https://doi.org/10.5061/dryad.98sf7m0q8

Anterior pituitary cells are highly active with regards to protein synthesis and secretion, processes which depend heavily on mitochondrial ATP production and functional endoplasmic reticula. It is well known that obesity adds an allostatic overload to tissues, requiring them to adapt to inflammation and oxidative stress. Therefore, we hypothesized that the pituitary is highly vulnerable to the stress of high fat diet-induced weight gain. In this study, we utilized a 10-15 week high fat diet (HFD, 60%) plus a thermoneutral housing paradigm, testing both male and female FVB.129P mice. We quantified serum hormones and cytokines, characterized the metabolic phenotype, and defined changes in the pituitary transcriptome using single-cell RNA-seq. Weight gain was significant by 3 weeks in HFD mice, and by 10 weeks all HFD groups had gained 20 g.  HFD females (15 weeks) had increased energy expenditure and decreased activity. All HFD groups showed increases in serum leptin, Il-6, resistin, MCP-1, and TNFα. HFD males had increased insulin; both HFD males and females had increased TSH, and HFD females had decreased serum prolactin and growth hormone (GH) pulse amplitude.  Pituitary scRNA-seq revealed modest or no changes in pituitary cell gene expression in the different cell types from HFD males after 10 or 15 weeks or HFD females after 10 weeks. However, females exposed to a HFD for 15 weeks showed significant numbers of differentially expressed genes in lactotropes and stem cells. Pathway analyses identified a reduction in pathways that supported protein translation, ribosome biogenesis, and oxidative phosphorylation, indicating mitochondrial dysfunction. Collectively, these studies reveal that pituitary cells from males are more resilient to the oxidative stress of obesity than females and identify the most vulnerable pituitary cell populations in females.

Description of the data and file structure

This supplemental data set includes supporting figures and Tables as cited in the primary manuscript. This includes Author generated pituitary single cell RNA sequencing data comparing control and high fat diet fed mice at 10 and 15 weeks of treatment (see Gene Expression Omnibus (GEO) under the accession number cited in the primary manuscript).

Supplementary Figure 1. ANCOVA analysis of female and male energy expenditure

Supplementary Figure 2. Comprehensive Lab Animal Monitoring (CLAMS): 10-week HFD

Supplementary Figure 3. Glucose Tolerance Test in males after 15 weeks on normal or high fat diet

Supplemental Table 1. Single cell RNA sequencing of pituitaries from control and high fat diet treated mice - top ten genes in each identified cluster. 

Supplemental Table 2. Differentially expressed genes in each pituitary cell cluster from males exposed to HFD for 10 weeks

Supplemental Table 3. Differentially expressed genes in clusters from male pituitaries following a HFD for 15 weeks

Supplemental Table 4. Differentially expressed genes in each pituitary cell cluster from females exposed to HFD for 10 weeks

Supplemental Table 5. Lactotrope cluster from female pituitaries exposed to HFD for 15 weeks

Supplemental Table 6. Differentially expressed genes in female pituitary stem cell clusters exposed to HFD for 15 weeks

Supplemental Table 7. Differentially expressed genes in the lactotrope cell cluster from males exposed to HFD for 15 weeks vs age matched controls

Supplemental Table 8. GO analysis of biological processes impacted by all down-regulated DEGs in the female lactotrope cluster that are reduced at adjusted p<0.055 following a HFD for 15 weeks

Supplemental Table 9. GO analysis of biological processes impacted by all up-regulated DEGs in the female lactotrope cluster that are increased at adjusted p<0.055 following a HFD for 15 weeks

Supplemental Table 10. Differentially expressed genes in the pituitary stem cell cluster from males exposed to HFD for 15 weeks vs age matched controls

Supplemental Table 11. GO analysis of biological processes impacted by all down-regulated DEGs in the female stem cell cluster that are reduced at adjusted p<0.055 following a HFD for 15 weeks

Supplemental Table 12. GO analysis of biological processes impacted by all up-regulated DEGs in the female stem cell cluster that are reduced at adjusted p<0.055 following a HFD for 15 weeks