Adrenergic Signaling in Pancreatic Islet Development

Diabetes mellitus is a disease that has been rapidly increasing in prevalence worldwide, with more than 9% of the world’s adult population currently diagnosed, and an additional 4-5% thought to be living with diabetes without a diagnosis. Diabetes is caused by dysregulation of blood glucose homeostasis. A key regulator of blood glucose homeostasis is insulin secretion from β-cells that reside in pancreatic islets of Langerhans. While much is known about the intrinsic factors that control β-cell development, less is known about how the interaction of β-cells with other non-pancreatic cell types controls development and adult function. Recently, our lab discovered that sympathetic nerves that innervate the pancreas are essential for pancreatic islet development and adult islet function (Borden et al. 2013). These nerves secrete the neurotransmitter norepinephrine, a potent activator of adrenergic signaling. During development, the mouse pancreas expresses all nine of the adrenergic G-protein coupled receptors (GPCRs), including those most responsive to epinephrine, a hormone secreted from the adrenal gland that is distributed throughout the body through the circulatory system. This led us to hypothesize that adrenergic signaling may play a role in instructing endocrine pancreas development, beyond the commonly studied roles in acute, physiological stress responses. Here, we identify a sex-specific requirement for the β2-adrenergic receptor (Adrb2) in regulating Vegf-a production and vascularization during islet development. Loss of Adrb2 from islet β-cells enhances Vegf-a and insulin expression in β-cells, and results in hyper-vascularized islets in neonatal mice, specifically in females. Despite elevated insulin synthesis, adult mutant females, but not males, exhibited glucose intolerance and impaired glucose-stimulated insulin secretion (GSIS), in part, due to alterations in genes involved in calcium regulation and exocytosis in islets. Metabolic and molecular phenotypes in adult Adrb2-deficient mice were fully rescued by the blockade of Vegf-a signaling and correction of islet hyper-vascularization during development. Consistent with a predominant role for Adrb2 during development, glucose homeostasis was unaffected by adult-specific deletion of Adrb2. Together, these findings uncover a negative regulatory pathway that functions in a sex-specific manner during islet formation to control long-term glucose metabolism by restraining excessive vascular growth

Zebrafish as a model for apolipoprotein biology: comprehensive expression analysis and a role for ApoA-IV in regulating food intake

Improved understanding of lipoproteins, particles that transport lipids throughout the circulation, is vital to developing new treatments for the dyslipidemias associated with metabolic syndrome. Apolipoproteins are a key component of lipoproteins. Apolipoproteins are proteins that structure lipoproteins and regulate lipid metabolism through control of cellular lipid exchange. Constraints of cell culture and mouse models mean that there is a need for a complementary model that can replicate the complex in vivo milieu that regulates apolipoprotein and lipoprotein biology. Here, we further establish the utility of the genetically tractable and optically clear larval zebrafish as a model of apolipoprotein biology. Gene ancestry analyses were implemented to determine the closest human orthologs of the zebrafish apolipoprotein A-I (apoA-I), apoB, apoE and apoA-IV genes and therefore ensure that they have been correctly named. Their expression patterns throughout development were also analyzed, by whole-mount mRNA in situ hybridization (ISH). The ISH results emphasized the importance of apolipoproteins in transporting yolk and dietary lipids: mRNA expression of all apolipoproteins was observed in the yolk syncytial layer, and intestinal and liver expression was observed from 4–6 days post-fertilization (dpf). Furthermore, real-time PCR confirmed that transcription of three of the four zebrafish apoA-IV genes was increased 4 hours after the onset of a 1-hour high-fat feed. Therefore, we tested the hypothesis that zebrafish ApoA-IV performs a conserved role to that in rat in the regulation of food intake by transiently overexpressing ApoA-IVb.1 in transgenic larvae and quantifying ingestion of co-fed fluorescently labeled fatty acid during a high-fat meal as an indicator of food intake. Indeed, ApoA-IVb.1 overexpression decreased food intake by approximately one-third. This study comprehensively describes the expression and function of eleven zebrafish apolipoproteins and serves as a springboard for future investigations to elucidate their roles in development and disease in the larval zebrafish model

Retrotransposons Are the Major Contributors to the Expansion of the Drosophila ananassae Muller F Element.

The discordance between genome size and the complexity of eukaryotes can partly be attributed to differences in repeat density. The Muller F element (∼5.2 Mb) is the smallest chromosome in Drosophila melanogaster, but it is substantially larger (>18.7 Mb) in D. ananassae To identify the major contributors to the expansion of the F element and to assess their impact, we improved the genome sequence and annotated the genes in a 1.4-Mb region of the D. ananassae F element, and a 1.7-Mb region from the D element for comparison. We find that transposons (particularly LTR and LINE retrotransposons) are major contributors to this expansion (78.6%), while Wolbachia sequences integrated into the D. ananassae genome are minor contributors (0.02%). Both D. melanogaster and D. ananassae F-element genes exhibit distinct characteristics compared to D-element genes (e.g., larger coding spans, larger introns, more coding exons, and lower codon bias), but these differences are exaggerated in D. ananassae Compared to D. melanogaster, the codon bias observed in D. ananassae F-element genes can primarily be attributed to mutational biases instead of selection. The 5' ends of F-element genes in both species are enriched in dimethylation of lysine 4 on histone 3 (H3K4me2), while the coding spans are enriched in H3K9me2. Despite differences in repeat density and gene characteristics, D. ananassae F-element genes show a similar range of expression levels compared to genes in euchromatic domains. This study improves our understanding of how transposons can affect genome size and how genes can function within highly repetitive domains

Retrotransposons Are the Major Contributors to the Expansion of the Drosophila ananassae Muller F Element

The discordance between genome size and the complexity of eukaryotes can partly be attributed to differences in repeat density. The Muller F element (∼5.2 Mb) is the smallest chromosome in Drosophila melanogaster, but it is substantially larger (>18.7 Mb) in D. ananassae. To identify the major contributors to the expansion of the F element and to assess their impact, we improved the genome sequence and annotated the genes in a 1.4-Mb region of the D. ananassae F element, and a 1.7-Mb region from the D element for comparison. We find that transposons (particularly LTR and LINE retrotransposons) are major contributors to this expansion (78.6%), while Wolbachia sequences integrated into the D. ananassae genome are minor contributors (0.02%). Both D. melanogaster and D. ananassae F-element genes exhibit distinct characteristics compared to D-element genes (e.g., larger coding spans, larger introns, more coding exons, and lower codon bias), but these differences are exaggerated in D. ananassae. Compared to D. melanogaster, the codon bias observed in D. ananassae F-element genes can primarily be attributed to mutational biases instead of selection. The 5′ ends of F-element genes in both species are enriched in dimethylation of lysine 4 on histone 3 (H3K4me2), while the coding spans are enriched in H3K9me2. Despite differences in repeat density and gene characteristics, D. ananassae F-element genes show a similar range of expression levels compared to genes in euchromatic domains. This study improves our understanding of how transposons can affect genome size and how genes can function within highly repetitive domains