ULTRACYTOCHEMISTRY OF THE CELL MEMBRANE : I. THE DEMONSTRATION OF INTRAMEMBRANOUS AMINO GROUPS IN RABBIT ERYTHROCYTIC MEMBRANE BY THE HYDROQUINONE-TETRANITROBLUE TETRAZOLIUM SYSTEM

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Amino groups in rabbit erythrocytic membrane were ultracytochemically demonstrated using the hydroquinone-tetranitro blue tetrazolium system, Reaction products were observed within the membrane, which was immediately external to the phosphotungstic acid-positive zone, in the form of small globules or particles with generally a diameter of 3.0-9.0 nm, aligning in one or several rows. It was found by use of a goniometer stage that the membrane area with several rows of particles corresponded in reality to the tangentially cut area of that with one row of particles. The detected intramembranous amino groups seemed to have originated from proteins, probably globular proteins, rather than from carbohydrates or lipids, since reaction products detected were found within the membrane and resisted through extraction of specimens in organic solvents as well as digestion by phospholipase C, but were not found in specimens treated with protease. These findings are of great interest in relation to intramembranous particles observed in freeze-fractured or freeze-etched membranes of various cells and also to the hypothesis of the fluid mosaic model of the cell membrane.
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