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Regular Article |
a Department of Anatomy and Physiology, Royal Veterinary and Agricultural University, 1870 Frederiksberg C, Denmark
b Konstantin the Philosopher University, Nitra, Slovak Republic
c Research Institute of Animal Production, Nitra, Slovak Republic
d Veterinary University, Vienna, Austria
e Department of Biotechnology, Institut für Tierzucht und Tierverhalten, Mariensee, (FAL), Neustadt, Germany
f Precision Therapeutics, Pittsburgh, Pennsylvania 15213
g Universität Bonn, Institut für Tierzuchtwissenschaft, Germany
ABSTRACT
Ribosomal RNA genes are transcribed in the nucleolus. The formation of this organelle after fertilization is essential for embryonic protein synthesis and viability. We have examined nucleolus formation in in vivo-derived porcine embryos by light microscopical autoradiography following 20 min of 3H-uridine incubation, transmission electron microscopy (TEM), and immunocytochemical localization by confocal laser scanning microscopy of key nucleolar proteins involved in rRNA transcription (nucleolin, upstream binding factor, topoisomerase I, and RNA polymerase I) and processing (fibrillarin, nucleophosmin). During the first two postfertilization cell cycles, TEM revealed fibrillar spheres as the most prominent intranuclear entity of the blastomeres. Fibrillogranular nucleoli were established during the third cell cycle. Initially, fibrillar centers, a dense fibrillar component, and a granular component were formed on the surface of the fibrillar spheres. At the same time, autoradiographic labeling over the nucleoplasm and in particular the nucleoli was detected for the first time. The nucleolar proteins were, in general, not immunocytochemically localized to the presumptive nucleolar compartment until late during the third or early during the fourth cell cycle.
1 This project was supported by the Bundesministerium für Wissenschaft und Verkehr der Republik Österreich GZ 45.023/1-III/B/8/98, Danish Agricultural and Veterinary Research Council, and through an Alexander von Humboldt fellowship.
2 Correspondence: Poul Hyttel, Department of Anatomy and Physiology, Royal Veterinary and Agricultural University Groennegaardsvej 7, DK-1870 Frederiksberg C, Denmark. FAX: 45 3528 2547; poh{at}kvl.dk
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