Expression Of Ciliary Neurotrophic Factor Research Articles

Dose-Dependent Neuroprotective Effect of Ciliary Neurotrophic Factor Delivered via Tetracycline-Regulated Lentiviral Vectors in the Quinolinic Acid Rat Model of Huntington's Disease

The ability to regulate gene expression constitutes a prerequisite for the development of gene therapy strategies aimed at the treatment of neurologic disorders. In the present work, we used tetracycline (Tet)-regulated lentiviral vectors to investigate the dose-dependent neuroprotective effect of human ciliary neurotrophic factor (CNTF) in the quinolinic acid (QA) model of Huntington's disease (HD). The Tet system was split in two lentiviruses, the first one containing the CNTF or green fluorescent protein (GFP) cDNAs under the control of the Tet-response element (TRE) and a second vector encoding the transactivator (tTA). Preliminary coinfection study demonstrated that 63.8% +/- 2.0% of infected cells contain at least two viral copies. Adult rats were then injected with CNTF- and GFP-expressing viral vectors followed 3 weeks later by an intrastriatal administration of QA. A significant reduction of apomorphine-induced rotations was observed in the CNTF-on group. In contrast, GFP-treated animals or CNTF-off rats displayed an ipsilateral turning behavior in response to apomorphine. A selective sparing of DARPP-32-, choline acetyltransferase (ChAT)-, and NADPH-d-positive neurons was observed in the striatum of CNTF-on rats compared to GFP animals and CNTF-off group. Enzyme-linked immunosorbent assay (ELISA) performed on striatal samples of rats sacrificed at the same time point indicated that this neuroprotective effect was associated with the production of 15.5 +/- 4.7 ng CNTF per milligram of protein whereas the residual CNTF expression in the off state (0.54 +/- 0.02 ng/mg of protein) was not sufficient to protect against QA toxicity. These results establish the proof of principle of neurotrophic factor dosing for neurodegenerative diseases and demonstrate the feasibility of lentiviral-mediated tetracycline-regulated gene transfer in the brain.

Retinoic acid enhances leukemia inhibitory factor expression in OLN-93 oligodendrocytes

Brain injuries trigger physiological reactions which are mediated by a number of cytokines including interleukin-6 (IL-6), ciliary neurotrophic factor (CNTF), and leukemia inhibitory factor (LIF). Astrocytes and microglia, the protagonists in these traumatic responses, are known to secrete a variety of paracrine signals. Oligodendrocytes are involved as well and constitute another possible source of cytokines. Here we show the expression of IL-6, CNTF, and LIF in OLN-93 cells, derived from rat oligodendrocyte primary cultures. While differential gene transcription after injury has been described for many cytokines, the regulation of these physiological responses is unknown in many instances. Recent experiments indicate that the transcriptional activator retinoic acid (RA) plays a role in peripheral nerve regeneration. Transcripts of the retinoic acid receptors and the retinoid X receptors were also detected in OLN-93 oligodendrocytes. Using quantitative RT-PCR, we have therefore investigated the effect of RA on the expression of neuropoietic cytokines in these cells. Treatment with 1 microM all- trans RA for 24 h increased the mRNA concentration of LIF by a factor of 3.1 ( P<0.01). In contrast, RA had no significant effect on the expression of CNTF. The results suggest RA as a possible regulator of cytokine signaling in the CNS.

