Light-Mediated Enhancement of Glucose-Stimulated Insulin Release of Optogenetically Engineered Human Pancreatic Beta-Cells

Enhancement of glucose-stimulated insulin secretion (GSIS) in exogenously delivered pancreatic β-cells is desirable, for example, to overcome the insulin resistance manifested in type 2 diabetes or to reduce the number of β-cells for supporting homeostasis of blood sugar in type 1 diabetes. Optogenetically engineered cells can potentiate their function with exposure to light. Given that cyclic adenosine monophosphate (cAMP) mediates GSIS, we surmised that optoamplification of GSIS is feasible in human β-cells carrying a photoactivatable adenylyl cyclase (PAC). To this end, human EndoC-βH3 cells were engineered to express a blue-light-activated PAC, and a workflow was established combining the scalable manufacturing of pseudoislets (PIs) with efficient adenoviral transduction, resulting in over 80% of cells carrying PAC. Changes in intracellular cAMP and GSIS were determined with the photoactivation of PAC in vitro as well as after encapsulation and implantation in mice with streptozotocin-induced diabetes. cAMP rapidly rose in β-cells expressing PAC with illumination and quickly declined upon its termination. Light-induced amplification in cAMP was concomitant with a greater than 2-fold GSIS vs β-cells without PAC in elevated glucose. The enhanced GSIS retained its biphasic pattern, and the rate of oxygen consumption remained unchanged. Diabetic mice receiving the engineered β-cell PIs exhibited improved glucose tolerance upon illumination compared to those kept in the dark or not receiving cells. The findings support the use of optogenetics for molecular customization of the β-cells toward better treatments for diabetes without the adverse effects of pharmacological approaches

Table_1_Physiological and molecular mechanism of ginger (Zingiber officinale Roscoe) seedling response to salt stress.pdf

We used ‘Shannong No.1’ experimental material to simulate higher salt concentration in ginger and analyzed the physiological responses of different parts of ginger seedlings under salt stress. The results showed that salt stress led to a significant decrease in fresh and dry weight of ginger, lipid membrane peroxidation, increased sodium ion content and enhanced activity of antioxidant enzymes. Compared with the control, the overall plant dry weight of ginger under salt stress decreased by about 60%, and the MDA content in roots, stems, leaves, and rhizomes increased by 372.27%, 184.88%, 291.5%, and 171.13%, respectively, and the APX content increased by 188.85%, 165.56%, 195.38%, and 40.08%, respectively. After analysis of the physiological indicators, it was found that the roots and leaves of ginger were the most significantly changed parts. We analyzed the transcriptional differences between ginger roots and leaves by RNA-seq and found that they jointly initiated MAPK signaling pathways in response to salt stress. By combining physiological and molecular indicators, we elucidated the response of different tissues and parts of ginger to salt stress during the seedling stage.</p

Additional file 1 of Photosynthetic characteristics and chloroplast ultrastructure of welsh onion (Allium fistulosum L.) grown under different LED wavelengths

Additional file 1 Table S1. Growth and development of Welsh onions under different light conditions. Table S2. Photosynthetic parameters of Welsh onions under different light conditions and RuBPCase activity of Welsh onions under different light conditions. Table S3. Chlorophyll fluorescence parameters of Welsh onions under different light conditions. Table S4. Leaf anatomy and chloroplast ultrastructure of Welsh onions under different light conditions

Relationship between soil water potential and soil relative water content.

Relationship between soil water potential and soil relative water content.</p

Soil physical and chemical properties in the greenhouse.

Soil physical and chemical properties in the greenhouse.</p

Additional file 4 of Analyses of the photosynthetic characteristics, chloroplast ultrastructure, and transcriptome of apple (Malus domestica) grown under red and blue lights

Additional file 4: Table S4. The primers used for RT-qPCR

The effects of different irrigation systems on the quality of lettuce including soluble protein, soluble sugar and vitamin C content.

The effects of different irrigation systems on the quality of lettuce including soluble protein, soluble sugar and vitamin C content.</p

Additional file 2 of Analyses of the photosynthetic characteristics, chloroplast ultrastructure, and transcriptome of apple (Malus domestica) grown under red and blue lights

Additional file 2: Table S2. Differentially expressed genes between blue/red and white light-treated apple seedlings

Effects of different irrigation systems on soil temperature at 10cm soil layer at SS (seedling stage),RS (rosette stage), HS (heading stage) in spring or autumn.

Effects of different irrigation systems on soil temperature at 10cm soil layer at SS (seedling stage),RS (rosette stage), HS (heading stage) in spring or autumn.</p

Effects of water irrigation systems on absolute amount of water distribution and water distribution through the plant.

Effects of water irrigation systems on absolute amount of water distribution and water distribution through the plant.</p