Quantitative studies of superior cervical sympathetic ganglia in a variety of primates including man. I. The ratio of preganglionic fibers to ganglionic neurons

Estimates of the number of ganglionic neurons of superior cervical sympathetic ganglia and the number of preganglionic axons in the trunks just caudal to these ganglia were obtained from a sample of primates that included: man, chimpanzee, baboon, stump-tailed macaque, rhesus monkey, and squirrel monkey. The number of ganglionic neurons ranged from 63,625 in a squirrel monkey ganglion to 1,041,652 neurons in a human ganglion. Estimates of the number of preganglionic fibers varied between 2,285 in a cervical sympathetic trunk of a squirrel monkey and 12,008 in a human specimen. The resulting ratios of preganglionic fibers to ganglionic neurons ranged from 1:28 in a squirrel monkey ganglion to 1:196 in a human ganglion. The data reported in this study reveal considerable variation in the ratio of pre- to post-ganglionic neurons, and as was noted in regard to the number of cells in the ganglion, the ratios of ganglionic to preganglionic neurons appear to increase as a function of body size. In contrast, the number of preganglionic fibers does not increase as strikingly with body size, but varies greatly in the same species. The resulting ratio between the two orders of neurons is, therefore, less predictable than the number of ganglionic neurons in any given ganglion.     

Functional and morphologic organization of stomach vegetative innervation

The functional and morphological organization of stomach vegetative innervation was studied. It is shown, that the sympathetic nerve strengthens vagal stimulation of motor activity of various departments of a stomach. In realization of the phenomenon participate preganglionic serotoninergic fibres transmitting excitation on ganglionic serotoninergic neurons. It is established, that stimulation of a wandering nerve results to degranulation of serotoninergic EC-cells carrying on the surface ganglionic 5-HT3,4-receptors. The irritation of a wandering nerve results in reduction of volume of EC-cells in older age.     

Effects of preganglionic denervation and postganglionic axotomy on acetylcholine receptors in the chick ciliary ganglion

The regulation of nicotinic acetylcholine receptors (AChRs) in chick ciliary ganglia was examined by using a radiolabeled anti-AChR mAb to quantitate the amount of receptor in ganglion detergent extracts after preganglionic denervation or postganglionic axotomy. Surgical transection of the preganglionic input to the ciliary ganglion in newly hatched chicks caused a threefold reduction in the total number of AChRs within 10 d compared with that present in unoperated contralateral control ganglia. Surgical transection of both the choroid and ciliary nerves emerging from the ciliary ganglion in newly hatched chicks to establish postganglionic axotomy led to a nearly 10-fold reduction in AChRs within 5 d compared with unoperated contralateral ganglia. The declines were specific since they could not be accounted for by changes in ganglionic protein or by decreases in neuronal survival or size. Light microscopy revealed no gross morphological differences between neurons in operated and control ganglia. A second membrane component of cholinergic relevance on chick ciliary ganglion neurons is the alpha-bungarotoxin (alpha-Bgt)-binding component. The alpha-Bgt-binding component also declined in number after either postganglionic axotomy or preganglionic denervation, but appeared to do so with a more rapid time course than did ganglionic AChRs. The results imply that cell-cell interactions in vivo specifically regulate both the number of AChRs and the number of alpha-Bgt-binding components in the ganglion. Regulation of these neuronal cholinergic membrane components clearly differs from that previously described for muscle AChRs.     

Development of adrenergic neurons in uiuo: inhibition by ganglionic blockade

T he N ormal biochemical maturation of postsynaptic adrenergic neurons in mouse and rat superior cervical ganglion depends upon an intact preganglionic innervation (B lack , H endry and I versen , 1971a, 1972; T hoenen , S aner and K eitler , 1972). In recent studies tyrosine hydroxylase, the rate-limiting enzyme in norepinephrine biosynthesis (L evitt , S pector , S joerdsma and U denfriend , 1965), with localization to adrenergic neurons in the ganglion (B lack , H endry and I versen , 1971b), was used to monitor maturation of these cells. The developmental increase in tyrosine hydroxylase activity occurred simultaneously with the appearance of ganglionic synapses and was prevented by transection of the preganglionic nerve trunk (B lack , H endry and I versen , 1971a). These observations suggest that presynaptic cholinergic nerve terminals regulate the biochemical development of postsynaptic neurons in the superior cervical ganglion. The mechanism(s) by which presynaptic cholinergic terminals regulate postsynaptic development has not been elucidated. Such trans-synaptic regulation may be dependent on normal impulse transmission and/or may involve other unidentified, trophic factors. The results presented in the present communication suggest that normal development of ganglionic tyrosine hydroxylase activity is dependent on depolarization of postsynaptic adrenergic neurons.     

