In Vitro and In Vivo Analysis of Neuronal Polarity

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2024 Aug 12

PMID 39134847

Authors

Mini Jose

Affiliations

Soon will be listed here.

Abstract

Neuronal development is characterized by the unidirectional flow of signal from the axon to the dendrites via synapses. Neuronal polarization is a critical step during development that allows the specification of the different neuronal processes as a single axon and multiple dendrites both structurally and functionally, allowing the unidirectional flow of information. Along with extrinsic and intrinsic signaling, a whole network of molecular complexes involved in positive and negative feedback loops play a major role in this critical distinction of neuronal processes. As a result, neuronal morphology is drastically altered during establishment of polarity. In this chapter, we discuss how we can analyze the morphological alterations of neurons in vitro in culture to assess the development and polarity status of the neuron. We also discuss how these studies can be conducted in vivo, where polarity studies pose a greater challenge with promising results for addressing multiple pathological conditions. Our experimental model is limited to rodent hippocampal/cortical neurons in culture and cortical neurons in brain tissues, which are well-characterized model systems for understanding neuronal polarization.

References

Dotti C, Banker G . Experimentally induced alteration in the polarity of developing neurons. Nature. 1987; 330(6145):254-6. DOI: 10.1038/330254a0. View

Schelski M, Bradke F . Neuronal polarization: From spatiotemporal signaling to cytoskeletal dynamics. Mol Cell Neurosci. 2017; 84:11-28. DOI: 10.1016/j.mcn.2017.03.008. View

Toriyama M, Shimada T, Kim K, Mitsuba M, Nomura E, Katsuta K . Shootin1: A protein involved in the organization of an asymmetric signal for neuronal polarization. J Cell Biol. 2006; 175(1):147-57. PMC: 2064506. DOI: 10.1083/jcb.200604160. View

Yoshimura T, Arimura N, Kawano Y, Kawabata S, Wang S, Kaibuchi K . Ras regulates neuronal polarity via the PI3-kinase/Akt/GSK-3beta/CRMP-2 pathway. Biochem Biophys Res Commun. 2005; 340(1):62-8. DOI: 10.1016/j.bbrc.2005.11.147. View

Cheng P, Song A, Wong Y, Wang S, Zhang X, Poo M . Self-amplifying autocrine actions of BDNF in axon development. Proc Natl Acad Sci U S A. 2011; 108(45):18430-5. PMC: 3215064. DOI: 10.1073/pnas.1115907108. View

Shelly M, Lim B, Cancedda L, Heilshorn S, Gao H, Poo M . Local and long-range reciprocal regulation of cAMP and cGMP in axon/dendrite formation. Science. 2010; 327(5965):547-52. DOI: 10.1126/science.1179735. View

Leterrier C, Clerc N, Rueda-Boroni F, Montersino A, Dargent B, Castets F . Ankyrin G Membrane Partners Drive the Establishment and Maintenance of the Axon Initial Segment. Front Cell Neurosci. 2017; 11:6. PMC: 5266712. DOI: 10.3389/fncel.2017.00006. View

Leterrier C, Dubey P, Roy S . The nano-architecture of the axonal cytoskeleton. Nat Rev Neurosci. 2017; 18(12):713-726. DOI: 10.1038/nrn.2017.129. View

Leterrier C, Potier J, Caillol G, Debarnot C, Rueda Boroni F, Dargent B . Nanoscale Architecture of the Axon Initial Segment Reveals an Organized and Robust Scaffold. Cell Rep. 2015; 13(12):2781-93. DOI: 10.1016/j.celrep.2015.11.051. View

10.

Jose M, Sivanand A, Channakeshava C . Membrane Cholesterol Is a Critical Determinant for Hippocampal Neuronal Polarity. Front Mol Neurosci. 2021; 14:746211. PMC: 8566733. DOI: 10.3389/fnmol.2021.746211. View

11.

Miyata T, Kawaguchi A, Okano H, Ogawa M . Asymmetric inheritance of radial glial fibers by cortical neurons. Neuron. 2001; 31(5):727-41. DOI: 10.1016/s0896-6273(01)00420-2. View

12.

Nadarajah B . Radial glia and somal translocation of radial neurons in the developing cerebral cortex. Glia. 2003; 43(1):33-36. DOI: 10.1002/glia.10245. View

13.

Namba T, Funahashi Y, Nakamuta S, Xu C, Takano T, Kaibuchi K . Extracellular and Intracellular Signaling for Neuronal Polarity. Physiol Rev. 2015; 95(3):995-1024. DOI: 10.1152/physrev.00025.2014. View

14.

Banker G, Goslin K . Developments in neuronal cell culture. Nature. 1988; 336(6195):185-6. DOI: 10.1038/336185a0. View

15.

Meyer-Dilhet G, Courchet J . In Utero Cortical Electroporation of Plasmids in the Mouse Embryo. STAR Protoc. 2020; 1(1):100027. PMC: 7357676. DOI: 10.1016/j.xpro.2020.100027. View

16.

Binley K, Ng W, Tribble J, Song B, Morgan J . Sholl analysis: a quantitative comparison of semi-automated methods. J Neurosci Methods. 2014; 225:65-70. DOI: 10.1016/j.jneumeth.2014.01.017. View