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• W2025169862 abstract "OBJECTIVE: The SMAD signal transducer proteins mediate signaling by members of the TGF-beta superfamily of cytokines, including activin, TGF-beta, and BMP. TGF-beta superfamily members are known to control folliculogenesis and ovulation through direct effects on granulosa cells in the developing follicles, and signal mainly through two different SMAD pathways. However, the roles of individual SMAD proteins in ovarian function, and in particular their influence in fertility, are only beginning to be elucidated. Therefore, we set out to determine if the SMAD transcription factors are expressed in human granulosa cells.DESIGN: Laboratory experimental study.MATERIALS AND METHODS: This study was approved by the institutional review board (IRB) of our institute. Granulosa cells were obtained from consecutive patients undergoing in vitro fertilization (IVF). Granulosa cells were obtained from follicle aspirates after the cumulus-oocyte complexes were removed. The granulosa cells were separated from red blood cells by centrifugation and Percoll gradient prior to RNA extraction. Total RNA was extracted from the granulosa cells using the RNeasy Mini Kit (Qiagen) and cDNA was synthesized using the iScript cDNA Synthesis kit (Bio-Rad Laboratories). ChromaTaq™ DNA Polymerase (Denville Scientific Inc.) was then used to perform the polymerase chain reaction (PCR) using the primers for the SMAD transcription factors SMAD1, SMAD2, SMAD3, SMAD4 and SMAD5. Positive and negative controls were performed for each primer. After PCR, gel electrophoresis was performed to assess the presence of an amplified product.RESULTS: All SMAD family members tested, SMAD1, SMAD2, SMAD3, SMAD4 and SMAD5, were expressed in the granulosa cells of all patients undergoing IVF, regardless of the cause of their infertility.CONCLUSIONS: The SMAD signal transducer proteins play differing roles in mediating signaling by members of the TGF-beta superfamily in the ovary, and may thus influence folliculogenesis and fertility. In particular, primordial germ cell development is impaired in Smad1 and Smad5 knockout mice, and Smad3 deficient mice exhibit impaired folliculogenesis and reduced fertility. We have now shown that this class of signal transduction factors is expressed in luteinized granulosa cells in the human, and thus may also play important roles in folliculogenesis and fertility in humans. OBJECTIVE: The SMAD signal transducer proteins mediate signaling by members of the TGF-beta superfamily of cytokines, including activin, TGF-beta, and BMP. TGF-beta superfamily members are known to control folliculogenesis and ovulation through direct effects on granulosa cells in the developing follicles, and signal mainly through two different SMAD pathways. However, the roles of individual SMAD proteins in ovarian function, and in particular their influence in fertility, are only beginning to be elucidated. Therefore, we set out to determine if the SMAD transcription factors are expressed in human granulosa cells. DESIGN: Laboratory experimental study. MATERIALS AND METHODS: This study was approved by the institutional review board (IRB) of our institute. Granulosa cells were obtained from consecutive patients undergoing in vitro fertilization (IVF). Granulosa cells were obtained from follicle aspirates after the cumulus-oocyte complexes were removed. The granulosa cells were separated from red blood cells by centrifugation and Percoll gradient prior to RNA extraction. Total RNA was extracted from the granulosa cells using the RNeasy Mini Kit (Qiagen) and cDNA was synthesized using the iScript cDNA Synthesis kit (Bio-Rad Laboratories). ChromaTaq™ DNA Polymerase (Denville Scientific Inc.) was then used to perform the polymerase chain reaction (PCR) using the primers for the SMAD transcription factors SMAD1, SMAD2, SMAD3, SMAD4 and SMAD5. Positive and negative controls were performed for each primer. After PCR, gel electrophoresis was performed to assess the presence of an amplified product. RESULTS: All SMAD family members tested, SMAD1, SMAD2, SMAD3, SMAD4 and SMAD5, were expressed in the granulosa cells of all patients undergoing IVF, regardless of the cause of their infertility. CONCLUSIONS: The SMAD signal transducer proteins play differing roles in mediating signaling by members of the TGF-beta superfamily in the ovary, and may thus influence folliculogenesis and fertility. In particular, primordial germ cell development is impaired in Smad1 and Smad5 knockout mice, and Smad3 deficient mice exhibit impaired folliculogenesis and reduced fertility. We have now shown that this class of signal transduction factors is expressed in luteinized granulosa cells in the human, and thus may also play important roles in folliculogenesis and fertility in humans." @default.
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• W2025169862 date "2008-09-01" @default.
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• W2025169862 title "Expression of smads in human granulosa cells" @default.
• W2025169862 doi "https://doi.org/10.1016/j.fertnstert.2008.07.1282" @default.
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