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Background: Estrogens play a key role in the human male reproductive system. Estrogen receptors (ERs: ERα and ERβ) was detected in germ cells from spermatogonia to spermatozoa.

Objective: Tto evaluate the different estrogen receptors expressions on the sperm in infertile males using a new technique and apply that on infertile patient with varicocele.

Materials and Methods: This study included 200 males, divided into two equal groups. Group1  included  fertile males, and group II included infertile males tested according to presence of varicocele with different grades which were classified according to Dubin-Amelar clinical classification of varicocele (1970), 1 m pellet was examined for the expression of the ERα and ERβ using immunocytochemistry stain. The stained sperm was processed in Photoshop program. The estrogen receptors were visualized after magnification and increased resolution in addition using a yellow filter. Percentage of stained sperm with different location in relation to the unstained sperm was counted.

Results: There was a strong expression of ER β on the sperm in fertile males while ERα expression very reduced in fertile males and almost absent in all infertile males. Also, there was a statistically significant decrease in both ERα and β expression in infertile males than fertile males and in varicocele.

Conclusion: ER technical estimation using immunoassay was better than any other techniques assay because ER could be present, but not function due to dislocation and deposition into wrong position.

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