Differential labeling of the cell surface of single ciliary ganglion neurons in vitro.

During development, parasympathetic ciliary ganglion neurons arise from the neural crest and establish synaptic contacts on smooth and striate muscle in the eye. The factors that promote the ciliary ganglion pioneer axons to grow toward their targets have yet to be determined. Here, we show that glial cell line-derived neurotrophic factor (GDNF) and neurturin (NRTN) constitute target-derived factors for developing ciliary ganglion neurons. Both GDNF and NRTN are secreted from eye muscle located in the target and trajectory pathway of ciliary ganglion pioneer axons during the period of target innervation. After this period, however, the synthesis of GDNF declines markedly, while that of NRTN is maintained throughout the cell death period. Furthermore, both in vitro and in vivo function-blocking of GDNF at early embryonic ages almost entirely suppresses ciliary axon outgrowth. These results demonstrate that target-derived GDNF is necessary for ciliary ganglion neurons to innervate ciliary muscle in the eye. Since the down-regulation of GDNF in the eye is accompanied by down-regulation of GFRα1 and Ret, but not of GFRα2, in innervating ciliary ganglion neurons, the results also suggest that target-derived GDNF regulates the expression of its high-affinity coreceptors.

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Choroid tissue supports the survival of ciliary ganglion neurons in vitro

Journal of Neuroscience ◽

10.1523/jneurosci.13-07-03143.1993 ◽

1993 ◽

Vol 13 (7) ◽

pp. 3143-3154 ◽

Cited By ~ 8

Author(s):

LA Wentzek ◽

CW Bowers ◽

L Khairallah ◽

G Pilar

Keyword(s):

Ciliary Ganglion ◽

Ciliary Ganglion Neurons ◽

Ganglion Neurons

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Neph1 regulates steady-state surface expression of Slo1 Ca2+-activated K+ channels: different effects in embryonic neurons and podocytes

AJP Cell Physiology ◽

10.1152/ajpcell.00354.2009 ◽

2009 ◽

Vol 297 (6) ◽

pp. C1379-C1388 ◽

Cited By ~ 19

Author(s):

Eun Young Kim ◽

Yu-Hsin Chiu ◽

Stuart E. Dryer

Keyword(s):

Steady State ◽

Cell Surface ◽

Splice Variants ◽

Surface Expression ◽

Immunoglobulin Superfamily ◽

Ciliary Ganglion ◽

Whole Cell ◽

Differentiated Cells ◽

Ciliary Ganglion Neurons ◽

Ganglion Neurons

Large-conductance Ca2+-activated K+ (BKCa) channels encoded by the Slo1 gene are often components of large multiprotein complexes in excitable and nonexcitable cells. Here we show that Slo1 proteins interact with Neph1, a member of the immunoglobulin superfamily expressed in slit diaphragm domains of podocytes and in vertebrate and invertebrate nervous systems. This interaction was established by reciprocal coimmunoprecipitation of endogenous proteins from differentiated cells of a podocyte cell line, from parasympathetic neurons of the embryonic chick ciliary ganglion, and from HEK293T cells heterologously expressing both proteins. Neph1 can interact with all three extreme COOH-terminal variants of Slo1 (Slo1VEDEC, Slo1QEERL, and Slo1EMVYR) as ascertained by glutathione S-transferase (GST) pull-down assays and by coimmunoprecipitation. Neph1 is partially colocalized in intracellular compartments with endogenous Slo1 in podocytes and ciliary ganglion neurons. Coexpression in HEK293T cells of Neph1 with any of the Slo1 extreme COOH-terminal splice variants suppresses their steady-state expression on the cell surface, as assessed by cell surface biotinylation assays, confocal microscopy, and whole cell recordings. Consistent with this, small interfering RNA (siRNA) knockdown of endogenous Neph1 in embryonic day 10 ciliary ganglion neurons causes an increase in steady-state surface expression of Slo1 and an increase in whole cell Ca2+-dependent K+ current. Surprisingly, a comparable Neph1 knockdown in podocytes causes a decrease in surface expression of Slo1 and a decrease in whole cell BKCa currents. In podocytes, Neph1 siRNA also caused a decrease in nephrin, even though the Neph1 siRNA had no sequence homology with nephrin. However, we could not detect nephrin in ciliary ganglion neurons.

