长薄鳅外周血细胞的显微结构和细胞化学特征研究

长薄鳅外周血细胞可分为红细胞、中性粒细胞、单核细胞、淋巴细胞和血栓细胞.在数量上,中性粒细胞、单核细胞、淋巴细胞和血栓细胞占白细胞总数的百分比分别是17.06%、5.83%、28.16%和48.94%.细胞化学染色显示所有白细胞均含有糖原物质,所有红细胞均不含酸性磷酸酶,中性粒细胞、单核细胞、淋巴细胞和血栓细胞均含有酸性磷酸酶.非特异件酯酶染色显示单核细胞呈阳性反应,中性粒细胞、淋巴细胞和血栓细胞均为部分呈阳性反应.所有细胞的碱性磷酸酶、过氧化物酶、苏丹黑显色反应均呈阴性.     

Light- and Electronmicroscopic Observations on the Blood Cells of the Atlantic Hagfish,Myxine glutinosa (L.)

In the blood of Myxine glutinosa three cell lines may be distinguished: erythrocytes, granulocytes and agranulocytes. The erythrocytes are remarkably large, oval and nucleated. They are well defined in all their developmental stages by a characteristic micropinocytosis. They originate from blast cells which proliferate in the circulating blood. The blast cells probably form from agranulate stem cells of the intestinal myeloid tissue. The granulocytes constitute about half of the leucocytes. They are neutrophilic with a lobated nucleus. The granulocytopoiesis takes place in the intestinal myeloid tissue. The agranulocytes mainly include two cell types, termed spindle cells and lymphocyte-like cells. These cell types, however, transform into each other. Macrophages occur essentially in the peritoneal cavity rarely in the blood. Transition forms between macrophages and granulocytes may exist. The blood also contains cells which on morphological grounds have been termed thrombocytes. Whether these cells are identical with those necessary for clotting of the blood remains to be proved. With the exception of erythroblasts, the different lines of blast cells are difficult to identify and distinguish from each other. Possibly all lines of blood cells originate from agranulate lymphocyte-like stem cells, most of which are produced by the intestinal myeloid tissue.     

Some observations on the stained blood cellular elements of channel catfish, Ictalurus punctatus

Slides were prepared from blood of channel catfish, Ictalurus punctatus , and evaluated. The cellular blood elements observed were thrombocytes, lymphocytes, neutrophils, monocytes, eosinophils, basophils, granular anucleate bodies, erythrocytes. In channel catfish, thrombocytes, lymphocytes and neutrophils were the predominate leucocytes. Erythrocytes were observed in various stages of development. Smudge cells were observed in all smear preparations. The variations found between the observations in this study and information provided in the literature suggest the need for establishing a standardized terminology.     

团头鲂外周血细胞的显微结构及细胞化学特征

采用常规瑞氏染色和细胞化学染色方法对团头鲂(Megalobrama amblycephala)外周血细胞的显微结构及细胞化学特征进行了观察。在团头鲂外周血细胞中可区分出六类细胞: 红细胞、淋巴细胞、单核细胞、嗜中性粒细胞、嗜酸性粒细胞和血栓细胞。其中淋巴细胞是除红细胞外含量最多的细胞, 其次分别为血栓细胞、单核细胞、嗜中性粒细胞、嗜酸性粒细胞。成熟红细胞多为卵圆形, 表面光滑, 胞核呈椭圆形或圆形, 染色质较为致密; 淋巴细胞多呈圆形, 胞质较少, 胞核常偏位; 单核细胞多为圆形, 胞核呈圆形或椭圆形, 胞质内可见空泡状结构; 嗜中性粒细胞近似圆形, 胞核常偏于细胞一侧, 呈分叶状、肾形或椭圆形, 核质界限清晰; 嗜酸性粒细胞一般为圆形, 胞核为肾形或椭圆形, 胞质中充满紫红色颗粒; 血栓细胞形态多样, 主要有椭圆形、纺锤形、长杆状和泪滴形, 核质比较大。淋巴细胞呈α-醋酸萘酚酯酶(ANAE)阳性, 呈过碘酸-雪夫(PAS)、氯乙酸AS-D萘酚酯酶(AS-DCE)弱阳性, 呈苏丹黑B(SBB)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)及过氧化物酶(POX)阴性; 单核细胞呈POX、ACP强阳性, PAS、SBB、AS-DCE和ANAE为阳性, 呈AKP阴性; 嗜中性粒细胞除PAS和ANAE为弱阳性外, 其他染色结果和单核细胞相同; 嗜酸性粒细胞呈POX、ANAE强阳性, SBB、ACP阳性, PAS及AS-DCE则为弱阳性, 呈AKP阴性; 血栓细胞呈PAS、AS-DCE及ANAE弱阳性, 呈SBB、ACP、AKP及POX阴性。团头鲂外周血细胞的显微结构及细胞化学特征与其他鱼类具有相似之处, 但亦有其明显的物种特异性。该研究结果可作为监测团头鲂健康状态的依据, 为其养殖及病理诊断提供基础资料。     

