Histochemical Glycogen Studies on Echinostoma revolutum
| Autor: | Bernard Fried, Marshall D. Kramer |
|---|---|
| Rok vydání: | 1968 |
| Předmět: |
chemistry.chemical_classification
Glycogen Cuticle Periodic acid–Schiff stain Biology biology.organism_classification Polysaccharide Andrology chemistry.chemical_compound Chorioallantoic membrane Biochemistry chemistry Parenchyma Fasciola hepatica Parasitology Echinostoma revolutum Ecology Evolution Behavior and Systematics |
| Zdroj: | The Journal of Parasitology. 54:942 |
| ISSN: | 0022-3395 |
| DOI: | 10.2307/3277125 |
| Popis: | Histochemical studies were made to determine if Echinostoma revolutum depletes its glycogen reserves in a non-nutrient medium and resynthesizes the polysaccharide when placed in a nutrient medium or on the chick chorioallantois. The periodic acid Schiff technique on lipid-free cryostat sections of adult worms revealed that glycogen was depleted primarily from the parenchyma and musculature during a 72-hr period of starvation. Uterine eggs contained glycogen which was not depleted during starvation. The vitellaria, cuticle, and subcuticle did not contain glycogen. Flukes starved for 48 hr and then incubated on the chick chorioallantois for 48 hr resynthesized lost glycogen, but not to the same extent as those maintained in Tyrode's-glucose after starvation. Von Brand and Mercado (1961) demonstrated histochemically a rapid utilization of glycogen reserves in Fasciola hepatica starved for 24 hr, and a resynthesis of the polysaccharide when worms were incubated in a nutrient medium after starvation. Histochemical studies were made to determine if Echinostoma revolutum depletes its glycogen reserves in a non-nutrient medium as rapidly as F. hepatica, and resynthesizes the polysaccharide when placed in a nutrient medium or on the chick chorioallantois. Results are here reported. MATERIALS AND METHODS Nineteen-day-old E. revolutum obtained from laboratory-infected domestic chicks were separated into 6 groups of 5 or 6 worms each. Group 1 flukes were untreated controls, while groups 2, 3, and 4 were starved 24, 48, and 72 hr, respectively. During the inanition period worms were maintained individually at 37 to 38 C in 10 to 15 ml of Tyrode's (Paul, 1960) without glucose, and with antibiotics (1,000 units/ml penicillin and 200 /g/ml streptomycin). Group 5 flukes, starved for 48 hr, were then maintained an additional 48 hr in Tyrode's plus 0.1% glucose and antibiotics, whereas, group 6 flukes were incubated on the chick chorioallantois for 48 hr following the 48-hr starvation period. Worms were sectioned on a cryostat at 8 u, fixed in formalin fumes, immersed in ether for lipid extraction and stained for polysaccharide by the periodic acid Schiff (PAS) procedure. Control slides digested with 0.5% diastase were stained similarly to determine the glycogen nature of the polysaccharide. Since lipids interfere with the visualization of the final product they were extracted with ether. In paraffin sections it is assumed that most lipids are removed during treatment. The rapidity of the PAS procedure on cryostat sections and the fact that no appreciable loss of detail was observed at Received for publication 7 June 1968. * Supported by NIH Grant AI-06835. the tissue or organ level justified this procedure for polysaccharide histochemistry. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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