Analysis of Structural and Molecular Events Associated with Adult Rat Optic Chiasm and Nerves Demyelination and Remyelination; Possible Role for 3rd Ventricle Proliferating Cells

Multiple sclerosis frequently affects the optic apparatus, particularly optic chiasm and nerves. Here, we have reported the structural and molecular characteristics of remyelination in the adult rat optic chiasm and nerves. Moreover, considering the proximity of optic chiasm and 3rd ventricle, we have tried to determine if proliferating cells residing in 3rd ventricle region are able to migrate in response to experimental demyelination of the optic chiasm. Following local demyelination by lysolecithin, remyelination pattern in longitude of optic chiasm and proximal nerves was investigated using myelin staining and marker genes expression. Furthermore, cell tracing was carried out using BrdU labeling of proliferating cells prior to gliotoxin injection. Morphometric analysis revealed that demyelination was considerable on days 7 and 14 and an incomplete remyelination occurred on day 28 post-lesion. Interestingly, myelin repair was more evident in the caudal part of chiasm, compared to rostral part and proximal optic nerves. Following chiasm and nerve demyelination, trains of BrdU+ cells were seen near the 3rd ventricle which subsequently moved to lesion site. Nestin was significantly up-regulated in 3rd ventricle surroundings. At the lesion site, Nogo-A gene expression was significantly decreased on days 7 and 14 post lesion, while Olig2, nestin, and GFAP expression was increased on day 7. The changes were then reversed by the time. Myelin repair in optic chiasm seems to be mediated by endogenous progenitors and stem cells. Adult 3rd ventricle proliferating cells may play a role in this context by mobilization into the demyelinated chiasm.     

Electromagnetic Field Stimulation Potentiates Endogenous Myelin Repair by Recruiting Subventricular Neural Stem Cells in an Experimental Model of White Matter Demyelination

Electromagnetic fields (EMFs) may affect the endogenous neural stem cells within the brain. The aim of this study was to assess the effects of EMFs on the process of toxin-induced demyelination and subsequent remyelination. Demyelination was induced using local injection of lysophosphatidylcholine within the corpus callosum of adult female Sprague-Dawley rats. EMFs (60 Hz; 0.7 mT) were applied for 2 h twice a day for 7, 14, or 28 days postlesion. BrdU labeling and immunostaining against nestin, myelin basic protein (MBP), and BrdU were used for assessing the amount of neural stem cells within the tissue, remyelination patterns, and tracing of proliferating cells, respectively. EMFs significantly reduced the extent of demyelinated area and increased the level of MBP staining within the lesion area on days 14 and 28 postlesion. EMFs also increased the number of BrdU- and nestin-positive cells within the area between SVZ and lesion as observed on days 7 and 14 postlesion. It seems that EMF potentiates proliferation and migration of neural stem cells and enhances the repair of myelin in the context of demyelinating conditions.     

成年大鼠脑创伤后脑组织碱性成纤维细胞生长因子表达的变化

目的:研究液压冲击性脑损伤后成年大鼠脑组织碱性成纤维细胞生长因子(bFGF)表达时程和空间分布的变化。方法:制作液压冲击性脑损伤模型,免疫组织化学法动态检测内源性bFGF的变化。结果:在正常脑组织中,bFGF免疫活性低水平表达,且主要位于胶质细胞核及神经元细胞质中。液压冲击伤后bFGF免疫活性增强,聚集核周边。此外,部分bFGF免疫活性聚集于神经元周围间隙中半定量分析显示液压冲击伤后3d,大脑皮质bFGF阳性细胞数量较正常对照组升高4.8倍;伤后7 d,bFGF免疫活性达高峰,伤后30 d消失。结论:液压冲击伤后,损伤的胶质细胞和神经元暂短地合成bFGF,并以旁分泌和自分泌形式起作用。     

