Diverse Modes of Axon Elaboration in the Developing Neocortex

Carlos Portera-Cailliau,Robby M Weimer,Pico Caroni,Karel Svoboda,Vincenzo De Paola,Joshua R Sanes

doi:10.1371/journal.pbio.0030272

Abstract

The development of axonal arbors is a critical step in the establishment of precise neural circuits, but relatively little is known about the mechanisms of axonal elaboration in the neocortex. We used in vivo two-photon time-lapse microscopy to image axons in the neocortex of green fluorescent protein-transgenic mice over the first 3 wk of postnatal development. This period spans the elaboration of thalamocortical (TC) and Cajal-Retzius (CR) axons and cortical synaptogenesis. Layer 1 collaterals of TC and CR axons were imaged repeatedly over time scales ranging from minutes up to days, and their growth and pruning were analyzed. The structure and dynamics of TC and CR axons differed profoundly. Branches of TC axons terminated in small, bulbous growth cones, while CR axon branch tips had large growth cones with numerous long filopodia. TC axons grew rapidly in straight paths, with frequent interstitial branch additions, while CR axons grew more slowly along tortuous paths. For both types of axon, new branches appeared at interstitial sites along the axon shaft and did not involve growth cone splitting. Pruning occurred via retraction of small axon branches (tens of microns, at both CR and TC axons) or degeneration of large portions of the arbor (hundreds of microns, for TC axons only). The balance between growth and retraction favored overall growth, but only by a slight margin. Given the identical layer 1 territory upon which CR and TC axons grow, the differences in their structure and dynamics likely reflect distinct intrinsic growth programs for axons of long projection neurons versus local interneurons.

Highlights

In the developing mammalian brain, axons elaborate to form complex arbors that innervate specific target cells
We present our studies on the development of TC and CR axonal arbors in layer 1
green fluorescent protein (GFP) expression levels were sufficient for high-contrast in vivo two-photon imaging of individual axons in superficial cortical layers as early as postnatal day 3 (P3), and increased further thereafter (Figure 1)

Summary

Introduction

In the developing mammalian brain, axons elaborate to form complex arbors that innervate specific target cells. In the peripheral and central nervous systems, axons have an early phase of rapid growth, followed by a period of activitydependent pruning [1,2,3,4,5,6,7,8]. Time-lapse in vivo imaging has been critical for understanding axonal growth and refinement in lower vertebrates [14,15]. Relatively little is known about how axonal projections are refined within their target regions, at the level of a single cortical layer or column, or about the exact time course of growth and pruning. Even less is known about the elaboration of interneuron axonal arbors

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