Elsevier

Neuroscience

Volume 99, Issue 4, 23 August 2000, Pages 761-772
Neuroscience

In vivo localization and characterization of functional ciliary neurotrophic factor receptors which utilize JAK-STAT signaling

https://doi.org/10.1016/S0306-4522(00)90236-7Get rights and content

Abstract

The ciliary neurotrophic factor receptor is critically involved in embryonic motor neuron development. Postnatally, it may contribute to neuronal maintenance and regeneration. In addition, pharmacological stimulation of the receptor may slow the progression of several neurodegenerative disorders. The widespread nervous system expression of ciliary neurotrophic factor receptor components and the effects of low ciliary neurotrophic factor concentrations on a wide variety of cells in culture combine to suggest that functional ciliary neurotrophic factor receptors are expressed by many classes of neurons in vivo. However, the in vivo signaling properties and distribution of functional ciliary neurotrophic factor receptors have not been directly determined. We developed a novel in vivo assay of functional ciliary neurotrophic factor receptors which revealed that, in the adult nervous system, cranial and spinal motor neurons are very sensitive to ciliary neurotrophic factor and display a rapid, robust increase in phospho-STAT3 in their dendrites, cell bodies and nuclei, which is specifically blocked by the ciliary neurotrophic factor receptor antagonist, AADH-CNTF. In distinct contrast, several other classes of ciliary neurotrophic factor receptor expressing neurons fail to increase phospho-STAT3 levels following ciliary neurotrophic factor treatment, even when ciliary neurotrophic factor is applied at high concentrations. Leukemia inhibitory factor and epidermal growth factor elicit the same cell-type-dependent pattern of phospho-STAT3 increases. Responsive and non-responsive neurons express comparable levels of STAT3.

Therefore, in vivo ciliary neurotrophic factor receptor-initiated STAT3 signal transduction is regulated in a very cell-type-dependent manner. The present data suggest that at least some of this regulation occurs at the STAT3 tyrosine phosphorylation step. These unexpected results also suggest that other forms of receptor-initiated STAT3 signal transduction may be similarly regulated.

Section snippets

Experimental procedures

Adult, male (Sprague–Dawley, Harlan) rats (250–350 g) were anesthetized with xylazine/ketamine (1 ml/kg). CNTF (recombinant rat; PeproTech, Inc.), dissolved in 2 μl of isotonic saline, with 0.005% bovine serum albumin, was stereotaxically injected over an 8.5-min period with a Hamilton syringe (5 μl, 33-gauge needle) that was left in place for an additional 5 min after the injection. Stereotaxic coordinates (relative to bregma) were as follows: cerebellum (AP −10.3 mm, ML −3.0 mm, DV −3.5 mm), cerebral

A novel in vivo assay of functional ciliary neurotrophic factor receptors: characterization of receptors in facial motor neurons

In vitro studies indicate that a major form of CNTF receptor signal transduction involves the essential phosphorylation of STAT3 at Tyr705, which leads to its homodimerization, or heterodimerization with STAT1, and its subsequent activity as a transcription factor.24., 52., 56. We injected CNTF into various regions of the rat brain and spinal cord in order to visualize this signal transduction in the intact nervous system. Fixed tissue sections from the injected regions were

In vivo identification and characterization of somatic and dendritic ciliary neurotrophic factor receptor responses in motor neurons

In the experiments reported here, we developed and utilized an assay of in vivo CNTF receptor responses which immunohistochemically visualizes CNTF-induced STAT3 phosphorylation. The first series of experiments characterized facial motor neuron CNTF receptors. Later experiments demonstrated the same pattern of responding in other cranial motor neurons and spinal motor neurons. In all cases, CNTF elicited a large increase in pSTAT3 staining in nuclei, cell bodies and dendrites. This CNTF-induced

Acknowledgements

We thank Kevin Anderson, Dinali Fernando, Gregory Schrimsher and Don Walker for advice and support, and Alfred Chung and Jennifer Vaughn for technical assistance. This work was supported by NIH grants NS35224 and NS39127 (A.J.M.), the State of Florida Brain and Spinal Cord Injury Rehabilitation Trust Fund (A.J.M.) and the European Commission Biotechnology RTD Programme grant BI04-CT96-0433 (R.L.).

