HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal My Folders Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Markkula, M. Articles by Makarevich, A. Search for Related Content PubMed PubMed Citation Articles by Markkula, M. Articles by Makarevich, A. Agricola Articles by Markkula, M. Articles by Makarevich, A.

Ratio of Proliferating Cell Nuclear Antigen-Positive Nuclei to Total Cell Number Is Higher in Day 7 Than in Day 8 Vitrified In Vitro-Produced Bovine Embryos¹

^a Agricultural Research Centre of Finland, Animal Production Research, FIN-31600 Jokioinen, Finland ^b University of Turku, Department of Biology, FIN-20014 Turku, Finland ^c Research Institute of Animal Production, SK-94992 Nitra, Slovak Republic

ABSTRACT

The aim of the present study was to find a reliable functional criterion for the evaluation of the proliferation potential of bovine in vitro-produced embryos. We used immunocytochemical detection of proliferating cell nuclear antigen (PCNA) combined with propidium iodide (PI) staining and subsequent confocal laser scanning microscopy together with routine morphological evaluation under a stereomicroscope to study fresh Day 7, 8, and 9, and cryopreserved Day 7 and 8 embryos. The ratio of PCNA/PI-positive nuclei was equal in fresh Day 7 and Day 8 embryos and significantly lower in Day 9 embryos. In general, Day 7 embryos tolerated the cryopreservation treatments better than Day 8 embryos. Vitrification in normal straws was especially detrimental to Day 8 embryos. In fresh Day 7 and 8 embryos, the PCNA results were in agreement with stereomicroscopic evaluation. However, in Day 9 fresh and in Day 7 and 8 treated embryos, the missing PCNA revealed disorders that were not observed under morphological evaluation. PCNA immunocytochemistry is an effective method to obtain information about the functional state of nuclei. The ratio of PCNA-positive nuclei can provide more information and numerical data about the developmental potential of bovine embryos after cryopreservation.

FOOTNOTES

First decision: 2 November 2000.

¹ This work was supported by the Ministry of Agriculture and Forestry, Finland and Tekes, the National Technology Agency, Finland.

² Correspondence. FAX: 358 3 4188 8618; merja.markkula{at}mtt.fi

This article has been cited by other articles:

				A.V. Makarevich and M. Markkula Apoptosis and Cell Proliferation Potential of Bovine Embryos Stimulated with Insulin-Like Growth Factor I During In Vitro Maturation and Culture Biol Reprod, February 1, 2002; 66(2): 386 - 392. [Abstract] [Full Text] [PDF]