Cumulus and granulosa cell markers of oocyte and embryo quality

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Cumulus and granulosa cell markers of oocyte and embryo quality Asli Uyar, Ph.D., Saioa Torrealday, M.D., Emre Seli, M.D. Fertility and Sterility Volume 99, Issue 4, Pages 979-997 (March 2013) DOI: 10.1016/j.fertnstert.2013.01.129 Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Oocyte specific genes studied in mouse that are required for folliculogenesis. FIGα: factor in the germline, alpha; NOBOX: newborn ovary homeobox protein; SOHLH1: spermatogenesis and oogenesis specific basic helix-loop-helix 1; SOHLH2: spermatogenesis and oogenesis specific basic helix-loop-helix 2; LHX8: lim homebox protein 8; GDF-9: growth differentiation factor-9; EPAB: embryonic poly(A) binding protein; ZFP2: zinc finger protein 2; and BMP-15: bone morphogenic protein-15. Scale bars represent 10 μm. Reproduced with permission, from Guzeloglu-Kayisli et al. Biochemical Journal 2012;446:47–58 (10). Fertility and Sterility 2013 99, 979-997DOI: (10.1016/j.fertnstert.2013.01.129) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Illustration of the three steps (denaturation, primer annealing, and strand extension) involved in PCR resulting in the exponential amplification of PCR products assuming the theoretical efficiency of 2. Fertility and Sterility 2013 99, 979-997DOI: (10.1016/j.fertnstert.2013.01.129) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Graphs representing the qRT-PCR results: (A) Hypothetical amplification plots of a series of 10-fold dilutions with arbitrary fluorescence units. (Technical replicates were not shown in the figure.) (B) The standard curve obtained from the Ct values of the diluted samples. Fertility and Sterility 2013 99, 979-997DOI: (10.1016/j.fertnstert.2013.01.129) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Major steps of a typical microarray experiment. (A) Total RNA is extracted from the follicular cells samples, and mRNA is reverse transcribed into cDNA. Amplified and biotin-labeled cRNA is obtained through in vitro transcription. (B) Fragmented cRNA samples are hybridized to microarray slide. Labeled targets bind to their complementary oligonucleotides attached in the microscopic probes. The array is then washed and scanned to obtain the fluorescent image, which is further processed to get the intensity values for data analysis. (C) Relative gene expression values are first normalized to eliminate the possible nonbiological variations. Then statistical analyses are performed to identify the differentially expressed genes. Two sample methods and related graphs for analysis and visualization of microarray data are shown: PCA for visualization of the expression data by projection into a reduced dimensional space where each point in the graph represents a single sample; and HeatMap analysis for hierarchical clustering of samples and genes based on the differential expression observed. Fertility and Sterility 2013 99, 979-997DOI: (10.1016/j.fertnstert.2013.01.129) Copyright © 2013 American Society for Reproductive Medicine Terms and Conditions