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Regulation of neuronal K(+) currents by target-derived factors: opposing actions of two different isoforms of TGFbeta

Development ◽

10.1242/dev.126.18.4157 ◽

1999 ◽

Vol 126 (18) ◽

pp. 4157-4164

Author(s):

J.S. Cameron ◽

L. Dryer ◽

S.E. Dryer

Keyword(s):

Functional Expression ◽

Developmental Expression ◽

Ciliary Ganglion ◽

Target Tissue ◽

Intraocular Injection ◽

Ciliary Ganglion Neurons ◽

Ganglion Neurons ◽

K Currents

The developmental expression of macroscopic Ca(2+)-activated K(+) currents in chick ciliary ganglion neurons is dependent on an avian ortholog of TGFbeta1, known as TGFbeta4, secreted from target tissues in the eye. Here we report that a different isoform, TGFbeta3, is also expressed in a target tissue of ciliary ganglion neurons. Application of TGFbeta3 inhibits the functional expression of whole-cell Ca(2+)-activated K(+) currents evoked by 12 hour treatment with either TGFbeta1 or beta-neuregulin-1 in ciliary ganglion neurons developing in vitro. TGFbeta3 had no effect on voltage-activated Ca(2+) currents. A neutralizing antiserum specific for TGFbeta3 potentiates stimulation of Ca(2+)-activated K(+) currents evoked by a target tissue (iris) extract in cultured ciliary ganglion neurons, indicating that TGFbeta3 is an inhibitory component of these extracts. Intraocular injection of TGFbeta3 causes a modest but significant inhibition of the expression of Ca(2+)-activated K(+) currents in ciliary ganglion neurons developing in vivo. Further, intraocular injection of a TGFbeta3-neutralizing antiserum stimulates expression of Ca(2+)-activated K(+) currents in ciliary ganglion neurons developing in vivo, indicating that endogenous TGFbeta3 regulates the functional expression of this current. The normal developmental expression of functional Ca(2+)-activated K(+) currents in ciliary ganglion neurons developing in vivo is therefore regulated by two different target-derived isoforms of TGFbeta, which produce opposing effects on the electrophysiological differentiation of these neurons.

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Parameters of neuritic growth from ciliary ganglion neurons in vitro: influence of laminin, schwannoma polyornithine-binding neurite promoting factor ciliary neuronotrophic factor

Developmental Brain Research ◽

10.1016/0165-3806(85)90133-6 ◽

1985 ◽

Vol 17 (1-2) ◽

pp. 75-84 ◽

Cited By ~ 34

Author(s):

George E. Davis ◽

Marston Manthorpe ◽

Silvio Varon

Keyword(s):

Ciliary Ganglion ◽

Neuritic Growth ◽

Ciliary Ganglion Neurons ◽

Ganglion Neurons ◽

Ciliary Neuronotrophic Factor

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Changes in the number of chick ciliary ganglion neuron processes with time in cell culture.

The Journal of Cell Biology ◽

10.1083/jcb.104.2.363 ◽

1987 ◽

Vol 104 (2) ◽

pp. 363-370 ◽

Cited By ~ 20

Author(s):

L W Role ◽

G D Fischbach

Keyword(s):

Cell Culture ◽

Time Course ◽

Culture Conditions ◽

Ciliary Ganglion ◽

Intact Cells ◽

Dissociated Cell Culture ◽

Ciliary Ganglion Neurons ◽

Ganglion Neurons

The purpose of this study was to describe the shape of chick ciliary ganglion neurons dissociated from embryonic day 8 or 9 ganglia and maintained in vitro. Most of the neurons were multipolar during the first three days after plating, with an average of 6.0 processes extending directly from the cell body. The neurons became unipolar with time. The remaining primary process accounted for greater than 90% of the total neuritic arbor. This striking change in morphology was not due to the selective loss of multipolar cells, or to an obvious decline in the health of apparently intact cells. The retraction of processes was neither prevented nor promoted by the presence of embryonic muscle cells. Process pruning occurred to the same extent and over the same time course whether the cells were plated on a monolayer of embryonic myotubes or on a layer of lysed fibroblasts. Process retraction is not an inevitable consequence of our culture conditions. Motoneurons dissociated from embryonic spinal cords remained multipolar over the same period of time. We conclude that ciliary ganglion neurons breed true in dissociated cell culture in that the multipolar-unipolar transition reflects their normal, in vivo, developmental program.

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