CNTF is up‐regulated in astrocytes during spinal cord remyelination and stimulates FGF‐2 production

FGF‐2 is a potent mitogen and survival factor for oligodendrocyte progenitors in vitro, and it is induced in astrocytes in conjunction with successful remyelination during recovery from murine hepatitis virus (MHV) induced spinal cord demyelination. However, the factor(s) responsible for inducing FGF‐2 after demyelination in astrocytes are unknown. Here we used in vivo and in vitro paradigms to assess whether ciliary neurotrophic factor (CNTF) could be responsible for FGF‐2 induction during remyelination. In mice recovering from MHV‐induced demyelination, increased CNTF expression coincided with spinal cord remyelination. Immunohistochemistry demonstrated that CNTF protein was present within astrocytes surrounding and within remyelinating lesions. After directly injecting CNTF into the spinal cord, FGF‐2 mRNA increased in spinal cord astrocytes. Furthermore, CNTF induced FGF‐2 mRNA over 2.5 fold in cultured mouse spinal cord astrocytes as determined by kinetic RT–PCR analysis. Although IL‐1β has been implicated in FGF‐2 production, IL‐1β mRNA was not detected during remyelination, nor did IL‐1β induce FGF‐2 mRNA from mouse spinal cord astrocytes in vitro. Altogether, these results strongly implicate CNTF as an upstream positive regulator of FGF‐2 production in astrocytes during early remyelination.Acknowledgements: Supported by NMSS Award #RG 3837 (SWL) and NS 33316 (RCA).

CNTF is specifically expressed in developing rat pineal gland and eyes.

Ciliary neurotrophic factor (CNTF) attracts considerable attention because it supports survival and differentiation of various types of neurons and glial cells in vitro. Although CNTF functions as a moderate neurotrophic factor in mature motor neurons, its role in embryonic development remains unknown. Here, we found a specific CNTF expression in the rat pineal gland and eyes during embryonic development. In vitro, neonatal rat pineal extract including CNTF supported the survival of neonatal sympathetic neurons, which innervate pineal glands immediately after birth.

Development of Normal and Injury-induced Gene Expression of aFGF, bFGF, CNTF, BDNF, GFAP and IGF-I in the Rat Retina

Focal mechanical injury to the retina substantially increases basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF) mRNA expression, accompanied by a transient increase in FGFR-1 mRNA, and this response is thought to protect photoreceptors near the injury site from inherited and light-induced retinal degenerations. We have now examined retinal gene expression of the principal survival factors involved in the response to injury in normal rats as a function of postnatal age both in normal and injured retinas. Sprague–Dawley rats were injured in one eye by needle incision through the retina at postnatal day (P) 10, 22, 35, 60, 90, 120 and 180. The other eye was uninjured and served as the control. Retinas were taken 1 day post-injury. Northern blot analysis was performed to determine the mRNA expression of the following factors and receptors: bFGF and acidic fibroblast growth factors (aFGF) and FGF receptor-1 (FGFR-1); CNTF and CNTF receptor alpha (CNTFR-alpha); brain-derived neurotrophic factor (BDNF) and its receptor trk B; and insulin-like growth factor-I (IGF-I) and IGF-I receptor (IGF-IR); glial fibrillary acidic protein (GFAP) and opsin. In the uninjured control eyes, mRNA expression of most of the factors increased with postnatal age, with little expression at P10 and maximal expression levels reached at P22 (opsin), P35 (aFGF), P60 (BDNF) or P90 (bFGF, FGFR-1, CNTF and GFAP). In contrast, IGF-1 mRNA rapidly decreased from a high level of expression at P10 to about 55% of that level by P22, reaching a stable 45–50% of the P10 level at P35 and thereafter. The response to injury of bFGF, FGFR-1, CNTF and GFAP mRNAs increased with postnatal age. Unexpectedly, only minimal increases in bFGF, FGFR-1, CNTF and GFAP over those seen in the control eyes were observed before P35. Thereafter, the increase of bFGF mRNA after injury reached a maximum of three-fold at P60, maintained this level to P120, and slightly decreased to 2.5-fold by P180. Expression of FGFR-1 mRNA showed a maximum increase of 2.6-fold at P90. Expression of CNTF and GFAP mRNAs followed a time course similar to that of bFGF. Mechanical injury did not alter the mRNA levels of aFGF, BDNF, IGF-I, and receptors, CNTFR-alpha, trk B and IGF-IR. These data show that the response to injury is minimal at early postnatal ages but increases with age and peaks at P60–90 for most potential survival factors.

