Effects of glutaraldehyde fixative osmolarities on smooth muscle cell volume, and osmotic reactivity of the cells after fixation

Lee, R. M. K. W.; McKenzie, Roderick C.; Kobayashi, Ken; Garfield, R. E.; Forrest, James B.; Daniel, E. E.

doi:10.1111/j.1365-2818.1982.tb00324.x

Cited by 57 publications

(34 citation statements)

References 23 publications

(34 reference statements)

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“…Greater shrinkage of cells was observed with GO treatment, consistent with results described for other cell types (Hillman and Deutsch, 1978;Lee, 1984;Lee et al, 1982). This shrinkage was observed Fig.…”

Section: Discussionsupporting

confidence: 93%

“…Hillman and Deutsch (1978) reported a 20-30% decrease in tissue cross-sectional areas of rat brain with glutaraldehyde fixation and ethanol dehydration. Similarly, cell volume changes of 10-30% were reported in cultured rat smooth muscle tissue (Lee et al, 1982). These area and volume differences related to chemical fixation need to be understood and minimized so that comparisons between fixed tissue and living tissue may be made.…”

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confidence: 94%

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Morphology of human neutrophils: A comparison of cryofixation, routine gluteraldehyde fixation, and the effects of dimethyl sulfoxide

Gilbert

Parmley

1998

Anat. Rec.

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Cell shape and density are critical to the evaluation of neutrophil function and/or activation. Dimethyl sulfoxide-cryofixation-freeze-substitution processing (DCF) instantly preserves cell processes and ultrastructural elements with fewer artifacts than routine chemical fixation with glutaraldehyde and postfixation osmium tetroxide (GO). This study morphometrically examined density-separated neutrophils to assess differences in DCF and GO processing procedures and studied the effect of dimethyl sulfoxide followed by GO fixation (DGO) on morphology. Fifteen consecutive neutrophils were analyzed using computerized planimetry for differences in DCF v. GO treatments (n = 4) and DGO v. GO treatments (n = 4). Cryofixed and DGO-fixed cells were significantly rounder than GO cells which had a more irregular surface with membrane projections. The cell volume of GO cells was 27-30% smaller than in DCF or DGO processing, while the surface area was similar. The increased volume in DCF and DGO cells did not appear to be due to abnormal cell swelling, since membranes, nuclear envelope, and mitochondrial cristae were more intact than in GO cells. Preservation of mitochondria as well as endocytic caveolae with a subplasmalemmal coating was best in DCF samples, moderate in DGO, and poorest in GO. Morphometric data showed that the nuclear compartment was 22% smaller, while the cytoplasm (and its associated compartments) was 29% smaller in GO compared to DCF-processed neutrophils. This was consistent with the more dense cytoplasm in GO cells. Pretreatment of neutrophils with dimethyl sulfoxide (DMSO) resulted in volume preservation and improved the morphology of GO fixation. In summary, DCF appears to be an excellent method for preserving neutrophil membranes and cytoplasmic organelles (particularly mitochondria), and prevents a number of artifacts caused by routine GO fixation. Morphology can also be improved by using DMSO in conjunction with GO.

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Section: Discussionsupporting

confidence: 93%

mentioning

confidence: 94%

Morphology of human neutrophils: A comparison of cryofixation, routine gluteraldehyde fixation, and the effects of dimethyl sulfoxide

Gilbert

Parmley

1998

Anat. Rec.

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“…Moreover, Lee and co-workers reported that hyperosmotic medium led to contractions of VSMR after their fixation with glutaraldehyde [44]. It should be underlined, however, that despite the different mechanisms of force generation in shrunken and swollen SMC, their contractions were equally sensitive to actin cytoskeleton disruption.…”

Section: +mentioning

confidence: 96%

Vascular Smooth Muscle Contraction Evoked by Cell Volume Modulation: Role of the Cytoskeleton Network

Кольцова

Гусакова

Anfinogenova

et al. 2008

Cell Physiol Biochem

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Previously, we reported that hyposmotic swelling evoked transient vascular smooth muscle cell (SMC) contraction that was completely abolished by L-type Ca²⁺ channel blockers. In contrast, sustained contraction revealed in hyper- and isoosmotically-shrunken SMCs was insensitive to L-type channel blockers and was diminished in Ca²⁺-free medium by only 30-50%. Several research groups reported cell volume-dependent cytoskeleton network rearrangements. This study examines the role of cytoskeleton proteins in cell volume-dependent contraction of endothelium-denuded vascular smooth muscle rings (VSMR) from the rat thoracic aorta. Hyperosmotic shrinkage and hyposmotic swelling were triggered by modulation of medium osmolality; isosmotic shrinkage was induced by VSMR transfer from hypo- to isosmotic medium. The relative content of globular (G) and fibrillar (F) actin was estimated by fluorescence microscopy. Hyperosmotic shrinkage and hyposmotic swelling led to elevation of the F-actin/G-actin ratio by 2.5- and 1.8-fold respectively. Contraction of shrunken and swollen VSMR was insensitive to modulators of microtubules such as vinblastine, colchicine and docetaxel. Microfilament disassembly by cytochalasin B resulted in dramatic attenuation of the maximal amplitude of contraction of hyperosmotically-shrunken and hyposmotically-swollen VSMR, and almost completely abolished the contraction triggered by isosmotic shrinkage. These data suggest that both L-type Ca²⁺ channel-mediated contraction of swollen vascular SMC and Ca²⁺_o-insensitive contractions of shrunken cells are triggered by reorganization of the microfilament network caused by elevation of the F-actin/G-actin ratio.

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“…1976;Schroeder-Henning, 1981;Mathieu et al. 1977;Lee. 1982], Our results support the latter point of view.…”

Section: Discussionmentioning

confidence: 99%

Effect of Osmolarity of the Fixative on the Ultrastructure of Preimplantation Rabbit Embryos

Hegele‐Hartung

Piegsa

Fischer³

1989

Cells Tissues Organs

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In an attempt to clarify whether or not glutaraldehyde molecules contribute to the effective osmotic pressure of the fixative solution the ultrastructure of noncultured and in vitro-cultured day 1 3 and 4 rabbit embryos was evaluated. Total osmolarity of the fixative solution (200–800 mosm) was only varied by changing the aldehyde concentration, whereas the vehicle osmolarity (145 mosm) remained unchanged. Optimum preservation in all embryonic stages was obtained when total osmolarity of the fixative solution was 285–340 mosm. Higher (480–800 mosm) or lower (250 mosm) osmolarities of the fixative solution led to alterations mainly in mitochondria and smooth-surfaced endoplasmic reticulum. Shrinkage of cells and condensation of the cytoplasm occurred only occasionally. Compared with early cleavage stage embryos blastocysts were generally more susceptible to hyperton and hypoton fixative solutions. In vitro culture for 24 h per se did not have any influence on the fixation.

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Effects of glutaraldehyde fixative osmolarities on smooth muscle cell volume, and osmotic reactivity of the cells after fixation

Cited by 57 publications

References 23 publications

Morphology of human neutrophils: A comparison of cryofixation, routine gluteraldehyde fixation, and the effects of dimethyl sulfoxide

Morphology of human neutrophils: A comparison of cryofixation, routine gluteraldehyde fixation, and the effects of dimethyl sulfoxide

Vascular Smooth Muscle Contraction Evoked by Cell Volume Modulation: Role of the Cytoskeleton Network

Effect of Osmolarity of the Fixative on the Ultrastructure of Preimplantation Rabbit Embryos

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