Abstract
Several lines of controversial evidence concerning estrogen receptor β (ERβ) remain to be solved because of the unavailability of specific antibodies against ERβ. The recent validation analysis identified a monoclonal antibody (PPZ0506) with sufficient specificity against human ERβ. However, the specificity and cross-reactivity of PPZ0506 antibody against ERβ proteins from laboratory animals have not been confirmed. In the present study, we aimed to validate the applicability of PPZ0506 to rodent studies. The antibody exhibited specific cross-reactivity against mouse and rat ERβ proteins in immunoblot and immunocytochemical experiments using transfected cells. In immunohistochemistry for rat tissue sections, PPZ0506 showed immunoreactive signals in the ovary, prostate, and brain. These immunohistochemical profiles of rat ERβ proteins in rat tissues accord well with its mRNA expression patterns. Although the antibody was reported to show the moderate signals in human testis, no immunoreactive signals were observed in rat testis. Subsequent RT-PCR analysis revealed that this species difference in ERβ expression resulted from different expression profiles related to the alternative promoter usage between humans and rats. In conclusion, we confirmed applicability of PPZ0506 for rodent ERβ studies, and our results provide a fundamental basis for further examination of ERβ functions.
Highlights
Ovarian steroid hormones, estrogens, play an important role in signal transduction pathways across various organs in females and males [1]
We demonstrated that the anti-human estrogen receptor β (ERβ) monoclonal antibody, PPZ0506, reacted with human, mouse, and rat ERβ proteins in immunoblot and immunocytochemical experiments using transfected cells
The recent discovery of a well validated monoclonal antibody (PPZ0506) against human ERβ proteins has prompted attempts to solve the conflicting problems in ERβ research
Summary
Estrogens, play an important role in signal transduction pathways across various organs in females and males [1]. Two subtypes of ERs, ERα and ERβ ( termed NR3A1 and NR3A2 [2], and with the symbols ESR1 and ESR2, respectively), belong to the nuclear steroid receptor superfamily and function as ligand-inducible transcription regulators. They are widely expressed in a wide variety of organs and exhibit distinct expression patterns. Alternative promoter usage of ERα genes has been analyzed [4], and our comparative studies of these systems reported conserved and species-specific genomic structures and expression profiles among humans, mice, and rats [5,6,7,8,9]. Few studies have reported on species differences in patterns of alternative ERβ gene promoter usage
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