I’m looking into mouse lines for labeling of sensory, motor and autonomic neurons.
For sensory, Advillin-Cre has been a go-to, but it’s specificity has been called into question
https://pubmed.ncbi.nlm.nih.gov/30221190-advillin-is-expressed-in-all-adult-neural-crest-derived-neurons/
For autonomic, I’ve seen TH-Cre used, but TH is also in C-LTMRs.
Heart rate is under the precise control of the autonomic nervous system. However, the wiring of peripheral neural circuits that regulate heart rate is poorly understood. Here, we developed a clearing-imaging-analysis pipeline to visualize innervation...
For motor, one can use ChAT-Cre, although some autonomic should also have ChAT.
Is there any method or mouse line that can specifically label a single population?
@tberta @esypek @liz @thicunha
Collecting some responses from Twitter. Check it out.
tberta
December 16, 2019, 2:54pm
3
Pirt-cre is a good alternative to Advillin, see images of DRG and skin tissues obtained recently in our lab.
.
1 Like
Oooh that’s pretty! Did you check spinal cord and other areas? Is Pirt also in sympathetic and motor?
tberta
December 16, 2019, 3:38pm
5
A PhD student is characterizing these mice right now, and I will keep you posted. However, we obtained these mice from Qin Liu and she may have more information - just knock at her door!
We used Nav1.8-Cre mice to specifically target C-fibers, for deleting NMDA receptors:
JA McRoberts, HS Ennes, JC Marvizón, MS Fanselow, EA Mayer and B Vissel,
Neuroscience , Jan 2011 13
The role of NMDA receptors (NMDARs) expressed by primary afferent neurons in nociception remains controversial. The aim of this study was to develop mice with a tissue selective knockdown of NMDARs in these neurons and to evaluate their behavioral responses to different types of painful stimuli. Mice with floxed NMDAR NR1 subunit gene (fNR1) were crossed with mice expressing Cre recombinase under the control of the peripherin promotor (Prph-Cre). Male Prph-Cre+ floxed NR1 mice were compared to Cre- littermates. Both quantitative RT/PCR and Western blotting indicated a ∼75% reduction in NR1 expression in dorsal root ganglia (DRG) extracts with no effect on NR1 expression in spinal cord, brain or the enteric nervous system. Immunocytochemistry with antibodies to NR1 revealed decreased staining in all size classes of DRG neurons. NMDA produced a detectable increase in [Ca2+]i in 60% of DRG neurons cultured from Cre- mice, but only 15% of those from Cre+ mice. Furthermore, the peak [Ca2+]i responses were 64% lower in neurons from Cre+ mice. There was no significant difference between Cre+ and Cre- mice in response latencies to the hotplate or tail withdrawal tests of thermal nociception, nor was there a difference in withdrawal thresholds to mechanical stimuli of the tail or paw. However, compared to Cre- littermates, Cre+ knockdown mice had a 50% decrease in the phase 2 response to formalin injection (P<0.001). There was no effect on phase 1 responses. These results suggest that NMDA receptors expressed by primary afferent nerves play an important role in the development of sensitized pain states.
A Severino, W Chen, JK Hakimian, BL Kieffer, C Gaveriaux-Ruff, W Walwyn and JCG Marvizón,
Pain , Aug 2018
The latent sensitization model of chronic pain reveals that recovery from some types of long-term hyperalgesia is an altered state in which nociceptive sensitization persists but is suppressed by the ongoing activity of analgesic receptors such as μ-opioid receptors (MORs). To determine whether these MORs are the ones present in nociceptive afferents, we bred mice expressing Cre-recombinase under the Nav1.8 channel promoter (Nav1.8cre) with MOR-floxed mice (flMOR). These Nav1.8cre/flMOR mice had reduced MOR expression in primary afferents, as revealed by quantitative PCR, in situ hybridization, and immunofluorescence colocalization with the neuropeptide calcitonin gene-related peptide. We then studied the recovery from chronic pain of these mice and their flMOR littermates. When Nav1.8cre/flMOR mice were injected in the paw with complete Freund adjuvant they developed mechanical hyperalgesia that persisted for more than 2 months, whereas the responses of flMOR mice returned to baseline after 3 weeks. We then used the inverse agonist naltrexone to assess ongoing MOR activity. Naltrexone produced a robust reinstatement of hyperalgesia in control flMOR mice, but produced no effect in the Nav1.8/flMOR males and a weak reinstatement of hyperalgesia in Nav1.8/flMOR females. Naltrexone also reinstated swelling of the hind paw in flMOR mice and female Nav1.8cre/flMOR mice, but not male Nav1.8cre/flMOR mice. The MOR agonist DAMGO inhibited substance P release in flMOR mice but not Nav1.8cre/flMOR mice, demonstrating a loss of MOR function at the central terminals of primary afferents. We conclude that MORs in nociceptive afferents mediate an ongoing suppression of hyperalgesia to produce remission from chronic pain.
1 Like
Hi! Cool work with the Nav1.8 -Cre line! I’m thinking about a pan-sensory neuronal marker, and I wonder if NaV1.8 or peripherin may be good options. Based on your work, do you think these might fit the bill? If so, do you have a good NaV1.8 antibody that perhaps works in humans as well as mice?
Are you asking about the Cre lines or IHC?
Check out this paper:
A vast diversity of salient cues is sensed by numerous classes of primary sensory neurons, defined by specific neuropeptides, ion channels, or cytoskeletal proteins. Recent evidence has demonstrated a correlation between the expression of some of...
If you just want to label all the neurons in the DRG, fluorescent Nissl stain (Neurotrace) works pretty well:https://www.thermofisher.com/order/catalog/product/N21480?us&en#/N21480?us&en
Some people use TUBB3 to label all neurons in the DRG@tberta
Depends on what you’re trying to achieve.
See the recent paper from @sshiers for all these markers in humans
