Relative expression of insulin like growth factor I (IGFI) and follicle stimulating hormone receptor (FSHR) in follicles and ovarian tissue from Bos primigenius indicus (Nelore)
Keywords:Semi-quantitative RT-PCR, Gene expression, Cattle
AbstractThe improvement of techniques to maximize reproductive potential of females needs the whole comprehension of the controlling mechanisms of follicular development. An alternative for this purpose is the quantification of relative gene expression from those genes involved in recruitment, selection and follicular development, using reverse transcriptase - polymerase chain reaction (RT - PCR). The present study aimed to quantify relative gene expression from insulin-like growth factor I (IGF-I) and follicle stimulating hormone receptor (FSHR) in cattle (Bos primigenius indicus), using as internal control the gliceraldheyde 3 phosphate dehydrogenase (GAPDH) gene. Ovaries in different estral cycle stages were obtained from slaughtered animals. Total RNA from follicles and ovarian tissue was purified and the RT-PCR conditions were standardized. PCR products were analyzed in ethydium bromide agarose gels and the bands submitted to densitometric analysis. Exponential amplification curves were constructed and the method's validation was performed using regression analysis to determine the amplification coefficient (E) for each of the three genes. Relative expression for each gene was calculated using the formula described by Prelle et al.12. In every sample there was gene expression detected for each gene showing differences related to cycle temperature. Amplification coefficient (E) was similar between control gene (GAPDH) and IGFI, independently of the class analyzed. IGFI linear amplification could be related to the constitutive characteristic of this protein since the transcripts are not dependents of FSH levels. It was observed difference in FSHR mRNA expression between the classes analyzed. It could be due to the variation of the receptor number in granulosa cells for each different phase of estral cycle. Semi-quantitative RT-PCR has a large application in biotechnology since it is useful to help us the better understanding of follicular dynamics. However these studies must be conducted using other genes in order to provide new clues for the physiology of folliculogenesis.
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