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Gene expression profiling of preovulatory follicle in the buffalo cow: effects of increased IGF-I concentration on periovulatory events.

Rao JU, Shah KB, Puttaiah J, Rudraiah M - PLoS ONE (2011)

Bottom Line: Thus, further experiments were conducted to verify the effects of increased intrafollicular IGF-I levels on the expression of genes associated with the above mentioned processes.The results indicated that increased intrafollicular concentrations of IGF-I caused changes in expression of genes associated with steroidogenesis (StAR, SRF) and apoptosis (BCL-2, FKHR, PAWR).These results taken together suggest that onset of gonadotropin surge triggers activation of various biological pathways and that the effects of growth factors and peptides on gonadotropin actions could be examined during preovulatory follicle development.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore, India.

ABSTRACT
The preovulatory follicle in response to gonadotropin surge undergoes dramatic biochemical, and morphological changes orchestrated by expression changes in hundreds of genes. Employing well characterized bovine preovulatory follicle model, granulosa cells (GCs) and follicle wall were collected from the preovulatory follicle before, 1, 10 and 22 h post peak LH surge. Microarray analysis performed on GCs revealed that 450 and 111 genes were differentially expressed at 1 and 22 h post peak LH surge, respectively. For validation, qPCR and immunocytochemistry analyses were carried out for some of the differentially expressed genes. Expression analysis of many of these genes showed distinct expression patterns in GCs and the follicle wall. To study molecular functions and genetic networks, microarray data was analyzed using Ingenuity Pathway Analysis which revealed majority of the differentially expressed genes to cluster within processes like steroidogenesis, cell survival and cell differentiation. In the ovarian follicle, IGF-I is established to be an important regulator of the above mentioned molecular functions. Thus, further experiments were conducted to verify the effects of increased intrafollicular IGF-I levels on the expression of genes associated with the above mentioned processes. For this purpose, buffalo cows were administered with exogenous bGH to transiently increase circulating and intrafollicular concentrations of IGF-I. The results indicated that increased intrafollicular concentrations of IGF-I caused changes in expression of genes associated with steroidogenesis (StAR, SRF) and apoptosis (BCL-2, FKHR, PAWR). These results taken together suggest that onset of gonadotropin surge triggers activation of various biological pathways and that the effects of growth factors and peptides on gonadotropin actions could be examined during preovulatory follicle development.

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Related in: MedlinePlus

Expression of genes associated with cell survival, apoptosis and blood coagulation in periovulatory follicle.(A) Microarray fold expression changes at 1 and 22 h post peak LH surge in granulosa cells for few of the selected up and down regulated genes associated with specific biological process like cell survival, apoptosis and blood coagulation are represented as bar graph. (B) The genes selected by microarray analysis were also subjected to qPCR analysis and the fold expression changes for granulosa cells and follicular wall collected at different time points are represented in bar graphs. Bar for each gene represents the mean ± SEM fold expression change values at each time point (n = 3 animals). Bars with different alphabets above them are significantly different (p<0.05). More details on the analysis are provided in the results section.
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pone-0020754-g007: Expression of genes associated with cell survival, apoptosis and blood coagulation in periovulatory follicle.(A) Microarray fold expression changes at 1 and 22 h post peak LH surge in granulosa cells for few of the selected up and down regulated genes associated with specific biological process like cell survival, apoptosis and blood coagulation are represented as bar graph. (B) The genes selected by microarray analysis were also subjected to qPCR analysis and the fold expression changes for granulosa cells and follicular wall collected at different time points are represented in bar graphs. Bar for each gene represents the mean ± SEM fold expression change values at each time point (n = 3 animals). Bars with different alphabets above them are significantly different (p<0.05). More details on the analysis are provided in the results section.

Mentions: Figure 7 shows qPCR fold change in expression pattern with corresponding microarray data of selected genes associated with cell survival, apoptosis and blood coagulation. Expression of BCL-2 gene was up regulated post peak LH surge with higher expression at 10 h time point. However, BCL-2 expression in the follicle wall did not change significantly (Fig. 7B). Expression of FKHR and PAWR in GCs were significantly down regulated after the onset of peak LH surge, while expression of PAWR was down regulated at 10 and 22 h time points and expression of FKHR was up regulated at 10 h in the follicle wall (Fig. 7B).


