Optimization of whole-brain rabies virus tracing technology for small cell populations
Roelofs, Theresia J.M.; Menting-Henry, Shanice; Gol, Lieke M.; Speel, Annelijn M.; Wielenga, Vera H.; Garner, Keith M.; Luijendijk, Mieneke C.M.; Hennrich, Alexandru A.; Conzelmann, Karl Klaus; Adan, Roger A.H.
(2021) Scientific Reports, volume 11, issue 1
(Article)
Abstract
The lateral hypothalamus (LH) is critically involved in the regulation of homeostatic energy balance. Some neurons in the LH express receptors for leptin (LepRb), a hormone known to increase energy expenditure and decrease energy intake. However, the neuroanatomical inputs to LepRb-expressing LH neurons remain unknown. We used rabies virus tracing
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technology to map these inputs, but encountered non-specific tracing. To optimize this technology for a minor cell population (LepRb is not ubiquitously expressed in LH), we used LepRb-Cre mice and assessed how different titers of the avian tumor virus receptor A (TVA) helper virus affected rabies tracing efficiency and specificity. We found that rabies expression is dependent on TVA receptor expression, and that leakiness of TVA receptors is dependent on the titer of TVA virus used. We concluded that a titer of 1.0–3.0 × 107 genomic copies per µl of the TVA virus is optimal for rabies tracing. Next, we successfully applied modified rabies virus tracing technology to map inputs to LepRb-expressing LH neurons. We discovered that other neurons in the LH itself, the periventricular hypothalamic nucleus (Pe), the posterior hypothalamic nucleus (PH), the bed nucleus of the stria terminalis (BNST), and the paraventricular hypothalamic nucleus (PVN) are the most prominent input areas to LepRb-expressing LH neurons.
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Keywords: Animals, Avian Proteins/genetics, Connectome/methods, Female, Genetic Vectors/administration & dosage, Helper Viruses/genetics, Hypothalamus/cytology, Male, Mice, Mice, Transgenic, Microscopy, Fluorescence, Molecular Imaging/methods, Neurons/metabolism, Rabies virus/genetics, Receptors, Leptin/analysis, Receptors, Virus/genetics, Septal Nuclei/cytology, Stereotaxic Techniques, Journal Article, Research Support, Non-U.S. Gov't
ISSN: 2045-2322
Publisher: Nature Publishing Group
Note: Funding Information: This work was supported by the European Union Seventh Framework Program (FP/2007-2013) [Grant number 607310 (Nudge-it)]. The funding source had no involvement in the collection, analysis, or interpretation of the data, or in the writing of this article. We thank Inge Wolterink for assistance with vector injections. Publisher Copyright: © 2021, The Author(s).
(Peer reviewed)