Friedrich Miescher and the discovery of DNA

Over the past 60 years, DNA has risen from being an obscure molecule with presumed accessory or structural functions inside the nucleus to the icon of modern bioscience. The story of DNA often seems to begin in 1944 with Avery, MacLeod, and McCarty showing that DNA is the hereditary material. Within 10 years of their experiments, Watson and Crick deciphered its structure and yet another decade on the genetic code was cracked. However, the DNA story has already begun in 1869, with the young Swiss physician Friedrich Miescher. Having just completed his education as a physician, Miescher moved to Tübingen to work in the laboratory of biochemist Hoppe-Seyler, his aim being to elucidate the building blocks of life. Choosing leucocytes as his source material, he first investigated the proteins in these cells. However, during these experiments, he noticed a substance with unexpected properties that did not match those of proteins. Miescher had obtained the first crude purification of DNA. He further examined the properties and composition of this enigmatic substance and showed that it fundamentally differed from proteins. Due to its occurrence in the cells' nuclei, he termed the novel substance "nuclein"--a term still preserved in today's name deoxyribonucleic acid.     

Cell types in peripheral blood of the nurse shark: An approach to structure and function

Ultrastructural and functional studies were carried out on nurse shark (Ginglymostoma cirratum) peripheral blood cells in order to identify cells of definitive morphology and specific function. Along with erythrocytes and thrombocytes, four morphologically distinct leucocytes are recognized in peripheral blood: two types of granulocytes, the ‘eosiniphil’ and the ‘granulocyte’, and two mononuclear agranulocytic cells, one resembling mammalian macrophage and monocyte, the other resembling mammalian lymphocyte. Also present in peripheral circulation are blast-like cells and mitotic cells. In vitro phagocytosis was demonstrated by the monocyte-macrophage and the granulocyte while thrombocytes, eosinophils and lymphocytes showed no phagocytic activity in the system studied. It is stressed that care must be used in drawing functional analogies between blood cells of a mammal and an elasmobranch on the basis of morphological similarity alone.     

Identification of peripheral blood leucocytes of the dogfish (Scyliorhinus canicula L.) by electron microscopy

Dogfish peripheral blood leucocytes were examined by electron microscopy after the injection of colloidal carbon. The cells were classified as lymphocytes, plasma cells, monocytes, thrombocytes and granulocytes. The granulocytes were further classified into four types according to the structure of their granules. Monocytes, thrombocytes and two types of the granulocytic cells were phagocytic.     

Haematology of the turbot, Psetta maxima (L.): ultrastructural, cytochemical and morphological properties of peripheral blood leucocytes

Optimum procedures for fish handling and sample processing for use when employing haematological parameters as health indicators in turbot, Psetta maxima (L.), have been established. We found thrombocytes to be the most abundant blood cell, representing approximately 52% of circulating leucocytes (lymphocytes, 40.8%; granulocytes, 5.6%; monocytes, 1.6%; total number of leucocytes=1.3 × 10⁵ ml⁻¹; packed cell volume=22.7%). The light- and electron-microscopical characteristics of these cell types are described, together with their cytochemical properties using Sudan Black B, Periodic Acid Schiff, Non-specific Esterase, and Acid and Alkaline Phosphatase. Turbot thrombocytes showed a high degree of shape alterations when observed in live preparations using phase contrast microscopy, while ultrastructural observations following the in vitro uptake of carbon particles supported an active process of phagocytosis by the thrombocytes, rather than passive entrapment. The lymphocytes of turbot are structurally similar to mammalian lymphocytes with the highest nuclear:cytoplasmic ratio of all the leucocytes observed. Small lymphocytes predominated, large lymphocytes forming less than 1% of the total white blood cell population. The most frequent granulocyte type was a neutrophil-like cell with an eccentric nucleus, only rarely seen in segmented form. In vitro uptake of carbon particles by granulocytes was not observed under the conditions of the experiment, although turbot granulocytes are capable of phagocytosis under different circumstances. These are discussed, along with other physiological and technical factors which can influence the blood parameter findings in fish.     

