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. 2014 Jun;224(6):732-6.
doi: 10.1111/joa.12183. Epub 2014 Apr 2.

Identification of G protein-coupled estrogen receptor in human and pig spermatozoa

V Rago  1 , F Giordano, E Brunelli, D Zito, S Aquila, A Carpino
Affiliations

Identification of G protein-coupled estrogen receptor in human and pig spermatozoa

V Rago et al. J Anat. 2014 Jun.

Abstract

Estrogens are known to influence functional properties of mammalian spermatozoa inducing rapid responses through the classical estrogen receptors (ERα and ERβ). Recently, the G protein-coupled estrogen receptor (GPER) has been identified as mediator of fast non-genomic estrogen effects in different cells. This work investigated the expression of GPER in human and pig spermatozoa using immunofluorescence, Western blot analysis and RT-PCR. GPER was found to be confined to the mid-piece of human sperm cells, whereas it was detected in the acrosomal region, the equatorial segment and the mid-piece of pig spermatozoa. Furthermore, in the male gametes of both species, the immunoblots of sperm extracts revealed a band at ~42 kDa, consistent with the GPER molecular weight, and RT-PCR detected the GPER transcripts. This is the first report demonstrating the expression of GPER in human and pig mature sperm cells and evidencing its species-specific cellular localization.

Keywords: G protein-coupled estrogen receptor; estrogens; human spermatozoa; pig spermatozoa.

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Figures

Figure 1
Figure 1
Expression of GPER in human spermatozoa (A) Green immunofluorescence labeling in the sperm mid piece (arrow heads). Sperm heads show the iodure propidium red stain. The insert show the control spermatozoa. (B) Iimmunoblots of protein extracts from the six human sperm samples (lane 1–6), positive control (lane C). β actin as loading control. (C) RT-PCR results in two representative human sperm samples (lane 1–2), negative control (lane-), and markers (lane M). GAPDH (450 bp) is the internal control.
Figure 2
Figure 2
Expression of GPER in pig spermatozoa (A) Green immunofluorescence in the mid-piece (arrowhead), in the equatorial segment (short arrow) and in the acrosome (long arrow) of sperm cells. Sperm heads show the propidium iodide red stain. The insert show the control spermatozoa. (B) Immunoblots of protein extracts from six pig sperm samples (lane 1–6), positive control (lane C), β-actin as loading control. (C) RT-PCR results in two representative pig sperm samples (lane 1–2), negative control (lane-), and markers (lane M). GAPDH (450 bp) is the internal control.
See this image and copyright information in PMC

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