Freeze-substitution and X-ray microprobe analysis of amine-storage organelles of rat platelets after treatment with reserpine.
Freeze-substitution was successfully used to determine the elemental composition of the amine storage organelles of rat platelets after treatment with reserpine. Conventional electron microscopy of platelets fixed with glutaraldehyde and OsO4 revealed many empty vesicles indicating the liberation of 5-hydroxytryptamine from the amine storage organelles. However, the organelles were totally filled with an osmiophilic material in platelets freeze-substituted with OsO4-acetone. Freeze-substitution of platelets with pure acetone without OsO4 revealed also the presence of an electron-opaque material in the organelles. The emission spectra of X-ray generated by the spot analysis of the organelles exhibited Na-K alpha, Mg-K alpha, P-K alpha, K-K alpha, and Ca-K alpha. A semi-quantitative comparison of elemental concentrations in the amine storage organelles between reserpine-dosed rats and normal intact controls showed that K was significantly higher in controls, while the content of other elements was similar.
Daimon
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Department of Anatomy, School of Medicine, Teikyo University, Tokyo, Japan.
D000588:Amines / Q000378:metabolism*; D000818:Animals; D001792:Blood Platelets / Q000737:chemistry* / Q000187:drug effects / Q000648:ultrastructure; D002118:Calcium / Q000032:analysis; D004577:Electron Probe Microanalysis; D017110:Freeze Substitution / Q000379:methods; D005976:Glutaral; D008274:Magnesium / Q000032:analysis; D008297:Male; D008670:Metals / Q000032:analysis*; D008854:Microscopy, Electron; D015388:Organelles / Q000737:chemistry* / Q000187:drug effects / Q000648:ultrastructure; D009993:Osmium Tetroxide; D010758:Phosphorus / Q000032:analysis; D011188:Potassium / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D017208:Rats, Wistar; D012110:Reserpine / Q000494:pharmacology*; D012964:Sodium / Q000032:analysis; D016707:Tissue Fixation / Q000379:methods
D000588:Amines; D008670:Metals; D010758:Phosphorus; D012110:Reserpine; D012964:Sodium; D008274:Magnesium; D009993:Osmium Tetroxide; D011188:Potassium; D002118:Calcium; D005976:Glutaral