Cytochemical observations of hemolymph cells during coagulation in the crayfish, orconectes virilis

Abstract

Smears of Orconectes virilis hemolymph were prepared by fixation in 10% formalin immediately upon withdrawal, and after an interval of 1, 5, or 15 minutes. These smears were then stained by a variety of histochemical means designed to identify a material which is released from both hyaline and granular cells during the coagulation process. Based upon reactions to various carbohydrate, lipid and protein tests, this material appears to consist of a glycoor mucwprotein, and its localization and activity coincide with the "fibrin ferment" described by Hardy (1892). Its release is probably initiated from the hyaline cells, followed shortly thereafter by a similar activity of the granulocytes. While it cannot be ascertained by the methods used in this study whether or not this material participates actively in clotting, its release is nevertheless coincidental with coagulation.

Haemocytes in Haemolymph Coagulation of Arthropods

“…The granules are positive to high iron diamine (HID) test, stained purple after Alcian-blue PAS technique, red brown after HID -PAS test, Pyronin-methyl green after RNA -ase treatment, non-reactive to Millon's and dihydroxy-dinapthyl sulphide tests (Costin,I 975). Histochemical characteristics of the granules of granular haemocytes of various other arthropods also confirm their periodate-reactive acid mucopolysaccharide nature (Wood et al, 1971 ;Johnston et al, 1973;Ravindranath, 1973, 1 9 7 5~;Franqois, 1974;Bauchau et al, 1975). I n addition to granules, the cytoplasm of granular haemocytes also contained numerous Oil Red-positive, sudanophilic droplets (Costin, 1975).…”

“…(iii) Histochemistry. Histochemical characterization of granules of granular haemocytes has been carried out by a number of investigators (Hoffmann, 1967;Wood et al, 1971 ;Johnston, Elder & Davies, 1973;Ravindranath, 1973Ravindranath, ,1975aFranqois, 1974;Bauchau, De Brouwer, Passelecq-Gerin & Mengeot, 1975 ;Costin, 1975). In Locusta migratoria, the granules exhibited amylase-resistant PAS-positivity, pyridine-labile sudanophilia (Hoffmann, 1967).…”

Fine Structure of the Haemocytes of the Indian White Shrimp, Fenneropenaeus Indicus (H. Milne Edwards, 1837)

“…Based on cytochemical observations, Wood & Visentin (1967) and Wood et al (1971), described a hyaline cell, a small granulocyte, and a large granulocyte in Orconectes virilis (Hagen, 1870). Stang-Voss (1971) described only one cell type in Astacus astacus, the amoebocyte.…”

“…They have been reported to be involved in haemostasis (Bang, 1971;Wood et al, 1971;Bauchau & De Brouwer, 1974), ecdysis (Bauchau & Plaquet, 1973;Vacca & Fingerman, 1975), and storage and transport of glycogen and other nutrients (Johnston et al, 1973) besides production of haemocyanin (Stang-Voss, 1971). The important role of haemocytes in defense mechanisms (phagocytosis, encapsulation, nodule formation, wound repair, and coagulation) has also been recognized by several authors (Wood et al, 1971;Sinderman, 1971;Fontaine & Lightner, 1974;Johnson & Bodammer, 1975;Stewart, 1975;Johnson, 1976;Durliat, 1985;and Söderhäll & Cerenius, 1992). Morphological studies on blood cells of Crustacea, however, are scarce, so that it is difficult to identify particular haemocyte types involved in defence reactions.…”

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“…Hyaline cells initiate haemolymph coagulation and hardening of the exoskeleton after moulting (Vacca and Fingerman, 1983;Omori et al, 1989). Granulocytes play diverse roles, being involved in phagocytosis, agglutination, coagulation, encapsulation and storage of haemocyanin and glycoproteins (Wood and Visentin, 1967;Busselen, 1970;Stang-Voss, 1971;Wood et al, 1971;Ravindranath, 1980). Light microscopic and ultrastructural investigations with regard to the morphological and functional aspects of haemocytes have been extensively studied in economically important marine decapods.…”