Neurofilament-like and glial fibrillary acidic protein-like immunoreactivities in rat and guinea-pig sympathetic ganglia in situ and after perturbation

Summary The presence of neurofilament (NF)-like and glial fibrillary acidic protein (GFAP)-like immunoreactivities was studied in sympathetic ganglia of adult rats and guinea pigs during normal conditions and after perturbation. In the superior cervical ganglion (SCG) of normal rats, many ganglion cells and nerve fibers show NF immunoreactivity. Some of these nerve fibers disappear after preganglionic decentralization of SCG; this indicates the presence of a mixture of preand postganglionic NF-positive nerves in the ganglion. Cuts in both preand postganglionic nerves result in a marked increase in GFAP immunoreactivity in SCG, whereas NF immunoreactivity increases in nerve cell bodies after preganglionic cuts. Only a few ganglion cells show NF immunoreactivity in the normal SCG of guinea pig. All intraganglionic NF-positive nerves are of preganglionic origin; decentralization abolishes NF immunoreactivity in these nerve fibers. The inferior mesenteric ganglion, the hypogastric nerves and colonic nerves in guinea pigs contain large numbers of strongly NF-immunoreactive nerve fibers.When the SCG of adult rat is grafted to the anterior eye chamber of adult rat recipients, both ganglionic cell bodies and nerve fibers, forming on the host iris from the grafted ganglion, are NF-positive. As only the perikarya of these neurons normally exhibit NF immunoreactivity, and the terminal iris arborizations are NF-negative, it appears that the grafting procedure causes NF immunoreactivity to become more widespread in growing SCG neurons.     

Tonic impulsation in preganglionic and postganglionic sympathetic fibers of mammals

A study of the tonic electrical activity of nerves containing preganglionic and postganglionic fibers in the superior cervical and stellate sympathetic ganglia of cats and rabbits has shown that this activity consists of groups of spikes synchronous with the pulse or respiration, and occurs on a background of irregular low-amplitude impulses. The frequency of spikes is higher (250/sec) in nerves containing preganglionic fibers than in those containing postganglionic fibers (100/sec). Groups of spikes in a nerve containing preganglionic fibers correspond in some preparations to groups of spikes of lower frequency in a nerve containing postganglionic fibers of the same ganglion; in other preparations, this correspondence was lacking, apparently due to the absence of synaptic contacts between those groups of pre- and postganglionic neurons whose activity was recorded. Neurons send axons to different nerves (cardiac and vertebral) of the stellate ganglion discharged synchronously in some preparations, and asynchronously in others. Where synchronization was observed, the neurons discharged in rhythm with cardiac contractions.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 1, No. 3, pp. 303–308, November–December, 1969.     

Neuronal organization of the left celiac ganglion in the cat

Unit responses of the isolated left celiac ganglion to stimulation of various nerves of the solar plexus were studied by intracellular microelectrode recording in cats before and after degeneration of the preganglionic fibers. The resting potential of the ganglionic neurons was ?62.2±2.9 mV and the amplitude of the spike potential 72.4±3.2 mV. The spike was followed by after-hyperpolarization with a mean amplitude of 24% of the spike amplitude and a duration of between 25 and 180 msec. A characteristic feature of the ganglion was the presence of orthodromic unit responses to stimulation of peripheral nerve fibers of the solar plexus. The higher threshold of activation of the neurons by peripheral fibers than by preganglionic fibers and the preservation of orthodromic unit responses to stimulation of peripheral nerves after degeneration of the preganglionic fibers are evidence that the peripheral reflex arc is closed in this ganglion. Neurons of the left celiac ganglion are divided into three groups. Only preganglionic fibers of the splanchnic nerve with different properties converge on the neurons of the first group (the most numerous); only afferent fibers of peripheral nerves converge on the neurons of the third group (the least numerous); both types of fibers terminate on neurons of the second group. This convergence may lie at the basis of the mechanism of the centrifugal and peripheral reflex interaction in the ganglion for coordinated visceral activity.     

Electrophysiological investigation of pathways in the middle cervical sympathetic ganglion of the turtle

Single unit responses in the middle cervical sympathetic ganglion ofEmys orbicularis to stimulation of other nerves and changes in these responses during the action of sympathetic blocking agents on the ganglion were investigated. The results showed that some fibers of the cervical sympathetic trunk of the turtle are interrupted in this ganglion. Postganglionic fibers pass out of the ganglion and enter the lateral branch and the sympathetic trunk. Other fibers pass through the ganglion without interruption and, together with postganglionic fibers, leave the ganglion in the cervical sympathetic trunk in a cranial direction. The velocity of conduction of excitation along the preganglionic fibers is between 4–3 and 2–1.5 m/sec and along the postganglionic fibers between 4–2.6 and 0.7–0.5 m/sec (fibers of types B₂ and C). Synaptic delay in the fast-conducting fibers averages 6.6 msec. Preganglionic fast-conducting fibers form synaptic contacts on neurons with type B₂ axons, while preganglionic slow-conducting fibers form contacts on neurons with type C axons. Terminals of two preganglionic fibers differing very slightly in their threshold of excitability, and probably constituting the same group, converge on some neurons.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukranian SSR, Kiev. Translated from Neirofiziologiya, Vol. 4, No. 1, pp. 83–89, January–February, 1972.     

