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Retrograde Tract-Tracing “Plus”: Adding Extra Value to Retrogradely Traced Neurons

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Neural Tracing Methods

Part of the book series: Neuromethods ((NM,volume 92))

Abstract

Classical neuroanatomical tract-tracing methods have formed the basis for most of our current understanding of brain circuits. However, to obtain a deeper knowledge of the main operational principles of the brain, the simple delineation of brain connectivity is not sufficient. This particularly holds true in regard to the analysis of connections within the diseased brain, for instance, the study of a number of major neurological disorders through the use of animal models. In other words, the information gathered from tract-tracing techniques is often too static, and recent findings in the fields of neurophysiology, receptor mapping, and neuroimaging (among others) need to be integrated within the context of structural data of brain connectivity as seen with neuroanatomical tracing techniques. During the past few years, our laboratory has pioneered a number of combinations of retrograde tracers with in situ hybridization, analyzing the changes in mRNA expression levels within brain circuits of interest. More recently, we have succeeded in combining a number of tract-tracing methods with a newly introduced technique known as in situ proximity ligation assay (PLA). The PLA technique is particularly well suited for the analysis of protein-protein interactions. This combination of methods enabled us to elucidate unequivocally the presence of GPCR heteromers within identified brain circuits and we strongly believe that this will soon become a popular approach in the field. Here we provide readers with a landscape view of these approaches, together with step-by-step protocols so that these methods may be easily reproduced even by inexperienced users.

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Acknowledgments

It is a great pleasure to acknowledge the continuous support received from colleagues, laboratory team members, and technicians. To mention just a few team members: Alberto J. Rico, Iria González-Dopeso, Salvador Sierra, Mónica Pérez-Manso, Pedro Barroso-Chinea, Natasha Luquin, Virginia Gómez-Bautista, Iciar P. López, Lorena Conte-Perales, Nancy Gonzalo, María Castle, and Elena Erro. Furthermore, I am also particularly indebted to my technician Elvira Roda who joined the laboratory more than 10 years ago and who still contributes continuously with new ideas and improvements. Moreover, it is also worth recognizing the extensive training that I received in the Department of Neuroanatomy and Neuropharmacology at the Amsterdam Vrije Universiteit under the guidance and mentorship of Floris G. Wouterlood, as well as from the laboratory technicians Barbara Jorritsma-Byham, Annaatje Pattiselanno, and Peter Goede. They all made my stay in Amsterdam (1992 and 1996) an easy, pleasant, and fruitful experience. Supported by grants from the Ministerio de Economía y Competitividad (BFU2012-37907, SAF2008-03118-E and SAF39875-C02-01), Eranet-Neuron (Heteropark), CiberNed (CB06/05/0006), Departamento de Salud, Gobierno de Navarra, and UTE project/Foundation for Applied Medical Research (FIMA). Salary for S.S. is partially supported by a grant from Mutual Médica.

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  1. Laboratory of Basal Ganglia Neuromorphology, Department of Neurosciences, Center for Applied Medical Research (CIMA and CIBERNED), University of Navarra, Pio XII Avenue 55, Pamplona, 31008, Spain

    José L. Lanciego

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  1. José L. Lanciego
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Correspondence to José L. Lanciego .

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  1. Baylor College of Medicine, Houston, Texas, USA

    Benjamin R. Arenkiel

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Lanciego, J.L. (2015). Retrograde Tract-Tracing “Plus”: Adding Extra Value to Retrogradely Traced Neurons. In: Arenkiel, B. (eds) Neural Tracing Methods. Neuromethods, vol 92. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1963-5_3

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  • DOI: https://doi.org/10.1007/978-1-4939-1963-5_3

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  • Publisher Name: Humana Press, New York, NY

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