Peptidergic inputs to sympathetic preganglionic neurons

This paper presents data showing that the sympathetic autonomic areas of the cat thoracolumbar spinal cord contain nerve terminals and fibres with immunoreactivity for at least seven neuropeptides. The distribution in the intermediolateral cell column of the terminals and fibres which contain enkephalin-, neuropeptide Y-, neurotensin-, substance P-, and neurophysin II-like immunoreactivity (ENK, NPY, NT, SP, and NP2, respectively) suggests that these peptides are involved in more generalized functions of the autonomic nervous system. On the other hand, peaks in density of immunoreactivity at certain levels suggest that different levels of influence of sympathetic preganglionic neurons by the various peptides may occur along the length of the thoracolumbar cord. The distribution of terminals and fibres containing somatostatin- and oxytocin-like immunoreactivity (SS and OXY) suggests that these peptides may be part of specific pathways to particular sympathetic preganglionic neurons. The possible sources of the terminals and fibres containing ENK, NPY, NT, SS, and SP include the spinal cord and supraspinal areas, whereas the source of these structures with OXY and NP2 is most likely supraspinal. The data suggest that coexistence of peptides and interactions between structures containing different neuropeptides occur in the spinal autonomic areas. It is speculated that neuropeptides have an important role to play in the regulation of the cardiovascular division of the autonomic nervous system.     

Effects of ciliary neurotrophic factor (CNTF) and depolarization on neuropeptide expression in cultured sympathetic neurons.

We examined the effects of ciliary neurotrophic factor (CNTF) and depolarization, two environmental signals that influence noradrenergic and cholinergic function, on neuropeptide expression by cultured sympathetic neurons. Sciatic nerve extract, a rich source of CNTF, increased levels of vasoactive intestinal peptide (VIP), substance P, and somatostatin severalfold while significantly reducing levels of neuropeptide Y (NPY). No change was observed in the levels of leu-enkephalin (L-Enk). These effects were abolished by immunoprecipitation of CNTF-like molecules from the extract with an antiserum raised against recombinant CNTF, and recombinant CNTF caused changes in neuropeptide levels similar to those of sciatic nerve extract. Alterations in neuropeptide levels by CNTF were dose-dependent, with maximal induction at concentrations of 5-25 ng/ml. Peptide levels were altered after only 3 days of CNTF exposure and continued to change for 14 days. Depolarization of sympathetic neuron cultures with elevated potassium elicited a different spectrum of effects; it increased VIP and NPY content but did not alter substance P, somatostatin, or L-Enk. Depolarization is known to block cholinergic induction in response to heart cell conditioned medium and we found that it blocked the induction of choline acetyltransferase (ChAT) and peptides by recombinant cholinergic differentiation factor/leukemia inhibitory factor (CDF/LIF). In contrast, it did not antagonize the effects of CNTF on either ChAT activity or neuropeptide expression. Thus, while CNTF has effects on neurotransmitter properties similar to those previously reported for CDF/LIF, the actions of these two factors are differentially modulated by depolarization, suggesting that the mechanisms of cholinergic and neuropeptide induction for the two factors differ. In addition, in contrast to CDF/LIF, CNTF did not alter levels of ChAT, VIP, substance P, or somatostatin in cultured dorsal root ganglion neurons. These observations indicate that CNTF and depolarization affect the expression of neuropeptides by sympathetic neurons and provide evidence for an overlapping yet distinct spectrum of actions of the two neuronal differentiation factors, CNTF and CDF/LIF.     

