First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea (original) (raw)
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First cytochemical study of haemocytes from the crab Carcinus aestuarii (Crustacea, Decapoda)
European Journal of Histochemistry, 2010
For the first time, a morphological study of haemocytes from the crab Carcinus aestuarii was carried out by means of light microscopy and differing cytochemical assays. Analysis of haemocyte size frequency distribution (performed by means of a Coulter Counter) revealed the presence of two distinct haemocyte fractions in C. aestuarii haemolymph, depending on cell size. The first fraction was of about 3-5 µm in diameter and 30-50 fL in volume, the second was of about 6-12 µm in diameter and over 200 fL in volume. Mean cell diameter and volume were 8.20±1.7 µm and 272.30±143.5 fL, respectively. Haemocytes observed under light microscope were distinguished in three cell types: granulocytes (28%; 11.94±1.43 µm in diameter) with evident cytoplasmic granules, semigranulocytes (27%; 12.38±1.76 µm in diameter) with less granules than granulocytes, and hyalinocytes (44%; 7.88±1.6 µm in diameter) without granules. In addition, a peculiar cell type was occasionally found (about 1%): it was 25-30 µm in diameter and had a great vacuole and a peripheral cytoplasm with granules. Granulocyte and semigranulocyte granules stained in vivo with Neutral Red, indicating that they were lysosomes. Giemsa's dye confirmed that granulocytes and semigranulocytes were larger than hyalinocytes. Pappenheim's panoptical staining and Ehrlich's triacid mixture allowed to distinguish granule-containing cells (including semigranulocytes) in acidophils (64%), basophils (35%) and neutrophils (1%). Hyalinocytes showed always a basophilic cytoplasm. Haemocytes were positive to the PAS reaction for carbohydrates, even if cytoplasm carbohydrate distribution varied among cell types. Lastly, lipids were found on cell membrane and in cytoplasm of all haemocyte types in the form of black spots produced after Sudan Black B staining. The morphological characterisation of C. aestuarii haemocytes by light microscopy was necessary before performing both ultrastructural and functional studies of circulating cells.
International Journal of Fisheries and Aquaculture, 2013
For the first time haematological studies on freshwater crab P. masoniana (Henderson) has been carried out in Jammu, India. The aim of studies were to determine various type of hemocytes, their total and differential count as well. Based on morphological studies of hemocytes under microscope three types of haemocytes were identified viz., Hyalinocytes (H), Semigranulocytes (SG) and Granulocytes (G). These cells were varied with respect to their proportion, Semigranulocytes being the heighest (54%) follow by Granulocytes (30%) while Hyalinocytes were least (16%) among three cells.
Scylla tranquebarica revealed the presence of prohaemocytes, granular haemocytes, cystocytes, plasmatocytes and podocytes. The diagnostic characteristics of these cells conformed to the descriptions given by earlier authors in decapod crustaceans and insects. The spherule cells, adipohemocytes and oenocytoids have been found to be absent. Anucleate cells have been identified by both light and transmission electron microscopic observations. The discernibility of such anulceated cells added a new dimension to the concept of hemocytes origin and differentiation. The existence of anucleate granulocytes is construed to be a tenable phenomenon and situation considering the stem cells property that they undergo both symmetric and asymmetric divisions to maintan cell population in organisms. The total hemocyte count and the differential count indicated the defence potential of the species.
Crustacean haemocytes and haematopoiesis
Aquaculture, 2000
Crustacean haemocytes play important roles in the host immune response including recognition, phagocytosis, melanization, cytotoxicity and cell-cell communication. Classification of the haemocyte types in decapod crustaceans is based mainly on the presence of cytoplasmic granules into hyaline cells, semigranular cells, and granular cells. Each cell type is active in defence reactions, for example; in crayfish, the hyaline cells are chiefly involved in phagocytosis, the semigranular cells are the cells active in encapsulation, while the granular cells participate in Ž . storage and release of the prophenoloxidase proPO system and cytotoxicity. The haematopoietic tissue has been described in several crustacean decapod species and shown to be the haemocyteproducing organ. Tentative stem cells have been shown to be present in this tissue. Using in situ hybridization, we demonstrated that proPO is not present in the haematopoietic tissue of crayfish which suggests that protein expression is different between circulating haemocytes and the cells in the haematopoietic tissue. q
Journal of FisheriesSciences. …, 2009
In the research, haemocyte types existing in hemolymph (the liquid consisting of blood and lymph) of Blue Crab's (Callinectes sapidus Rathbun, 1896) which was captured in Akyatan Lagoon in the North East of Mediterranean region, their amounts in ml and this cells relations with genital and carapace length were defined. Three types cell were identified at the end of the research and these are hyaline, granule and semigranule. The total amount of haemocyte at average ml in female was calculated as 242.300 ±6.113x10 4 , hyaline 25.722 ±0.916x10 4 , semigranule 125.817 ±4.847x10 4 and the amount of granule was calculated as 91.078 ±3.047x10 4 whereas in males total haemocyte was calcuated as 216.434 ±4.778x10 4 , hyaline was 21.447±0.609x10 4 , semigranule was 112,355±3,058x10 4 and the amount of granule haemocyte was calculated as 82.632 ±2.080x10 4 . In total (male and female) semigranule was dominated with 55% and respectively granule with 31% and hyaline haemocyte with 14 % were dominated. Total amount of haemocyte, hyaline, semigranule and granule haemocyte in females whose average carapace length is 60,715 mm, carapace width is 106,368 mm and weight is 147,884 g was found higher than males whose carapace length is 72.161 mm, carapace width is 126.391 mm and weight is 273.719 g (p<0.05).
