Jennifer K. Ness

Perinatal Hypoxia-Ischemia Induces Apoptotic and Excitotoxic Death of Periventricular White Matter Oligodendrocyte Progenitors

Hypoxia-ischemia (HI) is a leading cause of white matter damage, a major contributor to cerebral palsy in premature infants. Preferential white matter damage is believed to result from vulnerability of the immature oligodendrocyte (the pro-OL) to factors elevated during ischemic damage, such as oxygen free radicals and glutamate. In order to determine whether pro-OLs undergo apoptotic death after HI, we analyzed periventricular white matter OLs in P7 rats 4, 12 and 24 h after HI to analyze the time course and mode of cell death. DNA fragmentation was seen at 12 and 24 h of recovery after HI, representing a 17-fold increase over control. In addition, caspase-3 activation was found in NG2+ pro-OLs at 12 h. Electron-microscopic analysis of cell death in the white matter revealed a transition from early necrotic deaths to hybrid cell deaths to classical apoptosis between 4 and 24 h of recovery from HI. The delayed time course of apoptosis in pro-OLs supports the feasibility of interventions to improve clinical outcomes for newborns surviving birth asphyxia.

Insulin-like Growth Factor I, but Not Neurotrophin-3, Sustains Akt Activation and Provides Long-Term Protection of Immature Oligodendrocytes from Glutamate-Mediated Apoptosis

Glutamate toxicity is a major contributor to death of oligodendroglia in diverse CNS disorders. The goal of these studies was to investigate the mechanisms of glutamate toxicity and trophic factor protection of the immature pro-oligodendroblast (pro-OL). Glutamate induced time- and dose-dependent DNA fragmentation and caspase-3 activation in pro-OLs. IGF-I or NT-3, but not CNTF, prevented apoptosis of pro-OLs by 24 h via a PI3-kinase-dependent pathway; however, only IGF-I protected pro-OLs from glutamate toxicity through 48 h. Long-term protection of pro-OLs by IGF-I was correlated with sustained activation of Akt while NT-3 activation of Akt was transient. The differential ability of IGF-I and NT-3 to maintain Akt activation was due to differences in receptor activation and stability. In the presence of NT-3, TrkC phosphorylation and protein expression decreased significantly while activation of the IGF-IR was maintained in the pro-OLs in the presence of IGF-I.

Astrocytes produce CNTF during the remyelination phase of viral-induced spinal cord demyelination to stimulate FGF-2 production.

Multiple sclerosis is characterized by multiple lesions with selective loss of myelin and oligodendrocytes, leading to deficits of sensation and movement, as well as cognitive disabilities. Consequently, a major research endeavor is to identify strategies to enhance oligodendrocyte regeneration and remyelination. FGF-2 is a potent mitogen for OPCs, and it is induced in astrocytes in animal models of demyelinating diseases in conjunction with successful remyelination. However, the factors responsible for inducing FGF-2 after demyelination in astrocytes are unknown. Here we show that CNTF mRNA and protein increase coincident with spinal cord remyelination in mice recovering from MHV-induced demyelination. We identify CNTF within astrocytes surrounding and within remyelinating lesions, and show that CNTF increases FGF-2 ligand and receptor mRNAs in spinal cord after direct application. Furthermore, we show that CNTF increases FGF-2 mRNA approximately 2.5-fold in cultured mouse spinal cord astrocytes. Altogether, these results strongly implicate CNTF as an important cytokine in demyelinating disease and as an upstream regulator of FGF-2 production in astrocytes during early remyelination.

IGF-I prevents glutamate-mediated bax translocation and cytochrome C release in O4+ oligodendrocyte progenitors.

Oligodendroglial death due to overactivation of the AMPA/kainate glutamate receptors is implicated in white matter damage in multiple CNS disorders. We previously demonstrated that glutamate induces caspase‐3 activation and death of the late oligodendrocyte progenitor known as the pro‐oligodendroblast (pro‐OL) via activation of the AMPA/kainate glutamate receptors. We also demonstrated that IGF‐I had the unique ability to sustain activation of Akt in the pro‐OL and provide long‐term protection of these cells from glutamate‐mediated apoptosis. The goal of these studies was to investigate the mechanisms of glutamate toxicity and IGF‐I‐mediated survival in the pro‐OL. IGF‐I prevented glutamate‐induced loss of mitochondrial membrane potential, cytochrome c release, and caspase‐9 activation. In contrast to IGF‐I mediated survival mechanisms in neurons, IGF‐I had no effect on the influx or recovery of intracellular calcium levels or on levels of major pro‐ and anti‐apoptotic molecules including Bax or Bcl‐xL. Rather, IGF‐I prevented the glutamate‐induced translocation of Bax to the mitochondria. Moreover, IGF‐I prevented caspase‐3 activation in pro‐OLs as long as 8 h after exposure of the cells to glutamate, suggesting that delayed activation of IGF‐I‐mediated survival pathways can block glutamate‐mediated apoptosis in pro‐OLs. The results of these experiments define the mechanisms by which glutamate kills oligodendrocyte progenitor cells and by which IGF‐I blocks glutamate‐induced apoptosis in these cells. The data also demonstrate that IGF‐I disrupts the glutamate‐mediated apoptotic pathway in the pro‐OL through mechanisms that are distinct from its survival‐promoting actions in neurons.

