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Immunoreactivities of androgen receptor, estrogen receptors, p450arom, p450c17 proteins in wild ground squirrels ovaries during the nonbreeding and breeding seasons.

Li X, Zhang H, Sheng X, Li B, Zhou J, Xu M, Weng Q, Watanabe G, Taya K - J Ovarian Res (2012)

Bottom Line: However, the immunoreactivities of ERa and ERb were both significantly reduced in the nonbreeding season compared to the breeding season.The positive stains of FSHR and LHR were found in the granulosa cells and theca cells of the ovaries of the breeding and nonbreeding seasons.In addition, the Western blotting results of FSHR and LHR showed a significant reduction in the nonbreeding season compared with the breeding season.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Biological Science and Technology, Beijing Forestry University, Beijing, 100083, China. qiangweng@bjfu.edu.cn.

ABSTRACT
The aim of this study was to elucidate the regulatory role of androgen in the follicular development of wild female ground squirrels. Immunohistochemical staining of FSHR, LHR, P450c17, P450arom, androgen receptor (AR), estrogen receptors (ERa and ERb) were executed in ovaries of female ground squirrels from both breeding and nonbreeding seasons. In addition, total ovarian proteins were extracted from the ovaries of squirrels from breeding and nonbreeding seasons, and Western blot analysis were performed in order to probe for FSHR, LHR, P450c17, P450arom, AR, ERa and ERb. The results of immunohistochemical staining and Western blotting of P450c17 showed that there was no significant difference between the breeding and nonbreeding seasons. It was found that granulosa cells expressed P450arom during the breeding season. In contrast, there was no positive staining of P450arom in the nonbreeding season. There was no significant difference in immunoreactivity of AR between the breeding and nonbreeding seasons. However, the immunoreactivities of ERa and ERb were both significantly reduced in the nonbreeding season compared to the breeding season. The positive stains of FSHR and LHR were found in the granulosa cells and theca cells of the ovaries of the breeding and nonbreeding seasons. In addition, the Western blotting results of FSHR and LHR showed a significant reduction in the nonbreeding season compared with the breeding season. These findings suggested that androgen might be predominantly converted into estrogen in order to regulate the follicular development via binding of estrogen receptors during the breeding season, whereas androgen might predominantly directly bind androgen receptor to regulate the follicular development during the nonbreeding season in the ovaries of wild female ground squirrels.

No MeSH data available.


The immunolocalization of P450c17 and P450arom in the ovary of wild female ground squirrels. The immunolocalization of P450c17 (a, b, c) and P450arom (d, e, f) in the ovary of wild female ground squirrels during the breeding and nonbreediang seasons. The breeding season divided into two stages, the large follicle stage (a, d) and the small follicle (b, e). g, negative control. IC, interstitial cell; TC, theca cell; GC, granulosa cell. Bars =50μm.
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Figure 2: The immunolocalization of P450c17 and P450arom in the ovary of wild female ground squirrels. The immunolocalization of P450c17 (a, b, c) and P450arom (d, e, f) in the ovary of wild female ground squirrels during the breeding and nonbreediang seasons. The breeding season divided into two stages, the large follicle stage (a, d) and the small follicle (b, e). g, negative control. IC, interstitial cell; TC, theca cell; GC, granulosa cell. Bars =50μm.

Mentions: Immunoreactivities of steroidogenic enzymes (P450c17 and P450arom) were detected in the ovaries during the breeding and nonbreeding seasons (Figure 2). The positive staining of P450c17 was localized in the theca cells in the ovaries of the breeding and nonbreeding seasons (Figure 2a, b, c). Meanwhile, immunostaining of P450arom was detected in granulosa cells only in the ovaries of the breeding season (Figure 2)d and e. No immunostaining was detected in control sections when normal rabbit serum was substituted for the primary antibody (Figure 2g). The immunolocalization of AR was observed in theca cells, granulosa cells and interstitial cells in the ovaries of both the breeding and nonbreeding seasons (Figure 3a, b and c). However, seasonal variance in the immunolocalization of ERa and ERb was very similar: they were both present in granulosa cells, theca cells and interstitial cells in the ovaries of the breeding (Figure 3d, e, g and h) and nonbreeding seasons (Figure 3f and i). Negative controls did not exhibit any staining (Figure 3j). The positive stains of FSHR and LHR were found in the granulosa cells and theca cells in the ovaries of both the breeding and nonbreeding seasons (Figure 4a, b, c and d). No immunostaining was detected in negative control sections when normal rabbit serum was used instead of the primary antibody (Figure 4e).