A comparison of internal standards for plant cytophotometry

Plant cell nuclei were compared with chicken erythrocyte nuclei for use as internal standards for microspectrophotometry. The amount of DNA per nucleus and the coefficient of variation for measurement of individual nuclei were determined for cells from dormant embryos of Pinus taeda and Pinus coulteri, from onion root tips and from chicken erythrocytes. The chicken erythrocytes had the least variability and thus were best suited for use as a standard. Onion root tips were least suitable, with a coefficient of variation 2 1/2 times that of erythrocytes. Although onion root tips have been used as an internal standard in other studies, their mitotic activity, in contrast with the nonreplication of DNA of mature erythrocytes, is reflected in a broad distribution of nuclei with values in the 2C-4C range. Coulter pine mature embryos were at the 3C level, whether dry or hydrated, while loblolly pine embryos were in the 2C state. This confirms previous reports. The coefficient of variability for the pine embryo cells was 1 1/2 times that of erythrocytes for nonhydrated seeds and twice the erythrocyte value for hydrated seeds. The larger 2C values for pine (26 pg for P. taeda and 17 pg for P. coulteri) are closer to values expected for many plant species than the 3 pg level of the chicken erythrocytes. Dormant P. taeda embryo cells (2C) are suggested as an alternative where the experimental material has large DNA values and/or chicken erythrocytes are difficult to procure. Large sample size is recommended for the plant materials if they are to be used as internal standards in Feulgen cytophotometry.     

Observations on the ultrastructure of nucleated erythrocytes and thrombocytes,with particular reference to the structural basis of their discoidal shape

Summary The marginal band of nucleated erythrocytes in the toadfish is found, in electron micrographs, to be composed of about twenty-five microtubules approximately 200 Å in diameter. These form a bundle that encircles the erythrocyte just beneath the plasma membrane. These observations support the interpretation of Meves 1904, that this relatively stiff equatorial band may contribute to the maintenance of the discoid shape of nucleated erythrocytes in fish, amphibians, reptiles and birds.Similar microtubules form an annular bundle encircling the nucleus in fish thrombocytes. The number of tubular elements involved here is in excess of one hundred and they are located deep to the ectoplasmic layer instead of immediately beneath the plasmalemma. The term endoplasmic ring is therefore proposed for this structure.Comparative observations on nucleated erythrocytes of various species are presented showing that the density and fine structure of the material occupying the interchromosomal areas of the nucleus, always matches the cytoplasm and is related to the hemoglobin concentration of the species. These ultrastructural observations are consistent with the optical absorption and biochemical findings of other investigators indicating the presence of intranuclear hemoglobin in nucleated erythrocytes. Crystalline order is occasionally found in electron micrographs of the hemoglobin rich areas of the nucleus in toadfish erythrocytes but is not found in the cytoplasm.This research was supported by grant G-12916 of the National Science Foundation.     

Studies on the reticulo-endothelial system of the dogfish,Scyliorhinus canicula

Summary Clearance and subsequent localisation of a range of materials, including colloidal carbon, latex beads, sheep erythrocytes, bacteria and dextran were followed in the lesser spotted dogfish, Scyliorhinus canicula. It was found that two populations of peripheral blood leucocytes — monocytes and thrombocytes, but not granulocytes — were involved in clearance of the circulation. In the case of carbon, this material was cleared from the plasma after 12 h, and both the colloid-containing thrombocytes and monocytes disappeared from circulation by 8 weeks post injection. Upon injection of some of the materials, and particularly bacteria, a settling out of monocytes containing phagocytosed material was seen in the secondary lamellae and cavernous bodies of the gills. Large clumps of monocytes were found in the gills as early as 30 min post injection and these increased in size for up to one week, after which they gradually dispersed. The lining cells of the cavernous body, known as CB cells, were also responsible for the sequestration of carbon, latex beads and probably erythrocytes, but dextran and bacteria were not internalised. The origin, functions and phylogenetic significance of the CB cells are discussed.     

Catecholamines and the abundance of blood cells in a fresh water tropical teleost (Colisa fasciatus) in relation to cold-shock.

