By Geoffrey Bourne (Eds.)

ISBN-10: 0121192547

ISBN-13: 9780121192549

ISBN-10: 0323162010

ISBN-13: 9780323162012

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Example text

C. MITOCHONDRIA 1. Isolation and Identification of Mitochondria T h e isolation of mitochondria was first reported by Bensley and Hoerr (1934) from suspensions of guinea pig liver disrupted in isotonic saline. T h e details of this procedure were not published until consider­ ably later (Hoerr, 1943), however, and in the meantime Claude (1941) described the isolation of "large granules" which were believed to be mixtures of mitochondria and secretory granules. In a reinvestigation of the latter work, it was found by Hogeboom et al.

Adsorption or absorp­ tion, leakage of materials from subcellular organelles, morphological alterations, and redistribution are among those that might be men­ tioned. All of these problems can, however, be studied in a positive 22 WALTER C. SCHNEIDER AND EDWARD L. KUFF manner and need not serve as an a priori basis for dismissing cell frac­ tionation as a cytochemical tool. T h e question of morphological and cytological alteration should be considered first because it is the first grounds on which cell frac­ tionation can be criticized and is the most susceptible to direct exami­ nation.

Other instances which might be cited to show how adsorption was recognized include the enzymes deoxyribonuclease, ribonuclease, and fumarase (Schneider and Hogeboom, 1952b; Kuff, 1954). A consideration of the quantitative aspects of the intracellular dis­ tribution of substances can also have a decided bearing upon whether they are interpreted to be adsorbed or not. For example, if it can be shown that a large fraction of the total enzymatic activity of a tissue is associated with a subcellular component after its isolation, adsorption would seem unlikely, especially if the enzymatic activity was tightly bound by the subcellular element and retained during numerous wash­ ings in various media designed to remove adsorbed material.

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Cytology and Cell Physiology by Geoffrey Bourne (Eds.)

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