Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 May 21;12:3020. doi: 10.1038/s41467-021-23294-7

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 9 — a TUJ1 (cyan) and OPRM1 (magenta) IF in day 22 H3.3.1 EMLO. High magnification images provided (bottom-left) with multi-dimensional view of gut tube (bottom-right). b Top: OPRM1 expression in apical aspect of neural rosettes and basal neurons. Bottom: TFAP2α expression at the basal aspect of GABAergic rosettes. c Fluo-4 AM in dissociated EMLO cultures. High magnification image with example soma ROI (right). d 2D adherent spinal sensory neurons (top, TUJ1 in blue, BRN3A in magenta) and spinal motor neurons (bottom, TUJ1 in green, Nkx-6.1 in red) used as controls for opioid-responsive firing. IF of MOR was performed in N = 3 separate H3.3.1 EMLO formation experiments with similar results. e Examples of calcium transients quantified in dissociated EMLO neuronal soma using Fluo-4 AM. Shown are no calcium transients (none), one transient (single), or multiple transients (multiple) within a 1.5 min acquisition window. f Histogram of percent of neurons with multiple calcium transients. EMLO-dissociated cultures were compared with spinal sensory neurons (positive control) and spinal motor neurons (negative control). Baseline firing in BrainPhys (blue) was compared to addition of 1 μM DAMGO (tan), followed by 10 μM naloxone in the same cultures (light blue). n = number of neurons quantified. Statistical p values are as follows from left-to-right on the plot: **p = 0.0031, **p = 0.002, n.s. p = 0.766; ***p = 0.001, *p = 0.014, n.s. p = 0.972; n.s. p = 0.593, n.s. p = 0.64, n.s. p = 0.94). Violin plot statistics are as follows: EMLO-derived (BrainPhys: max = 47, min = 13, median = 29, q1 = 29, q3 = 39; BrainPhys + DAMGO: max = 32, min = 7, median = 11, q1 = 9, q3 = 15; BrainPhys + DAMGO + naloxone: max = 47, min = 32, median = 27, q1 = 21, q3 = 39); spinal sensory (BrainPhys: max = 48, min = 27, median = 37, q1 = 28, q3 = 46; BrainPhys + DAMGO: max = 16, min = 6, median = 10, q1 = 6, q3 = 14; BrainPhys + DAMGO + naloxone: max = 59, min = 7, median = 36, q1 = 19, q3 = 44); spinal motor (BrainPhys: max = 35, min = 15, median = 32; BrainPhys + DAMGO: max = 38, min = 23, median = 31, q1 = 25, q3 = 35; BrainPhys + DAMGO + naloxone: max = 38, min = 14, median = 29, q1 = 22, q3 = 34). Data reported as (mean ± s.e.m.). Individual scale bars provided.