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