Effect of different fixatives on Con A surface receptors of mouse peritoneal macrophages

Abstract

The effects of glutaraldehyde, formaldehyde, or osmium tetroxide fixation on the number of labeled Con A surface receptors on mouse peritoneal macrophages were compared. Gold-labeled Con A receptors were found to be isolatedly arranged and evenly distributed on cell surfaces independent of the fixative used. Only cells preincubated with Con A and subsequently fixed by osmium tetroxide showed arrangement of labeled receptors in clusters. Significant differences were found in the number of Con A receptors per cell depending on the fixative used. The fluorescence intensity of FITC-Con A staining was detected spectrophotometrically, the characteristic X-rays of gold-labeled Con A receptors were determined by means of electron beam-induced X-ray microanalysis. The experimental results obtained both at light and electron microscopic level pointed to formaldehyde being the best fixative also for this purpose.

A novel mechanism of murine hepatocyte death inducible by Concanavalin A

“…For the determination of lectin binding, cells were fixed with formaldehyde and washed several times with saline contammg 15 mM phosphate, pH 7.4 (PBS). Fixation was shown not to alter the number of binding sites in accordance with data presented by Peschke et al (14). After incubation of the cells with solutions of fluorescein-Iabeled Con A in PBSI2% bovine serum albumin (BSA) for 60 min at 37°C, they were washed several times with cold PBS/2% BSA and then further incubated for 90 min (37 CC) in a solution containing 50 mM a-methylmannoside (amm), 5 mM EDTA and 2% BSA.…”

“…The plant lectin Con A binds to surface glycoproteins and glycolipids of many cell types, e.g. leukocytes (14,15), hepatocytes (16) and a large number of transformed and nontransformed cell lines (17,IS). Moreover, it may induce a distinct metabolic response after binding to structures such as the insulin receptor on the surface of adipocytes (19,20) or to molecules encoded by the major histocompatibility complex in mice (15).…”

Concanavalin A for in vivo glucose sensing: A biotoxicity review

“…This mitogenicity has been associated with cytokine expression and secretion (Ito et al, 1984;diSabato et al, 1989). ConA has also been found to bind to surface glycoproteins and glycolipids of many cell types including leukocytes (Pink et al, 1983;Peschke et al, 1990), keratonicytes (Schaumberger-Lever, 1990), hepatocytes (McMillan et al, 1984), and a large number of transformed and nontransformed cell lines (Ozanne and Sambrook, 1971;Cline and Livingston, 1971). ConA has been shown to directly stimulate B cells to synthesize DNA and to proliferate (Andersson et al, 1972;Gunther et al, 1973).…”

Concanavalin A promotes angiogenesis and proliferation in endothelial cells through the Akt/ERK/Cyclin D1 axis

“…1984 ; Peschke et al. 1990 ; Schaumburg-Lever 1990 ). Thus far, Con A has been generated rising attention for its anti-proliferative and antitumor activities towards various types of cancer cells.…”