Striatal medium-sized spiny neurons: identification by nuclear staining and study of neuronal subpopulations in BAC transgenic mice.

Matamales, Miriam, Bertran-Gonzalez, Jesus, Salomon, Lucas, Degos, Bertrand, Deniau, Jean-Michel, Valjent, Emmanuel, Hervé, Denis and Girault, Jean-Antoine (2009) Striatal medium-sized spiny neurons: identification by nuclear staining and study of neuronal subpopulations in BAC transgenic mice.. PLoS One, 4 3: e4770.1-e4770.11. doi:10.1371/journal.pone.0004770


Author Matamales, Miriam
Bertran-Gonzalez, Jesus
Salomon, Lucas
Degos, Bertrand
Deniau, Jean-Michel
Valjent, Emmanuel
Hervé, Denis
Girault, Jean-Antoine
Title Striatal medium-sized spiny neurons: identification by nuclear staining and study of neuronal subpopulations in BAC transgenic mice.
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2009-03
Sub-type Article (original research)
DOI 10.1371/journal.pone.0004770
Open Access Status DOI
Volume 4
Issue 3
Start page e4770.1
End page e4770.11
Total pages 11
Place of publication San Francisco United States
Publisher Public Library of Science
Language eng
Formatted abstract
Precise identification of neuronal populations is a major challenge in neuroscience. In the striatum, more than 95% of neurons are GABAergic medium-sized spiny neurons (MSNs), which form two intermingled populations distinguished by their projections and protein content. Those expressing dopamine D1-receptors (D1Rs) project preferentially to the substantia nigra pars reticulata (SNr), whereas those expressing dopamine D2- receptors (D2Rs) project preferentially to the lateral part of the globus pallidus (LGP). The degree of segregation of these populations has been a continuous subject of debate, and the recent introduction of bacterial artificial chromosome (BAC) transgenic mice expressing fluorescent proteins driven by specific promoters was a major progress to facilitate striatal neuron identification. However, the fraction of MSNs labeled in these mice has been recently called into question, casting doubt on the generality of results obtained with such approaches. Here, we performed an in-depth quantitative analysis of striatal neurons in drd1a-EGFP and drd2-EGFP mice. We first quantified neuronal and non-neuronal populations in the striatum, based on nuclear staining with TO-PRO-3, and immunolabeling for NeuN, DARPP-32 (dopamine- and cAMP-regulated phosphoprotein Mr~32,000), and various markers for interneurons. TO-PRO-3 staining was sufficient to identify MSNs by their typical nuclear morphology and, with a good probability, interneuron populations. In drd1a-EGFP/drd2-EGFP double transgenic mice all MSNs expressed EGFP, which was driven in about half of them by drd1a promoter. Retrograde labeling showed that all MSNs projecting to the SNr expressed D1R and very few D2R (<1%). In contrast, our results were compatible with the existence of some D1R-EGFP-expressing fibers giving off terminals in the LGP. Thus, our study shows that nuclear staining is a simple method for identifying MSNs and other striatal neurons. It also unambiguously confirms the degree of segregation of MSNs in the mouse striatum and allows the full exploitation of results obtained with BAC-transgenic mice.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Queensland Brain Institute Publications
 
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Created: Wed, 30 Jul 2014, 09:44:36 EST by J Bertran Gonzalez on behalf of Clem Jones Centre for Ageing Dementia Research