Protease Activity Assay in Fly Intestines

Nawrot-Esposito, Marie-Paule; Loudhaief, Rihab; Gallet, Armel

doi:10.21769/bioprotoc.2560

Cited by 2 publications

(1 citation statement)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Intestinal protease activity assay was performed as described in [ 43 ] with eight mid-third instar larval midguts per condition. Lipids were assayed as described in [ 44 ].…”

Section: Methodsmentioning

confidence: 99%

Bacillus thuringiensis Bioinsecticides Induce Developmental Defects in Non-Target Drosophila melanogaster Larvae

Nawrot-Esposito

Babin

Pasco

et al. 2020

Insects

Self Cite

View full text Add to dashboard Cite

Bioinsecticides made from the bacterium Bacillus thuringiensis (Bt) are the bestselling bioinsecticide worldwide. Among Bt bioinsecticides, those based on the strain Bt subsp. kurstaki (Btk) are widely used in farming to specifically control pest lepidopteran larvae. Although there is much evidence of the lack of acute lethality of Btk products for non-target animals, only scarce data are available on their potential non-lethal developmental adverse effects. Using a concentration that could be reached in the field upon sprayings, we show that Btk products impair growth and developmental time of the non-target dipteran Drosophila melanogaster. We demonstrate that these effects are mediated by the synergy between Btk bacteria and Btk insecticidal toxins. We further show that Btk bioinsecticides trigger intestinal cell death and alter protein digestion without modifying the food intake and feeding behavior of the larvae. Interestingly, these harmful effects can be mitigated by a protein-rich diet or by adding the probiotic bacterium Lactobacillus plantarum into the food. Finally, we unravel two new cellular mechanisms allowing the larval midgut to maintain its integrity upon Btk aggression: First the flattening of surviving enterocytes and second, the generation of new immature cells arising from the adult midgut precursor cells. Together, these mechanisms participate to quickly fill in the holes left by the dying enterocytes.

show abstract

“…Intestinal protease activity assay was performed as described in [ 43 ] with eight mid-third instar larval midguts per condition. Lipids were assayed as described in [ 44 ].…”

Section: Methodsmentioning

confidence: 99%

Bacillus thuringiensis Bioinsecticides Induce Developmental Defects in Non-Target Drosophila melanogaster Larvae

Nawrot-Esposito

Babin

Pasco

et al. 2020

Insects

Self Cite

View full text Add to dashboard Cite

show abstract

Determination of Chromatin Accessibility in Drosophila Midgut Enterocytes by in situ 5mC Labeling

Bitman-Lotan¹,

Rincón‐Arano²,

Raz³

et al. 2019

BIO-PROTOCOL

View full text Add to dashboard Cite

Regulation of gene expression involves dynamic changes in chromatin organization, where in many cases open chromatin structure correlates with gene activation. Several methods enable monitoring changes in chromatin accessibility, including ATAC-seq, FAIRE-seq, MNase-seq and DNAseseq methods, which involve Next-generation-sequencing (NGS). Focusing on the adult Drosophila differentiated gut enterocytes (ECs) we used a sequencing-free method that enables visualizing and semi-quantifying large-scale changes in chromatin structure using in vitro methylation assay with the bacterial CpG Methyltransferase, M. Sssl, that determine chromatin accessibility. In brief, as CpG methylation is minimal in differentiated somatic Drosophila cells, we used the bacterial M. SssI enzyme to methylate CpG dinucleotides in situ depending on their chromatin accessibility. The methylated dinucleotides are detected using 5mCytosine monoclonal antibody and nuclei are visualized microscopically. Thus, the 5mC method enables to monitor large-scale chromatin changes in heterogenic cellular tissue focusing on the cell type of interest and without the need for cell purification or NGS.

show abstract

Protease Activity Assay in Fly Intestines

Cited by 2 publications

References 8 publications

Bacillus thuringiensis Bioinsecticides Induce Developmental Defects in Non-Target Drosophila melanogaster Larvae

Bacillus thuringiensis Bioinsecticides Induce Developmental Defects in Non-Target Drosophila melanogaster Larvae

Determination of Chromatin Accessibility in Drosophila Midgut Enterocytes by in situ 5mC Labeling

Contact Info

Product

Resources

About