IGF-I and NT-3 Signaling Pathways in Developing Oligodendrocytes: Differential Regulation and Activation of Receptors and the Downstream Effector Akt

A previous study from our laboratory demonstrated differences in the ability of insulin-like growth factor-I (IGF-I) and neurotrophin-3 (NT-3) to promote survival of pro-oligodendroblasts (pro-OLs) against glutamate-mediated apoptosis. In the current study, we tested whether submaximal concentrations of NT-3 would maintain receptor tyrosine kinase TrkC activation and Akt phosphorylation and thus promote long-term survival of the pro-OL against glutamate. Our results demonstrate that NT-3 at any concentration sufficient to activate the TrkC receptor results in a transient phosphorylation of the receptor and of Akt due, in part, to downregulation of the Trk receptor. In contrast, even submaximal IGF-I concentrations maintain long-term Akt activation and prevent glutamate-mediated apoptosis in pro-OLs. In addition, we also present data showing that IGF-I and NT-3 differentially activate their receptors and Akt depending on the maturational stage of the oligodendrocyte.

Lck tyrosine kinase mediates β1-integrin signalling to regulate Schwann cell migration and myelination

The interaction between laminin and β1-integrin on the surface of Schwann cells regulates Schwann cell proliferation, maturation and differentiation. However, the signalling mediators that fine-tune these outcomes are not fully elucidated. Here we show that lymphoid cell kinase is the crucial effector of β1-integrin signalling in Schwann cells. Lymphoid cell kinase is activated after laminin treatment of Schwann cells, while downregulation of β1-integrin with short interfering RNAs inhibits lymphoid cell kinase phosphorylation. Treatment of Schwann cells with a selective lymphoid cell kinase inhibitor reveals a pathway that involves paxillin and CrkII, which ultimately elevates Rac-GTP levels to induce radial lamellipodia formation. Inhibition of lymphoid cell kinase in Schwann cell-dorsal root ganglion cocultures and dorsal root ganglions from Lck−/− mice show a reduction of Schwann cell longitudinal migration, reduced myelin formation and internode length. Finally, Lck−/− mice exhibit delays in myelination, thinner myelin with abnormal g-ratios and aberrant myelin outfoldings. Our data implicate lymphoid cell kinase as a major regulator of cytoskeletal dynamics, migration and myelination in the peripheral nervous system.

Nuc-ErbB3 regulates H3K27me3 levels and HMT activity to establish epigenetic repression during peripheral myelination

Nuc‐ErbB3 an alternative transcript from the ErbB3 locus binds to a specific DNA motif and associates with Schwann cell chromatin. Here we generated a nuc‐ErbB3 knockin mouse that lacks nuc‐ErbB3 expression in the nucleus without affecting the neuregulin‐ErbB3 receptor signaling. Nuc‐ErbB3 knockin mice exhibit hypermyelination and aberrant myelination at the paranodal region. This phenotype is attributed to de‐repression of myelination associated gene transcription following loss of nuc‐ErbB3 and histone H3K27me3 promoter occupancy. Nuc‐ErbB3 knockin mice exhibit reduced association of H3K27me3 with myelination‐associated gene promoters and increased RNA Pol‐II rate of transcription of these genes. In addition, nuc‐ErbB3 directly regulates levels of H3K27me3 in Schwann cells. Nuc‐ErbB3 knockin mice exhibit significant decrease of histone H3K27me3 methyltransferase (HMT) activity and reduced levels of H3K27me3. Collectively, nuc‐ErbB3 is a master transcriptional repressor, which regulates HMT activity to establish a repressive chromatin landscape on promoters of genes during peripheral myelination. GLIA 2016;64:977–992

IGF-I and Brain Growth: Multifarious Effects on Developing Neural Cells and Mechanisms of Action

Numerous investigators have provided data supporting an essential role for IGF-I in growth of the brain. IGF-I contributes to multiple processes during brain development, including neural cell survival, proliferation, differentiation and maturation. The IGF type I receptor (IGF-IR) is present on all cell types in the brain, and IGF-I has known actions on neural stem and progenitor cells as well as neurons and glia. IGF-I is highly expressed throughout the brain during development, and its expression is retained in the meninges and in many cell types in the adult brain. While IGF-I has multiple actions on developing neural cells, very few studies have addressed the mechanisms or pathways by which IGF-I mediates these multiple effects. The goal of this chapter is to briefly review data on IGF-I in the developing brain and then to discuss more recent studies that focus on the mechanisms for its varied actions.