Structure of peripheral synapses: autonomic ganglia

Final motor neurons in sympathetic and parasympathetic ganglia receive synaptic inputs from preganglionic neurons. Quantitative ultrastructural analyses have shown that the spatial distribution of these synapses is mostly sparse and random. Typically, only about 1%-2% of the neuronal surface is covered with synapses, with the rest of the neuronal surface being closely enclosed by Schwann cell processes. The number of synaptic inputs is correlated with the dendritic complexity of the target neuron, and the total number of synaptic contacts is related to the surface area of the post-synaptic neuron. Overall, most neurons receive fewer than 150 synaptic contacts, with individual preganglionic inputs providing between 10 and 50 synaptic contacts. This variation is probably one determinant of synaptic strength in autonomic ganglia. Many neurons in prevertebral sympathetic ganglia receive additional convergent synaptic inputs from intestinofugal neurons located in the enteric plexuses. The neurons support these additional inputs via larger dendritic arborisations together with a higher overall synaptic density. There is considerable neurochemical heterogeneity in presynaptic boutons. Some synapses apparently lack most of the proteins normally required for fast transmitter release and probably do not take part in conventional ganglionic transmission. Furthermore, most preganglionic boutons in the ganglionic neuropil do not form direct synaptic contacts with any neurons. Nevertheless, these boutons may well contribute to slow transmission processes that need not require conventional synaptic structures.     

Development of synaptic transmission at autonomic synapses in vitro revealed by cytochrome oxidase histochemistry

We have studied the development of synaptic transmission by innervating sympathetic neurons in vitro and monitoring synaptic activity with both physiological recording and cytochrome oxidase histochemistry. The onset of synaptic transmission was reflected in increased cytochrome oxidase reaction product within individual neurons. Within 24 hours of co-culture, relatively low frequency suprathreshold potentials were recorded in approximately 20% of the innervated neurons. At this stage the cytochrome oxidase activity of innervated neurons, as assessed by optical density of the histochemical reaction product, was increased twofold compared with uninnervated neurons. Over the next 2-4 days of innervation, changes in the pattern and extent of synaptic activity and superthreshold events were accompanied by a net fourfold increase in cytochrome oxidase activity levels compared with noninnervated neurons. The increase in density of cytochrome oxidase reaction product observed after innervation was reversed completely by blockade of synaptic transmission. Differences in the efficacy of synaptic input provided to the sympathetic neurons by appropriate versus inappropriate presynaptic sources was determined by co-culturing sympathetic neurons with explants that contained either preganglionic neurons or somatic motor neurons. Although sympathetic neurons innervated by motor neuron explants had increased levels of cytochrome oxidase activity compared with noninnervated controls, the density of cytochrome oxidase reaction product was even greater in sympathetic neurons innervated by preganglionic explants. We conclude that both the onset of innervation of sympathetic neurons as well as the subsequent maturation of synaptic function is directly reflected in graded increases in cytochrome oxidase reaction product.     

New synapses associated with the granule-containing cells of rat sympathetic ganglia

The synapses of the rat superior cervical sympathetic ganglion were studied with both conventional and ultrastructural histochemical methods. Besides the cholinergic synapses polarized from preganglionic fibers to sympathetic ganglion neurons, two morphologically and functionally different types of synapses were observed in relation to the small granule-containing (catecholamine-containing) cells of the rat superior cervical ganglion. The first type is an efferent adrenergic synapse polarized from granule-containing cells to the dendrites of the sympathetic ganglion neurons. This type of synapse might mediate the inhibitory effects (slow inhibitory postsynaptic potentials) induced by catecholamines on the sympathetic neurons. The second type is a reciprocal type of synapse between the granule-containing cells and the cholinergic preganglionic fibers. Through such synapses, these cells could exert a modulating effect on the excitatory preganglionic fibers. Therefore, we propose that these cells, through their multiple synaptic connections, exhibit a local modulatory feedback system in the rat sympathetic ganglia and may serve as interneurons between the preganglionic and postganglionic sympathetic neurons.     

