Experimental manipulation of cell surface to affect cellular recognition mechanisms

Takeichi, Masatoshi; Ozaki, Hiroki S.; Tokunaga, Ken; Okada, Tamotsu

doi:10.1016/0012-1606(79)90016-2

Cited by 100 publications

(32 citation statements)

References 15 publications

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“…2 can selectively remove either Ca2+-independent or Ca2"-dependent CAMs (also known as cadherins) from the surface of Chinese hamster V79 lung cells, depending on the presence or absence of Ca2", and on the concentration of trypsin (30). In the presence of Ca2 , cadherins assume a three-dimensional configuration that renders their cleavage sites inaccessible to trypsin, while Ca2+-independent CAMs are digested (TC treatment).…”

Section: Methodsmentioning

confidence: 99%

Tumor necrosis factor-alpha modifies adhesion properties of rat islet B cells.

Cirulli

Halban²,

Rouiller³

1993

J. Clin. Invest.

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The characteristic three-dimensional cell type organization of islets of Langerhans is perturbed in animal models of diabetes, suggesting that it may be important for islet function. Rat islet cells in culture are able to form aggregates with an architecture similar to native islets (pseudoislets), thus providing a good model to study the molecular basis of islet architecture and its role in islet function. Sorted islet B cells and non-B cells were permanently labeled with two different fluorescent dyes (DiO and DiI), mixed, and allowed to form aggregates during a 5-d culture in the presence or absence of TNF-a (100 U/ml), a cytokine suggested to be implicated in the early physiological events leading to insulin-dependent diabetes mellitus. Confocal microscopy of aggregates revealed that TNF-a reversibly perturbs the typical segregation between B and non-B cells. Insulin secretion, was altered in the disorganized aggregates, and returned towards normal when pseudoislets had regained their typical architecture. The homotypic adhesion properties of sorted B and non-B cells cultured for 20 h in the presence or absence of TNF-a were studied in a short term aggregation assay. TNF-a induced a significant rise in Ca2"-independent adhesion of B cells (from 24±1.1% to 44.3+1.2%; n = 4, P <0.001). These findings raise the possibility that the increased expression of Ca2"-independent adhesion molecules on B cells leads to altered islet architecture, which might be a factor in the perturbation of islet function induced by TNF-a.

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Section: Methodsmentioning

confidence: 99%

Tumor necrosis factor-alpha modifies adhesion properties of rat islet B cells.

Cirulli

Halban²,

Rouiller³

1993

J. Clin. Invest.

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“…In addition to the Chinese hamster lung cell line V79 and embryonic chick neural retina, several neuronally derived cell lines have also been suggested to have multiple adhesive systems (24,27). The relationship of the adhesive systems described here to those characterized previously by our (2,23) and other (9,17,29,30) laboratories remains to be elucidated . We are at present analyzing CaT and chymotrypsin-dispersed cells serologically and by radioiodination to identify components unique to each of the two adhesive systems.…”

Section: Discussionmentioning

confidence: 88%

“…The data presented here are largely in agreement with the studies of Takeichi et al . (29) and Urushihara et al . (34), although there are significant differences .…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Enzymatic dissection of embryonic cell adhesive mechanisms.

Grunwald¹,

Geller²,

Lilien³

1980

The Journal of Cell Biology

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In this paper we describe a kinetic assay for cell adhesion which measures the formation of cell clusters . Cluster formation is dependent on both calcium and protein synthesis, two parameters essential for the formation of histotypic aggregates .We also describe modifications of the standard method for trypsinization of tissues which result in populations of single cells that appear to bear intact and functional cell surface adhesive systems . These modifications involve the use of chymotrypsin and the inclusion of calcium during enzyme digestion of tissues with trypsin and chymotrypsin .Using the cluster formation assay and the modified tissue dissociation techniques, we demonstrate the presence of two functionally distinct adhesive systems operating among embryonic chick neural retina cells . These two systems differ in proteolytic sensitivity, protection by calcium against proteolysis, dependence on calcium for function and morphogenetic potential. Cells possessing one of these intact adhesive systems are capable of extensive morphogenetic interactions in the absence of protein synthesis .Since the classic studies of Townes and Holtfreter (31), the reaggregation of cells derived from embryonic tissues has served as a model system for analysis of morphogenetic cell interactions (see references 13, 19, and 32 for reviews) . Two problems are encountered in such studies : the need for quantitative assays that measure adhesions relevant to the formation of histotypic aggregates and the inability to dissociate most embryonic avian or mammalian tissues into single cells without the use of proteolytic enzymes .The magnitude of the first problem is reflected in the many assays that have been developed to measure intercellular adhesion (reviewed in references 13 and 14) . Two discrepant facts emerge when the reaggregation of trypsin-dispersed single cells is monitored . Whereas the formation of his-766 totypic aggregates is dependent on protein synthesis (20) and the presence of divalent cations (18), neither of these dependencies is usually reflected in assays that measure the kinetics of intercellular adhesion (reference 12 and this paper) . Furthermore, Roth has shown that the initial adhesions between trypsin-dispersed cells are nonspecific (22) . Thus, the relevance of these initial adhesions is questionable with respect to mechanisms of tissue reconstruction and, presumably, morphogenesis .Most efforts directed at avoiding the use of proteolytic enzymes have relied on the use of chelating agents combined with mechanical shear (5, 11) or a recovery period after trypsinization (3,22) . The first approach generally results in low cell yields and, as Weiss (35) has pointed out, uncer-J . CELL BIOLOGY

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“…To identify the molecules responsible for the CID aggregation of BHK and pyBHK cells, we used anti E-V79 antiserum, an antiserum originally raised for the study of adhesion molecules in Chinese hamster V79 cells (10). This use was based on the fact that the CIDS's in various cells can cross-react to a degree (17). In fact, Fab fragments prepared from anti E-V79 antiserum inhibited the aggregation of LTE-BHK and LTE-pyBHK cells.…”

Section: Discussionmentioning

confidence: 99%