Basic Fibroblast Growth Factor in the Adrenal Gland

The importance of trophic agents for the development and maintenance of neurons and their presence in mesenchyme-derived neuronal target organs such as muscle is well exemplified by the protein nerve growth factor (NGF) and its synthesis in target areas of sympathetic and sensory nerves. Stringent conceptualization of target organ-regulated neuronal maintenance would imply that neurons were able to provide trophic support to their presynaptic counterparts. We present data suggesting that basic fibroblast growth factor (bFGF), a mitogen and trophic factor for several neuron populations in vitro, may be such a protein involved in retrograde trophic neuron - neuron interaction. Basic FGF or a closely related protein is present in the adrenal medulla and its sympathetic neuron-like chromaffin cells. A polyclonal antibody specific for bFGF recognizes an 18 kD band in Western blots of bFGF-enriched bovine adrenal medulla extracts and immunostains isolated bovine chromaffin cells. This antibody also blocks the bFGF-like activity present in adrenal medullary extracts and chromaffin granule extracts that both promote in vitro survival of embryonic chick ciliary ganglionic neurons. Furthermore, like bFGF, the soluble proteins of bovine chromaffin granules are mitogenic for cultured bovine aorta endothelial cells. Electrothermal unilateral destruction of the adrenal medulla causes the disappearance of 25% of Nissl-stained neurons in the ipsilateral intermediolateral column (IML) of the spinal cord between levels Th7 and L1, which contains the preganglionic neurons projecting to the adrenal medulla. Substitution of the adrenal medulla by gel foams soaked with bFGF prevents neuron losses in the IML. The effects are specific in that NGF and cytochrome C are ineffective. Our results suggest that bFGF is located in chromaffin cells and maintains target-deprived autonomic spinal cord neurons, thus possibly acting as an interneuronal trophic messenger in vivo.     

Alterations in Fibroblast Growth Factor Receptor Expression Following Brain Injury

Traumatic injuries to the central nervous system result in astrogliosis and the formation of a dense scar at the site of the wound. Basic fibroblast growth factor (bFGF) has mitogenic and morphogenic effects on astrocytes, and an interaction between bFGF and its receptor is likely to play a role in astrogliosis. We examined trauma-induced changes in the spatial and temporal expression of FGF receptor (FGFR) in adult rats over a 28-day period following a stereotaxic lesion through the cortex and hippocampus. Immunohistochemistry and image analysis were used to evaluate the changes. Antibody characterization studies strongly suggested that staining represented FGFR 1, but did not rule out possible cross-reactivity with FGFR 2 or 3. Double immunohistochemistry for FGFR and glial fibrillary acidic protein demonstrated that mature astrocytes expressed FGFR. Expression was increased on astrocytes adjacent to the wound cavity by Day 2 postlesion. Staining increased further through Day 10 and decreased to control values by Day 28, except for a sustained increase in staining of reactive astrocytes immediately adjacent to the wound cavity. Basic FGF was detected in the nuclei of cells staining for FGFR, suggesting that FGFR-expressing astrocytes also contained bFGF. These data demonstrate a time course for astrocyte expression of FGFR that precedes and parallels the time course for astrocyte hypertrophy. Our observations suggest that endogenous bFGF, acting directly on FGFR-expressing astrocytes, may contribute to astrogliosis.     

Nerve Growth Factor and Basic Fibroblast Growth Factor Protect Cholinergic Neurons Against Quinolinic Acid Excitotoxicity in Rat Neostriatum

In the present work we have characterized a possible mechanism leading to the early survival of neostriatal cholinergic neurons after quinolinic acid injection. Different doses of quinolinic acid were injected in rat neostriatum and two different parameters were analysed 7 days after the lesion: choline acetyltransferase (ChAT) activity and nerve growth factor (NGF) levels. We have observed that ChAT activity decreased (until 68 nmol quinolinic acid) and NGF levels increased (until 34 nmol quinolinic acid) in a dose-dependent manner. In order to characterize the time-course of the lesion on NGF levels and ChAT activity, and the possible protective effect of NGF and basic fibroblast growth factor (bFGF) on cholinergic neurons, we have used the quinolinic acid dose (68 nmol) at which the first decrease of ChAT activity was observed. ChAT activity and NGF levels showed different patterns of response to quinolinic acid injection, since the maximal effect was reached at 1 day for ChAT activity and at 2 days for NGF levels. NGF or bFGF simultaneously injected with quinolinic acid (68 nmol) completely prevented the decrease in ChAT activity in a dose-dependent manner but NGF was more effective than bFGF. Furthermore, differences observed in ChAT activity after NGF but not bFGF treatment were correlated with changes in the number of ChAT immunoreactive cells. Finally, we have also observed that, although bFGF alone was not able to modify NGF levels, bFGF simultaneously injected with quinolinic acid produced an increase of NGF levels higher than that observed after quinolinic acid injection alone. Our results show that NGF and bFGF protect striatal cholinergic neurons against quinolinic acid injury, and bFGF is able to potentiate the increase of NGF after the lesion, suggesting a cooperative action between different trophic factors in neuronal protection after excitotoxic injury. Thus, administration of trophic factors may be relevant in the prevention and treatment of neurodegenerative disorders, such as Huntington's disease.     

