Incidence of apoptosis and transcript abundance in bovine follicular cells is associated with the quality of the enclosed oocyte
THERIOGENOLOGY
Authors: Janowski, D.; Salilew-Wondim, D.; Torner, H.; Tesfaye, D.; Ghanem, N.; Tomek, W.; El-Sayed, A.; Schellander, K.; Hoelker, M.
Abstract
The close contact and interaction between the oocyte and the follicular environment influence the establishment of oocyte developmental competence. Moreover, it is assumed that apoptosis in the follicular cells has a beneficial influence on the developmental competence of oocytes. The aim of this study was to investigate whether bovine oocytes with varied developmental competence show differences in the degree of apoptosis and gene expression pattern in their surrounding follicular cells (cumulus and granulosa cells). Oocytes and follicular cells from follicles of 3 to 5 mm in diameter were grouped as brilliant cresyl blue (BCB)+ and BCB- based on glucose-6-phosphate dehydrogenase (G6PDH) activity in the ooplasm by BCB staining. In the follicular cells initial, early and late apoptotic events were assessed by analyzing caspase-3 activity, annexin-V and TUNEL, respectively. Global gene expression was investigated in immature oocytes and corresponding follicular cells. BCB+ oocytes resulted in a higher blastocyst rate (19.3%) compared to the BCB- group (7.4%, P < 0.05). Moreover, the analysis of apoptosis showed a higher caspase-3 activity in the follicular cells and an increased degree of late apoptotic events in granulosa cells in the BCB + compared with the BCB- group. Additionally, the global gene expression profile revealed a total of 34 and 37 differentially expressed genes between BCB + and BCB- cumulus cells and granulosa cells, respectively, whereas 207 genes showed an altered transcript abundance between BCB+ and BCB- oocytes. Among these, EIF3F, RARRES2, RNF34, ACTA1, GSTA1, EIF3A, VIM and CS gene transcripts were most highly enriched in the BCB + oocytes, whereas OLFM1, LINGO1, ALDH1A3, PTHLH, BTN3A3, MRPS2 and PPM1K were most significantly reduced in these cells. Therefore, the follicular cells enclosing developmentally competent oocytes show a higher level of apoptosis and a different pattern of gene expression compared to follicular cells enclosing non-competent bovine oocytes. (C) 2012 Elsevier Inc. All rights reserved.
Changes of Chemerin Production in Obese Patients with Different States of Carbohydrate Metabolism
BIOCHEMISTRY MOSCOW-SUPPLEMENT SERIES B-BIOMEDICAL CHEMISTRY
Authors: Vasilenko, M. A.; Kirienkova, E. V.; Skuratovskaia, D. A.; Zatolokin, P. A.; Mironyuk, N. I.; Litvinova, L. S.
Abstract
Chemerin is an adipose tissue mediator involved in the regulation of many processes, including lipogenesis, inflammatory responses, etc. The role of chemerin in the development of insulin resistance still needs better understanding. The aim of the study was to investigate chemerin production in obese patients with different states of carbohydrate metabolism. The study included 155 patients with diagnosed obesity and 34 patients with overweight. The control group 1 consisted of 43 conditionally healthy donors without obesity. For comparison of the research data on evaluation of tissue-specific mRNA expression of the genes IL-6, TNF-alpha, RARRES2, (encoding IL-6, TNF-alpha, and chemerin, respectively) control group 2 consisting of 30 non-obese was also included into this study. The relative level of mRNA expression of the genes IL-6, TNF-alpha and RARRES2 (encoding IL-6, TNF-alpha and chemerin, respectively) was carried out using real time PCR. Concentrations of IL-6, TNF-alpha, and chemerin were measured in serum/plasma using an enzyme-linked immunosorbent assay (ELISA). Significant differences were found in the plasma level of chemerin and tissue-specific patterns of RARRES2 gene expression in obese patients; these changes depended on the degree of obesity and the state of carbohydrate metabolism. Opposite associations between RARRES2 gene expression and expression TNF-alpha and IL-6 genes have been recognized in adipose tissues of different localization: in obese patients (BMI <= 40 kg/m(2)) without type 2 diabetes mellitus (DM2) they were negative, while in obese patients with DM2 diabetes they were positive. The recognized correlations between the plasma content of chemerin and the expression level of its gene in biopsies with various parameters of carbohydrate and lipid metabolism, and proinflammatory molecules indicate chemerin involvement in metabolic and immune processes in obesity.