Intraperitoneal injections of epinephrine (0.05 microng/g) elicited characteristic changes in the abundance of circulating leucocytes at selected time intervals (viz., 3, 15 and 27 min. and, later, at intervals of 48 minutes up to 363 min. and at 12 and 24 h) post-injection in Colisa. Leucocytosis was evident as 15, 27 and 75 min, and tendency towards leucocytosis was observed at 123 and 267 min.; at 3, 171, 219, 315 and 363 min. and at 12 and 24 h, the total leucocyte counts for the experimentals and controls were not significantly different. No such corresponding significant changes were observed in the abundance of circulating erythrocytes or thrombocytes. Leucocyte sequence elicited by epinephrine was also apparent in Colisa which had been exposed to a temperature of 2 degrees C for one minute. Pretreatment with phenoxybenzamine (3.0 mg/l aquarium water), an alpha-adrenergic antagonist, abolished the cold-shock leucocytic phases, except 27 min. leucocytosis which emerged unaffected. Exogenous norepinephrine (2 microng/g) and isoproterenol (0.1 microng/g) failed to elicit any significant change in the number of circulating leucocytes, erythrocytes, or thrombocytes. Clearly adrenergic mechanisms are involved in the cold-shock leucocyte stress sequence. The results suggest alpha-activating role of epinephrine during the leucocyte stress syndrome in Colisa and, apparently, epinephrine is leucocytic.     

Stage-Specific Expression and Localization of MENT, a Nuclear Protein Associated with Chromatin Condensation in Terminally Differentiating Avian Erythroid Cells

Polyclonal antibodies have been raised against a nonhistone protein (MENT) which has been previously shown to be associated with the repressed chromatin of mature chicken erythrocytes and to promote the in vitro condensation of chromatin of immature erythrocyte nuclei. Here we report that the expression pattern of MENT closely follows chromatin condensation in maturing arian erythrocytes of definitive and primary lineages. Accumulation of MENT correlates more strongly with chromatin condensation than does accumulation of histone H5. In addition to being present in erythrocytes, the protein was also found in neutrophil nuclei and an immunofluorescence reaction was observed with embryonic (nucleated) thrombocytes. MENT was not detected in other chicken tissues (brain, liver, testis). In intact erythrocytes, MENT immunofluorescence was found in foci close to the nuclear periphery, while in isolated, decondensed nuclei, the fluorescence signal was uniformly distributed. In neutrophil nuclei, containing approximately 10 times more MENT than adult erythrocytes, intense staining associated with the peripheral heterochromatin was observed. These findings are discussed in regard to a possible mechanism for chromatin condensation by MENT.     

版纳鱼螈外周血细胞观察

以濒危两栖动物版纳鱼螈(Ichthyophis bannanica)为材料,应用瑞氏-姬姆萨混合染色法与血细胞计数法观察并统计了版纳鱼螈各种外周血细胞的形态特征和数量比例.结果表明,版纳鱼螈的外周血液中红细胞数量较多,呈卵圆形、椭圆形、梭形和梨形,平均含量为2.57 ×105个/mm3.白细胞数量较少,多呈近圆形,平均含量为0.72×103个/mm3.白细胞中,淋巴细胞最多,其次为单核细胞、嗜中性粒细胞、嗜碱性粒细胞和嗜酸性粒细胞.血栓细胞数量较少,常数个集合在一起.同时,将此研究结果与鱼类、爬行类和其他两栖类的血细胞比较,进而探讨了版纳鱼螈的进化地位.     

A Comparison of Internal Standards for Plant Cytophotometry

Plant cell nuclei were compared with chicken erythrocyte nuclei for use as internal standards for microspectrophotometry. The amount of DNA per nucleus and the coefficient of variation for measurement of individual nuclei were determined for cells from dormant embryos of Pinus taeda and Pinus coulteri, from onion root tips and from chicken erythrocytes. The chicken erythrocytes had the least variability and thus were best suited for use as a standard. Onion root tips were least suitable, with a coefficient of variation 2 1/2 times that of erythrocytes. Although onion root tips have been used as an internal standard in other studies, their mitotic activity, in contrast with the nonreplication of DNA of mature erythrocytes, is reflected in a broad distribution of nuclei with values in the 2C-4C range. Coulter pine mature embryos were at the 3C level, whether dry or hydrated, while loblolly pine embryos were in the 2C state. This confirms previous reports. The coefficient of variability for the pine embryo cells was 1 1/2 times that of erythrocytes for nonhydrated seeds and twice the erythrocyte value for hydrated seeds. The larger 2C values for pine (26 pg for P. taeda and 17 pg for P. coulteri) are closer to values expected for many plant species than the 3 pg level of the chicken erythrocytes. Dormant P. taeda embryo cells (2C) are suggested as an alternative where the experimental material has large DNA values and/or chicken erythrocytes are difficult to procure. Large sample size is recommended for the plant materials if they are to be used as internal standards in Feulgen cytophotometry.     

