Responses of cultured neural retinal cells to substratum-bound laminin and other extracellular matrix molecules

Adler, Ruben; Jerdan, Janice A.; Hewitt, Alice

doi:10.1016/0012-1606(85)90124-1

Cited by 127 publications

(72 citation statements)

References 41 publications

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“…In that respect, cell adhesion to galectin-8 resembles cell adhesion to other ECM proteins, which is largely inhibited at low temperatures (compare Ref. 46). Cellular attachment and spreading on fibronectin involves an initial requirement for Cdc42 in the formation of filopodial protrusions and the subsequent involvement of both Cdc42 and Rac during cell spreading and organization of the actin cytoskeleton (47).…”

Section: Fig 11mentioning

confidence: 99%

Sustained Induction of ERK, Protein Kinase B, and p70 S6 Kinase Regulates Cell Spreading and Formation of F-actin Microspikes Upon Ligation of Integrins by Galectin-8, a Mammalian Lectin

Levy

Ronen

Bershadsky

et al. 2003

Journal of Biological Chemistry

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Galectin-8, a member of the galectin family of mammalian lectins, is a secreted protein that promotes cell adhesion and migration upon binding to a subset of integrins through sugar-protein interactions. Ligation of integrins by galectin-8 triggers a distinct pattern of cytoskeletal organization, including formation of F-actin-containing microspikes. This is associated with activation of integrin-mediated signaling cascades (ERK and phosphatidylinositol 3 kinase (PI3K)) that are much more robust and are of longer duration than those induced upon cell adhesion to fibronectin. Indeed, formation of microspikes is enhanced 40% in cells that overexpress protein kinase B, the downstream effector of PI3K. Inhibition of PI3K activity induced by wortmannin partially inhibits cell adhesion and spreading while largely inhibiting microspike formation in cells adherent to galectin-8. Furthermore, the inhibitory effects of wortmannin are markedly accentuated in cells overexpressing PKB or p70S6K (CHO PKB and CHO p70S6K cells), whose adhesion and spreading on galectin-8 (but not on fibronectin) is inhibited ϳ25-35% in the presence of wortmannin. The above results suggest that galectin-8 is an extracellular matrix protein that triggers a unique repertoire of integrin-mediated signals, which leads to a distinctive cytoskeletal organization and microspike formation. They further suggest that downstream effectors of PI3K, including PKB and p70 S6 kinase, in part mediate cell adhesion, spreading, and microspike formation induced by galectin-8. Extracellular matrix (ECM)1 proteins have important functions in providing structural integrity to tissues and in presenting proper environmental cues for cell adhesion, migration, growth, and differentiation (1-5). These functions rely on spatio-temporal expression of adhesive as well as anti-adhesive components of the ECM proteins (6). ECM proteins like fibronectin (7-9), collagen (10), and laminin (11) are best characterized, though other types of proteins, including mammalian lectins, also function as modulators of cell adhesion. Selectins mediate cell-cell interactions (12) through calciumdependent recognition of sialylated glycans (13, 14), whereas galectins, animal lectins that specifically bind ␤-galactoside residues (15), were implicated as modulators of cell-matrix interactions. Although lacking a signal peptide and found mainly in the cytosol, galectins are externalized by an atypical secretory mechanism (16) to regulate cell growth, cell transformation, embryogenesis, and apoptosis (reviewed in Refs. 17 and 18). In accordance with their proposed functions, galectins enhance or inhibit cell-matrix interactions (reviewed in Ref. 19).Cellular adhesion to extracellular matrix proteins is mediated by a diverse class of cell surface ␣␤ heterodimeric receptors known as integrins (2,20,21). In addition to mediating cell adhesion, integrins induce multiple signal transduction pathways that regulate cytoskeletal rearrangements, cell spreading, migration, differentiation, survival, and cell...

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Section: Fig 11mentioning

confidence: 99%

Sustained Induction of ERK, Protein Kinase B, and p70 S6 Kinase Regulates Cell Spreading and Formation of F-actin Microspikes Upon Ligation of Integrins by Galectin-8, a Mammalian Lectin

Levy

Ronen

Bershadsky

et al. 2003

Journal of Biological Chemistry

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“…Laminin provided a better substrate for retinal neurite growth and survival than type I collagen, fibronectin, or poly (DL-omithine) (results not shown), as indicated by others (Adler et al, 1985;Smalheiser et al, 1984). Many retinal neurons began developing neurite outgrowth during the first few hours on a laminin substrate, irrespective of the presence or absence of BDNF (Figs.…”

Section: Fetal Retinal Culturesmentioning

confidence: 51%

Brain-derived neurotrophic factor supports the survival of cultured rat retinal ganglion cells

