Related Experiment Videos
Identification of androgen receptor protein and 5alpha-reductase mRNA in human ocular tissues
E M Rocha1, L A Wickham, L A da Silveira
1Schepens Eye Research Institute and Department of Ophthalmology, Harvard Medical School, Boston, MA 02114, USA.
Background/Aims:
Androgens have been reported to influence the structural organisation, functional activity, and/or pathological features of many ocular tissues. In addition, these hormones have been proposed as a topical therapy for such conditions as dry eye syndromes, corneal wound healing, and high intraocular pressure. To advance our understanding of androgen action in the eye, the purpose of the present study was twofold: firstly, to determine whether tissues of the anterior and posterior segments contain androgen receptor protein, which might make them susceptible to hormone effects following topical application; and, secondly, to examine whether these tissues contain the mRNA for types 1 and/or 2 5alpha-reductase, an enzyme that converts testosterone to the very potent metabolite, dihydrotestosterone.
Methods:
Human ocular tissues and cells were obtained and processed for histochemical and molecular biological procedures. Androgen receptor protein was identified by utilising specific immunoperoxidase techniques. The analysis of type 1 and type 2 5alpha-reductase mRNAs was performed by the use of RT-PCR, agarose gel electrophoresis, and DNA sequence analysis. All immunohistochemical evaluations and PCR amplifications included positive and negative controls.
Results:
These findings show that androgen receptor protein exists in the human lacrimal gland, meibomian gland, cornea, bulbar and forniceal conjunctivae, lens epithelial cells, and retinal pigment epithelial cells. In addition, our results demonstrate that the mRNAs for types 1 and 2 5alpha-reductase occur in the human lacrimal gland, meibomian gland, bulbar conjunctiva, cornea, and RPE cells.
Conclusion:
These combined results indicate that multiple ocular tissues may be target sites for androgen action.
Insights
Androgen receptor protein and key enzymes for its activation are present in multiple human ocular tissues. This suggests that various parts of the eye may respond to androgen hormones.
Area of Science:
- Ophthalmology
- Endocrinology
- Molecular Biology
Background:
- Androgens impact ocular tissue structure, function, and pathology.
- Androgens are explored for treating dry eye, corneal healing, and glaucoma.
- Understanding androgen action in the eye is crucial for therapeutic development.
Purpose of the Study:
- To detect androgen receptor protein in human ocular tissues.
- To identify the presence of 5alpha-reductase (types 1 and 2) mRNA in ocular tissues.
- To assess potential androgen target sites within the eye.
Main Methods:
- Human ocular tissues and cells were analyzed using histochemistry and molecular biology.
- Androgen receptor protein was identified via immunoperoxidase staining.
- 5alpha-reductase mRNA presence was determined using RT-PCR and DNA sequencing.
Main Results:
- Androgen receptor protein was found in the lacrimal gland, meibomian gland, cornea, conjunctivae, lens epithelium, and retinal pigment epithelium.
- mRNA for 5alpha-reductase types 1 and 2 was detected in the lacrimal gland, meibomian gland, conjunctiva, cornea, and retinal pigment epithelium.
- These findings confirm the expression of androgen-related molecules in diverse ocular tissues.
Conclusions:
- Multiple ocular tissues express androgen receptors and the enzymes necessary for androgen activation.
- These results indicate that various ocular tissues are potential targets for androgen action.
- This supports the investigation of androgens as a therapeutic strategy for ocular conditions.