Sequencing and annotated analysis of an Estonian human genome

In present study we describe the sequencing and annotated analysis of the individual genome of Estonian. Using SOLID technology we generated 2,449,441,916 of 50-bp reads. The Bioscope version 1.3 was used for mapping and pairing of reads to the NCBI human genome reference (build 36, hg18). Bioscope enables also the annotation of the results of variant (tertiary) analysis. The average mapping of reads was 75.5% with total coverage of 107.72 Gb. resulting in mean fold coverage of 34.6. We found 3,482,975 SNPs out of which 352,492 were novel. 21,222 SNPs were in coding region: 10,649 were synonymous SNPs, 10,360 were nonsynonymous missense SNPs, 155 were nonsynonymous nonsense SNPs and 58 were nonsynonymous frameshifts. We identified 219 CNVs with total base pair coverage of 37,326,300 bp and 87,451 large insertion/deletion polymorphisms covering 10,152,256 bp of the genome. In addition, we found 285,864 small size insertion/deletion polymorphisms out of which 133,969 were novel. Finally, we identified 53 inversions, 19 overlapped genes and 2 overlapped exons. Interestingly, we found the region in chromosome 6 to be enriched with the coding SNPs and CNVs. This study confirms previous findings, that our genomes are more complex and variable as thought before. Therefore, sequencing of the personal genomes followed by annotation would improve the analysis of heritability of phenotypes and our understandings on the functions of genome

The differential transcriptome and ontology profiles of floating and cumulus granulosa cells in stimulated human antral follicles

Communication between various ovarian cell types is a prerequisite for folliculogenesis and ovulation. In antral follicles gran-ulosa cells divide into two distinct populations of mural and cumulus granulosa cells (CGC), enveloping the antrum and surrounding the oocyte, respectively. Both cell types, with the mural compartment in excess, contribute to the floating granulosa cell (FGC) population in the follicular fluid. The aim of this study was to compare the transcriptomes of FGC and CGC in stimulated antral follicles obtained from 19 women undergoing IVF-ICSI procedure. FGC were obtained from follicular fluid during the follicle puncture procedure and CGC were acquired after oocyte denudation for micromanipulation. Gene expression analysis was conducted using the genome-wide Affymetrix transcriptome array. The expression profile of the two granulosa cell populations varied significantly. Out of 28 869 analysed transcripts 4480 were differentially expressed (q-value > 10-4) and 489 showed ≥2-fold difference in the expression level with 222 genes up-regulated in FGC and 267 in CGC. The transcriptome of FGC showed higher expression of genes involved in immune response, hematological system function and organismal injury, although CGC had genes involved in protein degradation and nervous system function up-regulated. Cell-to-cell signalling and interaction pathways were noted in both cell populations. Furthermore, numerous novel transcripts that have not been previously described in follicular physiology were identified. In conclusion, our results provide a solid basis for future studies in follicular biology that will help to identify molecular markers for oocyte and embryo viability in IVF. © The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: [email protected]

THE DIFFERENTIAL TRANSCRIPTOME AND ONTOLOGY PROFILES OF MURAL AND CUMULUS GRANULOSA CELLS IN STIMULATED HUMAN ANTRAL FOLLICLES

Communication between various cell types in the ovary is a prerequisite for successful folliculogenesis and ovulation. In human antral follicles, granulosa cells divide into 2 distinct cell populations: mural (enveloping the antrum, MGC) and cumulus granulosa cells (surrounding the oocyte, CGC). During infertility treatment using in vitro fertilization (IVF), granulosa cells can be retrieved during puncture of stimulated follicles offering an excellent opportunity for analysing their functional properties. The aim of this study was to compare the transcriptomes of MGC and CGC. Twenty infertile women undergoing IVF-ICSI treatment were enrolled. The MGC were obtained from follicular fluid and CGC were acquired after oocyte denudation before micromanipulation. Gene expression of both cell populations was analysed using a genome-wide transcription array. The expression profile of the 2 granulosa cell populations varied significantly: out of 28 869 transcripts, 4480 were differentially expressed (q-value < 10–4); 623 transcripts differed in their expression levels by at least 2-fold. The transcriptome of MGC showed higher expression of genes involved in immune regulation (toll-like receptors, IL18, IL17R). In CGC, pathways participating in intercellular interactions, tissue remodelling and protein degradation were more clearly distinguished (tenascin C, IGFBP5). Among the identified differentially expressed genes, several are involved in follicle development, oocyte maturation, or ovulatory processes. Our findings fit well with previously published data. The results provide a basis for future studies on intra- and intercellular signaling in the preovulatory follicle leading towards identifying methods for improving oocyte health, embryo selection, and ultimately IVF success rate

The Effect of Serum and Follicular Fluid Amyloid-Associated Protein Levels on in Vitro Fertilization Outcome in Patients with Polycystic Ovary Syndrome

In this study, we aimed to investigate serum and follicular fluid amyloid A protein levels in non-obese non-hyperandrogenic patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization (IVF) and IVF outcome. A total of 81 patients undergoing IVF treatment, 41 patients diagnosed as PCOS according to the Rotterdam criteria (group I) and 40 patients with the etiology of male factor infertility (group II), were included in the study. On the day of oocyte pickup, serum and follicular fluid samples were collected from all patients. Serum E2 level on the day of hCG (2849.93 +/- 541.54 vs. 2494.28 +/- 712.98) and total number of retrieved oocytes (13.73 +/- 3.57 vs. 10.53 +/- 4.07) were significantly higher in group I when compared to group II (p 0.05). No significant difference was found between two groups regarding the serum and follicular fluid amyloid A protein levels on the day of oocyte retrieval (p > 0.05).Wo