生物可降解聚合物结合碱性成纤维细胞生长因子对内皮细胞粘附的研究

目的 探讨利用可降解聚己内酯接枝肝素材料,体外负载碱性成纤维细胞生长因子,观察其对于内皮细胞粘附的影响。方法 采用可降解聚己内酯接枝肝素材料,电纺丝技术纺织血管支架,体外负载碱性成纤维细胞生长因子。采用低密度内皮细胞短期静态种植,观察负载细胞生长因子的可降解聚己内酯材料对内皮细胞的粘附影响。结果 利用可降解聚己内酯接枝肝素材料,电纺丝技术成功体外构建可降解血管支架,体外负载碱性成纤维细胞生长因子。内皮粘附实验证实,负荷生长因子的可降解支架材料,利于内皮细胞粘附。结论 可降解聚己内酯接枝肝素材料负荷碱性细胞生长因子(b-FGF)支架,利于内皮细胞粘附,可用于小口径组织工程血管的支架的研究。     

TGF-β1和bFGF在人脑胶质瘤的表达及意义

目的观察转化生长因子β1(TGF-β1)和碱性成纤维生长因子(bFGF)在人脑胶质瘤中的表达和分布,加深对它们在胶质瘤恶性转化中作用的认识。方法采用免疫组化方法,对临床经过石蜡包埋的35例胶质瘤和7例正常脑组织标本中TGF-β1和bFGF的表达进行定位和定性检测,并分析二者的相关性。结果TGF-β1和bFGF在高级别胶质瘤(Ⅲ-Ⅳ级)中表达较多,在低级别胶质瘤(Ⅰ-Ⅱ级)中表达较少,在正常脑组织中几乎不表达,两两比较差异有显著性(P<0.05),且二者在胶质瘤中的表达呈显著正相关(r=0.79,P<0.01)。结论TGF-β1和bFGF的协同作用可能在人脑胶质瘤细胞的恶性转化中具有重要意义。     

Pharmacological Induction of Nerve Growth Factor mRNA in Adult Rat Brain

Three structurally unrelated compounds, all of which induce nerve growth factor (NGF) in cell culture systems, were assessed for their ability to induce NGF mRNA in adult rat brain using a highly sensitive RNAse protection assay. Interleukin-1β (0.5-1 pmol) and 1,25-dihydroxyvitamin D₃ (25-25,000 pmol) were extremely potent inducers of NGF mRNA, being respectively at least 50,000 and 4000 times more potent than 4-methylcatechol. These compounds elicited an approximate twofold increase in NGF mRNA in both the hippocampus and cortex, without altering β-actin mRNA levels after a single intracerebroventricular injection. The duration of NGF induction was dependent on the compound administered. For example, the elevation of NGF mRNA elicited by interleukin-1β peaked at 8 h and lasted for at least 24 h. In contrast, the induction of NGF after 1,25-dihydroxyvitamin D₃ and 4-methylcatechol administration peaked between 4 and 8 h and was not apparent 24 h after injection. These results demonstrate induction of NGF mRNA in vivo by administration of physiological or pharmacological agents and differentiate these agents by potency and duration of action. Further, these findings indicate that pharmacological induction of NGF may be a viable strategy for the treatment of neurodegenerative disorders such as Alzheimer's disease.     

