This provides evidence that, during Drosophila oogenesis, the nurse cells undergo apoptosis. Apoptotic nuclei have also been detected in dumping-defective egg chambers, indicating that the cytoplasmic depletion of nurse cells is concurrent with but apparently not the cause of the process. Received: 12 December 1997 / Accepted: 6 January 1998  相似文献   

13.
Summary Intercellular bridges have been detected in ovarian follicle cells of Drosophila melanogaster. These bridges occur widely between follicle cells of previtellogenic chambers, while, in vitellogenic chambers, they become restricted to the columnar follicle cells. Usually, only one bridge is detectable between adjacent follicle cells, but a single cell may form two cytoplasmic continuities.The fine structure of the intercellular bridges is similar to that previously described in the development of Drosophila. The bridge wall consists of two layers of which the more external is more electron dense and thinner than the inner one.The role played by the intercellular bridges in the determination of a synchronous differentiation of the linked follicle cells is discussed in relation to the known behaviour of these cells in the secretion of the egg covering precursors.  相似文献   

14.
A number of years ago we reported that tight junctions between adjacent Sertoli cells subdivide the seminiferous epithelium into two compartments, basal and adluminal, thus forming the morphological basis of the blood-testis barrier. It is now generally believed that the special milieu created by the Sertoli cells in the adluminal compartment is essential for germ cell differentiation. In order to duplicate the compartmentalization that occurs in vivo, Sertoli cells were cultured in bicameral chambers on Millipore filters impregnated with a reconstituted basement membrane. Confluent monolayers of these cells were tall columnar (40–60 µ in height) and highly polarized. These Sertoli cell monolayers established electrical resistance that peaked when the Sertoli-Sertoli tight junctions developed in culture. In addition, the monolayers formed a permeability barrier to 3H-inulin and lanthanum nitrate. The bicameral chambers were utilized in a number of studies on protein secretion, and it was revealed that numerous proteens are secreted in a polarized manner. In another study, hormone- stimulated aromatase activity was measured in Sertoli cells grown on plastic culture dishes, plastic dishes coated with laminin or Matrigel, and in the bicameral chambers. Cell culture on basement membrane substrate decreased the FSH-dependent estrogen production. No estrogen production was observed when the Sertoli cells were cultured in the bicameral chambers. These results are in accord with the hypothesis that differentiated Sertoli cells lose their ability to metabolize androgen to estrogen in an hormone-dependent manner, whereas undifferentiated cells in culture, or in vivo, have a very active FSH-dependent aromatase activity. This bicameral culture system could serve as an important model system to examine various functions of Sertoli cells including interactions of Sertoli cells with germ, Leydig, and myoid cells.  相似文献   

15.
Distribution and density of the chloride cells in the newly hatched larvae of teleosts vary depending on species and environmental salinity at hatching. In the euryhaline freshwater ayu (Plecoglossus altivelis), chloride cells are concentrated in the skin posterior to the pectoral fins and gradually decrease in number toward the head and tail. In the stenohaline sea water flounder (Kareius bicoloratus), most chloride cells are localized at the inner membrane of gill chambers and in the skin near the openings of gill chambers, but only a few cells appear in the skin of the yolk sac. In the stenohaline freshwater carp (Cyprinus carpio), only a few small chloride cells are scattered in the body skin. The density and abundance of chloride cells appears to be correlated with the different requirements for osmoregulation in teleost larvae.  相似文献   

16.
A comparative cytological study was made of oogenesis in flies carrying various mutant alleles of the female sterile gene otu. It resides at 22.7 on the genetic map and within subdivision 7F of the cytological map of the X-chromosome. Each of the five ethyl methane sulfonate-induced mutations observed falls into one of three classes. In class 1, most mutant ovarioles lack germ cells; in class 2, most mutant ovarioles contain tumorous chambers; and in class 3 mutants, chambers occur that possess defective oocytes. The otu2 allele belongs to class 1; otu1 to class 2; and otu3, otu4, and otu5 to class 3. The mutations have no effects upon female viability or upon the viability and fertility of hemizygous males. Heterozygous females are fertile and have cytologically normal ovaries. In otu5 homozygotes, all ovarioles contain egg chambers, but oogenesis is prematurely terminated to produce a pseudo-stage 12 oocyte. Ovarioles from otu3 and from otu4 homozygotes contain both ovarian tumors and oocytes. Pseudonurse cells (PNC), which are cystocytes that have stopped dividing and have entered the nurse cell mode of development, are also abundant. PNCs contain polytene chromosomes. Since the homologs are paired, each nucleus has the haploid number of chromosomes. In chambers lacking an oocyte, the number of PNCs is less than the normal number of nurse cells. In chambers containing an oocyte, the number of accompanying nurse cells may be 15, or above or below normal. In vitellogenic chambers, the chromosomes in the nurse cells connected directly to the oocyte are more expanded than those in more distant nurse cells. The KA14 deficiency lacks the plus allele of otu. KA14 heterozygotes are fertile and have cytologically normal ovaries. When females carry KA14 and otu1, otu3, otu4, or otu5, 80% of their ovarioles are agametic. When females carry otu2 and one of the other mutant alleles, the ovarioles proceed further in development. So otu2 produces a product that has a beneficial effect on the test allele. When two different otu alleles are combined in a single fly, the phenotype of the hybrid ovary usually most resembles that of the ovary homozygous for the “stronger” allele (the otu mutant that allows oogenesis to proceed farthest). The results indicate that the product of the otu+ locus functions at least three different times during oogenesis; first to permit oogonia to proliferate, second to control the division and differentiation of germarial cystocytes, and third to facilitate the normal growth of the ooplasm. The gene product appears to be required in higher concentrations at each developmental period. The lesions produced by the mutations are thought to interfere with the stability or functioning of the gene product, and the ovarian phenotype produced by a given genotype depends upon the concentration of functional gene product available to the germ cells.  相似文献   

