Transcriptome landmarks of the functional maturity of rat beta-cells, from lactation to adulthood

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   Figure 1

   FACS isolation strategy and quantification of enriched beta-cell populations obtained from rats at different postnatal ages. (A) Gating strategy followed to get enriched beta-cell populations by fluorescence-activated cell sorting after primary insular cell culture from neonatal, suckling, weaning, and adult Wistar rats. (B) Insulin (green) immunostaining and nuclear counterstaining (blue) of sorted-cell populations obtained from rats of different ages. (C) Quantification of the beta-cell fraction contained in sorted populations. (D) Heat map and dendrogram that represent the global expression-level correlation among microarrays and the similarity among samples, respectively. Scale bar=40 μm.
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   Figure 2

   Basal and glucose-induced insulin secretion-related expression and physiologic changes observed in beta-cells and involved in their maturation process. (A) Enrichment plots of the Reactome regulation of insulin secretion and the Reactome integration of energy metabolism gene sets that show the running sum for each set within the beta-cell dataset (green line), and the leading-edge subset of enriched genes (vertical black lines). Heatmap of the leading-edge core genes based on their expression levels, and dendrogram depicting similarities among samples. (B) Representative images of the hemolytic immunoplaques produced by insulin secretion of neonate, suckling, weaning, and adult sorted beta-cells, incubated at basal (5.6 mM glucose; top panel) and stimulating (15.6 mM glucose; bottom panel) conditions. (C) Quantification of immunoplaque areas observed in basal (left graph) and stimulating (right graph) conditions. Red bars represent the mean+s.e.m. Scale bar=50μm. ⁺P<0.01 vs the previous age, *P<0.01 vs the same age at 5.6 mM glucose.
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   Figure 3

   Calcium channels expression and functional changes that contribute, in part, to the maturation of basal and glucose-induced insulin secretion observed in beta-cells. (A) Enrichment plot of the GO calcium channel activity gene set that show the running sum for the set within the beta-cell dataset (green line), and the leading-edge subset of enriched genes (vertical black lines). Heat map of the leading-edge core genes based on their expression levels and a dendrogram is depicting similarities among samples. (B) Representative recordings of global calcium channel currents observed in 20d and adult beta-cells. Arrows represent T-type calcium channel current. (C) Quantification of the beta-cell percentage with and without T-type calcium channel current (gray: percentage of cells without T-type calcium current; black: percentage of cells with T-type calcium current). A full colour version of this figure is available at http://dx.doi.org/10.1530/JME-16-0052.
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   Figure 4

   Cell-cycle-related expression and functional changes observed in beta-cells and involved in the transition from a proliferating phenotype to a quiescent one. (A) Enrichment plots of the Reactome cell cycle gene set that show the running sum for the set within the beta-cell dataset (green line), and the leading-edge subset of enriched genes (vertical black lines). Heatmap of the leading-edge core genes based on their expression levels and a dendrogram is depicting similarities among samples. (B) The gating strategy followed to assess beta-cell cycle after primary insular cell culture from Wistar rats at different ages. (C) Representative histograms of DNA content from beta-cells of suckling and adult rats. (Insets) Histogram zoom to show the S and G2/M phases. (D) Quantification of the percentage of beta-cells from different ages at each cell-cycle phase. ⁺P<0.05 vs the same cell cycle phase at 20d. A full colour version of this figure is available at http://dx.doi.org/10.1530/JME-16-0052.

• © 2016 Society for Endocrinology