Cellular and Subcellular Patterns of Expression of bFGF and CNTF in the Normal and Light Stressed Adult Rat Retina

This study compared the distributions in the normal and light stressed rat retina of the neuroprotective factors bFGF (basic fibroblast growth factor) and CNTF (ciliary neurotrophic factor). Albino Sprague–Dawley rats were raised in cyclic light and some were exposed to bright continuous light for 48hr, to induce light damage of photoreceptors. Their retinas were prepared as cryosections, immunolabelled with antibodies to bFGF and CNTF and analysed by confocal microscopy. Both factors were prominent in macroglial cells (astrocytes, Müller cells) and the retinal pigment epithelium (RPE). In the somas of these cells the distributions of the two factors were complementary, with bFGF concentrated in the nuclei and CNTF in the cytoplasm. Both factors were distributed along the processes of macroglial cells, in granular form. CNTF was not detected in neurones, but bFGF was consistently present in the cytoplasm of ganglion cell somas and, in regions of retina subject to stress, in the cytoplasm of photoreceptors. bFGF was not detected in the nuclei or processes of neurones. In retina stressed by light exposure or proximity to the anterior edge of the retina, the levels of bFGF and CNTF were up-regulated, without major changes in localization. Macroglial cells (Müller cells, astrocytes) play a major role in distributing bFGF and CNTF throughout the retina. The different localizations of the two factors within the somas of macroglial, RPE and photoreceptor cells, suggest that their protective actions are exerted by distinctive mechanisms.

Inhibition of Ras extracellular-signal-regulated kinase (ERK) mediated signaling promotes ciliary neurotrophic factor (CNTF) expression in Schwann cells.

Ciliary neurotrophic factor (CNTF) can prevent injury-induced motor neuron death. However, it is also evident that expression of CNTF in Schwann cells is suppressed during nerve regeneration. In this report, we have addressed the mechanism underlying the down-regulation of CNTF expression in injured nerves using a mouse Schwann cell line IMS32 and mouse sciatic nerve. In IMS32 cells, activation of the Ras extracellular-signal-regulated kinase (ERK) pathway by adenoviral vector-mediated expression of dominant active MEK1 did not alter a basal level of CNTF expression, whereas inhibition of the Ras-ERK pathway by using adenoviral vectors resulted in a marked increase in CNTF expression. This inverse relation between before and after axotomy was also observed in mouse sciatic nerve. In the axotomized sciatic nerve, the phosphorylated ERK was markedly increased; in contrast, the expression of CNTF was markedly decreased. These findings suggest that an inactive state of ERK is crucial for the CNTF expression in Schwann cells, and that activation of ERK following nerve injury critically influences the expression of CNTF. This might well explain why CNTF is highly expressed in quiescent Schwann cells in the peripheral nervous system, and also why CNTF is not abundant in axotomized nerves or cultured Schwann cells in which the proliferation signal is obviously active.

Upregulation of ciliary neurotrophic factor (CNTF) and CNTF receptor alpha in rat kidney with ischemia-reperfusion injury.

Ciliary neurotrophic factor (CNTF) is presumed to play a role as a survival factor in neuronal cells, but little is known about its role in the kidney. To investigate this, the expression of CNTF and CNTF receptor alpha (CNTFR alpha) was analyzed in the ischemic rat kidney. An ischemia/reperfusion (I/R) injury was induced by clamping both renal arteries for 45 min. Animals were killed at 1, 2, 3, 5, 7, 14, and 28 d after ischemia. The expression of CNTF and CNTFR alpha was monitored by reverse transcription-PCR, in situ hybridization, immunoblotting, immunohistochemistry, and electron microscopy. In sham-operated rat kidneys, CNTF expression was weak and limited to the descending thin limb of the loop of Henle. With I/R injury, CNTF mRNA and protein expressions were strikingly increased as compared with the sham-operated rat kidney, and the immunoreactivity of CNTF was mainly observed in the regenerating proximal tubules. The expression of CNTFR alpha mRNA was also increased after I/R injury, and its location and expression patterns were similar to the expression of CNTF. These findings suggest a possible role of CNTF as a growth factor during renal tubular repair processes after I/R injury and an autocrine or paracrine function of CNTF acting against CNTFR alpha.

