Summary
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.
Similar content being viewed by others
References
Alberts B, Bray D, Lewis J, Raff M, Roberts K, Watson JD (1983) Molecular biology of the cell. Garland Publishing, New York London
Aumüller G, Seitz J, Heyns W, Kirchner C (1985) Ultrastructural localization of uteroglobin immunoreactivity in rabbit lung and endometrium, and rat ventral prostate. Histochemistry 83:413–417
Beier HM (1966) Das Proteinmilieu in Serum, Uterus und Blastozysten des Kaninchens vor der Nidation. In: Beermann W (Chairman) Biochemie der Morphogenese, Colloquium, Deutsche Forschungsgemeinschaft, Konstanz, 28.–30.11.1966, pp 1–10
Beier HM (1967) Veränderungen am Proteinmuster des Uterus bei dessen Ernährungsfunktion für die Blastocyste des Kaninchens. Verh Dtsch Zool Ges Heidelberg 31:139–148
Beier HM (1968) Biochemisch-entwicklungsphysiologische Untersuchungen am Proteinmilieu für die Blastocystenentwicklung des Kaninchens (Oryctolagus cuniculus). Zool Jb Anat 85:72–190
Beier HM (1978) Physiology of uteroglobin. In: Spilman CH, Wilks JW (eds) Novel aspects of reproductive biology, vol 8, SP Medical and Scientific Books, New York London, pp 219–248
Beier HM, Mootz U, Kühnel W (1972) Asynchrone Eitransplantation während der verzögerten Uterussekretion beim Kaninchen. Proc VII, Congr Anim Reprod Artif Inserm, München, pp 1891–1892
Beier HM, Mootz U, Fischer B, Ströbele-Müller R (1983) Growth and differentiation of rabbit blastocysts in defined culture media. In: Beier HM, Lindner HR (eds) Fertilization of the human egg in vitro. Springer, Berlin Heidelberg New York, pp 371–386
Dannhorn DR, Kirchner C (1990) Uptake of tritiated uteroglobin by the endometrium of the rabbit during peri-implantation. Cell Tissue Res 259:519–528
Dannhorn DR, Henkel R, Kirchner C (1988) Synthese von Uteroglobin in der Blastozyste des Kaninchens? Autoradiographische und immunoytochemische Untersuchungen. Fertilität 4:223–226
Dannhorn DR, Wirth B, Kirchner C (1989) Purification of uteroglobin using monospecific antibodies coupled to divinylsulphone-activated agarose. J Immunol Methods 119:223–230
De Dure C (1975) Exploring cells with a centrifuge Science 189:186–194
Denker H-W, Gerdes H-J (1979) The dynamic structure of rabbit blastocyst coverings. I. Transformation during regular preimplantation development. Anat Embryol 157:15–34
Dreiner UH (1988) Immunhistochemische Lokalisation des Uteroglobins in der Blastozyste des Kaninchens. Dissertation, Medical Faculty of the RWTH Aachen
El-Banna AA, Daniel JC, Jr (1972) The effect of protein fractions from rabbit uterine fluids on embryo growth and uptake of nucleic acid and protein precursors. Fertil Steril 23:105–114
El-Etreby MF, Beier HM, Elger W, Mahrous AT, Töpert M (1983) Immunocytochemical localization of uteroglobin in the genital tract of the male rabbit. Cell Tiss Res 229:61–73
Fischer B (1987) Development retardation in cultured preimplantation rabbit embryos. J Reprod Fert 79:115–123
Fischer B, Denker H-W, Beier HM, Gerdes H-J (1983) Der Umbau der extrazellulären Blastozystenhüllen als ein empfindlicher Indikator für unphysiologische Milieubedingungen in der In-vitro-Kultur. Verh Anat Ges 77:427–429
Fischer B, Jung T, Hegele-Hartung C, Beier HM (1990a) Development of preimplantation rabbit embryos in uterine flushing supplemented culture media. Mol Reprod Dev 27:216–223
Fischer B, Mootz U, Denker H-W, Lambertz M, Beier HM (1990b) The dynamic structure of rabbit blastocyst coverings. III. Transformation of coverings under non-physiological developmental conditions. Anat Embryol (submitted)
Hastings RA, Enders AC (1974) Uptake of exogenous protein by the preimplantation rabbit. Anat Rec 179:311–330
