长吻鮠血细胞发生的研究

用Wright-Giemsa和PAS染色对长吻鮠头肾、肾脏、脾脏、肝脏等器官组织的涂片、印片染色观察发现,头肾、肾脏和脾脏是其主要造血器官。红细胞、粒细胞和淋巴细胞主要在肾脏和头肾中发生,其次是脾脏。单核细胞则主要在肾脏和脾脏中发生,头肾中也有少量单核细胞产生。肝脏中无原始型血细胞,可能不是其造血器官。红细胞的发育经历四个阶段,其胞体体积经历了由大到小,由小到大再变小的"两大两小"发育过程;粒细胞的发育经历五个阶段,其胞体体积均由大变小,双叶或多叶核的粒细胞可能是衰老的粒细胞亦即核的分叶是粒细胞衰老的标志;淋巴细胞和单核细胞的发育各经历了三个阶段,两者发育成熟过程中胞体体积均由大变小。巨噬细胞由单核细胞发育而来。原血细胞和部分早期幼稚血细胞可以进行有丝分裂,部分成熟红细胞和血栓细胞可以进行直接分裂。红细胞在整个发育过程中,PAS反应均呈阴性,各类白细胞的发育过程中,PAS反应由阴性到阳性并逐渐增强,这显示随着白细胞的逐渐发育成熟,细胞内糖原物质含量逐渐增多。     

Fine structure of melanocytes and macrophages in the Harderian gland of the mouse

The presence of dendritic cells containing melanin granules has been demonstrated employing silver impregnation and electron microscopy in the interstitial tissue of the Harderian gland of the mouse. Two types of melanocytes, either with or without the various developmental stages of melanin granules, were found in the gland. Cells with developing granules were more dendritic and contained a large number of cytoplasmic organelles. The other cells were ellipsoidal or slender in shape and contained few cytoplasmic organelles and a large number of fully melanized granules, but no developing granules. In general, the granules of the Harderian gland melanocytes resembled granules from other organs (particularly the skin of the eyelids). The general size range of the granules was 0.2-0.9 micron. Each granule was enclosed by a membrane. The Harderian gland macrophages contained fully pigmented melanin granules of various sizes. The granules were enclosed by a membrane either singly or in groups. Some of the melanin granules within the phagosomes showed signs of degradation, revealing the underlying matrix.     

Structural and functional characterisation of the blood cells of the bivalve mollusc,Scrobicularia plana

Light and electron microscopical studies were carried out in order to characterise the blood cells of the bivalve mollusc, Scrobicularia plana. Three types of haemocytes were recognised: eosinophilic granular haemocytes, basophilic granular haemocytes and basophilic agranular haemocytes. The eosinophilic granulocytes were vesicular and contained large granules whereas the basophilic granulocytes were found to contain small granules and glycogen 'lakes'. The basophilic agranular haemocytes were significantly smaller than the granular haemocytes and had a high nucleus to cytoplasm ratio. Functional characterisation of the blood cells identified activity for the lysosomal enzymes: acid phosphatase, beta-glucuronidase, non-specific esterase and arylsulphatase. There was also a weak staining reaction for phenoloxidase and peroxidase activities. Phagocytosis of Gram-positive bacteria was demonstrated by the haemocytes and antibacterial activity was shown by cell-free haemolymph. Assays to determine release of reactive oxygen species from the haemocytes did not detect any reactive oxygen generation.     

