Larval NMJ Developmental Analysis

Larvae were reared, dissected and processed as previously described with analysis of hemisegment A2 (Schuster et al., 1996 (link); Grice et al., 2015a (link),b (link)), and at least 23 NMJs per genotype per stage were scored. Ectopic branch contact analysis was performed using L3 larval hemisegments A1 to A3, and 14 flies per genotype were scored. For axonal bruchpilot (Brp) localization studies, the transverse nerve (TN) section bypassing muscle 6 and 7 in hemisegments A2 and A3 was used. For immunohistochemistry, anti-discs large (DLG, DSHB), anti-HRP (Jackson), anti-Bruchpilot (DSHB) and anti-HA (Santa Cruz) were all used at 1/100 in combination with secondary antibodies, AlexaFluor 488 goat anti-rabbit and AlexaFluor 633 goat anti-mouse (Invitrogen) at 1/1000. Z-stacks were taken using a Leica SP5 laser confocal microscope and analysis performed using ImageJ and Photoshop (Adobe).

Free full text: Click here

Grice S.J., Sleigh J.N, & Zameel Cader M. (2018). Plexin-Semaphorin Signaling Modifies Neuromuscular Defects in a Drosophila Model of Peripheral Neuropathy. Frontiers in Molecular Neuroscience, 11, 55.

Publication 2018

anti-HRP anti-HA AlexaFluor 488 goat anti-rabbit AlexaFluor 633 goat anti-mouse SP5 laser confocal microscope ImageJ

Antibodies Axonal Flies Genotype Goat Immunohistochemistry Larval Laser microscope Mouse Muscle Nerve Nmjs Rabbit

Corresponding Organization :

Other organizations : University of Oxford, Genomics (United Kingdom), University College London, Sobell House, John Radcliffe Hospital, MRC Weatherall Institute of Molecular Medicine

Top 5 similar protocols

Variable analysis

independent variables

Genotype

dependent variables

Number of neuromuscular junctions (NMJs) scored per genotype per stage
Ectopic branch contact analysis
Axonal bruchpilot (Brp) localization

control variables

Larval stage (L3)
Hemisegments (A1 to A3)

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!