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. 2020 Sep;47(3):180-185.
doi: 10.5653/cerm.2020.03573. Epub 2020 Sep 1.

Localization (and profiles) of tyrosinephosphorylated proteins in female reproductive organs of adult rats

Sudtida Bunsueb  1 , Nareelak Tangsrisakda  1   2 , Alexander T H Wu  3   4 , Sitthichai Iamsaard  1   2
Affiliations

Localization (and profiles) of tyrosinephosphorylated proteins in female reproductive organs of adult rats

Sudtida Bunsueb et al. Clin Exp Reprod Med. 2020 Sep.

Abstract

Objective: Tyrosine phosphorylation is an essential process in many biological systems, including the male reproductive system. The presence of tyrosine-phosphorylated (TyrPho) proteins has been well documented in male reproductive organs, but research in fertile females is still limited.

Methods: The ovary, oviduct, and uterus of adult female Sprague-Dawley rats in the estrus phase were used to localize TyrPho proteins using an immunohistochemical technique. These proteins were separated and their expression patterns were examined by sodium dodecyl sulfatepolyacrylamide gel electrophoresis and Western blot analysis, respectively.

Results: TyrPho proteins were localized in the cytoplasm of the oocyte except the antral fluid; in the granulosa cells, theca cells, and stromal cells of the ovary; at the apical surface of oviductal epithelial cells; and in the basal epithelium and submucosa of the uterine wall. Moreover, we found that 72-, 43-, and 28-kDa TyrPho proteins were localized in the ovary, while 170-, 55-, and 43-kDa proteins were localized in the oviduct. In the uterus, we detected four major bands, corresponding to 61-, 55-, 54-, and 43-kDa TyrPho proteins.

Conclusion: Given that these TyrPho proteins were found in major reproductive organs in the estrus phase, these proteins may play important roles in female fertility.

Keywords: Ovary; Oviduct, Phosphorylation; Rats; Uterus.

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Conflict of interest statement

Conflict of interest

No potential conflict of interest relevant to this article was reported.

Figures

Figure 1.
Figure 1.
Immunohistochemical micrographs stained with 3,3’-diaminobenzidine (DAB) demonstrating the localization of anti-phosphotyrosine antibody in the female reproductive organs (the mature follicles of the ovary, oviduct, and uterus; A-C). The negative control (D-F) was not treated with primary antibody. CyO, cytoplasm of the oocyte; GC, granulosa cell; ThC, theca cell; Ost, ovarian stroma cell; Epi, epithelial cell; Sub, submucosa.
Figure 2.
Figure 2.
Protein profiles (100 mg/lane) of the ovary (OVR; A), oviduct (OVD; B), and uterus (UTR; C). Proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with Coomassie blue staining and Western blotting with anti-phosphotyrosine antibody. EGF, epidermal growth factor (used as a positive control for tyrosine phosphorylation); BSA, bovine serum albumin (used as a negative control).
Figure 3.
Figure 3.
Summary of the localization of tyrosine-phosphorylated (TyrPho) proteins in the ovary, especially in the mature follicle (72-, 43-, and 28-kDa proteins), oviduct (170-, 55-, and 43-kDa proteins), and uterus (61-, 55-, 54-, and 43-kDa proteins), of adult female rats in the estrus phase.
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