Dissection and Lipid Droplet Staining of Oenocytes in <em>Drosophila</em> Larvae

Chuanxian Wei; Yan Yan; Xiaoli Miao; Renjie Jiao

doi:10.3791/60606

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae

Full Text

12,755 Views

05:37 min

December 28, 2019

DOI: 10.3791/60606-v

Chuanxian Wei*¹, Yan Yan*¹, Xiaoli Miao¹, Renjie Jiao^1,2,3

¹Sino-French Hoffmann Institute, School of Basic Medical Sciences,Guangzhou Medical University, ²The Second Affiliated Hospital of Guangzhou Medical University, ³The State Key Laboratory of Respiratory Disease,Guangzhou Medical University

Overview

This study presents a method for dissecting oenocytes and staining lipid droplets in Drosophila larvae, utilizing the BODIPY 493/503 fluorescent dye. It aims to investigate lipid droplet behavior under normal and stress conditions, facilitating insights into lipid metabolism in both insects and mammals.

Key Study Components

Research Area

Cell biology
Lipid metabolism
Insect physiology

Background

Oenocytes play a significant role in lipid storage and metabolism.
Understanding lipid droplet dynamics is crucial for insights into metabolic diseases.
Drosophila serves as a powerful model organism for genetic studies.

Methods Used

Dissection of oenocytes from Drosophila larvae
Use of BODIPY 493/503 for lipid droplet staining
Confocal microscopy for imaging

Main Results

Demonstrated the visualization of lipid droplets in oenocytes.
Showed an increase in lipid droplet numbers in response to starvation.
Provided a feasible protocol for studying lipid metabolism across various contexts.

Conclusions

This method enables researchers to investigate genetic and environmental impacts on lipid metabolism.
It contributes to broader biological understanding and research methodologies in lipid biology.

Frequently Asked Questions

What is the significance of oenocytes in Drosophila?

Oenocytes are crucial for lipid storage and metabolism, making them significant for studies on fat regulation.

How does starvation affect lipid droplet formation?

Starvation leads to an increase in the number of lipid droplets in oenocytes, indicating metabolic responses.

What technologies are utilized in this study?

The study employs dissection techniques, fluorescent staining, and confocal microscopy.

Can this method be applied to mammalian studies?

Yes, with minor modifications, this method can be adapted for mammalian lipid metabolism studies.

What are the conditions for egg laying in Drosophila?

150 virgin females and 75 males are used, kept in light-proof boxes at 25°C with 60% humidity.

How long does it take for larvae to develop?

The larvae develop for approximately 84 hours to reach the third instar stage.

What role does BODIPY 493/503 play in this research?

BODIPY 493/503 is a fluorescent dye used to specifically stain lipid droplets for imaging.

Presented here are detailed methods for the dissection and lipid droplet staining of oenocytes in Drosophila larvae using BODIPY 493/503, a lipid droplet-specific fluorescent dye.

This method of an oenocyte dissection and lipid droplet staining can be used to realize the appearance of lipid droplet in drosophila larvae oenocyte under normal and stressed conditions. This method provide a useful tool for studying lipid metabolism not only in insects but also in mammals with only a few minor modifications. To induce egg laying, place 150 virgin female flies and 75 males of the desired genotypes in an egg-laying bottle and place the bottle under a light-proof box in a 25 degrees Celsius incubator with 60%humidity.

After one hour, transfer the flies to a new bottle and let the flies lay eggs in the new bottle for one hour before removing the adults. Then, allow the eggs to develop into the third instar larvae for 84 hours at 25 degrees Celcius with a 12-hour light-dark cycle. Before initiating the starvation treatment, place an appropriately sized piece of filter paper into a six centimeter Petri dish and add one milliliter of PBS onto the paper to create a starvation chamber.

View the full transcript and gain access to thousands of scientific videos