3. Various Early-Life Stressors, The Same Target: The Developing Beta-Cell Mass
As abundantly illustrated in animal models, many early-life stressors such as maternal hyperglycaemia, undernutrition, overnutrition, hypercholesterolemia, corticosteroid therapy, uteroplacental insufficiency, or hypoxia trigger a beta-cell mass adaptive response in the fetus (Figure 2, Table 1).

3.1. Critical Windows for Adaptive Response to Early-Life Stressors
The development of the endocrine pancreas starts from a pool of common precursor cells that become progressively committed to the endocrine lineage under the control of a hierarchical network of transcription factors. During late fetal and early postnatal life, the beta-cell mass is determined by the recruitment of undifferentiated precursors, as well as the replication and apoptosis rates of the beta cells. Obviously, any disturbance of the environment of the endocrine cells at a specific developmental time-point, as it occurs in a perturbed intrauterine milieu, may modify the balance of controlling factors, thereby contributing to an adaptive beta-cell growth response which is metabolically appropriate on the short term. However, this adaptive response may turn to be detrimental if maintained on the long term, as it may foster beta-cell failure and diabetes later in life. We are largely ignorant of when programming may be initiated during development.
Preimplantatio
An early onset for programming was indicated, as maternal low-protein diet during only the preimplantation period of rat development (0–4 days after mating), before return to control diet for the remainder of the gestation, induced blastocyst abnormalities, and programming of postnatal growth rate and hypertension [102]. More specifically it was shown that preimplantation embryos collected from dams after 0–4 days of maternal low-protein diet displayed significantly reduced cell numbers, within the inner cell mass and trophectoderm lineages, apparently induced by a slower rate of cellular proliferation. The low-protein diet significantly reduced insulin and essential amino acid levels and increased glucose levels within maternal serum by day 4 of development. These data indicate that the mildly hyperglycemic and amino-acid-depleted maternal environment generated by undernutrition may act as an early mechanism of programming and initiate conditions of “metabolic stress,” restricting early embryonic proliferation and the generation of appropriately sized stem-cell lineages. In chemically or genetically obtained rat diabetes models in which maternal serum insulin depletion and hyperglycemia are induced, proliferation of inner cell mass or total cell numbers within blastocysts is inhibited [103, 104]. Therefore, the preimplantation embryo is particularly sensitive to metabolic modifications that may have programming consequences [105, 106], and one possibility is that it is the preimplantation embryo itself that is programmed. 
Postimplantation
Embryo transfer experiments may also help to dissociate the impact of the maternal environment in early (preimplantation) versus late gestation (postimplantation). We recently found that embryos (blastocysts) from a nondiabetic Wistar strain placed into a diabetic GK/Par uterus develop a reduced beta-cell mass which remains low on the long term [42]. Data with rat models of prenatal undernutrition [95] also illustrate that low-energy and low-protein diets that reduce the development of the beta-cell mass in both cases act at different critical time windows. The beta-cell mass is deficient in the low-energy pancreas because this diet reduces neogenesis, probably because of high glucocorticoid levels, rather than by impairing vascularisation and proliferation. Early gestation is thus a very sensitive period in this model. By contrast, pancreatic alterations take place at a later fetal stage in the low-protein model, and the beta-cell mass is deficient in this case because this diet reduces beta-cell vascularisation and proliferation without altering beta-cell differentiation [95]. 
Postnatal versus Prenata
Further support for the crucial impact of prenatal nutritional environment is the recent report that prenatal nutrient restriction in both male and female rats led to an inappropriate postnatal beta-cell mass formation attributed to a decrease in the rate of beta-cell replication and beta-cell neogenesis [93]. In contrast, male and female rats exposed to postnatal nutrient restriction alone (with normal prenatal nutrient exposure) were characterized by decreased pancreatic and body weights, but a weight-adjusted beta-cell mass higher compared to control animals [93]. Another illustration is offered by observations in normal rat pups reared artificially on a high-carbohydrate milk formula [107]: such alteration of nutrition, during the suckling period only, induced persistent adaptation of energy metabolism in adulthood (obesity, glucose intolerance, and impaired insulin secretion).

3.2. Molecular Mechanisms Mediating the Perinatal Beta-Cell Adaptive Response to Early-Life Stressors
Molecular mechanisms responsible for impaired beta-cell mass formation after IUCR or IUPR have come under investigation. 
First, it has been proposed that IUCR can result in a reduction of the embryonic beta-cell progenitor pool leading to inappropriate postnatal beta-cell formation. Stanger et al. [108] demonstrated that selective genetic reduction in the size of PDX-1+ pancreatic progenitors during the fetal period results in impaired beta-cell formation during the postnatal period with consequent development of glucose intolerance during adulthood. Consistent with this, maternal food restriction leads to significant reduction in PDX-1+ and neurogenin-3+ pancreatic precursors during embryonic development in rats, diminished postnatal beta-cell formation, and inability to expand beta-cell mass in response to pregnancy [47, 94]. The UPI model is also characterized by a permanent decrease in islet PDX-1 mRNA expression. This decrease has recently been shown to be due to progressive epigenetic silencing of the Pdx1 gene locus secondary to proximal promoter methylation [69, 109], and it may be responsible for the decreased rate of beta-cell replication and inappropriate postnatal beta-cell mass development [69, 110]. In the same way of thinking, studies have demonstrated that the maintenance of methylated histone H3 Lys4 by Set7/9, a member of the SET methyltransferase family, is crucial to Pdx1 activity in beta-cell lines [111–113]. This led to the hypothesis that Set7/9 may represent a novel chromatin-modifying protein that functions in part through its recruitment to target genes by cell-specific transcription factors such as Pdx1. Since then, a role of histone methyl transferases, particularly set7, has also been demonstrated in the sustained deleterious effects of chronic hyperglycemia on human microvascular endothelial cells [114]. Such an epigenetic change could potentially be involved in the deleterious effect of high glucose upon the fetal pancreas in the IUED models.