Gene expression profiling of preovulatory follicle in the buffalo cow: effects of increased IGF-I concentration on periovulatory events.

Rao JU, Shah KB, Puttaiah J, Rudraiah M - PLoS ONE (2011)

Expression of genes associated with cell survival, apoptosis and blood coagulation in periovulatory follicle.(A) Microarray fold expression changes at 1 and 22 h post peak LH surge in granulosa cells for few of the selected up and down regulated genes associated with specific biological process like cell survival, apoptosis and blood coagulation are represented as bar graph. (B) The genes selected by microarray analysis were also subjected to qPCR analysis and the fold expression changes for granulosa cells and follicular wall collected at different time points are represented in bar graphs. Bar for each gene represents the mean ± SEM fold expression change values at each time point (n = 3 animals). Bars with different alphabets above them are significantly different (p<0.05). More details on the analysis are provided in the results section.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3119055&req=5

pone-0020754-g007: Expression of genes associated with cell survival, apoptosis and blood coagulation in periovulatory follicle.(A) Microarray fold expression changes at 1 and 22 h post peak LH surge in granulosa cells for few of the selected up and down regulated genes associated with specific biological process like cell survival, apoptosis and blood coagulation are represented as bar graph. (B) The genes selected by microarray analysis were also subjected to qPCR analysis and the fold expression changes for granulosa cells and follicular wall collected at different time points are represented in bar graphs. Bar for each gene represents the mean ± SEM fold expression change values at each time point (n = 3 animals). Bars with different alphabets above them are significantly different (p<0.05). More details on the analysis are provided in the results section.
Mentions: Figure 7 shows qPCR fold change in expression pattern with corresponding microarray data of selected genes associated with cell survival, apoptosis and blood coagulation. Expression of BCL-2 gene was up regulated post peak LH surge with higher expression at 10 h time point. However, BCL-2 expression in the follicle wall did not change significantly (Fig. 7B). Expression of FKHR and PAWR in GCs were significantly down regulated after the onset of peak LH surge, while expression of PAWR was down regulated at 10 and 22 h time points and expression of FKHR was up regulated at 10 h in the follicle wall (Fig. 7B).

Bottom Line: Thus, further experiments were conducted to verify the effects of increased intrafollicular IGF-I levels on the expression of genes associated with the above mentioned processes.The results indicated that increased intrafollicular concentrations of IGF-I caused changes in expression of genes associated with steroidogenesis (StAR, SRF) and apoptosis (BCL-2, FKHR, PAWR).These results taken together suggest that onset of gonadotropin surge triggers activation of various biological pathways and that the effects of growth factors and peptides on gonadotropin actions could be examined during preovulatory follicle development.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore, India.

ABSTRACT
The preovulatory follicle in response to gonadotropin surge undergoes dramatic biochemical, and morphological changes orchestrated by expression changes in hundreds of genes. Employing well characterized bovine preovulatory follicle model, granulosa cells (GCs) and follicle wall were collected from the preovulatory follicle before, 1, 10 and 22 h post peak LH surge. Microarray analysis performed on GCs revealed that 450 and 111 genes were differentially expressed at 1 and 22 h post peak LH surge, respectively. For validation, qPCR and immunocytochemistry analyses were carried out for some of the differentially expressed genes. Expression analysis of many of these genes showed distinct expression patterns in GCs and the follicle wall. To study molecular functions and genetic networks, microarray data was analyzed using Ingenuity Pathway Analysis which revealed majority of the differentially expressed genes to cluster within processes like steroidogenesis, cell survival and cell differentiation. In the ovarian follicle, IGF-I is established to be an important regulator of the above mentioned molecular functions. Thus, further experiments were conducted to verify the effects of increased intrafollicular IGF-I levels on the expression of genes associated with the above mentioned processes. For this purpose, buffalo cows were administered with exogenous bGH to transiently increase circulating and intrafollicular concentrations of IGF-I. The results indicated that increased intrafollicular concentrations of IGF-I caused changes in expression of genes associated with steroidogenesis (StAR, SRF) and apoptosis (BCL-2, FKHR, PAWR). These results taken together suggest that onset of gonadotropin surge triggers activation of various biological pathways and that the effects of growth factors and peptides on gonadotropin actions could be examined during preovulatory follicle development.

Show MeSH
Related in: MedlinePlus