scholarly journals Combined Vital Dye Labelling and Catecholamine Histofluorescence of Transplanted Ciliary Ganglion Cells

1989 ◽  
Vol 1 (3-4) ◽  
pp. 113-128 ◽  
Author(s):  
John Sechrist ◽  
James N. Coulombe ◽  
Marianne Bronner-Fraser
Keyword(s):  
Neural Crest ◽  
Adrenal Medulla ◽  
Ganglion Cells ◽  
Large Cell ◽  
Cell Types ◽  
Sympathetic Ganglia ◽  
Variable Number ◽  
Ciliary Ganglion ◽  
Soma Size ◽  
Ganglion Neurons

We have utilized the carbocyanine dye, DiI, to label suspensions of dissociated ciliary ganglion cells removed from 6 to 12 day old quail embryos. Some of the cells were injected into the trunk somites of 2.5 - 3 day old chick embryos along pathways where neural crest cells migrate to form sensory and sympathetic ganglia, aortic plexuses and the adrenal medulla; the remainder of the cells were cultured to check their viability and the persistence of the DiI label. Embryos were incubated for 1 – 8 days post-injection, fixed in 4% paraformaldehyde/0.25% glutaraldehyde and processed for cryostat sectioning. DiI-labelled cells were readily identifiable in culture and in sections of embryos at all stages examined. Several cell types were identified, based on their morphology and soma size. These included cells with large cell bodies and bright DiI-labelling that appeared to be neurons and smaller, more weakly labelled cells that appeared non-neuronal. The latter presumably had divided several times, accounting for their reduced levels of dye. Many of the DiI-labelled cells were found in and around neural crest-derived sympathetic ganglia, aortic plexuses and adrenomedullary cords, but were rarely observed in dorsal root ganglia. The aldehyde fixative (Faglu mixture) used in this study reacts with catecholamines to form a bright reaction product in adrenergic cells including those in the sympathetic ganglia and the adrenal medulla. The catecholamine biproduct and the DiI in the same cell can easily be viewed with different fluorescent filter sets. A variable number of the DiI-labelled cells in these adrenergic sites contained catecholamines. Cells derived from younger 6 day ciliary ganglion dissociates exhibited detectable catecholamine neurotransmitters earlier and more frequently than those derived from 8 day embryos. The presence of cells exhibiting both bright DiI and catecholamine fluorescence is consistent with previous indications that post-mitotic ciliary ganglion neurons can undergo phenotypic conversion from cholinergic to adrenergic when transplanted to the trunk environment.

Brn3a is a transcriptional regulator of soma size, target field innervation and axon pathfinding of inner ear sensory neurons

Development ◽  
2001 ◽  
Vol 128 (13) ◽  
pp. 2421-2432 ◽  
Author(s):  
Eric J. Huang ◽  
Wei Liu ◽  
Bernd Fritzsch ◽  
Lynne M. Bianchi ◽  
Louis F. Reichardt ◽  
...  
Keyword(s):  
Inner Ear ◽  
Ganglion Cells ◽  
Substantial Reduction ◽  
Spiral Ganglion ◽  
Neurotrophin Receptor ◽  
Axon Pathfinding ◽  
Target Field ◽  
Afferent Fibers ◽  
Soma Size ◽  
Ganglion Neurons

The POU domain transcription factors Brn3a, Brn3b and Brn3c are required for the proper development of sensory ganglia, retinal ganglion cells, and inner ear hair cells, respectively. We have investigated the roles of Brn3a in neuronal differentiation and target innervation in the facial-stato-acoustic ganglion. We show that absence of Brn3a results in a substantial reduction in neuronal size, abnormal neuronal migration and downregulation of gene expression, including that of the neurotrophin receptor TrkC, parvalbumin and Brn3b. Selective loss of TrkC neurons in the spiral ganglion of Brn3a−/− cochlea leads to an innervation defect similar to that of TrkC−/− mice. Most remarkably, our results uncover a novel role for Brn3a in regulating axon pathfinding and target field innervation by spiral and vestibular ganglion neurons. Loss of Brn3a results in severe retardation in development of the axon projections to the cochlea and the posterior vertical canal as early as E13.5. In addition, efferent axons that use the afferent fibers as a scaffold during pathfinding also show severe misrouting. Interestingly, despite the well-established roles of ephrins and EphB receptors in axon pathfinding, expression of these molecules does not appear to be affected in Brn3a−/− mice. Thus, Brn3a must control additional downstream genes that are required for axon pathfinding.

scholarly journals Electron probe x-ray microanalysis of small granulated cells in rat sympathetic ganglia after sequential aldehyde and dichromate treatment.