Pathway specific expression of neuropeptides and autonomic control of the vasculature

In this article, we review the immunohistochemical evidence for the pathway-specific expression of co-existing neuropeptides in autonomic vasomotor neurons, and examine the functional significance of these expression patterns for the autonomic regulation of the vasculature. Most final motor neurons in autonomic vasomotor pathways contain neuropeptides in addition to non-peptide co-transmitters such as catecholamines, acetylcholine and nitric oxide. Neuropeptides also occur in preganglionic vasomotor neurons. The precise combinations of neuropeptides expressed by neurons in vasomotor pathways vary with species, vascular bed, and the level within the vascular bed. This applies to both vasoconstrictor and vasodilator pathways. There is a similar degree of variation in the expression of neuropeptide receptors in the vasculature. Consequently, the contributions of different peptides to autonomic vasomotor control are closely matched to the functional requirements of specific vascular beds. This arrangement allows for a high degree of precision in vascular control in normal conditions and has the potential for considerable plasticity under pathophysiological conditions.     

Neuronal organization of ganglia in the lumbar sympathetic chain in cats

The study of the neuronal organization of ganglia L₃–L₆ of the sympathetic chain in cats by intracellular recording showed that neurons of the ganglion can be divided into three main groups on the principle of sympathetic preganglionic fibers of different types converging on them. The most numerous group (66%) consists of neurons on which sympathetic preganglionic fibers of the B₁, B₂, and C groups (with conduction velocities of 12.0±0.7, 4.4±0.3, and 1.0±0.1 m/sec respectively) simultaneously converge, while the least numerous group (10%) is formed by neurons with only sympathetic preganglionic fibers of the C-group converging on them; an intermediate group (24%) consists of neurons activated by sympathetic preganglionic fibers of the B₁ and B₂ groups. The preganglionic fibers to the ganglionic neurons can mainly be traced from the rostral segments of the spinal cord through the white rami communicantes. Sympathetic preganglionic fibers activating the neurons also enter the ganglion through their own and caudally situated white rami communicantes. Neurons of the ganglion were found to receive a preganglionic (C input) run in the composition of the gray ramus communicans and caudal commissure; the remaining neurons send their axons evidently into visceral branches.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 9, No. 5, pp. 518–526, September–October, 1977.     

Sympathetic and sensory innervation of small intensely fluorescent (SIF) cells in rat superior cervical ganglion

The sympathetic ganglion contains small intensely fluorescent (SIF) cells derived from the neural crest. We morphologically characterize SIF cells and focus on their relationship with ganglionic cells, preganglionic nerve fibers and sensory nerve endings. SIF cells stained intensely for tyrosine hydroxylase (TH), with a few cells also being immunoreactive for dopamine β-hydroxylase (DBH). Vesicular acetylcholine transporter (VAChT)-immunoreactive puncta were distributed around some clusters of SIF cells, whereas some SIF cells closely abutted DBH-immunoreactive ganglionic cells. SIF cells contained bassoon-immunoreactive products beneath the cell membrane at the attachments and on opposite sites to the ganglionic cells. Ganglion neurons and SIF cells were immunoreactive to dopamine D2 receptors. Immunohistochemistry for P2X3 revealed ramified nerve endings with P2X3 immunoreactivity around SIF cells. Triple-labeling for P2X3, TH and VAChT allowed the classification of SIF cells into three types based on their innervation: (1) with only VAChT-immunoreactive puncta, (2) with only P2X3-immunoreactive nerve endings, (3) with both P2X3-immunoreactive nerve endings and VAChT-immunoreactive puncta. The results of retrograde tracing with fast blue dye indicated that most of these nerve endings originated from the petrosal ganglion. Thus, SIF cells in the superior cervical ganglion are innervated by preganglionic fibers and glossopharyngeal sensory nerve endings and can be classified into three types. SIF cells might modulate sympathetic activity in the superior cervical ganglion.     

Chemical profile of vagal preganglionic motor cells innervating the airways in ferrets: the absence of noncholinergic neurons.

