Further Studies on the Dissociation of Adult Mouse Tissue.

Rappaport, Catherine; Howze, Gwendolyn B.

doi:10.3181/00379727-121-30952

Cited by 21 publications

(3 citation statements)

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“…Some attempts were made to obtain a better suspension medium by varying its composition and by addition of citrate, sodium tetraphenylborate (Rappaport and Howze, 1966) proteolytic enzymes, and other solutes . However, none of these variations was clearly superior ; in most experiments we used the simple sucrose media described in Materials and Methods .…”

Section: Preparation Of Whole Cell Suspensions From Brainmentioning

confidence: 99%

Comparative Studies on Mitochondria Isolated From Neuron-Enriched and Glia-Enriched Fractions of Rabbit and Beef Brain

Hamberger

Blomstrand

Lehninger

1970

The Journal of Cell Biology

114

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Fractions enriched in neuronal and glial cells were obtained from dispersions of whole beef brain and rabbit cerebral cortex by large-scale density gradient centrifugation procedures. The fractions were characterized by appropriate microscopic observation. Mitochondria were then isolated from these fractions by differential centrifugation of their homogenates. The two different types of mitochondria were characterized with respect to certain enzyme activities, respiratory rate, rate of protein synthesis, and their buoyant density in sucrose gradients. The mitochondria from the neuron-enriched fraction were distinguished by a higher rate of incorporation of amino acids into protein, higher cytochrome oxidase activity, and a higher buoyant density in sucrose density gradients. Mitochondria from the glia-enriched fraction showed relatively high monoamine oxidase and Na+- and K+-stimulated ATPase activities. The rates of oxidation of various substrates and the acceptor control ratios did not differ appreciably between the two types of mitochondria. The difference in the buoyant density of mitochondria isolated from the neuron-enriched and glia-enriched cell fractions was utilized in attempts to separate neuronal and glial mitochondria from the mixed mitochondria obtained from whole brain homogenates in shallow sucrose gradients. The appearance of two peaks of cytochrome oxidase, monoamine oxidase, and protein concentration in such gradients shows the potential feasibility of such an approach.

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Section: Preparation Of Whole Cell Suspensions From Brainmentioning

confidence: 99%

Comparative Studies on Mitochondria Isolated From Neuron-Enriched and Glia-Enriched Fractions of Rabbit and Beef Brain

Hamberger

Blomstrand

Lehninger

1970

The Journal of Cell Biology

114

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“…EDTA is a widely used chelating agent for Ca ++ and Mg ++. TPB chelates K + (1) and has recently been used as a cell-dispersing agent (2). Our dispersing media contained EDTA and/or TPB in variously modified Tyrode's solutions (Table I).…”

Section: Methodsmentioning

confidence: 99%

Some Effects of Edta and Tetraphenylboron on the Ultrastructure of Mitochondria in Mouse Liver Cells

Harris

Leone

1966

The Journal of Cell Biology

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“…During the past two decades, workers attempting to obtain suspensions of isolated kidney cells have employed a variety of methods for the disruption of the tissue. These methods include treatment with trypsin (Dulbecco & Vogt, 1954;Youngner, 1954;Fenje, 1960;Wallace et al, 1961;Phillips, 1967), Aspergillin-O (Sabina et al, 1963), tetraphenylboron (Rappaport & Howze, 1966) and various chelating agents (Kalant & Young, 1957;Rappaport & Howze, 1966), as well as the purely mechanical methods of forcing kidney tissue through progressively smaller needles (Zaroff et al, 1961) or a stainless-steel screen and pipette (Phillips, 1967(Phillips, , 1968. Thimmappayya et al (1970) used a modified Potter-Elvehjem homogenizer with a soft rubber pestle in their mechanical method for isolating kidney cells.…”

Section: Preparation and Some Properties Of A Suspension Of Fragmentedmentioning

confidence: 99%

Preparation and some properties of a suspension of fragmented tubules from rat kidney

Dawson

1972

Biochemical Journal

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A method is described whereby short fragments of rat kidney tubule were obtained when kidney slices were gently dispersed by exposure to collagenase and hyaluronidase. When suspended in buffered saline the fragmented tubules respired actively over a period of several hours, the rate of oxygen consumption being proportional to the amount of cell protein. Oxygen uptake was stimulated by the addition of glucose, lactate, butyrate, alpha-oxoglutarate and other substrates and was decreased by the omission of Ca(2+) from the suspending medium. With alpha-oxoglutarate as the added substrate, dinitrophenol strongly stimulated oxygen uptake. Dinitrophenol had a less-marked stimulatory effect when glucose was the added substrate, and inhibited respiration in the absence of added substrate. Oligomycin inhibited respiration and this inhibition was partially reversed by dinitrophenol. Fragmented tubules synthesized glucose from lactate at a high rate but this capacity for gluconeogenesis was abolished by dinitrophenol and by physically damaging the cells.

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Further Studies on the Dissociation of Adult Mouse Tissue.

Cited by 21 publications

References 1 publication

Comparative Studies on Mitochondria Isolated From Neuron-Enriched and Glia-Enriched Fractions of Rabbit and Beef Brain

Comparative Studies on Mitochondria Isolated From Neuron-Enriched and Glia-Enriched Fractions of Rabbit and Beef Brain

Some Effects of Edta and Tetraphenylboron on the Ultrastructure of Mitochondria in Mouse Liver Cells

Preparation and some properties of a suspension of fragmented tubules from rat kidney

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