1977 ◽  
Vol 25 (4) ◽  
pp. 275-279 ◽  
Author(s):  
J D Lever ◽  
R M Santer ◽  
K S Lu ◽  
R Presley
Keyword(s):  
Adrenal Medulla ◽  
Electron Probe ◽  
Potassium Dichromate ◽  
Cell Types ◽  
Sympathetic Ganglia ◽  
Type Ii ◽  
Treatment Sequence ◽  
X Ray ◽  
Small Granulated Cells

Rat adrenal medulla and celiac-mesenteric sympathetic ganglia were fixed by a glutaraldehyde/formaldehyde-potassium dichromate-osmium treatment sequence and plastic-embedded. Fine sections were examined by electron probe x-ray microanalysis. Comparable peaks for chromium (Kalpha = 5.4 keV) were obtained from cytoplasmic fields containing membrane-bounded inclusion granules in both adrenomedullary noradrenaline cells and a type (type II) of sympathetic small granulated cell whose inclusion granules closely resemble those of the adrenomedullary noradrenaline cell. Chromium was not detected in granules within adrenomedullary adrenaline cells nor in two other sympathetic small granualted cell types. In no material was chromium detected in agranular cytoplasmic or nuclear fields. Since chromium binds to the Schiff monobase formed by glutaraldehyde and noradrenaline during fixation, we infer that noradrenaline is present in the granules of the type II sympathetic small granulated cell, as well as in adrenomedullary noradrenaline cells.

scholarly journals Migrating neural crest cells in the trunk of the avian embryo are multipotent

Development ◽  
1991 ◽  
Vol 112 (4) ◽  
pp. 913-920 ◽  
Author(s):  
S.E. Fraser ◽  
M. Bronner-Fraser
Keyword(s):  
Neural Crest ◽  
Neural Tube ◽  
Sympathetic Neurons ◽  
Morphological Characteristics ◽  
Neural Crest Cells ◽  
Cell Types ◽  
Sympathetic Ganglia ◽  
Avian Embryo ◽  
Developmental Potential ◽  
Trunk Neural Crest

Trunk neural crest cells migrate extensively and give rise to diverse cell types, including cells of the sensory and autonomic nervous systems. Previously, we demonstrated that many premigratory trunk neural crest cells give rise to descendants with distinct phenotypes in multiple neural crest derivatives. The results are consistent with the idea that neural crest cells are multipotent prior to their emigration from the neural tube and become restricted in phenotype after leaving the neural tube either during their migration or at their sites of localization. Here, we test the developmental potential of migrating trunk neural crest cells by microinjecting a vital dye, lysinated rhodamine dextran (LRD), into individual cells as they migrate through the somite. By two days after injection, the LRD-labelled clones contained from 2 to 67 cells, which were distributed unilaterally in all embryos. Most clones were confined to a single segment, though a few contributed to sympathetic ganglia over two segments. A majority of the clones gave rise to cells in multiple neural crest derivatives. Individual migrating neural crest cells gave rise to both sensory and sympathetic neurons (neurofilament-positive), as well as cells with the morphological characteristics of Schwann cells, and other non-neuronal cells (both neurofilament-negative). Even those clones contributing to only one neural crest derivative often contained both neurofilament-positive and neurofilament-negative cells. Our data demonstrate that migrating trunk neural crest cells can be multipotent, giving rise to cells in multiple neural crest derivatives, and contributing to both neuronal and non-neuronal elements within a given derivative.(ABSTRACT TRUNCATED AT 250 WORDS)

A transgene containing lacZ is expressed in primary sensory neurons in zebrafish

Development ◽  
1992 ◽  
Vol 115 (2) ◽  
pp. 421-426 ◽  
Author(s):  
T.A. Bayer ◽  
J.A. Campos-Ortega
Keyword(s):  
Trigeminal Ganglion ◽  
Ganglion Cells ◽  
Transgenic Fish ◽  
Cell Types ◽  
Single Copy ◽  
Reporter Construct ◽  
Lacz Expression ◽  
Promoter Sequences ◽  
Rous Sarcoma ◽  
Ganglion Neurons

