Response of purified chick motoneurons to myotube conditioned medium: Laminin is essential for the substratum-binding, neurite outgrowth-promoting activity

Calof, Anne L.; Reichardt, Louis F.

doi:10.1016/0304-3940(85)90197-1

Cited by 58 publications

(29 citation statements)

References 16 publications

(18 reference statements)

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“…As previously reported for the neurite-promoting activity found in conditioned media of muscle cells (Calof and Reichardt, 1985) we eould immunoprecipitate the activity in musele extract using antilaminin antiserum. Immunoblotting showed that Al did indeed contain laminin, both A and B chains of the molecule being deteeted.…”

Section: Discussionmentioning

confidence: 95%

“…Furthermore, an activity that stimulates neurite outgrowth from motor neurons has been identified and shown to have structural and antigenic similarities with the basement membrane protein, laminin (Calof and Reichardt, 1985). It has, however, been shown previously that organ explants can begin to produce neuronal survival factors when brought into culture, although no such activity was detectable in the intact tissue (Harper et al, 1980;Ebendal et aL, 1980;Barth et aL, 1984).…”

Section: Introductionmentioning

confidence: 97%

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Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture

Dohrmann

Edgar

Sendtner

et al. 1986

Developmental Biology

156

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The purpose of the experiments reported is to provide an unambiguous demonstration that embryonie skeletal muscle contains factors that act directly on embryonie spinal motor neurons both to support their survival and to stimulate the outgrowth of neurites. Cells of lumbar and brachial ventral spinal cords from 6-day-old chick embryos were separated by centrifugation in a two-step metrizamide gradient, and a motor neuron enriched fraction was obtained. Motor neurons were identified by retrogradely labeling with rhodamine isothiocyanate, and were enriched fourfold in the motor neuron fraction relative to unfractionated cells. In culture, the isolated motor neurons died within 3-4 days unless they were supplemented with embryonie chick skeletal muscle extract. Two functionally distinct entities separable by ammonium sulfate precipitation were responsible for the effects of muscle extracts on motor neurons. The 0-25% ammonium sulfate precipitate contained molecules that alone bad no effect on neuronal survival but when bound to polyornithine-coated culture substrata, stimulated neurite outgrowth and potentiated the survival activity present in muscle. Mostofthis activity was due to a laminin-like molecule being immunoprecipitated with antisera against laminin, and immunoblotting demonstrated the presence of both the A and B chains of laminin. A long-term survival activity resided in the 25-70% ammonium sulfate fraction, and its apparent total and specific activities were strongly dependent on the culture substrate. In contrast to the motor neurons, the cells from the other metrizamide fraction (including neuronal cells) could be kept in culture for a prolonged time without addition of exogenous factor(s).

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Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 97%

Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture

Dohrmann

Edgar

Sendtner

et al. 1986

Developmental Biology

156

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“…It is surprising to note that in spite of heparin-binding sites on both LN and FN (Yamada et al 1985) LN is predominantly associated with a sulfated proteoglycan, whereas FN is mainly present in CM as a noncomplexed entity. In previous investigations (Calof and Reichardt, 1985;Davis et al, 1985;Lander et al, 1985a1, it was suggested that a LN proteoglycan complex is associated with the NPA of a large number of different CMs. However, FN, which was present in CM in considerable amounts (for ref.…”

Section: Discussionmentioning

confidence: 98%

Astroglia‐released neurite growth‐inducing activity for embryonic hippocampal neurons is associated with laminin bound in a sulfated complex and free fibronectin

