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Isolation, Fixation, and Immunofluorescence Imaging of Mouse Adrenal Glands
Published on: October 2, 2018
Elevated luteinizing hormone receptor signaling or selenium treatment leads to comparable changes in adrenal
Jaroslaw Wieczorek1, Piotr Pawlicki2, Marta Zarzycka2
1University Centre of Veterinary Medicine JU-UA, University of Agriculture in Krakow, Mickiewicza 24/28, 30-059, Krakow, Poland.
Abstract:
The importance and regulation of adrenal androgen production and signaling are not completely understood and are scarcely studied. In addition, there is still a search for appropriate animal models and experimental systems for the investigation of adrenal physiology and disease. Therefore, the main objective of the study was to evaluate the effect of luteinizing hormone (LH) signaling and selenium (Se2+) exposure on androgen adrenal signaling via canonical androgen receptor (AR), and membrane androgen receptor acting as zinc transporter (zinc- and iron-like protein 9; ZIP9). For herein evaluations, adrenals isolated from transgenic mice with elevated LH receptor signaling (KiLHRD582G) and adrenals obtained from rabbits used for ex vivo adenal cortex culture and exposure to Se2+ were utilized. Tissues were assessed for morphological, morphometric, and Western blot analyses and testosterone and zinc level measurements.Comparison of adrenal cortex histology and morphometric analysis in KiLHRD582G mice and Se2+-treated rabbits revealed cell hypertrophy. No changes in the expression of proliferating cell nuclear antigen (PCNA) were found. In addition, AR expression was decreased (p < 0.001) in both KiLHRD582G mouse and Se2+-treated rabbit adrenal cortex while expression of ZIP9 showed diverse changes. Its expression was increased (P < 0.001) in KiLHRD582G mice and decreased (P < 0.001) in Se2+-treated rabbits but only at the dose 10 ug/100 mg/ tissue. Moreover, increased testosterone levels (P < 0.05) and zinc levels were detected in the adrenal cortex of KiLHRD582G mice whereas in rabbit adrenal cortex treated with Se2+, the effect was the opposite (P < 0.001).
Insights
Luteinizing hormone (LH) signaling and selenium (Se2+) impact adrenal androgen signaling differently. LH increased testosterone and zinc, while Se2+ decreased them, affecting androgen receptor (AR) and ZIP9 expression.
Area of Science:
- Endocrinology
- Adrenal Physiology
- Molecular Biology
Background:
- Adrenal androgen production and signaling are not fully understood.
- Appropriate animal models for adrenal research are needed.
- Investigating novel signaling pathways in adrenal cortex is crucial.
Purpose of the Study:
- To evaluate the effect of luteinizing hormone (LH) signaling on adrenal androgen signaling.
- To assess the impact of selenium (Se2+) exposure on adrenal androgen signaling.
- To explore the roles of the canonical androgen receptor (AR) and membrane androgen receptor ZIP9.
Main Methods:
- Utilized transgenic mice (KiLHRD582G) with elevated LH receptor signaling.
- Employed ex vivo rabbit adrenal cortex cultures exposed to Se2+.
- Performed histological, morphometric, Western blot, testosterone, and zinc level analyses.
Main Results:
- Cell hypertrophy observed in both KiLHRD582G mice and Se2+-treated rabbits.
- AR expression decreased in both models; ZIP9 expression varied.
- LH signaling increased adrenal testosterone and zinc; Se2+ decreased them.
Conclusions:
- LH and Se2+ differentially regulate adrenal androgen signaling pathways.
- AR and ZIP9 are key players in adrenal response to LH and Se2+.
- These findings contribute to understanding adrenal physiology and disease models.
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