Hastings RA, Enders AC, Schlafke S (1972) Permeability of the zona pellucida to protein tracers. Biol Reprod 7:288–296
Hegele-Hartung C, Beier HM (1985) Immunocytochemical localization of uteroglobin in the rabbit endometrium. Anat Embryol 172:295–301
Hegele-Hartung C, Beier HM (1986) Distribution of uteroglobin in the rabbit endometrium after treatment with an antiprogesterone (ZK 98.734): an immunocytochemical study. Hum Reprod 1:497–505
Hoffman LH, Olson GE (1980) Crystalline inclusions in the rabbit blastocyst. Exp Cell Res 127:1–14
Hoffman LH, Olson GE (1984) Crystalline inclusions in embryonic and maternal cells. In: Van Blerkom J, Motta PM (eds) Ultrastructure of reproduction. Martinus Nijhoff, Boston, pp 235–246
Kazemi M, Amann JF, Keisler DH, Ing NH, Roberts RM, Morgan G, Wooding FBP (1990) A progesterone-modulated, low-molecular weight protein from the uterus of the sheep is associated with crystalline inclusion bodies in uterine epithelium and embryonic trophectoderm. Biol Reprod 43:80–96
Kirchner C (1969) Untersuchungen an uterusspezifischen Glykoproteinen während der frühen Gravidität des Kaninchens Oryctolagus cuniculus. Wilhelm Roux' Arch Devel Biol 164:97–133
Kirchner C (1976) Uteroglobin in the rabbit. I. Intracellular localization in the oviduct, uterus and preimplantation blastocyst. Cell Tissue Res 170:415–424
Kirchner C (1979) Immunohistochemical localization of secretory proteins in the endometrial epithelium of the rabbit. Cell Tissue Res 199:25–36
Kirchner C, Seitz KA (1972) Elektronenmikroskopische Untersuchungen über die Blastozyste des Kaninchens vor der Implantation in Bezug auf ihre Wechselbeziehung zur uterinen Umgebung. Wilhelm Roux' Arch Entwicklungsmech Org 170:221–233
Krishnan RS, Daniel JC, Jr (1967) “Blastokinin”: inducer and regulator of blastocyst development in the rabbit uterus. Science 158:490–492
Maurer RR (1978) Advances in rabbit embryos. In: Daniel JC, Jr (ed) Methods in mammalian reproduction. Academic Press, New York, pp 259–272
Mayol RF, Longenecker DE (1974) Development of a radioimmunoassay for blastokinin. Endocrinology 95:1534–1542
Miele L, Cordella-Miele E, Mukherjee AB (1987) Uteroglobin structure, molecular biology, and new perspectives on its function as a phospholipase A2 inhibitor. Endocr Rev 8:474–490
Mootz U (1979) Rasterelektronenmikroskopische Untersuchungen an der implantationsbereiten Blastozyste des Kaninchens. Verh Anat Ges 73:435–444
Nieto A, Beato M (1980) Synthesis and secretion of uteroglobin in rabbit endometrial explants cultured in vitro. Mol Cell Endocr 17:25–39
Robinson DH, Kirk KL, Benos DJ (1989) Macromolecular transport in rabbit blastocysts: Evidence for a specific uteroglobin transport system. Mol Cell Endocr 63:227–237
Saxena SK, Sahib MK (1984) Uteroglobin synthesis by rabbit uterine endometrial explants. Indian J Biochem Biophys 21:47–50
Schlafke S, Enders AC (1973) Protein uptake by rat preimplantation stages. Anat Rec 175:539–560
Sternberger LA, Hardy PH, Cuculis JJ, Meyer GH (1970) The unlabeled antibody enzyme method of immunohistochemistry. J Histochem Cytochem 18:315–323
Stroband HWJ, Taverne N, Bogaard M vd (1984) The big blastocyst: its ultrastructure and uptake of protein macromolecules. Cell Tissue Res 235:347–356
Whitson GL, Murray FL (1974) Cell culture of mammalian endometrium and synthesis of blastokinin in vitro. Science 183:668–670
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hegele-Hartung, C., Dreiner, U. & Beier, H.M. Effect of in vitro culture on the dynamics of uteroglobin distribution in rabbit blastocysts. Anat Embryol 183, 119–128 (1991). https://doi.org/10.1007/BF00174392
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00174392