IL-6 up-regulates CNTF mRNA expression and enhances neurite regeneration.

Interleukin-6 (IL-6) is a neurotrophic cytokine, however, its direct effect on nerve regeneration has not been well characterized. We therefore examined the effect of IL-6 on neurite regeneration using the rat dorsal root ganglion. IL-6 significantly enhanced neurite regeneration from transected nerve terminals. We also examined the mRNA expression of IL-6 family cytokines and their receptors during the regeneration. The mRNA expressions of IL-6, IL-6 receptor, leukemia inhibitory factor (LIF), ciliary neurotrophic factor (CNTF) receptor alpha, and LIF receptor beta showed no significant differences by the addition of IL-6. In contrast, IL-6 enhanced the mRNA expression of gp130 and CNTF. In addition, CNTF significantly increased neurite regeneration when added exogenously. Our data suggest that IL-6 enhanced regeneration via up-regulating CNTF expression.

Adenovirus-Mediated Expression of Ciliary Neurotrophic Factor (CNTF) Rescues Axotomized Rat Retinal Ganglion Cells But Does Not Support Axonal Regeneration in Vivo

Rat optic nerve (ON) transection leads to mainly apoptotic cell death of about 85% of the retinal ganglion cell (RGC) population within 14 days after lesion. In the present study, we tested the effect of adenovirally delivered CNTF (Ad-CNTF) on survival and regeneration of axotomized adult RGCs in vivo. Single intravitreal Ad-CNTF injection led to stable CNTF mRNA and protein expression for at least 18 days and significantly enhanced RGC survival by 155% when compared to control animals 14 days after ON transection. ON stump application of Ad-CNTF also resulted in an increased number of surviving RGCs. Ad-CNTF injection led to better preservation of intraretinal RGC axons but did not support regeneration of axotomized RGCs into a peripheral nerve graft. Thus, adenovirus-mediated neurotrophic factor supply is a suitable approach for reducing axotomy-induced RGC death in vivo and may constitute a relevant strategy for clinical treatment of traumatic brain injury.

Upregulation of ciliary neurotrophic factor in reactive Müller cells in the rat retina following optic nerve transection

We have investigated the expression and cellular localization of ciliary neurotrophic factor (CNTF) in the rat retina following optic nerve transection. In the normal retina, CNTF immunoreactivity was restricted to profiles in the ganglion cell layer. Following optic nerve transection, immunoreactivity appeared in Müller cell somata and processes and its intensity increased between three and seven days post-lesion. Quantitative evaluation by immunoblotting confirmed that CNTF expression increased continuously up to 7 days after optic nerve transection (to 430% of control levels), but decreased again to 250% of controls at 4 weeks post-lesion. Our findings suggest that CNTF supplied by Müller cells may play a protective role for axotomized ganglion cells in the rat retina.