Composition and ultrastructure of elasmobranch granulocytes. III. Sharks (Lamniformes)

The peripheral blood granulocyte composition of five species of shark is given together with ultrastructural observations made on the epigonal organ and blood of Mustelus lenticulatus and spleen of Apristurus sp. Three granulocyte lineages occurred in Mustelus . Ultrastructurally, eosinophilic granulocytes contained granules that were very variable in size and contained parallel fibrillar arrays that were unidirectional in small granules, but which were often orientated in several directions in larger granules. This dense core material sometimes formed angular crystalloids.
Eosinophils had ovoid or irregular granules that were usually uniformly electron-dense, but some had an eccentric ovoid area of greater electron-density. Some eosinophils enter the blood where they are phagocytic, but others, in blood and epigonal organ, showed an unusual but characteristic process of disintegration, leaving a ragged cell full of cellular debris.
The third granulocyte type had features of mast cells and basophils of higher vertebrates and was tentatively identified as belonging to that lineage. The inter-relationships of these cells, and of them with granulocytes of other elasmobranchs, are discussed.     

Hemocytes of Bulinus truncatus rohlfsi (Mollusca: Gastropoda)

Two basic cell types occur in the hemolymph of Bulinus truncatus rohlfsi: granulocytes and hyalinocytes. Granulocytes are divided into three subtypes: (1) Granulocytes I, which account for 19% of the hemocytes, are small, young amoebocytes with 1–20 filopodia and small numbers of cytoplasmic granules, including some lysosomes; (2) granulocytes II, which account for 78% of the cells, are large, fully developed amoebocytes that possess 1–20 filopodia and many granules, both acidophilic and basophilic, including numerous lysosomes, phagosomes, and mitochondria; and (3) spent granulocytes, which are rare, have few filopodia, large accumulations of glycogen granules and prominent vacuoles in addition to lysosomes in the cytoplasm. These three subtypes of granulocytes probably represent ontogenetic stages within a single cell line. In addition, granulocytes with 40 or more filopodia and little ectoplasm, found in only 1 of 45 snails examined, probably reflect a pathologic condition. Hyalinocytes, which account for 3% of all hemocytes, are similar in size to mature granulocytes, but have few or no cytoplasmic granules and lack filopodia and glycogen granules. Total hemocyte concentration in hemolymph is 328,000 ± 188,000 cells/ml.     

Cellular composition of erythropoietic cell populations and aggregate cell cultures derived from early chick blastodiscs

Light and electron microscopy of suspensions of cells prepared by dispersing chick blastodiscs at primitive-streak and head-fold stages showed the presence of numerous yolk granules, yolk-rich endodermal cells and occasional presumed ecto- and mesodermal cells. Several cell fractions prepared from this suspension by sedimentation through discontinuous Ficoll gradients were of similar composition. No enrichment of any particular cell type which might account for either differential sedimentation or erythropoietic potential of the fractions could be recognized. Two fractions, EP 1 and EP 2, were cultured as cell aggregates on vitelline membranes. EP l produced highly organized blood islands containing developing erythrocyte cells, organized endothelium, fibroblasts, thrombocytes and occasional granulocytes. Blood islands derived from EP 2, on the other hand, contained essentially only aggregates of erythroblasts embedded in endoderm. It is tentatively suggested that EP 1 contains young multipotential hematopoietic precursors while EP 2 has only older blood-cell precursors committed to erythrocyte development. No cellular basis for the resolution of EP I into two complementary subfractions could be recognized.     

似刺鳊鮈外周血细胞显微结构和血清生化指标的初步研究

似刺鳊(鮈)(Paracanthobrama guichenoti)外周血细胞可分为红血细胞、嗜中性粒细胞、单核细胞及大、小淋巴细胞和血栓细胞,未发现嗜酸性和嗜碱性粒细胞.外周血液中可观察到少量未成熟的及正在分裂的红血细胞.白细胞中,血栓细胞体积最小,嗜中性粒细胞体积最大;数量上,血栓细胞最多,而大淋巴细胞则最少.似刺鳊(鮈)血清生化指标测定结果显示,谷丙转氨酶(ALT)、胆固醇(CHOL)和总蛋白(TP)变化范围较大,且谷草转氨酶(AST)的水平较高,似刺鳊(鮈)雄鱼的血液生化指标相应值均显著低于雌鱼(P<0.05).     

