Dexamethasone and sex regulate placental glucocorticoid receptor isoforms in mice
- 1School of Medical Science and Menzies Health Institute Queensland, Griffith University, Southport, Queensland, Australia
- 2School of Biomedical Sciences, and Child Health Research Centre, The University of Queensland, St Lucia, Queensland, Australia
- 3Mater Research Institute, Translational Research Institute, University of Queensland, Woolloongabba, Queensland, Australia
- Correspondence should be addressed to V L Clifton; Email: Vicki.clifton{at}mater.uq.edu.au
-
Figure 1
Representative Western blots of cytoplasmic (Cyto) and nuclear (Nuc) GR isoforms in placentae from saline-exposed male (open bars) and female (closed bars) mouse fetuses at embryonic day 14.5 (A). Densitometry was performed to measure the expression of each isoform for both the cytoplasmic (B) and nuclear (C) protein fraction. As protein levels of males and females were performed on different blots, data is presented as a ratio of each isoform to the total GR for each sample. Data are presented as mean ± s.e.m., n = 8–12 placentas from 10 litters per group,*P < 0.05 represents sex difference in GR isoform, #P < 0.05 indicates that the GR isoform in question is different to GRα-A.
-
Figure 2
Representative Western blots of cytoplasmic (Cyto) and nuclear (Nuc) GR isoforms in saline (open bars) and dexamethasone (closed bars) exposed placentae from male (A) and female (B) mouse fetuses at embryonic day 14.5. Densitometry was performed to measure the expression of each isoform for both the cytoplasmic (C and D) and nuclear (E and F) protein fractions. All samples from each sex were measured on three blots with a common sample on each membrane. As such total isoform expression was normalized to the average expression of the saline group for each isoform. Data are presented as mean ± s.e.m., n = 8–12 placentas from 10 litters per group, *P < 0.05 represents significant effect of dexamethasone exposure.
-
Figure 3
The effects of maternal dexamethasone (closed bars) on placental mRNA expression of target genes known to be regulated by GR isoforms in males (A) and females (B) compared to saline-exposed placentae (open bars). Data are presented as mean ± s.e.m., n = 7–11 placentas from 10 litters per group, *P < 0.05 represents significant effect of dexamethasone exposure. Ier3, immediate early response 3; Nfatc1, nuclear factor of activated T-cells 1; Sgk1, serum glucocorticoid kinase 1; Bcl2l11, Bcl-2-like protein 11; Cpeb1, cytoplasmic polyadenylation element-binding protein 1. Ier3 is a marker of regulation by any GR isoform, Nfatc1 is regulated specifically by GRα-A, Sgk1 and Bcl2l11 are regulated specifically by GRα-C and Cpeb1 is regulated by GRα-D isoforms.
-
Figure 4
Immunohistochemical staining of cleaved caspase three (brown DAB pigment) in placentae of male (A and C) and female fetuses (B and D) exposed to saline (A and B) or dexamethasone (C and D) during pregnancy (n = 3 per group). Isotype control sections are presented as inset images. All slides are counterstained with hematoxylin. The effects of maternal dexamethasone (closed bars) on placental cytoplasmic protein expression of cleaved caspase three in males (E) and females (F), BAX in males (G) and females (H) and BCL2 in males (I) and females (J) compared to saline-exposed placentae (open bars). Data is presented as mean ± s.e.m., *P < 0.05 represents significant effect of dexamethasone exposure. Representative blots contain n = 6 out of the n = 8–12 placentas from 10 litters per group analyzed.
-
Figure 5
The effects of maternal dexamethasone (closed bars) on placental protein carbonyl levels in males (A) and females (B), placental GPX protein concentrations in males (C) and females (D) and TXNRD1 protein concentrations in males (E) and females (F) compared to saline-exposed placentae (open bars). Data is presented as mean ± s.e.m., *P < 0.05 represents significant effect of dexamethasone exposure. Representative blots contain n = 6. n = 8–12 placentas from 10 Litters per group were analyzed for Western blotting and protein carbonyl ELISAs.
- © 2017 Society for Endocrinology