In order to screen for developmentally active chromosomal domains during zebrafish embryogenesis, we generated transgenic fish by microinjecting two different lacZ reporter constructs into fertilized eggs. Transgenic fish were screened among the progeny of injected fish (F0) crossed to non-injected fish. Groups of 15 to 20 progeny of each cross were tested for lacZ expression and/or transmission of injected sequences using PCR and Southern hybridizations. Progeny from 2 of 102 fish injected with supercoiled constructs containing Rous sarcoma virus promoter sequences showed apparently spatially regulated beta-galactosidase (beta-Gal) activity. However, we were not able to detect this reporter construct in DNA from fins of F1 fish. Injections of a linear reporter construct containing mouse heat-shock promoter sequences revealed transmission of injected sequences to F1 progeny in about 6% of cases (8 of 129 fish, tested with PCR). We found one lacZ-expressing line that showed a spatially and temporally restricted expression of lacZ and, therefore, features typical characteristics of “enhancer trap” lines. In this line, lacZ expression starts at 16 hours post-fertilization in trigeminal ganglion cells. At about 24 hours lacZ expression can be detected in trigeminal ganglion neurons and Rohon-Beard neurons, indicating that the development of these two cell types shows common features. The reporter gene has integrated as a single copy. The founder fish was mosaic: 19% of its offspring (3 of 16 tested animals) carried the reporter construct in their fins; about 51% (13 of 27 tested animals) of the progeny of F1 fish were beta-Gal positive indicating full hemizygosity.(ABSTRACT TRUNCATED AT 250 WORDS)

Catecholamine-, Indoleamine-, and GABA-containing cells in the chameleon retina

1995 ◽  
Vol 12 (4) ◽  
pp. 785-792 ◽  
Author(s):  
Mohamed Bennis ◽  
Claudine Versaux-Botteri
Keyword(s):  
Tyrosine Hydroxylase ◽  
Ganglion Cells ◽  
Amacrine Cells ◽  
Cell Types ◽  
Aminobutyric Acid ◽  
Retinal Cell ◽  
Gamma Aminobutyric Acid ◽  
First Report ◽  
Soma Size ◽  
Displaced Amacrine Cells

AbstractNeurons containing catecholamine, indoleamine, and gamma-aminobutyric acid (GABA) were identified by immunohistochemistry in the chameleon retina. Tyrosine hydroxylase (TH) and serotonin (5HT) were observed mostly in two subtypes of orthotopic amacrine cells differing in their soma size and process distribution within the IPL. Some labelled cells were displaced either to the IPL (5HT) or to the GCL (TH and 5HT). A multiplicity of retinal cell types contained GABA including cones, horizontal, amacrine, and ganglion cells. Our results confirmed those obtained in the retinas of other lizards except for the presence of interstitial and displaced amacrine cells containing TH or 5HT of which this is the first report.

Competition for survival among developing ciliary ganglion cells

1980 ◽  
Vol 43 (1) ◽  
pp. 233-254 ◽  
Author(s):  
G. Pilar ◽  
L. Landmesser ◽  
L. Burstein
Keyword(s):  
Cell Death ◽  
Ganglion Cells ◽  
Close Contact ◽  
Myoepithelial Cells ◽  
Ciliary Ganglion ◽  
Peripheral Target ◽  
Nerve Branch ◽  
Before And After ◽  
Soma Size ◽  
Hrp Transport