Johnson¹,

Barde²,

Schwab³

et al. 1986

J. Neurosci.

488

238

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Brain-derived neurotrophic factor (BDNF) is a small, basic protein purified from the mammalian brain that has been shown previously to support the survival of cultured spinal sensory neurons (Barde et al., 1982). In current studies, BDNF was tested for its ability to support the survival of cultured CNS cells isolated from the perinatal rat retina. Both immunofluorescent labeling of Thy-l and prior retrograde labeling with HRP were used as retinal ganglion cell markers in vitro. With embryonic day (E) 17 retinas, it was found that BDNF allowed the survival of a small subpopulation of neurons (about 7% of the cells plated at this age) identified by the immunofluorescent labeling of Thy-1. No detectable effects were seen when either the total number of cells or the number of tetanus toxin-positive neurons was measured. BDNF also had an effect on cultured neurons retrogradely labeled after HRP injections in the superior colliculi of neonatal rats. The BDNF-responsive population was therefore detected only in retinal cultures with specific markers and identified as consisting of retinal ganglion cells. These cells could be enriched about 80-fold by density gradient centrifugation, and purified ganglion cell cultures were shown to be responsive to BDNF. Whereas with El7 retinas, the number of surviving Thy-l positive neurons could be kept constant for at least 4 d, the survival of postnatal neurons was only transiently increased by BDNF.We conclude that in the retina, BDNF affects only the survival of ganglion cells in vitro by a direct action on these cells. The results are discussed in terms of target-derived neurotrophic support during development.The death of a substantial proportion of many different types of neurons is a common feature of the normal development of the vertebrate nervous system. It is generally believed to serve in the appropriate quantitative matching of a neuronal population with its respective terminal field .In vertebrates, evidence for the epigenetic regulation of neuronal cell death in the CNS as well as in the PNS has arisen from both target transplantation and ablation experiments. There is good evidence for molecular agents that serve to regulate neuronal cell death, the clearest being provided by the protein nerve growth factor (NGF). Treatment of developing mammals with antibodies against NGF results in the selective elimination of a large proportion of the peripheral sympathetic and sensory neurons. On the other hand, the treatment of animals with Received Jan. 7, 1986; revised Apr. 7, 1986; accepted Apr. 11, 1986. We wish to thank Dr. Hermann Rohrer for helpful suggestions and Mrs. C. Cap for technical assistance. This work was supported in part by fellowships from the Alexander von Humboldt Foundation, the Kempton Foundation and an NRSA from NIH, NINCDS (PHS-5F32-NSO7332-02X1) awarded to J.E.J.Correspondence should be addressed to Dr. James E. Thoenen and Barde, 1980). Results from in vitro assays have indicated that several other proteins with a similar function, but a differen...

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“…Axons change from fasciculated to unfasciculated growth which might suggest that different sub-strate molecules are used when sympathetic axons grow on the endfeet and on the plain basal lamina. The external limiting membrane contains several molecules which have previously been shown to be a substrate for axons from the central and peripheral nervous system, like laminin (Rogers et al, 1983;Manthorpe et al, 1983;Adler et a]., 1985), collagen (Halfter et al, 1983) or heparan sulfate proteoglycan (Hantaz-Ambroise et al, , 1987;Dow et al, 1988). These molecules remain in the basement membrane even after Triton-extraction.…”

Section: Discussionmentioning

confidence: 99%

Isolation, characterization, and substrate properties of the external limiting membrane from the avian embryonic optic tectum

Kröger

Niehörster

1990

J of Neuroscience Research

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The external limiting membrane of the avian embryonic optic tectum is isolated by mechanically separating the neuronal mesencephalon from the overlying mesenchymal tissue. The preparation consists of a basal lamina which is covered on its neural side by endfeet of neuroepithelial cells and has attached to it on its meningeal side a collageneous stroma, containing blood vessels. The external limiting membrane can be flat-mounted on a piece of nitrocellulose filter as mechanical support. It covers an area between 0.3 and 1 cm', depending on the age of the donor embryo. The endfeet can be removed together with all cellular components of the meninges by treatment with 2% Triton-X-100 or with distilled water. The basal lamina itself is approximately 80 nm thick and consists of two laminae rarae and a central lamina densa. Immunohistochemical staining reveals that the basal lamina in the embryo, after isolation and after detergent extraction of the isolated preparation, contains type IV collagen, nidogen, laminin, and low density heparan sulfate proteoglycan as do other basement membranes. Antibodies against the neural cell adhesion molecule (N-CAM), chondroitin sulfate proteoglycan, and fibronectin fail to stain the external limiting membrane, but these proteins were clearly identified in the blood vessel-containing meninges or in the optic tectum.The flat-mounted external limiting membrane preparation was used as substrate to culture several different neural tissues of central and peripheral origin. Explants of neural crest cells, dorsal root ganglia, and sympathetic ganglia can be cultured on the external limiting membrane. All explants grow well on the basal lamina preparations whether the endfeet are attached or detergent-extracted prior to explantation; however, neurite outgrowth from sympathetic ganglia is reduced in the presence of the endfeet. Although the endfoot-lined external limiting membrane represents at least part of the immediate environment encountered by retinal axons as they invade the optic tectum and despite its excellent properties as a substrate for retinal axons in vitro, cues guiding the orientation of axons were not detected in the flatmounted Preparation.

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Responses of cultured neural retinal cells to substratum-bound laminin and other extracellular matrix molecules

Cited by 127 publications

References 41 publications

Sustained Induction of ERK, Protein Kinase B, and p70 S6 Kinase Regulates Cell Spreading and Formation of F-actin Microspikes Upon Ligation of Integrins by Galectin-8, a Mammalian Lectin

Sustained Induction of ERK, Protein Kinase B, and p70 S6 Kinase Regulates Cell Spreading and Formation of F-actin Microspikes Upon Ligation of Integrins by Galectin-8, a Mammalian Lectin

Brain-derived neurotrophic factor supports the survival of cultured rat retinal ganglion cells

Isolation, characterization, and substrate properties of the external limiting membrane from the avian embryonic optic tectum

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