Viscoelasticity of spinal cord and meningeal tissues.

Compared to the outer dura mater, the mechanical behavior of spinal pia and arachnoid meningeal layers has received very little attention in the literature. This is despite experimental evidence of their importance with respect to the overall spinal cord stiffness and recovery following compression. Accordingly, inclusion of the mechanical contribution of the pia and arachnoid maters would improve the predictive accuracy of finite element models of the spine, especially in the distribution of stresses and strain through the cord's cross-section. However, to-date, only linearly elastic moduli for what has been previously identified as spinal pia mater is available in the literature. This study is the first to quantitatively compare the viscoelastic behavior of isolated spinal pia-arachnoid-complex, neural tissue of the spinal cord parenchyma, and intact construct of the two. The results show that while it only makes up 5.5% of the overall cross-sectional area, the thin membranes of the innermost meninges significantly affect both the elastic and viscous response of the intact construct. Without the contribution of the pia and arachnoid maters, the spinal cord has very little inherent stiffness and experiences significant relaxation when strained. The ability of the fitted non-linear viscoelastic material models of each condition to predict independent data within experimental variability supports their implementation into future finite element computational studies of the spine. STATEMENT OF SIGNIFICANCE: The neural tissue of the spinal cord is surrounded by three fibrous layers called meninges which are important in the behavior of the overall spinal-cord-meningeal construct. While the mechanical properties of the outermost layer have been reported, the pia mater and arachnoid mater have received considerably less attention. This study is the first to directly compare the behavior of the isolated neural tissue of the cord, the isolated pia-arachnoid complex, and the construct of these individual components. The results show that, despite being very thin, the inner meninges significantly affect the elastic and time-dependent response of the spinal cord, which may have important implications for studies of spinal cord injury.

On the route to mimic natural movements: synthesis and photophysical properties of a molecular arachnoid.

The synthesis, photoswitchability and NIR emitting properties of a novel π-extended pyrene derivative, peripherally decorated with four azobenzenyl-ethynyl legs, are reported.

Secretory meningioma, a rare meningioma subtype with characteristic glandular differentation: an histological and immunohistochemical study of 9 cases.

AIM: We studied the immunophenotype of 9 cases of secretory meningioma (SM), a rarely reported meningioma variant, characterized by small gland-like lumina filled with a PAS-positive, diastase-resistant substance. METHODS: Three samples of arachnoid tissue and 9 SMs were studied with a panel of monoclonal antibodies (MoAbs) against cytokeratins (Ck) 7,8, 20, vimentin, EMA, CEA and the mucin epitopes sialyl-Tn, Tn, CA19.9, CA125, against the BerEP4 and CD15; 3 colonic adenocarcinomas metastatic to the brain and a case of meningioma with metastasis from a gastric signet-ring cell carcinoma were also studied with the same panel of MoAbs for comparison. RESULTS: The 3 samples of arachnoid cap cells were positive only for EMA and vimentin in the supportive stroma. All 9 SMs resulted Ck7+, Ck8+, Ck20-, EMA+, CEA and mucin epitopes+, confirming at an immunohistochemical level the glandular differentiation. Notable exceptions were the negativity for BerEP4 and CD15 antigens. Conversely, the 3 metastatic colonic adenocarcinomas to the brain were Ck7-, Ck8+, Ck20+, CEA and mucin epitopes+, BerEP4 and CD15+; the gastric signet-ring cell carcinoma metastatic to a meningioma showed the same immunophenotype as the other metastatic adenocarcinomas with the exception of BerEP4 negativity. CONCLUSION: The different pattern of cytokeratin expression (Ck7+/Ck20- for SMs, and Ck7-/ Ck20+ for adenocarcinomas) and the negativity for BerEP4 and CD15 epitopes of SMs, could be relevant for the distinction between SMs and metastatic adenocarcinomas.

Giant cell arteritis in association with cerebral amyloid angiopathy: immunohistochemical and molecular studies.