Basic Fibroblast Growth Factor Opens Calcium-Permeable Channels in Quail Mesencephalic Neural Crest Neurons

In order to investigate the action of basic fibroblast growth factor (bFGF) in the nervous system, we have studied the ionic signals elicited by this peptide in cultured quail mesencephalic neural crest neurons using patch-clamp and cytofluorimetric techniques. In this preparation stimulation with bFGF induced, with a delay of some tens of seconds, an inward cationic current. Single-channel experiments provided evidence for the activation of a calcium-permeable channel. In single-cell cytofluorimetric measurements, a sustained rise in [Ca²⁺Ii was observed, which was dependent on the presence of external calcium. These events may play a role in the developmental effects of bFGF.     

Blockade of Basic Fibroblast Growth Factor Retards Recovery from Motor Cortex Injury in Rats

The endogenous expression of basic fibroblast growth factor (bFGF) was blocked by neutralizing antibodies following unilateral suction lesions of the motor cortex. Rats with control treatment (saline, goat IgG) after motor cortex lesions showed slow recovery of forelimb manipulatory abilities. Rats with blockade of bFGF expression showed little recovery. Anatomically, the control-treated lesioned rats showed an acute increase in bFGF and glial fibrillary acidic protein (GFAP) reactivity, and chronically they had normal dendritic arborization and spine density in layer V pyramidal cells in the remaining motor cortex. In contrast, rats treated with antibodies to bFGF showed little bFGF reactivity, normal GFAP reactivity, and atrophy of dendritic arbor and decreased spine density in layer V pyramidal cells. These results demonstrate the importance of endogenous bFGF release in processes related to functional recovery after cortical injury.     

Regional Increases in Brain-Derived Neurotrophic Factor and Nerve Growth Factor mRNAs During Amygdaloid Kindling, But Not in Acidic and Basic Fibroblast Growth Factor mRNAs

Summary: We studied mRNA levels for neurotrophic factors using the amygdaloid kindling model of epilepsy. One hour after stage 5 kindled seizures, there were four-to fivefold increases in brain-derived neurotrophic factor (BDNF) mRNA in rat dentate gyrus and perirhinal cortex. Nerve growth factor (NGF) mRNA levels were increased bilaterally in some (but not all) limbic areas. There were no detectable changes in acidic fibroblast GF (aFGF) mRNA or basic fibroblast GF (bFGF) mRNA for 24 h after the kindled seizures. During kindling, levels of BDNF mRNA in the dentate gyrus correlated with projection to generalized seizures, whereas NGF mRNA in the limbic regions continued to increase during seizure development. These results indicate that the induction of mRNAs for neurotrophic factors, especially for BDNF mRNA in the dentate gyrus, corresponds to the increases in metabolic and electrical ictal discharge associated with kindled seizures. The persistent increase observed in NGF mRNA may be related to enhanced synaptic efficacy during kindling, but aFGF and bFGF are presumed to have little relation to the kindling process.     

Neurotrophins and Basic Fibroblast Growth Factor Induce the Differentiation of Calbindin-Containing Neurons in the Cerebral Cortex

Lineage studies have recently shown that the expression of calcium-binding proteins in neurons of the cerebral cortex is not genetically programmed and is likely to be induced by external factors. Current hypotheses suggest that basic fibroblast growth factor (bFGF) and a number of neurotrophins play important roles in the proliferation and differentiation of cortical progenitor cells to a particular lineage. Using a dissociated cell culture system, we found that bFGF and the neurotrophins brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and nerve growth factor differentially affect the expression of the calcium-binding protein calbindin in selective neuronal subpopulations in the developing cerebral cortex. Specifically, BDNF and NT-3 greatly promoted the morphological differentiation of a relatively small, early-generated population of GABAergic neurons and induced the expression of calbindin in these cells. Furthermore, treatment with BDNF, NT-3, and bFGF produced an two- to threefold increase in the number of newly generated calbindin-positive neurons. The effect of bFGF was more striking in earlier (E14) than later (E16) ages, whereas the action of neurotrophins was independent of the age from which the cultures were prepared. Switching experiments combined with BrdU incorporation have suggested that NT-3 acts on postmitotic neurons rather than on proliferating progenitors to induce calbindin expression and that its action is mediated via trk receptors. Application of retroviral vectors in culture resulted in the presence of neuronal clones that were predominantly heterogeneous with regard to calbindin expression, suggesting, in agreement with our earlierin vivostudies, that the expression of this calcium-binding protein is not lineage dependent. Our results characterize the roles of BDNF, NT-3, and bFGF in the expression of calbindin in developing neocortical neurons.     

