Long-term effects of gestational diabetes mellitus on the pancreas of female mouse offspring

Figure 1 Experimental design, body weight, and blood glucose levels of female offspring born from control dams or dams with gestational diabetes mellitus. A: Diagram showing the experimental design and timeline; B and C: Female offspring born from dams treated with streptozotocin during pregnancy did not show significant changes in body weight or blood glucose compared to offspring born from dams treated with vehicle as controls; D: Scheme showing the region selected for quantification (splenic pancreas; square) of the neuronal, vascular, and inflammatory changes using immunohistochemistry and confocal microscopy. Data are shown as mean ± SEM, n = 5 per group. STZ: Streptozotocin; VEH: Vehicle.

Figure 2 Adult female offspring born from dams with streptozotocin-induced gestational diabetes mellitus have increased density of sensory and sympathetic nerve fibers innervating the pancreatic acinar cells. A and B: Representative confocal images of calcitonin gene-related peptide (CGRP) expressing small diameter peptidergic sensory nerve fibers; C and F: Significantly greater densities of CGRP⁺ and tyrosine hydroxylase⁺ (TH⁺) nerve fibers were found in offspring born from dams with GMD as compared to control offspring; D and E: Enzyme TH expressing postganglionic sympathetic nerve fibers in acinar pancreatic tissue (40 μm-thick) from offspring born of vehicle or streptozotocin injected dams; ^aP < 0.05 Student t test, n = 5 per group. Data are shown as mean ± SEM. CGRP: Calcitonin gene-related peptide; TH: Tyrosine hydroxylase; VEH: Vehicle; STZ: Streptozotocin.

Figure 3 Adult female offspring born from dams with streptozotocin-induced gestational diabetes mellitus have a greater density of blood vessels vascularizing the acinar cells and the pancreatic islets. A, B, D and E: Representative confocal images of vehicle showing the endocrine and exocrine pancreatic tissue immunostained for endomucin blood vessels from adult progeny born from dams treated with vehicle or streptozotocin (STZ); C and F: There was a significantly greater density of blood vessels positive to endomucin in the acinar cells and the islets of Langerhans of offspring mice born from mice treated with STZ as compared to control mice; G and H: 3D visualization of the islet of Langerhans from whole pancreatic tissue processed with a solvent-based clearing method. Three-dimensional images of a cleared pancreas show an increased intra-islet vasculature. ^aP < 0.05 Student t test, n = 4-5 per group. Data are shown as mean ± SEM. VEH: Vehicle; STZ: Streptozotocin.

Figure 4 Adult female offspring born from dams with streptozotocin-induced gestational diabetes mellitus have an increased number of macrophages in the pancreatic acinar cells, but not in the pancreatic islets. A, B, D, and E: Representative confocal images of pancreatic cells expressing CD68 (cluster of differentiation 68) for macrophage staining in acinar pancreatic tissue or pancreatic islets from offspring born of vehicle or streptozotocin (STZ) injected dams (40 μm-thick); C and F: Significantly greater number of macrophages were found in the acinar pancreatic tissue, but not in the pancreatic islets of offspring born from dams treated with STZ compared to control mice. ^aP < 0.05 Student t test, n = 4-5 per group. Data are shown as mean ± SEM. CD68: Cluster of differentiation 68; VEH: Vehicle; STZ: Streptozotocin.