In ferrets, we investigated the presence of choline acetyltransferase (ChAT), vasoactive intestinal peptide (VIP), and markers for nitric oxide synthase (NOS) in preganglionic parasympathetic neurons innervating extrathoracic trachea and intrapulmonary airways. Cholera toxin beta-subunit, a retrograde axonal transganglionic tracer, was used to identify airway-related vagal preganglionic neurons. Double-labeling immunohistochemistry and confocal microscopy were employed to characterize the chemical nature of identified airway-related vagal preganglionic neurons at a single cell level. Physiological experiments were performed to determine whether activation of the VIP and ChAT coexpressing vagal preganglionic neurons plays a role in relaxation of precontracted airway smooth muscle tone after muscarinic receptor blockade. The results showed that 1) all identified vagal preganglionic neurons innervating extrathoracic and intrapulmonary airways are acetylcholine-producing cells, 2) cholinergic neurons innervating the airways coexpress ChAT and VIP but do not contain NOS, and 3) chemical stimulation of the rostral nucleus ambiguus had no significant effect on precontracted airway smooth muscle tone after muscarinic receptor blockade. These studies indicate that vagal preganglionic neurons are cholinergic in nature and coexpress VIP but do not contain NOS; their stimulation increases cholinergic outflow, without activation of inhibitory nonadrenergic, noncholinergic ganglionic neurons, stimulation of which induces airway smooth muscle relaxation. Furthermore, these studies do not support the possibility of direct inhibitory innervation of airway smooth muscle by vagal preganglionic fibers that contain VIP.     

EFFECT OF GLUCOCORTICOIDS ON NERVE GROWTH FACTOR-MEDIATED ENZYME INDUCTION IN ORGAN CULTURES OF RAT SYMPATHETIC GANGLIA: ENHANCED RESPONSE AND REDUCED TIME REQUIREMENT TO INITIATE ENZYME INDUCTION

Abstract— After previous studies had shown that nerve growth factor produces a very similar change in the enzyme pattern of adrenergic neurons as does an increased activity of the preganglionic cholinergic nerves, the present experiments revealed that the nerve growth factor-mediated selective induction of TH and DBH is enhanced by glucocorticoids in a way similar to that mediated by acetylcholine via nicotinic receptors. Corticosterone (5 μM) produced not only an increase in the maximal response to NGF but shifted the concentration response curve of TH to NGF to the left. The potentiation effect was shown to be specific for glucocorticoids, since other steroid hormones like testosterone, β-estradiol and progesterone had no effect. Moreover, the glucocorticoid effect could be antagonized by cortexolone, suggesting an effect via glucocorticoid receptors. In addition to the potentiation of the nerve growth factor-mediated enzyme induction, glucocorticoids reduced the exposure time to NGF, necessary to initiate maximal TH induction, from 4 h to 10 min. The glucocorticoid potentiation of NGF-mediated specific enzyme induction is discussed in relation to the site and mechanism of action of NGF.     

Catecholaminergic primary sensory neurons: autonomic targets and mechanisms of transmitter regulation

Contrary to traditional teaching, mammalian primary sensory neurons may express catecholaminergic (CA) neurotransmitter characteristics in vivo. Sensory neurons in the nodose, petrosal, and dorsal root ganglia of rats express tyrosine hydroxylase, the rate-limiting enzyme in CA biosynthesis, and formaldehyde-induced CA fluorescence, in addition to other CA traits. These findings suggest that catecholamines may function as sensory as well as autonomic motor (e.g., sympathetic) neurotransmitters. Most CA cells in the petrosal ganglion project peripherally to the carotid body, which indicates a striking correlation between CA expression in sensory neurons and the pattern of sensory innervation. Inasmuch as petrosal ganglion afferents make synaptic contact with chemoreceptive glomus cells in the carotid body, it is likely that CA sensory neurons in the ganglion transmit chemoreceptor information to the brain stem. Comparison with sympathetic neurons indicates that some mechanisms of CA regulation, such as altered activity of tyrosine hydroxylase in response to depolarizing stimuli, are shared among sensory and traditional CA populations. Other mechanisms, including trophic regulation, appear to be distinct. Therefore, despite expression of common phenotypic traits, CA expression in diverse populations of peripheral neurons is not necessarily associated with a common repertoire of regulatory mechanisms.     

