Localisation of cannabinoid receptor 1 in rat dorsal root ganglion using in situ hybridisation and immunohistochemistry

doi:10.1016/S0306-4522(03)00200-8

Neuroscience

Volume 119, Issue 3, 4 July 2003, Pages 803-812

https://doi.org/10.1016/S0306-4522(03)00200-8 Get rights and content

Abstract

In this study we used in situ hybridisation and double-labelling immunohistochemistry to characterise cannabinoid receptor 1 (CB₁) expression in rat lumbar dorsal root ganglion (DRG) neurons.

Approximately 25% of DRG neurons expressed CB₁ mRNA and displayed immunoreactivity for CB₁. Sixty-nine percent to 82% of CB₁-expressing cells were also immunoreactive for neurofilament 200, indicative of myelinated A-fibre neurons, which tend to be large- and medium-sized DRG neurons (>600 μm²). Approximately 10% of CB1-expressing cells also expressed transient receptor potential vanilloid family ion channel 2 (TRPV2), the noxious heat-transducing channel found in medium to large lightly myelinated Aδ-fibre DRG neurons. Seventeen percent to 26% of CB₁-expressing cells co-stained using Isolectin B4, 9–10% for calcitonin gene-related peptide and 11–20% for transient receptor potential vanilloid family ion channel 1 (TRPV1), predominantly markers of small non-myelinated C-fibre DRG neurons (<600 μm²).

These findings suggest that whilst a wide range of DRG neuron phenotypes express CB₁, it is predominantly associated with myelinated fibres.

Section snippets

Experimental procedures

All experiments were performed using lumbar DRGs obtained from male Wistar rats (150–200 g) except for experiments performed on DRGs taken from CB1-knockout mice (CB₁^−/−) and wild type mice. CB₁^−/− mice and wild type mice were obtained from colonies maintained by Dr. D. Baker at the Institute of Neurology, London; the CB₁^−/− mice were derived from knockout-mice originally generated by Ledent et al. (1999). All experiments conformed to British Home Office regulations for laboratory animals. The

Distribution of cb₁ mrna

Both of the two in situ hybridisation probes used in this study produced identical results. For quantification a combination of the two probes was used to give a stronger signal. Addition of a 20-fold excess of unlabeled oligonucleotides to the hybridisation reaction mix effectively competed all specific binding of radiolabeled probe. Positive cells were indicated by clusters of silver grains, with specific labelling above background of CB₁ mRNA seen in 22.7±1.5% of the total cell profiles.

Discussion

In this study we have determined the expression profile of the CB₁ receptor in the DRG of naive rats using both in situ hybridisation and immunohistochemical techniques. In our results there appear to be some differences in the amount of co-expression of CB₁ with other markers using these two techniques detecting CB₁ mRNA and protein respectively (compare Table 1, Table 2). However, the general patterns and extent of co-expression are quite similar. Although we cannot rule out the possibility

Acknowledgements

The authors would like to thank Dr. D. Julius and Dr. Q. Yan for provision of antibodies to G.J.M. and Dr. D. Baker for provision of CB₁^−/− mice. G.J.M. would like to thank Dr. J. V. Priestley for support as well as encouragement throughout this project. This work is supported by grants from the Medical Research Council, the Association of Anaesthetists and the Wellcome Trust (grant 057058 awarded to M.R.E. and grant 058542 awarded to Dr. J. V. Priestley). Part of this study was presented at

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Localisation of cannabinoid receptor 1 in rat dorsal root ganglion using in situ hybridisation and immunohistochemistry

Abstract

Section snippets

Experimental procedures

Distribution of cb1 mrna

Discussion

Acknowledgements

Neuroscience

Neuroscience

Mol Cell Neurosci

Eur J Pharmacol

Mol Cell Neurol

Pain

Neurosci Lett

Neuroscience

Neuroscience

Brain Res

Neurosci Lett

Neurosci Lett

Pain

Neuroscience

Neurosci Lett

Neuron

Eur J Pharmacol

Pain

Neuroscience

J Biol Chem

Neuron

Neuroscience

mGlu5 receptors functionally expressed on peripheral sensory neurones mediate inflammatory hyperalgesia. Neuropharmacology

A distinct subgroup of small DRG cells express GDNF receptor components and GDNF is protective for these neurons after nerve injury

J Neurosci

Potent analgesic effects of GDNF in neuropathic pain states

Science

Distribution of cb₁ mrna