SS45779 – Targeting b3 MN, iii3 MN, hg1 MN

Our line:

Janelia image:

IS69306 – b1 mn driver line

Janelia image:

Retro-Tango control crossed with GF-Gal4 (BDSC: 79602), presynaptic neurons in magenta, giant fiber neurons in cyan and background staining in gray

Retro-Tango x GF-Gal4 from Sorkac et. al (2023)

rut and rad expression in b1 and b3 motor neurons, I looked for cells express 6 genes (rut, rad, FoxP, VGlut, AD TF, DBD TF) at the same time above certain threshold:

A: w1118 × dicer2; SNPFR-RNAi
B: w1118 × dicer2; H24-Gal4
C: dicer2; SNPFR-RNAi × dicer2; H24-Gal4

Larvae were placed in the center of an agar plate with plain agar on one side and salt-containing agar (1.5 M NaCl) on the other. After 3 minutes, larval distribution was quantified using a preference index under red light. All three groups showed significant salt avoidance, but no significant differences were observed between the groups.

On Wednesday I tested the “heatshock” group, using light (with the same intensity as the aversive stimulus in the joystick experiment: ~800lux) for 5 minutes before testing, to stress the flies.

Stressor: heat (5min; 7.4V; ~800lux)

Gr28bd+TrpA1(Co): control group

Gr28bd+TrpA1: experimental group

 
On Friday I tested flies who were maintained in a cold environment (18°C) during the ATR treatment (2 days).

Stressor: cold (18°C; 48h)

Gr28bd+TrpA1(Co): Control group

Gr28bd+TrpA1: Experimental group

p values: A red light <-> B red light = 0.39366; A blue light <-> B blue light: 0.68727; A red light <-> A blue light: 0.00136; B red light <-> B blue light: 0.02590

Flies 48 hours on RU486 wetted tissue

Flies 48 hours on RU486 wetted instant_food

Mean OMRs at start

Drosophila larvae from three groups (A, B, and C) were placed in the center of an agar plate containing plain agar on one side and salt-containing agar (1.5 M) on the other. After 3 minutes, larval distribution was quantified using a preference index under red light. All three groups showed a significant avoidance of salt. No significant differences in salt avoidance were observed between the groups.

Last week I started to test CsChrimson (NorpA, 20xUAS-Chrimson) flies with the joystick set up, to reproduce Daniels’ data on the positive control and to get used to the experimental set up.

There is a noticeable improvement (from my end) from the first two days of testing to the last two days.

These are the results of the first four days of testing. All in all, the positive control shows good avoidance. Althoug, there is a remarkable change in the error bars starting with the fifth period. Maybe due to exhaustion or any other effect that I don’t know of yet.