Matthiessen

Schmalenbach

Müller

1989

Glia

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Neurons from embryonic (E18) rat hippocampus were chosen to identify and characterize neurite growth-stimulating proteins accumulating in serum-free conditioned media (CM) obtained from primary or secondary cultures of cerebral astrocytes (less than 5% nonglial cells) using a quantitative cell culture bioassay. CM were fractionated by FPLC on an anion exchange column (Mono Q) and by gel filtration (Superose 6). Column fractions were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting and enzyme-linked immunosorbent assay (ELISA) using antibodies to laminin (LN) and fibronectin (FN). The neurite-promoting activity (NPA) was tested by incubating aliquots of the eluted fractions with poly-L-lysine precoated glass coverslips prior to addition of neurons suspended in chemically defined medium. We provide evidence that the NPA in astroglial CM could be assigned mainly to a negatively charged, highly sulfated LN complex consisting predominantly of the B-chains of LN and presumably a sulfated proteoglycan that was sensitive for chondroitinase and to a lower degree to heparinase degradation. In addition, a smaller proportion of the NPA was associated with uncomplexed LN and free FN. FN reached approximately 10 times the concentration of LN in astroglial CM. As revealed by immunofluorescence microscopy, both LN and FN are simultaneously expressed by cultured astrocytes; however, only the production of FN, measured by ELISA, increased during the time astrocytes were in culture, whereas the release of LN remained unchanged. We conclude that, besides the most active LN complex, FN bound to a polycationic matrix is able to induce neurite growth in hippocampal neurons in vitro.

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“…The first observation is that laminin, as a tissue culture substrate, is extremely potent in promoting and sustaining neurite outgrowth in vitro (Baron-van Evercooren et al, 1982;Manthorpe et al, 1983;Rogers et al, 1983;Edgar et al, 1984;Liesi et al, 1984a;Smallheiser et al, 1984;Calof & Reichardt, 1985). Furthermore, some neuronal growth cones possess surface membrane receptors that bind laminin (Horwitz et aI., 1985;Fig.…”

Section: Discussionmentioning

confidence: 94%

“…The evidence to support this idea comes from a variety of circumstantial observations. For instance, in cell culture, laminin has been shown to be the most effective of all substrates so far tested in supporting neurite outgrowth and elongation of neurons from both the CNS and PNS ( Baron-van Evercooren et al, 1982;Manthorpe et al, 1983;Rogers et al, 1983;Edgar et al, 1984;Faivre-Bauman et al, 1984;Liesi et aI., 1984a,b;Smallheiser et al, 1984;Calof & Reichardt, 1985). It is transiently expressed in early developing fibre tracts in the CNS in a form that is particulate and not associated with basal laminae (Cohen et al, 1987;Letourneau et al, 1988 but see Sosale et aI., 1988) and permanently present in regions of the CNS in which axons are added continuously throughout adult life, such as the rat olfactory system (Liesi, 1985b) and the goldfish optic nerve (Hopkins et al, 1985).…”

Section: Introductionmentioning

confidence: 96%

Transient expression of laminin immunoreactivity in the developing rat hippocampus

et al. 1989

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We have isolated and characterized a polyclonal antibody that recognizes the extracellular matrix glycoprotein laminin. In the developing rat hippocampus, in addition to staining the basal laminae of blood vessels and the glia limitans, this antibody detects a punctate deposit of non-basal lamina laminin that appears transiently, disappearing on E18. The punctate laminin is distributed throughout the hippocampus from the border of the ependyma and the neuropil to the pial surface, but is particularly concentrated in the presumptive molecular layer (marginal zone). Electron microscopy showed that the punctate laminin was in the extracellular space on the surfaces of cell bodies and, more commonly, their processes. In double-labelling experiments in which neurites were labelled with tau or MAP2 antibodies, there were occasional cases where neurites came into direct contact with punctate laminin. These observations suggest a role for laminin in hippocampal development and possibly in neurite outgrowth.

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Response of purified chick motoneurons to myotube conditioned medium: Laminin is essential for the substratum-binding, neurite outgrowth-promoting activity

Cited by 58 publications

References 16 publications

Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture

Muscle-derived factors that support survival and promote fiber outgrowth from embryonic chick spinal motor neurons in culture

Astroglia‐released neurite growth‐inducing activity for embryonic hippocampal neurons is associated with laminin bound in a sulfated complex and free fibronectin

Transient expression of laminin immunoreactivity in the developing rat hippocampus

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