Differential regulation of ciliary neurotrophic factor and its receptor in the rat hippocampus following transient global ischemia

To investigate a potential role of ciliary neurotrophic factor (CNTF) in transient global ischemia, we have studied the postischemic regulatory changes in the expression of CNTF and its receptor, the ligand-binding α-subunit (CNTFRα). Immunoblot analysis demonstrated CNTF levels were slightly upregulated already during the first day after ischemia and then increased markedly by more than 10-fold until 2 weeks postischemia. Immunoreactivity for CNTF became detectable 1 day after ischemia and was localized in reactive astrocytes. The intensity of the immunolabeling was maximal in CA1 during the phase of neuronal cell death (days 3–7 postischemia) and in the deafferented inner molecular layer of the dentate gyrus. Upregulation of CNTF expression was less pronounced in CA3 and absent in the stratum lacunosum moleculare and the outer molecular layer of the dentate gyrus and thus did not simply correlate with astroliosis as represented by upregulation of glial fibrillary acidic protein (GFAP). As shown by in situ hybridization, expression of CNTFRα mRNA was restricted to neurons of the pyramidal cell and granule cell layers in control animals. Following ischemia, reactive astrocytes, identified by double labeling with antibodies to GFAP, transiently expressed CNTFRα mRNA with a maximum around postischemic day 3. This astrocytic response was most pronounced in CA1 and in the hilar part of CA3. These results show that CNTF and its receptor are differentially regulated in activated astrocytes of the postischemic hippocampus, indicating that they are involved in the regulation of astrocytic responses and the neuronal reorganizations occurring after an ischemic insult.

Decrease in the Ciliary Neurotrophic Factor of the Spinal Cord in Amyotrophic Lateral Sclerosis

Ciliary neurotrophic factor (CNTF), a potent survival factor in spinal motoneurons of embryonic chick and rat, is currently being investigated in humans as a treatment for amyotrophic lateral sclerosis (ALS). However, its physiological and pathological activities in ALS remain unclear. We measured CNTF contents in the cervical enlargement of the spinal cord from 9 ALS patients and 12 age-matched control subjects using a sensitive enzyme-linked immunoassay. CNTF genotypes were determined by the polymerase chain reaction-restriction fragment length polymorphism method. In control subjects, there were 8 homozygotes and 4 heterozygotes, while in ALS patients there were 6 and 3, respectively. In both homozygotes and heterozygotes, CNTF expression in the spinal cord from ALS patients tended to decrease compared to control subjects. In homozygotes, the decrease was significant (p < 0.05). Concerning the regional concentrations of CNTF, in homozygotes, CNTF contents in the lateral corticospinal tract were markedly lower (p < 0.001) in ALS patients than in controls. The decrease in CNTF expression in the lateral corticospinal tract of the spinal cord from ALS patients may be a feature of ALS and could be related to motor neuron loss.

Expression of CNTF in Müller cells of the rat retina after pressure-induced ischemia.

We have investigated the expression and cellular localization of ciliary neurotrophic factor (CNTF) in the rat retina following ischemia induced by transiently increasing the intraocular pressure. In the normal retina, CNTF immunoreactivity was restricted to profiles in the ganglion cell layer. Following ischemia and reperfusion, immunoreactivity appeared in Müller cell somata and processes and its intensity increased between 1 day and 2 weeks post-lesion. Quantitative evaluation by immunoblotting confirmed that CNTF expression continuously increased up to 2 weeks after ischemic injury (to 600% of control levels), but had declined again to 250% of controls at 4 weeks post-lesion. Our findings suggest that CNTF supplied by Müller cells has a protective function for lesioned neurons following transient ischemia.

CNTF and its receptor subunits in human gliomas.

Ciliary neurotrophic factor (CNTF) promotes the survival of various neuronal cell populations. It is produced by astrocytes and influences the development and differentiation of glial cells. CNTF and related neuropoietic cytokines affect growth and differentiation of various neoplasms. Moreover, they induce the reactive transformation of astrocytes (gliosis) and influence growth and differentiation of neuroectodermal tumor cell lines in vitro. However, their role in gliomas is largely unknown. We studied the expression of CNTF and its receptor subunits in human astrocytomas and glioblastomas. In more than 95% of the tumors, CNTF transcripts were found by RNAase protection assay; in more than 80% of the cases, tumor cells were CNTF immunoreactive. CNTF receptor alpha (CNTFR alpha), the specific component of the tripartite CNTF receptor system, was detectable by Northern blot analysis in 80% of the cases. In situ hybridization revealed CNTFR alpha mRNA in the cytoplasm of neoplastic cells. Transcripts of the remaining two components of the CNTF receptor system, gp130 and LIFR beta, were found by Northern blotting in 83% and 70% of the tumors, respectively. Simultaneous expression of CNTF and all its receptor components was detected in approximately half of the tumors. These results indicate that CNTF and its receptor components are expressed by human glioma cells. The simultaneous expression of ligands and receptor subunits suggests that CNTF might act on human glioma cells via an auto- or paracrine mechanism.

