A quick and laidback way to detect the internal structure of the Drosophila eye: An alternative to cryosectioning.

Immunofluorescence techniques have been a great tool to chase the structure, localization, and function of many proteins within a cell. Drosophila eye is widely used as a model to answer various questions. However, the complex sample preparation and visualization methods restrict its use only with an expert's hand. Thus, an easy and hassle-free method is in need to broaden the use of this model even with an amateur's hand. The current protocol describes an easy sample preparation method using DMSO to image the adult fly eye. The brief description of sample collection, preparation, dissection, staining, imaging, storage, and handling has been described over here. For readers, the possible problems that might arise during the execution of the experiment have been described with their possible reason and solutions. The overall protocol reduces the use of chemicals and shortens the sample preparation time to only 3 h, which is significantly less in comparison to other protocols.

An infection of Enterobacter ludwigii affects development and causes age-dependent neurodegeneration in Drosophila melanogaster.

The effects of teeth-blackening bacteria Enterobacter ludwigii on the physiological system were investigated using the model organism Drosophila melanogaster. The bacteria were mixed with the fly food, and its effect was checked on the growth, development and behaviour of Drosophila. Microbes generate reactive oxygen species (ROS) within the haemolymph of the larvae once it enters into the body. The increased amount of ROS was evidenced by the NBT assay and using 2',7'-dichlorofluorescin diacetate dye, which indicates the mitochondrial ROS. The increased amount of ROS resulted in a number of abnormal nuclei within the gut. Besides that larvae walking became sluggish in comparison with wild type although the larvae crawling path did not change much. Flies hatched from the infectious larvae have the posterior scutellar bristle absent from the thorax and abnormal mechanosensory hairs in the eye, and they undergo time-dependent neurodegeneration as evidenced by the geotrophic and phototrophic assays. To decipher the mechanism of neurodegeneration, flies were checked for the presence of four important bioamines: tyramine, cadaverine, putrescine and histamine. Out of these four, histamine was found to be absent in infected flies. Histamine is a key molecule required for the functioning of the photoreceptor as well as mechanoreceptors. The mechanism via which mouth infectious bacteria E. ludwigii can affect the development and cause age-dependent neurodegeneration is explained in this paper.

Wood-Based Cellulose Nanofibrils: Haemocompatibility and Impact on the Development and Behaviour of Drosophila melanogaster.

Wood-based cellulose nanofibrils (CNF) offer an excellent scaffold for drug-delivery formulation development. However, toxicity and haemocompatibility of the drug carrier is always an important issue. In this study, toxicity-related issues of CNF were addressed. Different doses of CNF were orally administered to Drosophila and different tests like the developmental cycle, trypan blue exclusion assay, larva crawling assay, thermal sensitivity assay, cold sensitivity assay, larval light preference test, climbing behaviour, nitroblue tetrazolium (NBT) reduction assay, adult phenotype, and adult weight were conducted to observe the impact on its development and behaviour. A haemocompatibility assay was done on the blood taken from healthy Wistar rats. In Drosophila, the abnormalities in larval development and behaviour were observed in the behavioural assays. However, the cytotoxic effect could not be confirmed by the gut staining and level of reactive oxygen species. The larvae developed into an adult without any abnormality in the phenotype. The CNF did cause loss of weight in the adult flies and did not cause much toxicity within the body since there was no phenotypic defect. Hemolysis data also suggested that CNF was safe at lower doses, as the data was well within acceptable limits. All these results suggest that cellulose nanofibres have no significant cytotoxic effects on Drosophila. However, the developmental and behavioural abnormalities suggest that CNF may act as a behavioural teratogen.

Oral administration of graphene oxide nano-sheets induces oxidative stress, genotoxicity, and behavioral teratogenicity in Drosophila melanogaster.

The current study checks the effect of various concentrations of dietary graphene oxide (GO) nano-sheets on the development of Drosophila melanogaster. GO was synthesized and characterized by XRD, FTIR, FESEM, and TEM analytical techniques. Various concentrations of GO were mixed with the fly food and flies were transferred to the vial. Various behavioral and morphological as well as genetic defects were checked on the different developmental stages of the offspring. In the larval stage of development, the crawling speed and trailing path change significantly than the control. GO induces the generation of oxygen radicals within the larval hemolymph as evidenced by nitroblue tetrazolium assay. GO induces DNA damage within the gut cell, which was detected by Hoechst staining and within hemolymph by comet assay. Adult flies hatched after GO treatment show defective phototaxis and geotaxis behavior. Besides behavior, phenotypic defects were observed in the wing, eye, thorax bristles, and mouth parts. At 300 mg/L concentration, wing spots were observed. Altogether, the current study finds oral administration of GO which acts as a mutagen and causes various behavioral and developmental defects in the offspring. Here for the first time, we are reporting GO, which acts as a teratogen in Drosophila, besides its extensive medical applications.

The contribution of nanostructures towards the wing patterning of yellow Catopsilia pomona. How it differs from the lime?

Polyphenism, an adaptation to survive throughout the year, is shown by many butterflies including Catopsilia pomona. With the variation of seasons, different morphs were found. Among all the morphs, lime exists throughout the year whereas the yellow one is available only in the winter season. The current study deciphers the colouration mechanism of yellow morph using various microscopic and spectroscopic techniques. The scanning electron microscopy analysis reveals various types of scales on the dorsal as well as the ventral side. The shape of the cover scale varies from region to region. The fine structural arrangement of the scale like window, ridge, microrib, crossrib and pigments vary throughout the wing. The pigment present in the wing is pterin as evidenced from the shape and its isolation technique. Absorption spectroscopy further confirms the presence of various types of pterin within the wing. Scanning electron microscopy discloses the dense amount of pigments within the wing. The fine structural arrangement of the wing of yellow C. pomona is compared with the yellow region of the lime C. pomona. All together, the current study describes the fine structural arrangement of the wing of yellow C. pomona and the various types of pterin which contribute towards the wing colouration. The advantage of yellow morph over lime is also discussed in this paper.

Enhanced mechanical properties of hBN-ZrO2 composites and their biological activities on Drosophila melanogaster: synthesis and characterization.

In this study, six compositions in the system [x(h-BN)-(100 - x)ZrO2] (10 ≤ x ≤ 90) were synthesized by a bottom up approach, i.e., the solid-state reaction technique. XRD results showed the formation of a novel and main phase of zirconium oxynitrate ZrO(NO3)2 and SEM exhibited mixed morphology of layered and stacked h-BN nanosheets with ZrO2 grains. The composite sample 10 wt% h-BN + 90 wt% ZrO2 (10B90Z) showed outstanding mechanical properties for different parameters, i.e., density (3.12 g cm-3), Young's modulus (10.10 GPa), toughness (2.56 MJ m-3), and maximum mechanical strength (227.33 MPa). The current study further checked the in vivo toxicity of composite 10B90Z and composite 90B10Z using Drosophila melanogaster. The composite 10B90Z showed less cytotoxicity in this model, while the composite 90B10Z showed higher toxicity in terms of organ development as well as internal damage of the gut mostly at the lower concentrations of 1, 10, and 25 µg mL-1. Altogether, the current study proposes the composite 10B90Z as an ideal compound for applications in biomedical research. This composite 10B90Z displays remarkable mechanical and biological performances, due to which we recommend this composition for various biomedical applications.