High sucrose diet induces morphological, structural and functional impairments in the renal tubules of Drosophila melanogaster: A model for studying type-2 diabetes mediated renal tubular dysfunction.

Continuous feeding of high dietary sugar is strongly associated with type 2 diabetes (T2D) and its secondary complications. Diabetic nephropathy (DN) is a major secondary complication that leads to glomerular and renal tubular dysfunction. The present study is aimed to investigate the effects of chronic exposure of high sugar diet (HSD) on renal tubules. Malpighian tubules (MTs), a renal organ of Drosophila, were used as a model in the study. Feeding of HSD develops T2D condition in Drosophila. The MTs showed structural abnormalities in 20 days of HSD fed flies. Impaired insulin signaling, oxidative stress, enhanced levels of AGE-RAGE and induction of apoptosis were observed in the MTs of these flies. Further, altered expression of transporters, enhanced uric acid level and reduced fluid secretion rate confirmed the impaired function of MTs in these flies. RNA-seq and RT-PCR analyses in the MTs of HSD fed-and control-flies revealed the altered expression of candidate genes that regulate several important pathways including extracellular matrix (ECM), advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE), transforming growth factor ß (TGF-ß), galactose, starch and sucrose metabolism that are well known mediators of renal tubular dysfunction in DN patients. Disruption of insulin signaling in the MTs also causes renal tubular dysfunction similar to HSD fed flies. Overall, the study suggests that phenotypes observed in the MTs of HSD fed flies recapitulate several hallmarks of renal tubular dysfunction in DN patients. Therefore, we conclude that MTs of HSD fed flies may be used for deciphering the underlying mechanisms of T2D mediated renal tubular dysfunction.

Renal neuroendocrine control of desiccation and cold tolerance by Drosophila suzukii.

BACKGROUND: Neuropeptides are central to the regulation of physiological and behavioural processes in insects, directly impacting cold and desiccation survival. However, little is known about the control mechanisms governing these responses in Drosophila suzukii. The close phylogenetic relationship of D. suzukii with Drosophila melanogaster allows, through genomic and functional studies, an insight into the mechanisms directing stress tolerance in D. suzukii. RESULTS: Capability (Capa), leucokinin (LK), diuretic hormone 44 (DH44 ) and DH31 neuropeptides demonstrated a high level of conservation between D. suzukii and D. melanogaster with respect to peptide sequences, neuronal expression, receptor localisation, and diuretic function in the Malpighian tubules. Despite D. suzukii's ability to populate cold environments, it proved sensitive to both cold and desiccation. Furthermore, in D. suzukii, Capa acts as a desiccation- and cold stress-responsive gene, while DH44 gene expression is increased only after desiccation exposure, and the LK gene after nonlethal cold stress recovery. CONCLUSION: This study provides a comparative investigation into stress tolerance mediation by neuroendocrine signalling in two Drosophila species, providing evidence that similar signalling pathways control fluid secretion in the Malpighian tubules. Identifying processes governing specific environmental stresses affecting D. suzukii could lead to the development of targeted integrated management strategies to control insect pest populations. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effects of the cyclopeptide mycotoxin destruxin A on the Malpighian tubules of Rhodnius prolixus (Stål).

The production of peptide toxins by entomopathogenic fungi during the infection process plays critical roles in pathogenesis. To gain insight into the mechanism of action of these mycotoxins on insect internal organs, we have evaluated the effects of destruxin A, a cyclic peptide produced by Metarhizium anispliae, on Rhodnius prolixus Malpighian tubules measuring fluid secretion rate, transepithelial electrical potential (TEP), pH and ion composition of secreted fluid, and ATP content. Destruxin A dramatically inhibited fluid secretion rate on tubules stimulated by 5-Hydroxytryptamine (5-HT) or cAMP. The calculated IC(50) for destruxin A on 5-HT-stimulated tubules was 3 x 10(-7) M. Fluid secretion rate by Malpighian tubules exposed for 20 min to 10(-6) M destruxin A recovered completely when tubules were washed with saline; however, when tubules were exposed to 5 x 10(-6) M destruxin A the fluid secretion rate only partially recovered upon wash off. The use of Ca(2+)-free saline or addition of the calcium channel blocker CoCl(2) to the bathing saline did not interfere with the effects of destruxin A, and neither did the modification of intracellular calcium by TMB-8. Measurement of TEP of tubules challenged with 5-HT after preincubation for 10 min in saline containing 10(-6) M destruxin A showed that the second and third phases of the typical triphasic response to 5-HT were disrupted. Likewise, the positive shift in TEP in response to 5-HT in chloride-free bathing saline was significantly reduced when tubules were preincubated for 10 min in 10(-6) M destruxin A. The pH of the secreted fluid, but not the Na(+) or K(+) concentration, increased significantly when 5-HT-stimulated tubules were exposed to 10(-6) M destruxin A. The ATP content was not significantly different when tubules stimulated with 5-HT were exposed to destruxin A. Taken together, these results show that destruxin A, without interfering with the intracellular ATP production, strongly inhibits fluid secretion rate by the Malpighian tubules of R. prolixus. Changes in properties of the TEP suggest that one of the target sites for this peptide toxin might be associated with inhibition of the apical V-type H(+) ATPase of tubule cells.

Critical electrolyte concentration of heterochromatin and euchromatin in cells of starved animals.

The critical electrolyte concentration (CEC) of chromatin, considered to be the salt concentration at which the phenomenon of metachromasy is completely abolished, when using toluidine blue as the cationic dye and Mg2+ ions as inorganic cations, was determined for the euchromatin and chromocentral heterochromatin of cells from starved animals. The objective was to determine variations in CEC induced by the starvation stress, which could be related to structural alterations and/or to the composition of these chromatin types. The cells investigated were from Triatoma infestans Malpighian tubules and mouse liver. It was found that the CEC values were affected both by fasting and by refeeding, the effect varying according to chromatin type and the cell system under consideration.