17.
The tissue of glass sponges (Class Hexactinellida) is unique among metazoans in being largely syncytial, a state that arises during early embryogenesis when blastomeres fuse. In addition, hexactinellids are one of only two poriferan groups that already have clearly formed flagellated chambers as larvae. The fate of the larval chambers and of other tissues during metamorphosis is unknown. One species of hexactinellid, Oopsacas minuta, is found in submarine caves in the Mediterranean and is reproductive year round, which facilitates developmental studies; however, describing metamorphosis has been a challenge because the syncytial nature of the tissue makes it difficult to trace the fates using conventional cell tracking markers. We used three‐dimensional models to map the fate of larval tissues of O. minuta through metamorphosis and provide the first detailed account of larval tissue reorganization at metamorphosis of a glass sponge larva. Larvae settle on their anterior swimming pole or on one side. The multiciliated cells that formed a belt around the larva are discarded during the first stage of metamorphosis. We found that larval flagellated chambers are retained throughout metamorphosis and become the kernels of the first pumping chambers of the juvenile sponge. As larvae of O. minuta settle, larval chambers are enlarged by syncytial tissues containing yolk inclusions. Lipid inclusions at the basal attachment site gradually became smaller during the six weeks of our study. In O. minuta, the flagellated chambers that differentiate in the larva become the post‐metamorphic flagellated chambers, which corroborate the view that internalization of these chambers during embryogenesis is a process that resembles gastrulation processes in other animals.  相似文献   

18.
In monolayer cultures of guinea-pig bone marrow and spleen the development of discrete fibroblast colonies takes place on days 9–12. The linear increase in the number of colonies with increasing numbers of explanted cells and the distribution of male and female cells in mixed cultures support the view that fibroblast colonies are clones. The concentration of colony-forming cells in bone marrow and spleen is approximately 10-5. Bone marrow culture (but not spleen culture) fibroblasts are capable of spontaneous bone formation in diffusion chambers. Fibroblasts from both bone marrow and spleen cultures are inducible to osteogenesis in diffusion chambers in the presence of transitional epithelium.  相似文献   

19.
Summary Each choanocyte chamber of Petrosia ficiformis is formed by a slightly outpocked choanocyte epithelium and by a ring of three or four uniflagellated cone cells surrounding the apopyle. The apopyle opens into a small aphodus, which leads the water flow to larger excurrent canals. Pinacocytes of the incurrent canal system cover the basal surface of the choanocytes and separate them from the incurrent canals and the mesenchyme. The water flows into the chambers by pores in the pinacocyte cover and then through gaps between adjacent choanocytes. To our knowledge this is the first report of a leuconoid canal system in which choanocyte chambers are covered by a pinacocyte epithelium of the incurrent canal system that isolates the chambers from the mesenchyme. A future comprehensive revision of the types of canal systems in sponges seems to be necessary. Permanent affiliation: Department of Biology and Health Sciences, University of Hartford, West Hartford, CT 06117, USA  相似文献   

20.
The maturation of animal oocytes is highly sensitive to nutrient availability. During Drosophila oogenesis, a prominent metabolic checkpoint occurs at the onset of yolk uptake (vitellogenesis): under nutrient stress, egg chambers degenerate by apoptosis. To investigate additional responses to nutrient deprivation, we studied the intercellular transport of cytoplasmic components between nurse cells and the oocyte during previtellogenic stages. Using GFP protein-traps, we showed that Ypsilon Schachtel (Yps), a putative RNA binding protein, moved into the oocyte by both microtubule (MT)-dependent and -independent mechanisms, and was retained in the oocyte in a MT-dependent manner. These data suggest that oocyte enrichment is accomplished by a combination of MT-dependent polarized transport and MT-independent flow coupled with MT-dependent trapping within the oocyte. Under nutrient stress, Yps and other components of the oskar ribonucleoprotein complex accumulated in large processing bodies in nurse cells, accompanied by MT reorganization. This response was detected as early as 2 h after starvation, suggesting that young egg chambers rapidly respond to nutrient stress. Moreover, both Yps aggregation and MT reorganization were reversed with re-feeding of females or the addition of exogenous insulin to cultured egg chambers. Our results suggest that egg chambers rapidly mount a stress response by altering intercellular transport upon starvation. This response implies a mechanism for preserving young egg chambers so that egg production can rapidly resume when nutrient availability improves.  相似文献   

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