Giant cell arteritis (GCA) usually manifests as a transmural vascular infiltrate of mononuclear and multinucleated giant cells (MNGC). We describe six patients with GCA associated with severe cerebral amyloid angiopathy (CAA), all with cerebral hemorrhage or varying degrees of cerebral infarct, and histological evidence of Alzheimer's disease (cortical CAA often predominating over senile plaques and neurofibrillary tangles). One case showed mostly cortical involvement (with old microhemorrhages), and the others were primarily leptomeningeal (with involvement of the underlying cortex and extensive encephalomalacia of adjacent brain). Many vessels with CAA exhibited a pronounced adventitial and perivascular infiltrate of lymphocytes, histiocytes, and MNGC. Immunohistochemical staining showed deposition of beta/A4 peptide primarily in the thickened media of CAA vessels, and within the cytoplasm of MNGC--suggesting phagocytosis of insoluble peptide. Cystatin C antibody stained vascular amyloid and diffusely highlighted astrocytic and MNGC cytoplasm. HAM56-positive macrophages were frequently seen around amyloid-laden vessels. Anti-smooth muscle actin immunohistochemistry suggests the occurrence of medial destruction by amyloid, with relative preservation of intimal cells. Ultrastructural studies performed in one case confirmed the presence of intracytoplasmic amyloid in MNGC. The GCA seen in these cases of CAA most likely represents a foreign body response to amyloid proteins, causing secondary destruction of the vessel wall. DNA from brain tissues of five affected patients was examined to assess whether mutations were present in exon 17 of the APP gene or exon 2 of the cystatin C gene, a finding that might explain the foreign body giant cell response to amyloid proteins in these cases. However, restriction fragment mapping of amplified gene segments showed that previously described mutations were not present in these cases.

Prolactin receptors in human meningiomas: characterization and biological role.

Sixty cerebral meningioma specimens obtained at surgery from 34 female and 26 male patients were examined for the presence of prolactin (PRL) receptors. These were compared with normal arachnoid tissue from which these tumours arise. PRL receptors were detected in 61.7% of meningiomas whereas no PRL binding was found in samples of normal arachnoid tissue. No relationship was found when sex or histological findings were compared with the presence of PRL receptors. Receptor-positive tumours had saturable and high-affinity (Kd, 4.8 +/- 0.5 ng/ml) receptors with hormonal specificity for human PRL (hPRL) resembling that of other target tissues of PRL in man. The biological role of these receptors was investigated in primary cell cultures derived from meningioma tissue characterized for PRL receptor. When human PRL was added to the culture medium, in doses ranging from 1 to 200 ng/ml, a dose-dependent stimulation of 3H-thymidine incorporation was observed only in PRL-receptor positive tumours. The PRL concentrations required to produce a half-maximal effect ranged from 11 to 20 ng/ml and were quite close to the dissociation constant (Kd) of binding of PRL to its receptors. PRL also caused an increase of cell number compared with control with a significant effect after 3 and 4 days of culture. In conclusion, these findings indicate that a large number of human meningiomas express specific and functional receptors for PRL which are involved in mediating its proliferative effects.

Embryonic and postnatal expression of four gamma-aminobutyric acid transporter mRNAs in the mouse brain and leptomeninges.

The distribution of gamma-aminobutyric acid (GABA) transporter mRNAs (mGATs) was studied in mouse brain during embryonic and postnatal development using in situ hybridization with radiolabeled oligonucleotide probes. Mouse GATs 1 and 4 were present in the ventricular and subventricular zones of the lateral ventricle from gestational day 13. During postnatal development, mGAT1 mRNA was distributed diffusely throughout the brain and spinal cord, with the highest expression present in the olfactory bulbs, hippocampus, and cerebellar cortex. The mGAT4 message was densely distributed throughout the central nervous system during postnatal week 1; however, the hybridization signal in the cerebral cortex and hippocampus decreased during postnatal weeks 2 and 3, and in adults, mGAT4 labeling was restricted largely to the olfactory bulbs, midbrain, deep cerebellar nuclei, medulla, and spinal cord. Mouse GAT2 mRNA was expressed only in proliferating and migrating cerebellar granule cells, whereas mGAT3 mRNA was absent from the brain and spinal cord throughout development. Each of the four mGATs was present to some degree in the leptomeninges. The expression of mGATs 2 and 3 was almost entirely restricted to the pia-arachnoid, whereas mGATs 1 and 4 were present only in specific regions of the membrane. Although mGATs 1 and 4 may subserve the classical purpose of terminating inhibitory GABAergic transmission through neuronal and glial uptake mechanisms, GABA transporters in the pia-arachnoid may help to regulate the amount of GABA available to proliferating and migrating neurons at the sub-pial surface during perinatal development.

Absence of neuronal and glial proteins in human and rat leptomeninges in situ.

