RT Journal Article
T1 Benchmarking of tools for axon length measurement in individually-labeled projection neurons
A1 Rubio Teves, Mario
A1 Díez Hermano, Sergio
A1 Porrero, César
A1 Sánchez Jiménez, Abel
A1 Prensa Sepúlveda, Lucía
A1 Clascá, Francisco
A1 García Amado, María
A1 Villacorta Atienza, José Antonio
AB Projection neurons are the commonest neuronal type in the mammalian forebrain and their individual characterization is a crucial step to understand how neural circuitry operates. These cells have an axon whose arborizations extend over long distances, branching in complex patterns and/or in multiple brain regions. Axon length is a principal estimate of the functional impact of the neuron, as it directly correlates with the number of synapses formed by the axon in its target regions; however, its measurement by direct 3D axonal tracing is a slow and labor-intensive method. On the contrary, axon length estimations have been recently proposed as an effective and accessible alternative, allowing a fast approach to the functional significance of the single neuron. Here, we analyze the accuracy and efficiency of the most used length estimation tools—design-based stereology by virtual planes or spheres, and mathematical correction of the 2D projected-axon length—in contrast with direct measurement, to quantify individual axon length. To this end, we computationally simulated each tool, applied them over a dataset of 951 3D-reconstructed axons (from NeuroMorpho.org), and compared the generated length values with their 3D reconstruction counterparts. The evaluated reliability of each axon length estimation method was then balanced with the required human effort, experience and know-how, and economic affordability. Subsequently, computational results were contrasted with measurements performed on actual brain tissue sections. We show that the plane-based stereological method balances acceptable errors (~5%) with robustness to biases, whereas the projection-based method, despite its accuracy, is prone to inherent biases when implemented in the laboratory. This work, therefore, aims to provide a constructive benchmark to help guide the selection of the most efficient method for measuring specific axonal morphologies according to the particular circumstances of the conducted research.
PB Public Library of Science
SN 1553-734X
YR 2021
FD 2021-12-08
LK https://hdl.handle.net/20.500.14352/4909
UL https://hdl.handle.net/20.500.14352/4909
LA eng
NO Unión Europea. Horizonte 2020
NO Ministerio de Ciencia e Innovación (MICINN)
NO Fundación Bancaria “la Caixa"
DS Docta Complutense
RD 22 abr 2025