Successive Detection of Telomerase by IHC and Assessment of Telomere Length by Q-FISH in Paucicellular Cumulus Samples from Cumulus-Oocyte Complexes Obtained in Assisted Reproduction Programs

Abstract

This paper suggests an approach for the use of a single paucicellular histological sample to investigate two characteristics indicative of the cell’s functional potential: 1) the content of telomerase reverse transcriptase (TERT) and 2) the telomere length. An algorithm has been suggested for the successive detection of the catalytic telomerase subunit with immunohistochemical staining and assessment of telomere length with Q-FISH on the same set of cells. The described approach uses three incontestable advantages of the FISH technique, namely, the possibility of analyzing target sequences in individual cells and paucicellular samples, the performance of several successive hybridization rounds with different FISH probes on the same sample, and the examination of genomic regions consisting of repeats. This approach has been applied to assess the TERT content and telomere length in cumulus cells from human cumulus-oocyte complexes obtained from assisted reproduction programs. This approach provides an opportunity to examine the correlation between the TERT content and telomere length in cumulus cells surrounding the oocyte and the oocyte's capacity for fertilization, as well as the subsequent pre- and post-implantation development of the resulting embryo.