Morphological, cytochemical autoradiographic and quantitative studies of the cells in lymphocyte cultures with added tuberculin]

Lymphocyte culture experiments with purified tuberculin showed a transformation of small lymphocytes into medium sized, large and very large basophilic cells. During cell growth RNA synthesis and acid phosphatase activity increased. Some of the very large basophilic cells synthesized DNA. During the first and second day of culture experiment also medium sized and large weakly basophilic cells increased, which have a pale plasma and azurophilic granules. These cells did not synthesize DNA and showed only a low incorporation of 3H-uridine. The results of our experiments indicate a transformation of medium sized and large basophilic cells not synthesizing DNA into weakly basophilic (pale) cells showing azurophilic granules. In contrast to the lymphocyte culture experiments with tuberculin lymphocyte cultures without tuberculin showed no significant increase of medium sized, large and very large basophilic cells and of weakly basophilic cells.     

Ontogenetic changes of circulating erythroid cells and haemoglobin components in Esox lucius

Using light microscopy the morphology, the mitotic index and levels of erythroid cell types were detected from 48 h pike Esox lucius embryos before hatching to adult specimens. At the same developmental stages, the haemoglobins and globin chains expressed were electrophoretically characterized. The erythroid cells of the primitive generation were the most abundant from 48 h before hatching until 15–20 days after hatching, then their number decreased and only rare cells remained in the 3 month‐old juvenile specimens. These cells divided and differentiated in the blood and were substituted by the definitive erythrocyte series. As in other vertebrates, the immature cells of the two generations differed in morphological properties and in the synthetized haemoglobin. The circulating erythroid cells of the definitive population cell lineage were, at all differentiation stages, smaller than those of the primitive generation. The definitive erythrocytes appeared in blood smears of 7 days post‐hatching larvae, they increased rapidly and at 20 days they represented the predominant red blood cell population in the circulation of young pike. Electrophoretic analysis of haemolysates obtained from different developmental stages indicated the presence of distinct embryonic, larval and adult haemoglobins. The embryonic haemoglobins differed from those of the older larva and juvenile specimens and were detectable within the first week of post‐hatching development when only primitive erythrocytes were present in the blood.     

On the chromaffin cells in dog adrenal medulla; with special reference to the small granule chromaffin cells (SGC cells)

Small granule chromaffin cells (SGC cells) were identified in the adrenal medulla of adult dogs. They were small in size and usually showed a high nucleo-cytoplasmic ratio. Cytoplasmic projections were occasionally observed in some of these cells. They contained a variable number of small secretory granules with diameters ranging from 70 to 300 nm, but mostly from 100 to 200 nm. The densities of the secretory granules were variable, ranging from highly dense to less dense. These adrenal SGC cells were rich in free ribosomes and polysomes, but were relatively poor in other cell organelles. Chromaffin cells which were intermediate in their characteristics (IM cells) between the SGC cells and the typical A and N cells were also identified. These IM cells contained both highly electron dense and less dense granules in various proportions. The IM cells were classified into two subgroups, according to the proportions of adrenaline type granules and noradrenaline type granules. One group resembled A cells (IM-A cells) and the other resembled N cells (IM-N cells). Light microscopic histochemical studies of A cells stained with the ammoniacal silver solution demonstrated that they contained a small number of darkly stained granules. Electron microscopic cytochemistry revealed that the electron dense granuls in the SGC cells, IM cells and A cells reacted positively with both the potassium dichromate solution at pH 4.1 and the ammoniacal silver solution.     