Another mechanism proposed to explain reduced beta-cell formation after IUCR is related to prenatal glucocorticoid exposure. Administration of either dexamethasone or carbenoxolone (to inhibit 11 β-hydroxysteroid dehydrogenase type 2) to normal pregnant rats also causes fetal growth retardation and the adult offspring are hypertensive and hyperglycemic, with hyperactive hypothalamic-pituitary-adrenal axis [115]. Maternal undernutrition significantly increased both fetal and maternal corticosterone concentrations in rats [116]. Subsequently, maternal and/or fetal overexposure to glucocorticoids (via administration of dexamethasone) impairs both fetal and postnatal beta-cell formation in rodents and nonhuman primates [94, 117–119]. Seckl et al. [115] have shown that fetal corticosterone concentrations are inversely correlated with fetal insulin content and postnatal beta-cell formation in rats. Evidence suggests that glucocorticoids can exert a direct effect on the developing fetal pancreas via transcriptional modulation of transcription factors involved in beta-cell formation and differentiation [117]. Glucocorticoid receptors are present in the pancreas during embryonic development of rodents and humans [117], and glucocorticoids can bind to the Pdx1 promoter and thus suppress fetal endocrine cell differentiation [117]. Glucocorticoid treatment has been shown to significantly reduce fetal expression of key endocrine transcription factors such as Pdx1 and Pax6 but simultaneously increase expression of transcription factors that regulate development of the exocrine pancreas [119].
It has also been demonstrated that the UPI or the low-protein IUPR offspring experience increased oxidative stress and impaired mitochondrial function [96, 120]. The mitochondrial dysfunction was not limited to just the beta cell, as mitochondria from both the liver and skeletal muscle exhibit decreased oxidation of pyruvate, subsequently leading to the development of features commonly found in T2D [100, 121]. Also exposure to a Western-style diet before and during pregnancy (an IUEO model) alters the redox state as early as preimplantation development, leading to mild oxidative stress associated with inflammation. The finding that administration of antioxidants to the dam reverses oxidative stress and completely prevents the development of glucose intolerance and increased adiposity in the adult offspring suggests that oxidative stress plays an important role in the development of adiposity in this case [122]. Some studies in the low-protein IUPR model have demonstrated that oxidative stress is not limited to just mitochondrial DNA damage, but also to genomic DNA, impacting cell-cycle regulation and gene expression [123]. While DNA is being targeted throughout by ROS, there are particular regions that are known to be more sensitive to ROS-mediated damage, for example, telomeres. Telomeres comprise GC-rich repeats and are found at the ends of each chromosome. They are known to shorten with each cellular division and, hence, can act as a mitotic clock, registering the number of replicative divisions to have taken place within the cell. Investigations using an IUPR model have indeed reported a decrease in longevity in the offspring [123, 124] accompanied by reduction in mitochondrial antioxidant defences [96, 125] and telomere length in islets [125].
Pancreatic islet development has been shown to be influenced by a number of growth factors including the insulin-like growth factors, IGF-I and IGF-II whose expression in utero is regulated by nutrient and hormone concentrations. IUPR modifies expression of both IGF genes in a variety of fetal tissues. In an IUPR rat model with a decreased beta-cell mass and beta-cell replication and an increased rate of beta-cell apoptosis, gene expression for IGF-II but not IGF-I was found reduced in the fetal pancreas [126]. In a different IUPR model with more severe global food restriction which induced hyperinsulinemia and an increase in beta-cell mass in their fetuses [90], the fetal phenotype was unexpectedly associated with an increase in pancreatic IGF-I expression, islet IGF-1R [91], and IRS-2 [92]. In the fetal GK/Par rat exposed to mild hyperglycemia during gestation (a model of IUED), data from our group suggest that the beta-cell deficit (reduced by more than 50%) starts as early as fetal age E16 and reflects decreased beta-cell proliferation, a limitation of beta-cell neogenesis from precursors, and increased apoptosis of both beta cells and their precursors [86]. Notably, Pdx1 and Neurogenin3 expression were decreased on E18 but normally expressed on E13 [86]. Defective signalling through the Igf2/Igf1-R pathway may represent the primary instrumental anomaly since Igf2 and Igf1-R protein expressions are already decreased within the GK/Par pancreatic rudiment at E13, at a time when beta-cell mass (first wave of beta-cell expansion) is in fact normal [31]. Low levels of pancreatic Igf2 associated with beta-cell mass deficiency are maintained thereafter within the fetal pancreas [87]. Crossbreeding protocols between nondiabetic W and diabetic GK rats showed that, in late gestation (E18), pancreatic Igf2 protein expression was as low in GKmother/GKfather and Wmother/GKfather crosses as in GKmother/GKfather crosses [87]. These findings rather support the hypothesis that the pancreatic Igf2 anomaly in the GK diabetic model is linked to a genetic determinism. This view is also consistent with the results of genetic analyses that linked a locus containing the gene encoding Igf2 to diabetes in the GK rat [127]. The Igf2 gene is subjected to paternal genomic imprinting. However, because the Igf2 expression is similarly affected in fetuses, regardless of whether the father is W or GK [87], we cannot conclude with a simple change of Igf2 gene imprinting in the GK rat.
Finally, our understanding of the underlying mechanisms for reduced BCM in response to inappropriate perinatal nutrition is growing rapidly. However, the relative contribution of the many intrinsic and extrinsic factors which contribute to the adaptive response of the developing endocrine pancreas is still to be established.