References (64)

  • J. Mey et al.

    Intravitreal injections of neurotrophic factors support the survival of axotomized retinal ganglion cells in adult rats in vivo

    Brain Res.

    (1993)
  • Y. Misumi et al.

    The expressions of epidermal growth factor receptor mRNA and protein gene product 9.5 in developing rat brain

    Devl Brain Res.

    (1998)
  • T.E. Nelson et al.

    Altered physiology of Purkinje neurons in cerebellar slices from transgenic mice with chronic central nervous system expression of interleukin-6

    Neuroscience

    (1999)
  • Z. Sahenk et al.

    CNTF potentiates peripheral nerve regeneration

    Brain Res.

    (1994)
  • T.-C. Wen et al.

    Ciliary neurotrophic factor prevents ischemia-induced learning disability and neuronal loss in gerbils

    Neurosci. Lett.

    (1995)
  • S. Agarwala et al.

    Long-term protection of axotomized neurons in the dorsal lateral geniculate nucleus in the rat following a single administration of basic fibroblast growth factor or ciliary neurotrophic factor

    J. comp. Neurol.

    (1998)
  • K.D. Anderson et al.

    Ciliary neurotrophic factor protects striatal output neurons in an animal model of Huntington disease

    Proc. natn. Acad. Sci. USA

    (1996)
  • C. Bjorbaek et al.

    Activation of SOCS-3 messenger ribonucleic acid in the hypothalamus by ciliary neurotrophic factor

    Endocrinology

    (1999)
  • R.E. Clatterbuck et al.

    Ciliary neurotrophic factor prevents retrograde neuronal death in the adult central nervous system

    Proc. natn. Acad. Sci. USA

    (1993)
  • M.J. Comb

    Phospho-specific antibodies: new tools to study protein phosphorylation

    NEB Transcript

    (1995)
  • S. Davis et al.

    LIFR beta and gp130 as heterodimerizing signal transducers of the tripartite CNTF receptor

    Science

    (1993)
  • A. DiMarco et al.

    Identification of ciliary neurotrophic factor (CNTF) residues essential for leukemia inhibitory factor receptor binding and generation of CNTF receptor antagonists

    Proc. natn. Acad. Sci. USA

    (1996)
  • D.F. Emerich et al.

    Protective effect of encapsulated cells producing neurotrophic factor CNTF in a monkey model of Huntington’s disease

    Nature

    (1997)
  • T.A. Endo et al.

    A new protein containing an SH2 domain that inhibits JAK kinases

    Nature

    (1997)
  • N. Ghilardi et al.

    Defective STAT signaling by the leptin receptor in diabetic mice

    Proc. natn. Acad. Sci. USA

    (1996)
  • T. Hagg et al.

    Ciliary neurotrophic factor prevents degeneration of adult rat substantia nigra dopaminergic neurons in vivo

    Proc. natn. Acad. Sci. USA

    (1993)
  • A.W. Ham
  • D.J. Hilton et al.

    Twenty proteins containing a C-terminal SOCS box form five structural classes

    Proc. natn. Acad. Sci. USA

    (1998)
  • J.D. Houle et al.

    Changes occur in the ability to promote axonal regeneration as the post-injury period increases

    NeuroReport

    (1997)
  • N.Y. Ip et al.

    Ciliary neurotrophic factor enhances neuronal survival in embryonic rat hippocampal culture

    J. Neurosci.

    (1991)
  • N.Y. Ip et al.

    The neurotrophins and CNTF: two families of collaborative neurotrophic factors

    A. Rev. Neurosci.

    (1996)
  • N. Jain et al.

    Repression of Stat3 activity by activation of mitogen-activated protein kinase (MAPK)

    Oncogene

    (1998)
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