Phenotypic and cytologic studies of lymphoid cells and monocytes in primary culture of porcine bone marrow during infection of African swine fever virus

We have modeled in vitro infection of African swine fever virus (ASFV) in primary unstimulated cells of the porcine bone marrow and have studied the phenotypical changes in the population of porcine lymphoid cells by cytophotometry. Monocytes and large-sized lymphocytes completely vanished in 72 h of infection which is result of high sensitivity of those cells to ASFV. We describe DNA synthesis in monocytes at 24 h post infection. Cytophotometry of the uninfected cells revealed the few number of atypical lymphocytes and lymphoblasts after 72 h of cultivation; whereas in viral infected cultures, atypical cells appeared in large quantity (about 14%) with 24 h. Most of atypical lymphocytes and lymphoblasts had altered nucleus, and only a small number of atypical cells had additional nucleus. The cytophotometry of main and additional nuclei showed that DNA content didn't exceed diploid standard which indicates that the additional nuclei were consequence of fragmentation of nuclei in lymphocytes.     

DNA replication and H5 histone exchange during reactivation of chick erythrocyte nuclei in heterokaryons

Fusion of terminally differentiated chick erythrocytes (CE) with replicating quail myoblasts or established L6J1 rat myoblasts results in reactivation of DNA synthesis in the dormant CE nuclei and in suppression of DNA synthesis in the myoblast nuclei. The nuclei of primary quail myoblasts are more effectively inhibited than the nuclei of established rat myoblasts. Inhibition of DNA replication occurs not only by preventing G1 nuclei from entering S-phase but also by blocking nuclei in S-phase and by delaying nuclei in G2 from undergoing mitosis and starting a new DNA replication cycle. No inhibition of DNA synthesis could be observed when mouse erythrocytes, i.e., erythrocytes lacking nuclei, were fused with rat myoblasts to generate mouse-globin-containing L6J1 cybrids. — Reactivation of CE nuclei is associated with a loss of the tissuespecific H5 histone variant. Complete elimination of H5 histone, however, does not seem to be a necessary prerequisite for the initiation or completion of DNA replication in CE nuclei since H5 antigens are found on reactivated G1, S, and G2 nuclei.     

Changes in the amount and synthesis of DNA in the nuclei of large decidual cells in rats during their differentiation

Using cytophotometry of the Feulgen-stained nuclei, the quantity of DNA was measured in the nuclei of rat's large decidua cells (LDC) on tissue sections of the antimesometrial region within days 7-13 of gestation. The quantity of nuclear DNA was expressed in units of ploidy, the haploid DNA standard being the quantity of DNA in rat's spermatid nucleus. On different days of gestation, the nuclear DNA was seen to vary in cells located in different zones of decidua. The maximum DNA content was found in the LDC located on days 9-12 of gestation somewhat in the middle of the decidua thickness. On day 11, the quantity of nuclear DNA in these cells reached in average, 22c. The quantity of DNA in the nuclei of the least differentiated LDC located on the periphery of decidua never exceeded 4.9c, whereas that in the nuclei of the most differentiated LDC, located close to the embryo, varied from 2.9c to 9.3c. On days 10 and 11 of rat's false gestation, the maximum DNA contents in the nuclei were registered in the LDC located in the middle of the decidua thickness. 3H-thymidine incorporation into the nuclei of the most differentiated LDC located nearest to the embryo stopped starting from day 10 of gestation. Phenomena of lesser quantities of nuclear DNA in most differentiated LDC, compared to that in LDC in the previous steps of differentiation, are discussed.     

Biochemical and ultrastructural study of the sperm chromatin from Mytilus galloprovincialis

Protein composition and ultrastructure of the mature spermatozoa of the mussel Mytilus galloprovincialis were studied upon gradual decondensation of the nuclei with increasing NaCl concentration. Three types of protein were found, associated with the sperm DNA: (1) the sperm-specific proteins S1, S2 and S3 (80% of the acid-soluble proteins); (2) the four core histones (20%); (3) three non-histone proteins tightly bound to DNA (about 4 micrograms protein per 100 micrograms DNA). The sperm-specific protein S3 was the first to dissociate at about 0.5 M NaCl and electron micrographs of spread nuclei indicated its participation in the final compaction of the nucleus. Hypotonically treated sperm nuclei revealed the presence of 21-25 nm large granules irregularly scattered along some of the DNA fibers. These granules correspond to the 'superbeads' of histone-containing chromatins. The tightly bound non-histone proteins were represented by a triplet in the range 60-80 kD. They formed 30-60 nm large annular bodies holding DNA fibers and resisting high salt-detergent treatment.