Characterization of GH3 Cells Overexpressing Basic Fibroblast Growth Factor (FGF-2)

Basic fibroblast growth factor (FGF-2) is not only a potent mitogen for various cells but also a multifunctional factor with angiogenic and chemotactic activity, and the capacity to induce the synthesis of various proteinases and to modulate endocrine function. To clarify the role played by FGF-2 in the progression of pituitary tumor, we fused rat FGF-2 cDNA to the promoter SRα, consisting of the early promoter of SV40 and HTLV(I)-LTR, and we cotransfected GH₃ cells with pSV2-neo by an electroporation method. After selection by G418, we obtained 7 neomycin-resistant clones. Southern blot analysis of genomic DNA revealed the presence of transfected rat FGF-2 cDNA in 4 of the 7 clones. To measure FGF-2 molecules, we established a new immuno-fluorometric assay system, using 3 monoclonal antibodies against different portions of human FGF-2. This assay had a minimum sensitivity of 10 pg/ml and cross-reacted neither with acidic fibroblast growth factor (FGF-1) nor insulin-like growth factor 1 (IGF-1), even at a concentration of 100 ng/ml. Although FGF-2 was undetectable in the culture medium of any of the clones, the cell homogenate contained a significant amount of FGF-2 (7.2 ng/mg protein) in 1 of the 4 FGF-2-transfected clones (GH₃FGF(+)), whereas FGF-2 was not detected (<5.2 pg/mg protein) in the cell homogenates of either the parent GH₃ cells or the control cells transfected with pSV2-neo alone (GH₃FGF(?)). GH₃FGF(+) grew as adherent cells and formed epithelial sheets with a growth rate similar to that of control cells. The amount of prolactin(PRL) released by TRH was greater in GH₃FGF(+) than that in GH₃ or GH₃FGF(?). On the other hand, the sensitivity to SRIF was increased in GH₃FGF(+) compared with that in other clones. The findings of these in vitro studies indicate that FGF-2, if it is expressed in pituitary tumor cells, plays little if any role in cell growth but may modulate certain cell functions such as responsiveness to hormones.     

Expression of the Highly Polysialylated Neural Cell Adhesion Molecule During Postnatal Myelination and Following Chemically Induced Demyelination of the Adult Mouse Spinal Cord

We have investigated the expression of the highly polysialylated neural cell adhesion molecule in the mouse spinal cord during postnatal myelination and in the adult after chemically induced demyelination. By double immunohistochemistry, using a monoclonal antibody (anti-Men B) which specifically recognizes polysialic acid (PSA) units on neural cell adhesion molecule (N-CAM), and an anti-myelin basic protein, a caudorostral gradient of expression of PSA-NCAM was observed at postnatal day 1 (P1), which was inversely related to the gradient of myelination. At P7, PSA-NCAM labelling decreased relative to P1. In white matter, this decrease was correlated with the progression of myelination. PSA-NCAM immunoreactivity persisted in as yet unmyelinated structures, i.e. the corticospinal tract, the dorsomedial part of the ventral funiculus and the lateral funiculi, and decreased with the onset of myelination of these structures at P15. In the adult, PSA-NCAM expression remained in discrete structures, i.e. Iaminae I and II of the dorsal horn and lamina X around the central canal. The ependymal cells and the astrocyte endfeet under the meninges were also labelled. In addition, PSA-NCAM expression was reinduced on various cells and structures after lysolecithin-induced demyelination of the adult mouse spinal cord. At early times after demyelination, PSA-NCAM was expressed on glial cells around the lesion but also at a distance from this zone. Seven days after injection, cellular PSA-NCAM expression was found around but also within the lesion. This expression was totally abolished 15 days after injection. Double immunohistochemistry for PSA and cell-specific markers showed that the cells which expressed PSA-NCAM after demyelination were oligodendrocyte precursors, reactive astrocytes and Schwann cells. PSA-NCAM re-expression on all cell types was transient and ceased when myelin repair was accomplished. The spatial and temporal regulation of PSA-NCAM expression during development and after demyelination suggests a role for PSA-NCAM in glial plasticity during the myelination and remyelination processes.     