Haematology of swordtail, Xiphophorus helleri. III. Ultrastructural characterization and peroxidase activity localization of blood cells in kidney and spleen of Xiphophorus helleri

The morphology of blood cells in the kidney and spleen of Xiphophorus helleri was characterized by electron microscopy. Erythrocytes, thrombocytes, lymphocytes, granulocytes and monocytes/macrophages were identified. Thrombocytes were elongate and contained bundles of microtubules and a canalicular system. Lymphocytes were heterogeneous in morphology. Granulocytes were divided into two types, based on the light microscopic results, which were extended by the present electron microscopic study. Neutrophils and eosinophils differed in abundance, cell shape, morphology of the nucleus and granula. Neutrophils displayed a spherical to three-lobulated nucleus and different types of granula. Several intermediate forms of granula were observed. The complement of granules displayed a different composition in the cells. These findings suggest a maturation process of a single type of granulum rather than several types. Eosinophils also contain different types of granula, described as G1–G4. The cell shape was variable and nuclei were small and eccentric. Monocytes and macrophages frequently showed autophagocytotic figures. A peroxidase reaction was observed only in the granula of neutrophils.     

Composition and ultrastructure of elasmobranch granulocytes. II. Rays (Rajiformes)

The blood granulocyte composition of seven species of ray is given together with ultrastructural observations made on the epigonal organ and blood of Pavoraja spinifera and the spleen of a deepwater rajid skate. Under the light microscope three granulocyte types, eosinophils, eosinophilic granulocytes and neutrophilic granulocytes could be distinguished. At the EM level two granulocyte types were apparent, one with elongated granules containing longitudinal fibrils that consolidated to form an axial rod-like inclusion, and the other with large, spherical, uniformly electron-dense granules. Correlation of light and electron microscope observations indicated that the neutrophilic granulocytes with weakly basophilic, elongated granules become weakly eosinophilic, as eosinophilic granulocytes, and these in turn develop to eosinophils with granules containing axial rods. The other granulocyte type forms another population of eosinophils with spherical granules.
The inter-relationship of these granulocytes, the identification of eosinophilic granulocytes, or heterophils, as immature eosinophils, and the co-existence of two morphologically distinct eosinophil forms are discussed.     

Morphologic and immunologic characterization of human basophils developed in cultures of cord blood mononuclear cells

Selective growth of human basophilic granulocytes was obtained in suspension cultures of mononuclear cells from umbilical cord blood. Approximately 50 to 80% of nonadherent cells recovered from 2- to 3-wk-old cultures contained metachromatic granules, and these cells were identified as human basophilic granulocytes by electron microscopy. Histamine content of cultured human basophils was comparable to that in peripheral blood basophils. Cultured basophils bear 2.7 to 3.7 X 10(5) IgE receptors per cell that bind both human IgE and rodent IgE with comparable affinity. Average equilibrium constants of the receptors for human IgE and mouse IgE were 2.56 +/- 0.88 X 10(9) M-1 and 1.85 +/- 0.86 X 10(9) M-1, respectively. The cell-surface component of the IgE receptors on cultured basophils has a m.w. of 64,000. Cultured basophils could be passively sensitized with human IgE and mouse IgE monoclonal antibody, and sensitized basophils released characteristic cytoplasmic granules and both histamine and arachidonate upon challenge with either anti-human IgE or antigen. Incubation of cultured basophils with ionophore A23187 or F-Met-Leu-Phe resulted in histamine release. However, compound 48/80 failed to induce histamine release from the cells.     

The hematogenous origin of osteoclasts: experimental evidence from osteopetrotic (microphthalmic) mice treated with spleen cells from beige mouse donors

The excessive skeletal mass and reduced bone resorption characteristic of osteopetrosis in microphthalmic (mi) mice can be corrected by irradiation and transfer of spleen cells from a normal littermate. Osteoclasts in beige (bg) mice, a mutation without osteopetrosis, have giant lysosomal granules. These two facts were exploited to trace osteoclast lineage. Microphthalmic mice treated with whole-body irradiation and spleen cells from a beige donor resorbed the excessive skeletal mass and recovered from osteopetrosis. Furthermore, osteoclasts in treated mi mice had giant lysosomal granules and resembled those found in bg donors when examined by light and transmission electron microscopy. These data provide direct evidence for a hematogenous origin of osteoclasts in mammals.     

