Nigel P. Pringle

A role for platelet-derived growth factor in normal gliogenesis in the central nervous system

The bipotential progenitor cells (O-2A progenitors) that produce oligodendrocytes and type-2 astrocytes in the developing rat optic nerve are induced to proliferate in culture by type-1 astrocytes. Here, we show that the astrocyte-derived mitogen is platelet-derived growth factor (PDGF). PDGF is a potent mitogen for O-2A progenitor cells in vitro. Mitogenic activity in astrocyte-conditioned medium comigrates with PDGF on a size-exclusion column, competes with PDGF for receptors, and is neutralized by antibodies to PDGF. PDGF dimers can be immunoprecipitated from astrocyte-conditioned medium, and mRNA encoding PDGF is present in rat brain throughout gliogenesis. We propose that astrocyte-derived PDGF is crucial for the control of myelination in the developing central nervous system.

PDGF A chain homodimers drive proliferation of bipotential (O-2A) glial progenitor cells in the developing rat optic nerve.

The bipotential glial progenitor cells (O‐2A progenitors), which during development of the rat optic nerve give rise to oligodendrocytes and type 2 astrocytes, are stimulated to divide in culture by platelet‐derived growth factor (PDGF), and there is evidence that PDGF is important for development of the O‐2A cell lineage in vivo. We have visualized PDGF mRNA in the rat optic nerve by in situ hybridization, and its spatial distribution is compatible with the idea that type 1 astrocytes are the major source of PDGF in the nerve. We can detect mRNA encoding the A chain, but not the B chain of PDGF in the brain and optic nerve, suggesting that the major form of PDGF in the central nervous system is a homodimer of A chains (PDGF‐AA). PDGF‐AA is a more potent mitogen for O‐2A progenitor cells than is PDGF‐BB, while the reverse is true for human or rat fibroblasts. Fibroblasts display two types of PDGF receptors, type A receptors which bind to all three dimeric isoforms of PDGF, and type B receptors which bind PDGF‐BB and PDGF‐AB, but have low affinity for PDGF‐AA. Our results suggest that O‐2A progenitor cells possess predominantly type A receptors, and proliferate during development in response to PDGF‐AA secreted by type 1 astrocytes.

Oligodendrocyte Lineage and the Motor Neuron Connection

One of the more surprising recent discoveries in glial biology has been that oligodendrocytes (OLs) originate from very restricted regions of the embryonic neural tube. This was surprising because myelinating OLs are widespread in the mature central nervous system, so there was no reason to suspect that their precursors should be restricted. What we now know about early OL development suggests that they might have as much (or more) in common with ventral neurons—specifically motor neurons (MNs)—as with other types of glia. This has implications for the way we think about glial development, function, and evolution. In this article we review the evidence for a shared MN‐OL lineage and debate whether this is the only lineage that generates OLs. We decide in favour of a single embryonic lineage with regional variations along the anterior‐posterior neuraxis. GLIA 29:136–142, 2000.

Origins of Spinal Cord Oligodendrocytes: Possible Developmental and Evolutionary Relationships with Motor Neurons

Spinal cord oligodendrocytes develop from migratory glial progenitor cells that are generated by a small subset of neuroepithelial cells in the ventral part of the neural tube. Specification of these neuroepithelial oligodendrocyte precursors, in common with other ventral cells such as motor neurons, depends on morphogenetic signals from the notochord and/or floor plate. The ventrally derived signals can be mimicked in vitro by purified Sonic hedgehog (Shh) protein. Oligodendrocytes and motor neurons are induced over the same range of concentrations of Shh, consistent with the idea that Shh might specify a common precursor of motor neurons and oligodendrocytes. A lineage relationship between motor neurons and oligodendrocytes has previously been suggested by clonal analysis in the embryonic chick spinal cord. We propose a lineage diagram that connects oligodendrocytes and motor neurons and that takes into account the fact that motor neurons and oligodendrocyte precursors are generated at different times during development. Oligodendrocytes might originally have evolved to ensheath motor axons and facilitate a rapid escape response. If so, oligodendrocyte ontogeny and phylogeny might share a common basis.

Insulin-like growth factor I in cultured rat astrocytes: expression of the gene, and receptor tyrosine kinase.

Gene expression, receptor binding and growth‐promoting activity of insulin‐like growth factor I (IGF I) was studied in cultured astrocytes from developing rat brain. Northern blot analysis of poly(A)+ RNAs from astrocytes revealed an IGF I mRNA of 1.9 kb. Competitive binding and receptor labelling techniques revealed two types of IGF receptor in astroglial cells. Type I IGF receptors consist of alpha‐subunits (Mr 130,000) which bind IGF I with significantly higher affinity than IGF II, and beta‐subunits (Mr 94,000) which show IGF I‐sensitive tyrosine kinase activity. Type II IGF receptors are monomers (Mr 250,000) which bind IGF II with three times higher affinity than IGF I. Both types of IGF receptor recognize insulin weakly. DNA synthesis measured by cellular thymidine incorporation was stimulated 2‐fold by IGF I and IGF II. IGF I was more potent than IGF II, and both were significantly more potent than insulin. Our findings suggest that IGF I is synthesized in fetal rat astrocytes and acts as a growth promoter for the same cells by activation of the type I IGF receptor tyrosine kinase. We propose that IGF I acts through autocrine or paracrine mechanisms to stimulate astroglial cell growth during normal brain development.

