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Vitellogenesis and vitellocytes of the bothriocephalidean cestode Clestobothrium crassiceps (Rudolphi, 1819), a parasite of the teleost fish Merluccius merluccius (L., 1758), were studied by means of transmission electron microscopy (TEM) and cytochemistry. During vitellogenesis, four developmental stages were distinguished at the TEM level: (I) a stem cell stage of the gonial type; (II) an early differentiation stage, predominantly exhibiting lipid and protein synthetic activity; (III) an advanced differentiation or vitellocyte maturation stage, primarily exhibiting active glycogenesis still accompanied by an increase in lipid accumulation; and (IV) a mature vitellocyte stage. Vitellogenesis involves: (1) an increase in cell volume; (2) an extensive development of parallel, frequently concentrically arranged, cisternae of granular endoplasmic reticulum (GER) that produce dense, proteinaceous shell-gobules; (3) the development of Golgi complexes engaged in the packaging of this material; (4) an accelerated accumulation of unsaturated and saturated lipid droplets, along with their continuous enlargement and fusion; (5) the formation of individual β-glycogen particles and α-glycogen rosettes and their accumulation in the form of glycogen islands scattered among lipid droplets in the cytoplasm of maturing and mature vitellocytes; and (6) the rapid accumulation of large, saturated lipid droplets accompanied by dense accumulations of α- and β-glycogen along with proteinaceous shell-globules or shell-globule clusters in the peripheral layer during the advanced stage of vitellocyte maturation. Vitellogenesis in C. crassiceps generally resembles that previously described for three other bothriocephalideans, but differs from that of other cestode orders. Cytochemical staining with periodic acid-thiocarbazide-silver proteinate for glycogen indicates a strongly positive reaction for β-glycogen particles and α-glycogen rosettes, which form several large glycogen accumulations around the large, saturated lipid droplets of maturing and mature vitellocytes. Some hypotheses concerning the interrelationships between patterns of vitellogenesis, the possible modes of egg formation, embryonic development and life cycles in cestodes, and their phylogenetic implications are commented upon.
Ultrastructural and cytochemical characteristics of GER-bodies observed in the vitellocyte cytoplasm of the intrauterine eggs of the caryophyllidean cestode Wenyonia virilis are described. In this species GER-bodies were observed only in the cytoplasm of vitellocytes, surrounded by a newly formed egg-shell. They are composed of spherical areas of condensed, electron-dense cytoplasm which contains concentrically arranged parallel lamellae of granular endoplasmic reticulum (GER), forming characteristic balls of different sizes. Each GER-body is surrounded by numerous free ribosomes, polyribosomes, α-glycogen rosettes and large mitochondria. Results of cytochemical analysis by means of PATSC-SP test for polysaccharides indicated that glycogen is absent within the GER-bodies, however, a strongly positive reaction was observed only in large aggregations of α-glycogen rosettes and β-glycogen particles, localised usually near GER-bodies.
Ultrastructural evidence for early intraurerine embryonic development of Wenyonia virilis is presented. At the initial stage of egg formation, the fertilized oocyte or ovum is surrounded by numerous vitellocytes and newly formed eggshell. Individual vitellocytes undergo progressive fusion into a vitelline syncytium. During cleavage divisions, three types of blastomeres are formed: macromeres, mesomeres and micromeres. Two large macromeres contain a large nucleus with spherical nucleolus and numerous small heterochromatin islands dispersed in moderately electron-dense nucleoplasm. The granular cytoplasm shows a few large mitochondria. Medium-sized mesomeres contain a spherical nucleus with numerous heterochromatin islands, adjacent to the nuclear envelope, and a prominent electron-dense nucleolus. Their nuclei are embedded in granular cytoplasm with a few large and numerous small mitochondria and Golgi complexes. The small micromeres are characterized by presence of spherical nucleoli with large areas of highly condensed heterochromatin and a few islands of granular electron-lucent nucleoplasm. Their granular cytoplasm shows a few small lipid droplets and several spherical mitochondria. Majority of micromeres give rise to the hexacanth but many of them also undergo degeneration or apoptosis. Both mesomeres and macromeres are engaged in the formation of the oncospheral envelopes. The outer envelope is formed by a fusion of two macromeres whereas the inner envelope originates from a fusion of mesomeres. The intrauterine eggs of W. virilis usually contain an embryo at the early preoncopheral phase of development and possesses three primary envelopes: (1) thick eggshell; (2) thin cytoplasmic layer of the outer envelope and (3) inner envelope. Based on embryonic development, egg type and life-cycle characteristics, caryophyllideans tend to show closer affinities to spathebothriideans than to the former pseudophyllideans.
The first description of vitellogenesis in the Diphyllidea is presented in this paper. Though the type of vitellogenesis and mature vitellocyte in Echinobothrium euterpes appear to be unique among the Eucestoda, however, they somewhat resemble that observed in the two orders of the lower cestodes, Tetraphyllidea and Proteocephalidea. Vitellocyte maturation is characterized by: (1) an increase in cell volume; (2) extensive development of short, parallel, frequently concentric cisternae of GER that produce dense proteinaceous granules; (3) development of Golgi complexes engaged in packaging this material; (4) progressive formation of saturated lipid droplets; their continuous enlargement and fusion; (5) formation of small accumulations of glycogen particles scattered between and among lipid droplets in the cytoplasm of maturing vitellocytes; (6) concentration of dense proteinaceous granules in the peripheral layer of cytoplasm, around the cell plasma membrane; and (7) vacuolization of cytoplasm of mature vitellocytes accompanied by a rapid increase in its volume. A new, unreported type of dense proteinaceous granules, situated around the limiting plasma membranes of mature vitellocytes, is described. Vitellogenesis evidently differs from that with typical shell-globules and shell-globule clusters previously reported in other taxa of lower cestodes. Cytochemical staining with periodic acidthiosemicarbazide-silver proteinate for glycogen indicates a strongly positive reaction for glycogen particles between and around large unsaturated lipid droplets of the maturing and mature vitellocytes. Some hypotheses concerning the interrelationship between this pattern of vitellogenesis, possible mode of egg formation, embryonic development and diphyllidean life cycle, and their phylogenetic implications are drawn and discussed.
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