Hypothalamic Transforming Growth Factor β1 and Basic Fibroblast Growth Factor mRNA Expression is Modified During the Rat Oestrous Cycle

The present observations show that the mRNA levels of two growth factors, previously described to be involved in the control of neurones synthesizing the luteinizing hormone releasing hormone (LHRH) [i.e. transforming growth factor beta1 (TGFbeta1) and basic fibroblast growth factor (bFGF)], fluctuate in the hypothalamus of adult female rats during the oestrous cycle. In particular, the expression of TGFbeta1-mRNA shows a peak on the morning of the day of proestrus, which precedes the increased secretion of the two gonadotrophins that occurs on that day. In the case of bFGF, the peak is evident in the evening of the same day and is concomitant with that of the gonadotrophins. We evaluated the effects of ovariectomy and of exogenous oestrogens on the mRNA levels of these two growth factors in the hypothalamus. The data indicate that 3 weeks of ovariectomy are not able to change the hypothalamic messenger levels of the two growth factors considered, which remain at the levels found in diestrus 1, and that 17beta-oestradiol is able to induce a significant increase of both TGFbeta1- and of bFGF-mRNA levels in the hypothalamus of the ovariectomized rat. The present in vivo observations support the concept, previously proposed on the basis of in vitro data, that growth factors, such as TGFbeta1 and bFGF, play a role in the hypothalamic control of reproduction, and suggest that the control of LHRH dynamics involves a strict cooperation between gonadal steroids and growth factors.     

Critical Periods of Basic Fibroblast Growth Factor and Brain-Derived Neurotrophic Factor in the Development of the Chicken Cochleovestibular Ganglionin Vitro

The temporal roles of brain-derived neurotrophic factor (BDNF) and fibroblast growth factor-2 (FGF-2) in the development of sensory neurons have been studied in a cell culture preparation which models normal embryonic inner ear development (normocytic). Previous studies showed that FGF-2 stimulated migration and differentiation of ganglion cells for the first 2 daysin vitro,but after 5 days led to degeneration, implicating other factors in their later development. To see if BDNF could be such a factor, otocysts were explanted from white leghorn embryos at the time when ganglion cell precursors normally start migrating from the otic epithelium. Cultures were grown in a defined medium, either with or without human recombinant FGF-2 for 2 days or with BDNF. On Day 3, FGF-2 was replaced either with BDNF in defined medium or with defined medium only. Measurements of neuroblast migration and neurite outgrowth were made by time-lapse imaging in living cultures. In cultures receiving BDNF on Day 3, cell migration and neurite outgrowth from the explant increased for more than 3 weeks but not in cultures receiving only defined medium from Day 3. Cultures did not survive more than 3–4 days when receiving either BDNF in defined medium or defined medium alone from the first day. A neutralizing antibody to BDNF inhibited neuronal migration and neurite outgrowth, and it also blocked the effects of exogenous BDNF. BDNF did not enhance the effects of FGF-2 by interacting with it. These experiments defined a temporal sequence in which FGF-2 acts early in development, while BDNF affects a later stage.     

Association Study of Fibroblast Growth Factor 2 and Fibroblast Growth Factor Receptors Gene Polymorphism in Korean Ossification of the Posterior Longitudinal Ligament Patients

Objective

The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of fibroblast growth factor (FGF) 2 gene and fibroblast growth factor receptor (FGFR) genes are associated with ossification of the posterior longitudinal ligament (OPLL).

