Speaker Biography

Fatemeh Aminmarashi

Reproductive and fertility research center, Canada

Title: Granulosa cells exposed to fibroblast growth factor 8 and 18 reveal early onset of cell growth and survival

Fatemeh Aminmarashi
Biography:

Fatemeh Aminmarashi is a DVM ( doctor of veterinary medicine) from Iran and I have a master degree in reproduction biology (CRRF) Reproductive and fertility research center from canada, and my project it is about:

- Extract ovarian granulosa cells using various cell culture techniques
- Analyze the effects of various proteins on granulosa cells
- Identify apoptotic proteins using Western blotting and mass spectrometry.
- Extract ovarian granulosa cells using various cell culture techniques
- Analyze the effects of various proteins on granulosa cells
- Identify apoptotic proteins using Western blotting and mass spectrometry.

Abstract:

Fibroblast Growth factors (FGFs) are growth factors which have diverse biological activities including broad mitogenic and cell survival activities. FGFs constitute a large family of 22 distinct polypeptide growth factors varying in size from 17 to 34 kDa and have between 13 to 71% sequence homology. More specifically, FGF8 and FGF18 are both important modulator of granulosa cell functions. FGF8 and FGF18 are homologous factors which possess similar sequence homology, but we hypothesized they may interact to FGFRs differently leading to distinct effects, particularly on granulosa cell growth and induce proliferation following a short period of exposition. This study was performed to investigate the effects of FGF8 and FGF18 on ovine granulosa cells proteome. Ovine ovaries were obtained from adult sheep’s irrespective of stage of estrous cycle and were cultured using a standard protocol. Granulosa cells were harvested from follicles then, seeded and cultivated. After, they were exposed to 10 ng/mL of FGF8 or FGF18. Cell proteins were extracted, cysteine bonds were reduced and acetylated and proteins were digested with trypsin. Tryptic peptides were analyzed using a bottom-up proteomic approach, mass spectrometry and a label-free quantitation method. The results obtained revealed following treatment with FGF8 or FGF18 for 30 minutes, an important shift toward upregulation for the entire granulosa cell proteome was measured. Additionally, several proteins, including ATF1, STAT3, MAPK1, MAPK3, MAPK14, PLCG1, PLCG2, PKCA, PIK3CA, RAF1, GAB1 and BAG2 were significantly upregulated (> 1.5-fold; p < 0.01). Results are suggesting the activation of the MAPK/ERK pathway as expected. However, it is important to note that ATF1 and STAT3 are important transcription factors involved in cell growth, proliferation and survival and consequently can hamper or rescue the normal ovine reproductive system function. Both are strongly interacting with the MAPK/ERK pathway. They are directly involved in the growth, proliferation and mechanisms of cell survival. This very significant increase of STAT3 and ATF1 could have important consequences on the viability of the oocyte.

Keywords: Proteomics, mass spectrometry, ovary, granulosa cells, fibroblast growth factors, protein kinases, transcription factors, proliferation