ISOLATION OF RAT PITUITARY GRANULES AND THE STUDY OF THEIR BIOCHEMICAL PROPERTIES AND HORMONAL ACTIVITIES

A simple and rapid method is described for the isolation of the acidophilic and basophilic granules from the anterior pituitary gland of the rat. The method involves chromatography of pituitary particulates on columns of No. 545 Celite equilibrated and developed with 0.25 M sucrose. Mitochondria are retained quantitatively on the column. The granules and microsomes which are not retained on the Celite are further fractionated on a discontinuous density sucrose gradient and by differential centrifugation. Essentially homogeneous populations of acidophilic and basophilic granules were obtained as indicated by 1) extensive electron microscopic studies, 2) enzymatic determinations, and 3) fatty acid and RNA analyses on the granule pellets. Microfiltration studies indicated that the acidophilic granules were smaller than 450 mµ, but greater than 300 mµ in diameter. They were found, unlike the basophilic granules, to be partially stable to extraction with water, but were unstable on incubation in 1 mM EDTA at 37°. Magnesium ions were not detected in the granules. The acidophilic and basophilic granules contained, respectively, 5 and 8 per cent of the protein present in the whole homogenate. Extensive hormone studies showed that growth and lactogenic hormones were associated with the acidophilic granules, while thyroid-stimulating hormone and gonadotropin were associated with the basophilic granules. ACTH was not present in significant amounts in either of the granule fractions, but was localized in a particulate fraction which contained microsomes and small granules. The association of the pituitary hormones with specific granules and cell types is discussed.     

The blood cells of the Antarctic icefish Chaenocephalus aceratus Lönnberg: light and electron microscopic observations

Peripheral blood and haemopoietic tissues of spleen and kidney of the icefish, Chaenocephalus aceratus were examined using LM and EM techniques. The peripheral blood contained cellular elements from all the recognized cell lines usually seen in other teleost groups. Erythrocytes were very rare; when found, they were mature or senile and fragile. Thrombocytes of two morphologies, several cell types considered to be part of the lymphoid series and monocytes/macrophages were present. Two distinctive types of granulocytes also were found; their morphologies and granulation were so different from teleost granulocytes hitherto described that their identification was impossible.     

Electron microscopic study of the regeneration in vitro of rat peritoneal mast cells after histamine secretion

Summary Regeneration of rat mast cells was studied by TEM from 10 s to 48 h after secretion of histamine induced by compound 48/80. During the first 2 h, small intracellular cavities, formed during compound exocytosis and containing non-membrane-bound remnants of the granules, tended to coalesce, and after 2 h of incubation regeneration started. After 6 h, all the cavities had fused into one large central cavity which contained the remnants of the granules and remained open to the exterior during the entire period. The plasma membrane microfolds which disappeared just after secretion were reformed during regeneration. They were apparently involved in endocytotic-like activity and coated vesicles also appeared beneath the plasmalemma (membrane recycling?). The fate of the granule remnants in the cavity is unknown, as regeneration was not completed after 48 h which is the longest survival time obtained so far in ultrastructural studies of mast cell regeneration in vitro.     

Correlative Fine Structural,Behavioral, and Histochemical Analysis of Ascidian Blood Cells

Abstract The blood cells of a solitary ascidian, Halocynthia roretzi, were examined by electron microscopy (EM) with reference to their appearance by light microscopy (LM). In addition, their movement and stainability by vital dyes was observed by phase-contrast microscopy, and their stainability by Giemsa was also examined. Nine cell types were recognized: vacuolated cells, hyaline amoebocytes, small amoebocytes, granular amoebocytes, macrogranular cells, globular cells, lymphocyte-like cells, large basophilic cells and large granular cells. Vacuolated cells were found to possess various numbers of vacuoles containing strongly electron-dense materials and could be divided into at least three subgroups. Granular amoebocytes contained microfilaments and many granules of uniform size. Hyaline amoebocytes and small amoebocytes seemed to be specialized as phagocytes. Macrogranular cells and globular cells were not well characterized. In the blood of adult individuals, hemoblasts were rarely found, although lymphocyte-like cells were present. Each of two large cells, large basophilic cells and large granular cells, possessed novel granules or vacuoles, whose functions remain to be elucidated. The possible functions and relationships of these cells among various ascidian species are discussed.     