Fgfr3 expression by astrocytes and their precursors: evidence that astrocytes and oligodendrocytes originate in distinct neuroepithelial domains

The postnatal central nervous system (CNS) contains many scattered cells that express fibroblast growth factor receptor 3 transcripts (Fgfr3). They first appear in the ventricular zone (VZ) of the embryonic spinal cord in mid-gestation and then distribute into both grey and white matter — suggesting that they are glial cells, not neurones. The Fgfr3+ cells are interspersed with but distinct from platelet-derived growth factor receptor α (Pdgfra)-positive oligodendrocyte progenitors. This fits with the observation that Fgfr3 expression is preferentially excluded from the pMN domain of the ventral VZ where Pdgfra+ oligodendrocyte progenitors — and motoneurones — originate. Many glial fibrillary acidic protein (Gfap)- positive astrocytes co-express Fgfr3 in vitro and in vivo. Fgfr3+ cells within and outside the VZ also express the astroglial marker glutamine synthetase (Glns). We conclude that (1) Fgfr3 marks astrocytes and their neuroepithelial precursors in the developing CNS and (2) astrocytes and oligodendrocytes originate in complementary domains of the VZ. Production of astrocytes from cultured neuroepithelial cells is hedgehog independent, whereas oligodendrocyte development requires hedgehog signalling, adding further support to the idea that astrocytes and oligodendrocytes can develop independently. In addition, we found that mice with a targeted deletion in the Fgfr3 locus strongly upregulate Gfap in grey matter (protoplasmic) astrocytes, implying that signalling through Fgfr3 normally represses Gfap expression in vivo.

DORSAL SPINAL CORD NEUROEPITHELIUM GENERATES ASTROCYTES BUT NOT OLIGODENDROCYTES

There is evidence that oligodendrocytes in the spinal cord are derived from a restricted part of the ventricular zone near the floor plate. An alternative view is that oligodendrocytes are generated from all parts of the ventricular zone. We reinvestigated glial origins by constructing chick-quail chimeras in which dorsal or ventral segments of the embryonic chick neural tube were replaced with equivalent segments of quail neural tube. Ventral grafts gave rise to both oligodendrocytes and astrocytes. In contrast, dorsal grafts produced astrocytes but not oligodendrocytes. In mixed cultures of ventral and dorsal cells, only ventral cells generated oligodendrocytes, whereas both ventral and dorsal cells generated astrocytes. Therefore, oligodendrocytes are derived specifically from ventral neuroepithelium, and astrocytes from both dorsal and ventral.

Pax6 influences the time and site of origin of glial precursors in the ventral neural tube.

Neuroepithelial precursors in the ventral ventricular zone (VZ) of the spinal cord generate motor neurons (MNs) and interneurons, and then a subset of precursors starts to produce oligodendrocyte progenitors (OLPs). We show that OLPs originate in the ventral-most part of the Pax6-positive VZ, which at earlier times generates somatic (Isl2/Lim3-positive) MNs. In Small eye (Pax6-deficient) mice, the origin of OLPs is shifted dorsally and both OLPs and Isl2/Lim3 MNs are delayed. We suggest that somatic MNs and OLPs are generated sequentially from a common set of MN-OL precursors whose position in the VZ is influenced by Pax6. Neuron-glia fate switching might be a preprogrammed property of these precursors or a response to feedback from newly generated neurons. OLs developed normally in explants of Isl1(-/-) spinal cords, which lack MNs, arguing against feedback control and suggesting that the neuron-glia switch is an intrinsic developmental program in a specific subset of neural precursors.

Specification of CNS glia from neural stem cells in the embryonic neuroepithelium

All the neurons and glial cells of the central nervous system are generated from the neuroepithelial cells in the walls of the embryonic neural tube, the ‘embryonic neural stem cells’. The stem cells seem to be equivalent to the so-called ‘radial glial cells’, which for many years had been regarded as a specialized type of glial cell. These radial cells generate different classes of neurons in a position-dependent manner. They then switch to producing glial cells (oligodendrocytes and astrocytes). It is not known what drives the neuron–glial switch, although downregulation of pro-neural basic helix–loop–helix transcription factors is one important step. This drives the stem cells from a neurogenic towards a gliogenic mode. The stem cells then choose between developing as oligodendrocytes or astrocytes, of which there might be intrinsically different subclasses. This review focuses on the different extracellular signals and intracellular responses that influence glial generation and the choice between oligodendrocyte and astrocyte fates.

Ventral Neurogenesis and the Neuron-Glial Switch

In the developing spinal cord, neuroepithelial precursors at different positions along the dorsal-ventral axis generate distinct neuronal and glial subtypes. For example, one group of ventral precursors generates neurons followed by oligodendrocytes. A spate of recent articles, including several in this issue of Neuron, are devoted to the mechanisms governing neuronal and glial subtype specification in the ventral cord. We review these studies and discuss the nature of the ventral neuron-oligodendrocyte switch.