Immunoreactivities of androgen receptor, estrogen receptors, p450arom, p450c17 proteins in wild ground squirrels ovaries during the nonbreeding and breeding seasons.

Li X, Zhang H, Sheng X, Li B, Zhou J, Xu M, Weng Q, Watanabe G, Taya K - J Ovarian Res (2012)

The immunolocalization of P450c17 and P450arom in the ovary of wild female ground squirrels. The immunolocalization of P450c17 (a, b, c) and P450arom (d, e, f) in the ovary of wild female ground squirrels during the breeding and nonbreediang seasons. The breeding season divided into two stages, the large follicle stage (a, d) and the small follicle (b, e). g, negative control. IC, interstitial cell; TC, theca cell; GC, granulosa cell. Bars =50μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3473255&req=5

Figure 2: The immunolocalization of P450c17 and P450arom in the ovary of wild female ground squirrels. The immunolocalization of P450c17 (a, b, c) and P450arom (d, e, f) in the ovary of wild female ground squirrels during the breeding and nonbreediang seasons. The breeding season divided into two stages, the large follicle stage (a, d) and the small follicle (b, e). g, negative control. IC, interstitial cell; TC, theca cell; GC, granulosa cell. Bars =50μm.
Mentions: Immunoreactivities of steroidogenic enzymes (P450c17 and P450arom) were detected in the ovaries during the breeding and nonbreeding seasons (Figure 2). The positive staining of P450c17 was localized in the theca cells in the ovaries of the breeding and nonbreeding seasons (Figure 2a, b, c). Meanwhile, immunostaining of P450arom was detected in granulosa cells only in the ovaries of the breeding season (Figure 2)d and e. No immunostaining was detected in control sections when normal rabbit serum was substituted for the primary antibody (Figure 2g). The immunolocalization of AR was observed in theca cells, granulosa cells and interstitial cells in the ovaries of both the breeding and nonbreeding seasons (Figure 3a, b and c). However, seasonal variance in the immunolocalization of ERa and ERb was very similar: they were both present in granulosa cells, theca cells and interstitial cells in the ovaries of the breeding (Figure 3d, e, g and h) and nonbreeding seasons (Figure 3f and i). Negative controls did not exhibit any staining (Figure 3j). The positive stains of FSHR and LHR were found in the granulosa cells and theca cells in the ovaries of both the breeding and nonbreeding seasons (Figure 4a, b, c and d). No immunostaining was detected in negative control sections when normal rabbit serum was used instead of the primary antibody (Figure 4e).

Bottom Line: However, the immunoreactivities of ERa and ERb were both significantly reduced in the nonbreeding season compared to the breeding season.The positive stains of FSHR and LHR were found in the granulosa cells and theca cells of the ovaries of the breeding and nonbreeding seasons.In addition, the Western blotting results of FSHR and LHR showed a significant reduction in the nonbreeding season compared with the breeding season.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Biological Science and Technology, Beijing Forestry University, Beijing, 100083, China. qiangweng@bjfu.edu.cn.

ABSTRACT
The aim of this study was to elucidate the regulatory role of androgen in the follicular development of wild female ground squirrels. Immunohistochemical staining of FSHR, LHR, P450c17, P450arom, androgen receptor (AR), estrogen receptors (ERa and ERb) were executed in ovaries of female ground squirrels from both breeding and nonbreeding seasons. In addition, total ovarian proteins were extracted from the ovaries of squirrels from breeding and nonbreeding seasons, and Western blot analysis were performed in order to probe for FSHR, LHR, P450c17, P450arom, AR, ERa and ERb. The results of immunohistochemical staining and Western blotting of P450c17 showed that there was no significant difference between the breeding and nonbreeding seasons. It was found that granulosa cells expressed P450arom during the breeding season. In contrast, there was no positive staining of P450arom in the nonbreeding season. There was no significant difference in immunoreactivity of AR between the breeding and nonbreeding seasons. However, the immunoreactivities of ERa and ERb were both significantly reduced in the nonbreeding season compared to the breeding season. The positive stains of FSHR and LHR were found in the granulosa cells and theca cells of the ovaries of the breeding and nonbreeding seasons. In addition, the Western blotting results of FSHR and LHR showed a significant reduction in the nonbreeding season compared with the breeding season. These findings suggested that androgen might be predominantly converted into estrogen in order to regulate the follicular development via binding of estrogen receptors during the breeding season, whereas androgen might predominantly directly bind androgen receptor to regulate the follicular development during the nonbreeding season in the ovaries of wild female ground squirrels.

No MeSH data available.