Morphological alteration of peritoneal mast cells and macrophages in the mouse peritoneal cavity during the early phases of an allergic inflammatory reaction

We investigated the presence of mast cell granules in macrophages following an in vivo model of an allergic reaction. Injection of ovalbumin (100 microg) into the peritoneal cavity of sensitised mice produced a rapid (within 2 h) influx of neutrophils followed by a slower (after >4 h) eosinophil migration. Ovalbumin treatment induced a high incidence (approximately 50%) of mast cell degranulation compared to control phosphated-buffered saline-treated mice. The majority (approximately 90%) of peritoneal macrophages contained mast cell granules as early as 2 h post-ovalbumin, with lower values at later time-points, as determined by staining with Toluidine blue and Berberine sulphate. This was confirmed by electron microscopy which enabled us to identify the complex mast cell granule sub-structural components in macrophage phagosomes. In conclusion, we used histochemical and ultrastructural analyses to show that mast cell granules become internalised with macrophages during the early stages of an experimental allergic reaction.     

Histological evaluation of immunologically mediated tumor regression of the line 10 guinea pig hepatocarcinoma

The histology of immunologically mediated tumor regression was studied in the syngenic strain 2 guinea pig/line 10 hepatocellular carcinoma tumor system. Tumor regression was induced non-specifically by the intralesional injection of living Bacillus Calmette-Guérin (BCG) in 7-day-old established tumors (diameter 8-10 mm). In untreated line 10 tumors at day 7 a mild to moderate inflammatory reaction was present, which consisted mainly of small mononuclear cells; in addition large mononuclear cells and basophils were present. Intratumoral BCG-treatment induced a prominent increase in the inflammatory reaction due to an influx of small and large mononuclear cells and neutrophils. Small mononuclear cells were identified mainly as lymphocytes whereas large mononuclear cells belonged mainly to the macrophage line. Intratumoral administration of BCG resulted in a granulomatous reaction. A time-related decrease in the number of tumor cells and an increase in inflammation, associated with purulent lysis of the granulomatous tissue, was observed. Specific immune-mediated tumor rejection occurred in animals both after active immunization and after adoptive transfer of immune spleen cells. In actively immunized animals the tumor cells were rapidly rejected and from day 4 onwards no tumor cells could be detected at the injection site. Lymphocytes were the major component of the inflammatory reaction; large mononuclear cells were present to a lesser extent and basophilic granulocytes were regularly observed. After adoptive transfer of immunity with immune spleen cells given simultaneously with an intradermal innoculation of tumor cells, an essentially similar rejection reaction was found, although tumor cell rejection was delayed. Lymphocytes and large mononuclear cells were found in equal proportions, whereas basophilic granulocytes were always present in smaller numbers. After BCG-induced regression and in adoptively transferred immune rejection, a fibroblast component was more prominent than in untreated control tumors. This reaction tended to isolate smaller tumor cell areas into islets of decreasing sizes. In contrast with the fibroblast component of growing tumors, the proliferative pre-existing fibrous tissue in tumors undergoing regression or rejection showed a loosely arranged architecture and contained a marked cellular infiltrate. From the results of the present study it was concluded that the morphological expression of line 10 tumor rejection varies. Without immune cells, BCG is needed for the induction of a local inflammatory reaction, which was granulomatous in type and eventually led to complete tumor cell eradication.(ABSTRACT TRUNCATED AT 400 WORDS)