1. Functionally different subgroups, each innervating a different part of the peripheral target, were defined within the ciliary population of the avian ciliary ganglion by electrical stimulation of the various ciliary nerve branches. 2. Although neurons innervating defined parts of the peripheral target consistently sent their axons through certain nerves, the technique of retrograde horseradish peroxidase (HRP) transport showed that the ganglion cell bodies were not spatially grouped but distributed throughout the ganglion, both before and after the period of naturally occurring cell death. However, such neurons tended to be clustered into groups of two or greater. 3. Ciliary and choroid populations, however, were found to be for the most spatially separate and recognizable by location and soma size before the period of cell death. Choroid cells did not project out the ciliary nerves even prior to the cell death period, confirming previous observations of selective axon outgrowth in the two populations. 4. Competition for survival was demonstrated within the ciliary population by experimentally removing approximately two-thirds of the neurons by axotomy-induced cell death at stage 32-34 just prior to the normal cell death period. This reduction in the number of competing neurons resulted in rescue of approximately 40% of the neurons that would have died, as assessed both by the number of axon profiles in the remaining intact nerve branch, as well as the number of somata that could be retrogradely labeled from this nerve. 5. It was concluded that many of the neurons that are normally removed during the cell death period are not destined to die, but can be rescued by reducing the number of neurons competing for a limited supply of some aspect of the peripheral target. Further, the postulated interaction with the target was shown to occur relatively late, just prior to the onset of cell death. 6. At the time of the peripheral interaction, the target was found to consist primarily of myoepithelial cells, which had migrated into the target region following the arrival of the ciliary axons. The target per se, therefore, cannot be involved in the selective growth of ciliary axons to the appropriate region. Well-defined synapses were rare, although many axonal endings were observed in close contact with both myoepithelial cells and the sparser differentiated muscle fibers, which increased to account for 60% of the target by the end of the cell death period. 7. Competition was also found to retard the rate of neuronal maturation because intact axons in the partially axotomized ganglion developed more rapidly than control axons, as assessed by axon diameter, conduction velocity, and degree of glial ensheathment. 8. Finally, at least some of the neurons in the partially axotomized ganglion expanded to innervate the peripheral territory of the axotomized branches, suggesting that competition between neurons is involved in the establishment of the observed peripheral innervation pattern.

Isolation and characterization of slow, depolarizing responses of cardiac ganglion neurons in the crab, Portunus sanguinolentus

1979 ◽  
Vol 42 (4) ◽  
pp. 1000-1021 ◽  
Author(s):  
K. Tazaki ◽  
I. M. Cooke
Keyword(s):  
Ganglion Cells ◽  
Absolute Threshold ◽  
Large Cell ◽  
Complete Response ◽  
Resting Potential ◽  
Cardiac Ganglion ◽  
Isolation And Characterization ◽  
The Absolute ◽  
Driver Potential ◽  
Ganglion Neurons

1. Tetrodotoxin-resistant, active responses to depolarization of the large cardiac ganglion cells were studied in semi-isolated preparations from the crab, Portunus sanguinolentus. Impulse activity was monitored with extracellular electrodes, simultaneous recordings from two or three large cells were made with intracellular electrodes, and current was passed via a bridge or second intracellular electrode. Preparations were continuously perfused with saline containing 3 x 10(-7) M tetrodotoxin (TTX). 2. About 20 min after introduction of TTX, small-cell impulses and resultant EPSPs in large cells cease, while rhythmic, spontaneous bursting of large cells continues. A pacemaker depolarization between bursts and slow depolarizations underlying the impulse bursts are prominent at this time. Shortly after, spontaneous burst rate slows, and at ca. 25 min, the ganglion becomes electrically quiescent. 3. In the quiescent, TTX-perfused ganglion, injection of depolarizing current into any one of the large cells results in active responses. At current strengths of sufficient intensity and duration (e.g., 20 nA, 20 ms; 5 nA, 500 ms) to depolarize a large cell by ca. 10 mV from resting potential (-53 mV, avg), the graded responses become regenerative and of constant form, provided the stimulation rate is less thna 0.15/s. Such responses have been termed "driver potentials." At more rapid rates, thresholds are increased and responses reduced. 4. Driver potentials of anterior large cells reach peak amplitudes of ca. 20 mV (to -32 mV), have maximum rates of rise of 0.45 V/s and of fall of 0.2 V/s, and a duration of ca. 250 ms. They are followed by hyperpolarizing afterpotentials, a rapidly decaying one (1 s) to -58 mV, followed by a slowly decaying one (7.5 s), -55 mV. Responses of posterior large cells are smaller (16 mV) and slower; the site of active response may be at a distance from the soma. 5. The ability of elicit near-synchronous responses and the identity of amplitude and form of responses among anterior cells and of posterior cells, regardless of which cell receives depolarizing current, indicates that all cells undergo active responses and are stimulated by electrotonic spread of depolarization. 6. The responses involve a conductance increase since memses during a driver potential are much reduced. 7. Depolarization by steady current increases the absolute threshold, decreases the maximum depolarization of the peak, and slows rates of rise and fall. Hyperpolarization increases rates of rise and fall; the absolute value reached by the peak depolarization is unchanged. Hyperpolarization reduces the amplitude of the rapid after-potential relative to the displaced resting potential. 8. Hyperpolarizing current pulses imposed during the rise and peak of driver-potential responses are followed by redevelopment of a complete response. Sufficiently strong hyperpolarization can terminate a response. The current strength needed to terminate a response decreases the later during the response the pulse is given...

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