Vergleichende Untersuchungen �ber die histotopochemischen Qualit�ten der Tetrazolsalze MTT, Nitro-BT und TNBT in der Enzymhistochemie

Meier‐Ruge, W.

doi:10.1007/bf00306253

Cited by 16 publications

(1 citation statement)

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“…In these studies, the tissue slices were incubated for at least 3hr., during which time there was a considerable degree of autolysis, particularly of the brush border. Nitro-BT, the dye used by Dahlqvist & Brun (1962), was recently shown to be of very limited value in histochemical investigations, especially when autolysis occurred (Meier-Ruge, 1965). Thus, when considering all the observations on the intracellular distribution of invertase, there is more convincing evidence for a predominant localization of invertase in the brush border than for its occurrence in other subcellular structures.…”

Section: Discussionmentioning

confidence: 99%

Studies on the fractionation of mucosal homogenates from the small intestine

Hübscher¹,

West²,

Brindley³

1966

Biochemical Journal

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1. Homogenates of the mucosa of the small intestine of the guinea pig were separated by fractional sedimentation into seven different fractions. The enzymic properties of some of these subcellular fractions were compared with those obtained from the mucosa of the small intestine of the rabbit and cat. 2. The enzymic properties of the low-speed sediment (150OOg-min.) were investigated and it was shown that invertase and alkaline ribonuclease were predominantly located in this subcellular fraction, whereas alkaline phosphatase, aryl-amidase, acid phosphatase, acid ribonuclease and phosphoprotein phosphatase, though true constituents of this fraction, occurred to varying degrees in other subcellular structures also. 3. It was shown that the most probable source of the enzymic activities observed in the low-speed sediment was the brush border. Electron micrographs ofthe purified brush-border fraction indicated vesicles derived from the brush-border membrane. 4. A method is described for the fractionation of mucosal homogenates into a brush border-plus-nuclei fraction, a mitochondrial fraction, a microsomal fraction and a particle-free supematant. The fractions were shown to be relatively pure, as indicated by the distribution of invertase, DNA, succinate dehydrogenase, glucose 6-phosphatase and 6-phosphogluconate dehydrogenase. 5. Most of the activity of four lysosomal enzymes present in the nuclei-free homogenate was sedimented at 3750OOg-min., suggesting the occurrence of lysosomal particles in mucosal homogenates. 6. Further fractionation of the microsomal membranes into three fractions is described. The enzymic composition of the membrane fractions is given and discussed in relation to their structure as seen in electron micrographs.The fractionation of homogenates of the mucosa of the small intestine has been described for preparations from the rat (Schmidt,

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Section: Discussionmentioning

confidence: 99%