Caracterización de neuronas marcadas retrógradamente en ganglios de la raíz dorsal y nervios periféricos transparentes de ratón

• Bernal null, Laura ^[1] ; Roza, Carolina
  1. [1] Universidad de Alcalá

     Universidad de Alcalá

     Alcalá de Henares, España

     
• Localización: Dianas: revista de dianas terapéuticas, ISSN-e 1886-8746, Vol. 7, Nº. 1, 2018 (Ejemplar dedicado a: Actas del III Congreso de Señalización Celular, SECUAH 2018 / coord. por María José Carmena Sierra, Alberto Domingo Galán)
• Idioma: español
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• Resumen

  • Traditional molecular characterization of neurons is based on doing immunohistochemistry in frozen sections, but this technique is slow and time-consuming. The use of confocal microscopy in un-sectioned tissue samples is not posible due to light scattering, an issue overcome in transparent samples. Several protocols have been developed in the last years, but none of them is adapted to peripheral nervous tissue. We modified the previous protocols for lipids-clearing for a new low-cost passive lipids-clearance suitable for labs with basic equipment. First, we showed that this passive technique is able to make transparent different tissues from the peripheral nervous system (dorsal root ganglia and peripheral nerves) in a short period of time (10-12 days). We successfully stablished an immunohistochemistry protocol for transparent tissue. We test the suitability of different retrograde tracers (DiI, fluoro-ruby, fluoro-emerald, fluoro-gold and retrobeads) to remain in the tissue after lipid-clearing. Moreover, characterization of retrograde labelled neurons by immunohistochemistry (against the nociceptors’ markers isolectin-IB4 and calcitonin gene-related peptide) is also possible.