Purification of uteroglobin using monospecific antibodies coupled to divinylsulphone-activated agarose

Dannhorn, D. R.; Wirth, B.; Kirchner, C.

doi:10.1016/0022-1759(89)90400-6

Cited by 5 publications

(1 citation statement)

References 24 publications

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“…Quantitative uteroglobin measurements have been made with various techniques: Kirchner (1969) determined uteroglobin by scanning densitometry following gel electrophoresis, Saxena and Sahib (1984) by immunoprecipitation, and Dannhorn et al (1989) by competitive ELISA. The most sensitive technique, however, is the RIA, with its threshold of detection of 0.5 ng uteroglobin, as used in the present investigation.…”

Section: Discussionmentioning

confidence: 99%

Effect of in vitro culture on the dynamics of uteroglobin distribution in rabbit blastocysts

1991

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The purpose of this study was to investigate the localization and transport of uteroglobin in normal rabbit blastocysts (day 4-day 6 p.c.) and in those cultured for 6-48 h in vitro, using a specific radioimmunoassay and immunocytochemistry. The results of the radioimmunoassay showed that in day 4 p.c. blastocyst tissue (based on homogenate measurements) a significant decrease of the uteroglobin content started after only 6 h of culture in vitro. A significant concomitant rise of uteroglobin was observed in the culture medium after 12 h of in vitro culture. Using immunocytochemistry it was not possible to detect uteroglobin in any compartment of the non-cultured or in vitro cultured day 4 p.c. blastocysts. The efflux of uteroglobin down a concentration gradient was confirmed by the immunocytochemistry in non-cultured and in vitro cultured day 5 p.c. and day 6 p.c. blastocysts. Uteroglobin immunoreactions were mainly detected in non-cultured blastocysts (day 5 and 6 p.c.) in large vesicles of the trophoblast cells. In addition endocytotic vesicles at the inside of the apical membrane of trophoblast cells, some cell debris within the perivitelline space and the neozona were labelled. During in vitro culture of day 5 and 6 p.c. blastocysts, uteroglobin labelling in the coverings did not change. In non-cultured and cultured day 5 and 6 p.c. blastocysts neither the compartments of the embryoblast, the endoderm cells nor the blastocyst cavity showed any uteroglobin immunoreactions. After only 6 h of in vitro culture, uteroglobin immunoreactions were no longer found within the trophoblast cells. The reaction did not reappear during the course of in vitro culture up to 48 h, suggesting a complete lack of de novo synthesis of uteroglobin by blastocysts.

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Section: Discussionmentioning

confidence: 99%

Effect of in vitro culture on the dynamics of uteroglobin distribution in rabbit blastocysts

1991

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Biological activity of sulfoxides and sulfones

Kalir

1993

PATAI'S Chemistry of Functional Groups

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Uptake and accumulation of tritiated uteroglobin by day-6 rabbit blastocysts

Dannhorn

Kirchner

1990

Cell Tissue Res

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Uptake of uteroglobin (UGL) by day-6 rabbit blastocysts and the intracellular fate of this protein were studied by light- and electron-microscopic autoradiography, immunocytochemistry and acid-phosphatase cytochemistry. UGL, labelled with N-succinimidyl-(2-3-3H)-propionate, was administered to embryos in vitro for 25 min to 4 h. The kinetics, determined from light-microscopic autoradiographs, showed a continuous uptake of the labeled protein over a 4-h period of incubation. At the ultrastructural level, increasing numbers of silver grains and an intense UGL immunoreaction in protein vacuoles and crystalloid bodies of trophoblast cells indicated that 3H-UGL had accumulated in these organelles. The presence of crystalloid inclusions in protein vacuoles suggests their origin by a condensation of the protein content, including UGL. Lysosomes containing radioactivity were rarely found, suggesting a very low degradation rate of the 3H-UGL. Protein vacuoles and crystalloid bodies exhibited no acid phosphatase reaction. The enzyme was mainly found outside the basal and lateral cell membranes of trophoblast cells, and on the rough endoplasmic reticulum of endoderm cells.

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Purification of uteroglobin using monospecific antibodies coupled to divinylsulphone-activated agarose

Cited by 5 publications

References 24 publications

Effect of in vitro culture on the dynamics of uteroglobin distribution in rabbit blastocysts

Effect of in vitro culture on the dynamics of uteroglobin distribution in rabbit blastocysts

Biological activity of sulfoxides and sulfones

Uptake and accumulation of tritiated uteroglobin by day-6 rabbit blastocysts

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