48-hour access to this article
USD $30.00
USD $30.00
Effect of estradiol and IGF1 on glycogen synthesis in bovine uterine epithelial cells
in ReproductionIn brief
Glucose is an important nutrient for the endometrium and embryo during pregnancy. This study shows that estradiol (E2)/IGF1 signaling stimulates glycogen synthesis in the uterine epithelium of cows, which could provide glucose when needed.
Abstract
Glycogen storage in the uterine epithelium peaks near estrus and is a potential source of glucose for the endometrium and embryos. However, the hormonal regulation of glycogen synthesis in the uterine epithelium is poorly understood. Our objective was to evaluate the effect of E2 and insulin-like growth factor 1 (IGF1) on glycogenesis in immortalized bovine uterine epithelial (BUTE) cells. Treatment of BUTE cells with E2 (0.1–10 nM) did not increase glycogen levels. However, treatment of BUTE cells with IGF1 (50 or 100 ng/mL) resulted in a >2-fold increase in glycogen. To determine if the uterine stroma produced IGF1 in response to E2, bovine uterine fibroblasts were treated with E2, which increased IGF1 levels. Immunohistochemistry showed higher levels of IGF1 in the stroma on day 1 than on day 11, which coincides with higher glycogen levels in the uterine epithelium. Western blots revealed that IGF1 treatment increased the levels of phospho-AKT, phospho-GSKβ, hexokinase 1, and glycogen synthase in BUTE cells. Metabolomic (GC-MS) analysis showed that IGF1 increased 3-phosphoglycerate and lactate, potentially indicative of increased flux through glycolysis. We also found higher levels of N-acetyl-glucosamine and protein glycosylation after IGF1 treatment, indicating increased hexosamine biosynthetic pathway activity. In conclusion, IGF1 is produced by uterine fibroblasts due to E2, and IGF1 increases glucose metabolism and glycogenesis in uterine epithelial cells. Glycogen stored in the uterine epithelium due to E2/IGF1 signaling at estrus could provide glucose to the endometrium or be secreted into the uterine lumen as a component of histotroph.
Supplementary Materials
- Supplementary Figure 1 (A) Glycogen (50 μg/ml) or glycogen + glucose (both at 50 μg/ml) was dissolved in 30% KOH and measured via our glycogen assay. There was no difference in the concentration of glycogen detected. (B) Immunohistochemistry for IGF1 and IGF1R using antibodies or antibodies preincubated with the recombinant target protein. Scale bar = 100 μm.
- Supplementary Figure 2 (A) Images of bands produced by PCR for primers specific for WNT7A or FOXA2 in BUTE cells or samples of bovine endometrium (+Cont). WNT7A or FOXA2 are markers for the luminal or glandular epithelium, respectively. (B) Monolayers of BUTE cells after being treated with 0-10 μM picropodophyllotoxin (PPPT) or NVP-ADW742 (NVP) for 48 hours. Scale bar = 500 μM. (C) Western blot for phospho-AKT, AKT, and β actin in BUTE cells treated with 50 ng/ml IGF1, 0.1 μM picropodophyllotoxin, and 1 μM NVP-ADW742 as indicated for 30 minutes.
- Supplemental Table 1. Composition of αMEM media.
- Supplemental Table 2
Reproduction is committed to supporting researchers in demonstrating the impact of their articles published in the journal.
The two types of article metrics we measure are (i) more traditional full-text views and pdf downloads, and (ii) Altmetric data, which shows the wider impact of articles in a range of non-traditional sources, such as social media.
More information is on the Reasons to publish page.
| Sept 2018 onwards | Past Year | Past 30 Days | |
|---|---|---|---|
| Full Text Views | 176 | 176 | 9 |
| PDF Downloads | 187 | 187 | 10 |
Online ISSN: 1741-7899
Print ISSN: 1470-1626
Strengthening biomedical communities
to advance science and health
CONTACT US
Bioscientifica Ltd | Starling House | 1600 Bristol Parkway North | Bristol BS34 8YU | UK
Bioscientifica Ltd | Registered in England no 3190519
Powered by PubFactory