Journal of Invertebrate Pathology, 2012
Monoclonal antibodies (MAbs) to hemocytes of mud crab, Scylla serrata, were produced by immunizing mice with formalin-fixed hemocytes. Of the six MAbs produced, two (MAb 1D11 and MAb 1A2) reacted specifically with hemocyte proteins in western blot. MAb 1A2 showed strong immunofluorescent reaction with granular hemocytes and a weak reaction with semigranular cells. However, hyaline cells did not show any reaction. The MAbs also showed strong cross-reactivity with the hemocytes of different crab species but not with other crustaceans. The MAbs produced can be used as a marker for granular hemocytes of crabs.
Fish & Shellfish Immunology, 2008
This work presents the first detailed microscopic and functional analysis of the haemocytes of an abalone; the European Haliotis tuberculata. It is shown that in contrast to the situation in bivalves, only very few basophilic ''granulocytes'' could be found and exclusively with a histological stain. Neither flow cytometry, phase contrast observation nor transmission electron microscopy were able to detect any granular cells. The large majority of cells was constituted of ''hyalinocytes'', which could be sorted by flow cytometry, for the first time, into small (blast-like) and large cells. This permits a detailed analysis of haemocytes and especially of the lowly represented blast-like cells. The differences in haemolymph cell composition between bivalves and gastropods is reviewed in depth and discussed in view of the new data we present. Most of the abalone haemocytes analysed harbour many vacuoles, large glycogen deposits, lipid inclusions and acidic compartments. However, although the number of these ''inclusions'' was rather variable in between individual hyalinocytes, these experiments did not allow to discern subpopulations using these criteria, and the population appears more a v a i l a b l e a t w w w . s c i e n c e d i r e c t . c o m j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / f s i Fish & Shellfish Immunology (2008) 24, 400e411
Journal of Experimental Zoology, 1977
Localization of c l o t t a b l e protein (CP) i n hemocytes of the Japanese horseshoe crab, Tachypleus tridentatus, was investigated by a fluorescent antibody technique. minority of hemocytes (15%) were stained strongly and uniformly while those i n the majority of hemocytes (85%) were not. cytoplasmic matrix of the majority was stained with various degrees of intensity and t h a t of the minority was not. The two types of hemocytes which showed d i f f e r e n t i n t r a c e l l u l a r localization of CP might be i n d i f f e r e n t maturational stages o r m i g h t play d i f f e r e n t roles in blood coagulation.
Classification of Hemocytes from Four Crustaceans and Cross-Reactivity of Their Antisera
Journal of Shellfish Research, 2018
Common antigens and analyzed the hemocytes of the lady crab (Charybdis japonica) were studied. Chinese mitten crab (Eriocheir sinensis), Dungeness crab (Cancer magister), and red king crab (Paralithodes camtschaticus) using a laser scanning confocal microscope (LSCM), flow cytometry (FCM), Western blotting (WB), and transmission electron microscopy. In this paper analyzed hemocyte classification and immunological characteristics of the whole proteins in these species and their antisera. Four antisera positively cross-reacted, to varying degrees, with hemocytes of the four species. Two subpopulations were classified in C. japonica, P. camtschaticus, and E. sinensis and three in C. magister. The LSCM results showed that four antisera were positive with the hemocytes of the four crustacean species, respectively. Dot fluorescent signals on the hemocyte membranes show a cricoid intensive distribution. FCM analysis showed a positive rate of cross-reaction, in which the reaction of the antisera with its own hemocytes was higher than that with other speciesÕ hemocytes. The positive rate of granulocytes was always higher than that of hyalinocytes. Western blotting results showed that the four antisera mainly recognized antigens with molecular weights of about 75 or 80 kDa in the hemocytes of C. japonica, 80 or 90 kDa in the hemocytes of P. camtschaticus, 30 and 82 kDa in the hemocytes of E. sinensis, and 43 or 70 kDa in the hemocytes of C. magister. More lanes reacted between hemocytes of E. sinensis and C. magister and their antisera. The results indicate antigenic similarities among the hemocytes of the four crustacean species.
Cytometric investigations on hemocytes of the American oyster, Crassostrea virginica
Tissue and Cell, 1988
Pericardial hemolymph was obtained from American Oysters (Crassosrrea virginica) and the hemocytes characterized by flow cytometry. The cells were found to have a broad unimodal size distribution with a median diameter of 7 micrometers. Total protein measured by flow cytometric fluorescence of dansylated cells also revealed a broad unimodal distribution similar to that obtained for size. The proportion of hemocytes in each stage of the cell cycle was measured using DNA-specific DAPI fluorescence. Histograms showed a single peak representing the Go/G, population. There was no evidence of S or G2+M phases of the cell cycle, nor was polyploidy seen. The forward and orthogonal light scatter of lixed hemocytes showed no evidence of sub-populations on the basis of cytoplasmic granularity. Thus, in terms of these parameters, oyster hemocytes appear to represent a single population exhibiting graded cellular differences.