Methods

A total of 157 patients with OPLL and 222 controls were recruited for a case control association study investigating the relationship between SNPs of FGF2, FGFR1, FGFR2 and OPLL. To identify the association among polymorphisms of FGF2 gene, FGFR1, FGFR2 genes and OPLL, the authors genotyped 9 SNPs of the genes (FGF2 : rs1476217, rs308395, rs308397, and rs3747676; FGFR1 : rs13317 and rs2467531; FGFR2 : rs755793, rs1047100, and rs3135831) using direct sequencing method. SNPs data were analyzed using the SNPStats, SNPAnalyzer, Haploview, and Helixtree programs.

Results

Of the SNPs, a SNP (rs13317) in FGFR1 was significantly associated with the susceptibility of OPLL in the codominant (odds ratio=1.35, 95% confidence interval=1.01-1.81, p=0.048) and recessive model (odds ratio=2.00, 95% confidence interval=1.11-3.59, p=0.020). The analysis adjusted for associated condition showed that the SNP of rs1476217 (p=0.03), rs3747676 (p=0.01) polymorphisms in the FGF2 were associated with diffuse idiopathic skeletal hyperostosis (DISH) and rs1476217 (p=0.01) in the FGF2 was associated with ossification of the ligament flavum (OLF).

Conclusion

The results of the present study revealed that an FGFR1 SNP was significantly associated with OPLL and that a SNP in FGF2 was associated with conditions that were comorbid with OPLL (DISH and OLF).     

Immunohistological Localization of Tenascin in the Developing and Lesioned Adult Mouse Optic Nerve

To gain insight into the morphogenetic functions of the recognition molecule tenascin in the central nervous system, we have studied its localization in the developing and lesioned adult mouse optic nerve using light and electron microscopic immunocytochemistry. Since tenascin is a secreted molecule, we have analysed the tenascin-synthesizing cells in tissue sections of retinae and optic nerves by in situ hybridization. A weak and homogeneous tenascin immunoreactivity was detectable in the developing retinal nerve fibre layer and optic nerve of 14-day-old mouse embryos, the earliest developmental age investigated. In the optic nerve of neonatal and 1-week-old animals, a high number of tenascin messenger RNA (mRNA)-containing cells were present, and antibodies to tenascin labelled the surfaces of astrocytes and unmyelinated retinal ganglion cell axons. With increasing age, expression of tenascin in the optic nerve was down-regulated at the mRNA and protein levels. At the fourth postnatal week, blood vessels in the optic nerve and collagen fibrils in the vicinity of meningeal fibroblast-like cells still showed significant immunoreactivity, but the optic nerve tissue proper no longer did so. In adult animals, tenascin was no longer detectable in association with blood vessels located in the myelinated part of the optic nerve, and meninges were only weakly immunoreactive. Also, tenascin mRNA-containing cells were no longer detectable in the myelinated part of the adult mouse optic nerve and few labelled cells were found in the meninges. In the retina, ganglion cells contained no detectable levels of tenascin mRNA at any of the developmental ages analysed. No significant up-regulation of tenascin expression was seen in the nerve tissue proper of transected proximal (i.e. retinal) and distal (i.e. cranial) optic nerve stumps of adult mice during the first 4 weeks after lesioning, the time period studied. However, collagen fibrils associated with meningeal fibroblast-like cells and located near the lesion site became strongly tenascin-immunoreactive 2 days after lesioning. Also, some blood vessels at the lesion site became immunoreactive. We conclude that tenascin in the optic nerve is synthesized by glial cells and not by retinal ganglion cells. The detectability of tenascin at embryonic ages suggests that it may mediate neurite growth in vivo. The absence of a strong, lesion-induced up-regulation of tenascin expression in the regeneration-prohibitive mouse optic nerve contrasts with the lesion-induced pronounced up-regulation in the regeneration-permissive peripheral nervous system, and may indicate a functional involvement of tenascin in regenerative processes. The high tenascin positivity of collagen fibrils at early postnatal ages and after lesioning suggests that tenascin expression may be correlated with mitotic activity of the associated meningeal fibroblast-like cells. Finally, tenascin may be involved in the process of vascularization, since the molecule is associated with blood vessels in developing and adult lesioned, but not intact adult, optic nerves.