Alterations in the neurotrophic factors BDNF, GDNF and CNTF in the regenerating olfactory system.

Neurogenesis, axonal outgrowth and synapse formation are usually restricted to specific stages during central nervous system development, but the mature olfactory system maintains these capacities. The cycle of neuronal turnover can be experimentally induced by surgical ablation of the olfactory bulb (OB). We are interested in the growth factor regulation of these processes and the trophic role played by the target tissue, the OB. We studied the immunohistochemical expression of three neurotrophic factors, brain derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF) and ciliary neurotrophic factor (CNTF) in the rat olfactory neuroepithelium (ON) and OB and in target-deprived ON at 1, 3 and 12 weeks post unilateral bulbectomy. We found BDNF immunoreactivity (IR) was restricted to the basal cells and did not alter postbulbectomy. GDNF-IR was expressed by mature olfactory receptor neurons (ORNs), their axons and target cells in the OB in controls, but was absent from the ON postbulbectomy. Hence, the expression of GDNF by ORNs was found to be target-dependent. CNTF-IR was present in ORNs and their target cells in the OB, in basal cells and in some immature ORNs. Postbulbectomy, CNTF-IR was unaltered in the basal cells, and very low levels were detectable in maturing ORNs in the ON. Our results indicate that these three factors may contribute to the trophic regulation of this neuronal pathway in a coordinated fashion. Previous work has shown that BDNF promotes survival of ORNs in vitro, and TrkB expression has been found in both immature and mature ORNs. Hence, BDNF produced by basal cells may be acting locally on neurons expressing TrkB. Expression of CNTF by both the basal cells and the ORNs suggests that it may play an integral role in this neuronal differentiation pathway. Finally, the expression of GDNF exclusively by mature ORNs in the ON, its presence in the target cells in the OB and abolition of expression by bulbectomy, suggests that it may be target-derived. This provides a major mechanism by which the bulb could exert trophic influences on ORNs.

Increased ciliary neurotrophic factor expression in reactive astrocytes following spinal cord injury in the rat

Expression of ciliary neurotrophic factor (CNTF) was observed in reactive astrocytes in injured spinal cord of the adult rat. After unilateral incision of the dorsal funiculus at a midthoracic level, the rats were sacrificed on the day of postoperation (DPO) 3, 7, 14 or 28. Western blot analysis of spinal cord extracts showed that the intensity of the CNTF-immunoreactive band of 24 kDa was increased on DPO 3 and 7 and decreased on DPO 14. CNTF- or glial fibrillary acidic protein (GFAP)-like immunoreactivity was also increased progressively in the injured dorsal funiculus and adjacent gray matter. Double immunofluorescence histochemistry revealed that all CNTF-like immunoreactive cells showed GFAP-like immunoreactivity. The CNTF upregulation in the reactive astrocytes may play important roles in repair process after spinal cord injury.