Human leptomeningeal (arachnoid and pia mater) cells in culture have been demonstrated in replicated studies to express typical neuronal proteins such as neurofilament protein and neuron-specific enolase. In addition, they can express glial fibrillary acidic protein. The present study examines the possibility that neuronal and glial proteins might be present in rat and human leptomeningeal cells in situ. The neuronal proteins 160 kDa and 200 kDa neurofilaments, neuron-specific enolase and microtubule-associated protein 2 were, however, not immunolocalized in either the pia mater or arachnoid. Glial fibrillary acidic protein and galactocerebroside were also not detected, while fibronectin and vimentin immunoreactivities were robust in all layers of the leptomeninges. Together with the previously reported expression of some neuronal and astroglial markers in cultured human leptomeninges, these observations suggest that culture alters the properties of leptomeningeal cells.

Expression of NF2 gene product merlin in arachnoid villi and meningiomas.

Neurofibromatosis type 2 (NF2) gene encodes a novel 595 amino acid protein named merlin. Recently, Ruttledge et al demonstrated inactivation of NF2 gene in approximately 60% of sporadically occurring meningiomas. Merlin is thought to physiologically exist beneath the cell membrane, and to form a part of modulation in signal transduction, for example, information concerning contact inhibition. In NF2-related tumors, it is supposed that the mutation of merlin results in loss of this signal transduction leading to tumorigenesis. In this paper, we investigated the expression of NF2 gene product merlin in arachnoid villi and meningiomas. The immunohistochemical staining of merlin showed a striking contrast between arachnoid villi and meningiomas. In arachnoid cells, merlin was labeled in the whole cytoplasm, but not within the nuclei. In contrast, in meningiomas, immunoreactivity of merlin was mainly seen in the nuclei. These results suggest that arachnoid cells with normal merlin are capable of normal signal transduction, whereas meningioma cells with mutated merlin show impairment of signal transduction which may lead to tumorigenesis.

Expression of neural cell-adhesion molecule isoforms and epithelial cadherin adhesion molecules in 47 human meningiomas: correlation with clinical and morphological data.

Meningiomas are derived from arachnoidal cells exhibiting both epithelial and mesenchymal properties. The expression of epithelial cadherin and neural cell-adhesion molecule isoforms was examined in 47 meningioma specimens and normal arachnoid villi by using immunohistochemistry and immunoblot procedures. For immunohistochemistry, the percentage of immunostained surface versus counterstained surface and the intensity of labeling were quantified by image analysis and correlated with clinical and morphological data. Neural cell-adhesion molecule, 140-kDa, was expressed by all meningiomas examined and by cap cells and the fibrous capsule forming the arachnoid villi. Neural cell-adhesion-molecule immunostaining was correlated to tumor location, histological type, and grade. The expression of polysialylated isoforms was related to tumor location and grade, with a higher expression on convexity meningiomas and high-grade tumors. Epithelial cadherin was expressed with various intensity. In arachnoid villi, its expression was restricted to cap cells, but it was found in all meningiomas, whatever their histological type or grade. However, the expression was stronger in whorls and meningiotheliomatous areas versus fibroblastic areas. No correlation was observed with the tumor location or the grade. These data suggest that neural cell-adhesion molecule and epithelial cadherin play a role in the morphogenesis and histogenesis of human meningiomas.

Tenascin in the injured rat optic nerve and in non-neuronal cells in vitro: potential role in neural repair.

The distribution of tenascin was examined in the lesioned adult rat optic nerve and central nervous system (CNS) non-neuronal cells in vitro, by means of a double immunofluorescence technique. Tenascin-like immunoreactivity is localized to the leptomeninges and astrocytes that border the site of optic nerve transection. Anti-tenascin labeling was observed as early as 24 hours after transection, when it appeared as a fine interface between leptomeninges and neural tissue. The anti-tenascin labeling increased in the cells at this border zone during the next 2 weeks, and disappeared 18-21 days after transection. In vitro studies further confirmed that both astrocytes and leptomeningeal cells express tenascin as detected by immunofluorescence labeling with anti-tenascin antibodies. However, the pattern of immunolabeling associated with the two cell types differed. Astrocytes showed exclusively punctate labeling of the cell surface, while leptomeningeal cells showed mainly coarse, fibrillary, matrix-like deposits. Astrocytes and leptomeningeal cells remained segregated when cocultured. In these cultures, an increased amount of the fibrillary, matrix-like deposits of tenascin was also observed in the region of the interface between astrocytes and leptomeningeal cells when these two cell types contact each other. Given the antiadhesive and antispreading properties of tenascin, these in vivo and in vitro results suggest that tenascin might play a role in the initial segregation of leptomeningeal cells from neural tissue at the site of CNS trauma during the first 2 weeks after injury, i.e., prior to the formation of a fully differentiated glia limitans. Therefore, tenascin may influence the early stages in the formation of the glia limitans, and thus prevent the indiscriminate migration of leptomeningeal cells into CNS tissue after injury.