Coexistence of neuropeptides in sympathetic preganglionic neurons of the cat

Coexistence of four neuropeptides in sympathetic preganglionic neurons (SPN) was investigated immunohistochemically in cats after intrathecal administration of colchicine. Neurons were studied for the coexistence of all combinations of enkephalin-, neurotensin-, somatostatin-, and substance P-like immunoreactivity (ENK, NT, SS, and SP, respectively) in the intermediolateral cell column (IML), nucleus intercalatus (IC), and central autonomic area (CA). The results indicate that SP coexists with all three other peptides, SS coexists with NT and SP, and ENK coexists only with SP. In all cases, SPN which contained two peptides were found in the IML in almost all levels of the thoraco-lumbar cord. Much smaller numbers of SPN which contained two peptides (in the same combinations as above) were found in the IC and not all segments contained such neurons. In the CA, only one neuron was found which contained two peptides (SP/SS). The distribution of SPN containing two peptides suggests that these neurons may participate in more general functions of the autonomic nervous system and that they are not likely involved in the innervation of specific visceral organs.     

Peptidergic modulation of efferent sympathetic neurons in intrathoracic ganglia regulating the canine heart

When either substance P or vasoactive intestinal peptide was injected into an acutely decentralized intrathoracic sympathetic ganglion, short-lasting augmentation of cardiac chronotropism and inotropism was induced. These augmentations were induced before the fall in systemic arterial pressure occurred which was a consequence of these peptides leaking into the systemic circulation in enough quantity to alter peripheral vascular resistance directly. When similar volumes of normal saline were injected into an intrathoracic ganglion, no significant cardiac changes were induced. When substance P or vasoactive intestinal peptide was administered into an intrathoracic ganglion, similar cardiac augmentations were induced either before or after the intravenous administration of hexamethonium. In contrast, when these peptides were injected into an intrathoracic ganglion in which the beta-adrenergic blocking agent timolol (0.1 mg/0.1 ml of normal saline) had been administered no cardiac augmentation occurred. These data imply that in the presence of beta-adrenergic blockade intraganglionic administration of substance P or vasoactive intestinal peptide does not modify enough intrathoracic neurons to alter cardiac chronotropism and inotropism detectably. When neuropeptide Y was injected into an intrathoracic ganglion, no cardiac changes occurred. However, when cardiac augmentations were induced by sympathetic preganglionic axon stimulation these were enhanced following the intraganglionic administration of neuropeptide Y. As this effect occurred after timolol was administered into the ipsilateral ganglia, but not after intravenous administration of hexamethonium, it is proposed that the effects of neuropeptide Y are dependent upon functioning intrathoracic ganglionic nicotinic cholinergic synaptic mechanisms. Intravenous administration of either morphine or [D-ala2,D-leu5]enkephalin acetate did not alter the capacity of the preganglionic sympathetic axons to augment the heart when stimulated. Following the intravenous administration of naloxone, the positive inotropic cardiac responses induced by efferent preganglionic sympathetic axonal stimulation were enhanced minimally in control states and significantly following hexamethonium administration. Thus, it appears that enkephalins are involved in the modulation of intrathoracic ganglion neurons regulating the heart, perhaps via modification of beta-adrenergic receptors. Taken together these data indicate that substance P, vasoactive intestinal peptide, neuropeptide Y, or enkephalins modify intrathoracic ganglionic neurons which are involved in efferent sympathetic cardiac regulation.     

Mechanisms of inhibition of synaptic transmission in the sympathetic ganglia of rats with alloxan diabetes]

In experiments on the isolated superior cervical sympathetic ganglia of rats with alloxan diabetes rhythmic stimulation of preganglionic nerves was effected; summation presynaptic spikes and EPSPs of ganglionic neurons were registered. In rats with moderately severe alloxan diabetes progressive depression of rhythmic ganglion potentials was connected with suppression of the mediator emission to the impulse due to rapid exhaustion of its operational fraction. Rats with severe diabetes displayed also postsynaptic suppression of the ganglionic neurons. Dynamic characteristics of the transmitter turnover assessed on the basis of consideration of the successive patterns of posttetanic potentiation showed insignificant changes in the mediator output and a significant (by 38%) suppression of the mediator reserve per sec in comparison with control.