Ciliary neurotrophic factor stimulates nuclear hypertrophy and increases the GFAP content of cultured astrocytes

Studies using transgenic mice that overexpress ciliary neurotrophic factor (CNTF), direct injection of CNTF into brain parenchyma, and ectopic expression of CNTF by an adenoviral vector have demonstrated that CNTF activates astrocytes. Paradoxically, studies to date have failed to show an effect of CNTF on the expression of GFAP by cultured astrocytes. Therefore, the goal of this study was to use nuclear hypertrophy and GFAP expression as indices of glial activation to compare the responsiveness of forebrain type 1 and type 2 astrocytes to CNTF. As reported by others, CNTF did not increase GFAP in type 1 astrocytes; however, it rapidly increased their nuclear size by 20%. Nuclear hypertrophy was apparent within 4 h after CNTF exposure and persisted for at least 48 h. In contrast, type 2 astrocyte GFAP increased 2-fold over the course of 48 h of CNTF treatment. During this same treatment period type 2 astroglial nuclei enlarged by 25%. We conclude that CNTF stimulates both type 1 and type 2 astrocytes directly. Together with our in vivo studies (Levison et al., 1996: Exp. Neurol. 141: 256), these data support the concept that CNTF is responsible for many of the progressive astroglial changes that appear after CNS injury and disease.

Lesion‐induced changes in the expression of ciliary neurotrophic factor and its receptor in rat optic nerve

There is evidence that ciliary neurotrophic factor (CNTF) is involved in reactive changes following lesions of the nervous system. To investigate, whether differences in the regulation of CNTF and CNTF receptor α (CNTFRα) contribute to the differences in PNS and CNS responses to injury, we have studied their expression on the mRNA and protein level in the rat optic nerve following a crush lesion to compare them with the situation in peripheral nerve. Seven days after the lesion, CNTF mRNA and protein levels were markedly decreased at the lesion site, concommitant with the disappearance of GFAP- and CNTF-immunopositive astrocytes. CNTF levels in proximal and distal parts were less affected. This was in contrast to the situation in the PNS, where CNTF was downregulated at and distal to the lesion site. Different from other CNS regions, optic nerve astrocytes expressed CNTFRα mRNA under normal conditions. Following lesion, CNTFRα was reduced substantially only in distal and proximal parts of the optic nerve but continued to be expressed at high levels at the lesion site, suggesting that GFAP-negative, CNTF-responsive cells are present there. Our results suggest that differences in lesion-induced changes in the optic and sciatic nerve reflect differences in the response to injury of astrocytes and Schwann cells. In the light of the known actions of CNTF in inducing astrogliosis, the expression pattern observed in the optic nerve indicates that CNTF and CNTFRα are involved in glial scar formation in the lesion area. GLIA 23:239–248, 1998. © 1998 Wiley-Liss, Inc.

Lesion‐induced changes in the expression of ciliary neurotrophic factor and its receptor in rat optic nerve

There is evidence that ciliary neurotrophic factor (CNTF) is involved in reactive changes following lesions of the nervous system. To investigate, whether differences in the regulation of CNTF and CNTF receptor alpha (CNTFRalpha) contribute to the differences in PNS and CNS responses to injury, we have studied their expression on the mRNA and protein level in the rat optic nerve following a crush lesion to compare them with the situation in peripheral nerve. Seven days after the lesion, CNTF mRNA and protein levels were markedly decreased at the lesion site, concommitant with the disappearance of GFAP- and CNTF-immunopositive astrocytes. CNTF levels in proximal and distal parts were less affected. This was in contrast to the situation in the PNS, where CNTF was downregulated at and distal to the lesion site. Different from other CNS regions, optic nerve astrocytes expressed CNTFRalpha mRNA under normal conditions. Following lesion, CNTFRalpha was reduced substantially only in distal and proximal parts of the optic nerve but continued to be expressed at high levels at the lesion site, suggesting that GFAP-negative, CNTF-responsive cells are present there. Our results suggest that differences in lesion-induced changes in the optic and sciatic nerve reflect differences in the response to injury of astrocytes and Schwann cells. In the light of the known actions of CNTF in inducing astrogliosis, the expression pattern observed in the optic nerve indicates that CNTF and CNTFRalpha are involved in glial scar formation in the lesion area.