Expression of cell adhesion molecule E-cadherin in human arachnoid villi.

Calcium-dependent epithelial cell adhesion molecules designated as E-cadherin (also known as uvomorulin or L-CAM) were identified in human arachnoid villi by immunoblotting and immunocytochemical analyses using a monoclonal antibody HECD-1 raised against human mammary carcinoma MCF-7 cells. Immunoblot analysis showed that HECD-1 recognizes E-cadherin with a molecular weight of 124 kD. In all arachnoid cells of an arachnoid villus, E-cadherin was detected by immunolight microscopy within the cytoplasm rather than the cellular boundaries as seen in the control group. Furthermore, the extent of expression by immunolight microscopy varied from portion to portion. The expression was usually weak in the syncytial cluster which was ultrastructurally composed of tightly juxtaposed cells characterized by few extracellular cisterns and numerous cell junctions, while it was intense in the reticular cluster and the surface layer which were ultrastructurally characterized by abundant extracellular cisterns and smaller numbers of cell junctions. The cells of the reticular cluster and the surface layer contained more free ribosomes than those of the syncytial cluster. Immunoelectron microscopy showed that E-cadherin was localized not only to the opposing plasma membranes and the cytoplasm around the free ribosomes or the rough endoplasmic reticulum but also to the extracellular cisterns. As the expression of E-cadherin was closely related to the arachnoid cells adjacent to the cerebrospinal fluid pathway, it is suggested that, instead of the cell junctions, E-cadherin may play an important role in the flexible adhesion of arachnoid cells even in the presence of the cerebrospinal fluid.

Evidence for transforming growth factor-beta expression in human leptomeningeal cells and transforming growth factor-beta-like activity in human cerebrospinal fluid.

BACKGROUND: Little is known about the factors regulating growth and maintenance of human leptomeningeal cells. The influence of cerebrospinal fluid on these functions is also unknown. Possible mediators include the transforming growth factor-beta (TGF beta) family, three closely related peptides that regulate proliferation and numerous other physiologic processes in most mesenchymal cells. EXPERIMENTAL DESIGN: Expression of both mRNA and protein for TGF beta isoforms TGF beta 1, TGF beta 2, and TGF beta 3 as well as TGF beta-competing activity were evaluated in primary human leptomeningeal cultures by Northern blot analysis, immunohistochemistry, and a radioreceptor assay, respectively. TGF beta 1, TGF beta 2, and TGF beta 3 immunoreactivity was also evaluated in brain sections containing leptomeninges from which these cell cultures were established. An additional study analyzed human cerebrospinal fluid for TGF beta-like activity. RESULTS: Transcripts for TGF beta 1, TGF beta 2 and TGF beta 3 were detected in RNA from each of the eight leptomeningeal cultures. Significant TGF beta 1 immunoreactivity was detected in leptomeningeal tissue from five of eight cases. TGF beta 2 and TGF beta 3 immunostaining was seen in eight and seven of the cases, respectively. Similarly, cells cultured from these meninges exhibited variable TGF beta 1 and extensive TGF beta 2 and TGF beta 3 immunoreactivity. Radioreceptor assays of conditioned media from four cultures demonstrated significant latent TGF beta-like activity. TGF beta radioreceptor competing activity was also detected by radioreceptor assay in normal blood-free cerebrospinal fluid from 32 patients without neurological disease. In addition, pooled cerebrospinal fluid (from six additional patients) exhibited dose dependent TGF beta-like activity in the radioreceptor assay, stimulation of AKR-2B cell growth in soft agar and inhibition of growth in CCL-64 cell assays suggesting that cerebrospinal fluid contains TG beta-like activity. CONCLUSIONS: These findings suggest that the human leptomeninges synthesize TGF beta 1, TGF beta 2 and TGF beta 3 and secrete latent TGF beta s at least in vitro. Human cerebrospinal fluid may also contain TGF beta isoforms. Collectively, these observations raise the possibility that members of the TGF beta family contribute to biologic processes of the leptomeninges.