Cholinergic nature of the primary afferent vagus synapsed in cross anastomosed superior cervical ganglia.

The superior cervical ganglion was reinnervated by vagal afferent fibers following heterologous cross anastomosis between the superior cervical preganglionic trunk and the vagal trunk at the level of the supranodose ganglion in cats. The contractions of the nictitating membrane and the postganglionic action potentials from the external carotid sinus nerve in response to electrical stimulation of the vagal artificial preganglionic trunk in these operated cats were inhibited by treatment with tetraethylammonium and atropine. The choline acetyltransferase activities were measured by the radiometric method. The activities in cross anastomosed superior cervical ganglion were lower than those of normal superior cervical ganglion, but higher than those of chronically decentralized superior cervical ganglion. The activities in cross anastomosed nodose ganglion were lower than those of normal nodose ganglion, but higher than those of chronically decentralized superior cervical ganglion. These results further support the view that the primary afferent vagus artificially synapsed in the superior cervical ganglion is cholinergic.     

Electrophysiological investigation of the conduction of excitation in the pterygopalatine ganglion of the cat

When responses in some nerves of the pterygopalatine ganglion of the cat in situ to stimulation of its other nerves were recorded it was found that most fibers passing through the ganglion are continuous sympathetic postganglionic fibers (at least three groups). Most of the parasympathetic preganglionic fibers forming synapses on neurons of the ganglion constitute a group of fibers with the same threshold of excitation. Intracellular recording from single neurons of the pterygopalatine ganglion showed that stimulation of the Vidian nerve evokes orthodromic spike potentials in some neurons of the ganglion with a short latent period, and in others with a long latent period (2.5–6.0 and 10–44 msec, respectively). Evidently only fast-conducting fibers terminate synaptically on most neurons of the ganglion and only slow-conducting fibers on some of them. Recording from intact nerves of the pterygopalatine ganglion revealed no tonic activity in them. Microelectrode recording from single neurons of the ganglion showed that either the frequency of generation of spike potentials is relatively low (1–3/sec) or such potentials are absent altogether.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 8, No. 5, pp. 514–520, September–October, 1976.     

Electrical activity of the sympathetic ganglia during the depressor reflex

Tonic activity of neurons of the superior cervical sympathetic ganglion was recorded by the "sucrose gap" method and in the 4th and 5th lumbar sympathetic ganglia with the aid of focal nonpolarizing electrodes in acute experiments on anesthetized cats and rabbits. The preganglionic fibers of the ganglia were left intact. Stimulation of the depressor nerve not only sharply inhibited the tonic activity of the ganglia but also led to the appearance of electropositive potentials of 0.7 ± 0.2 mV in the superior cervical ganglion and 20–250 µV in the lumbar ganglia. The amplitude of this potential was unchanged by atropine (1 · 10^–6M). A similar effect occured without stimulation of the depressor nerve, after division of the preganglionic fibers or blocking of their conduction; it is attributed to the cessation of preganglionic tonic impulses which induce not only spikes, but also many EPSPs in neurons of the ganglion. Their frequency in the lumbar ganglia was 4/sec. Summation of these EPSPs leads to constant electronegativity of the ganglion surface relative to the postganglionic fibers, and its disappearance is recorded as a positive potential. Stimulation of the depressor nerve thus does not induce IPSPs in the ganglion; consequently, the inhibition of synaptic activity observed under these circumstances is located in the CNS and not in the ganglion.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 6, No. 5, pp. 519–524, September–October, 1974.     

Cytophotometric and electrophysiological analysis of the role of the celiac plexus in maintenance of temperature homeostasis during cold stress in rats

The role of the celiac plexus in maintenance of temperature homeostasis in rats exposed to cold stress was studied by histochemical, ultrastructural, and electrophysiological methods. Inhibition of efferent impulsation was found in the preganglionic (splanchnic) nerves and potentiation in postganglionic (superior mesenteric) nerves under the influence of short-term cold stress, leading to a state of mild hypothermia. During cooling of the animals after decentralization of the celiac plexus an increase was observed in the intensity of fluorescence, in the activity of enzymes of energy metabolism, and in hyperplasia of the ultrastructural formations responsible for protein synthesis and the energy supply of the cell. It is suggested that during cold stress, when the flow of efferent impulses along preganglionic nerves is considerably reduced the celiac plexus becomes the center regulating autonomic functions that are involved in the maintenance of temperature homeostasis.Institute of Physiology, Belorussian Academy of Sciences, Minsk. Translated from Neirofiziologiya, Vol. 24, No. 6, pp. 659–667, November–December, 1992.     

Acetylcholinesterase Molecular Forms in Chick Ciliary Ganglion: Pre- and Postsynaptic Distribution Derived from Denervation, Axotomy, and Double Section

Abstract: Four main molecular forms of acetylcholinesterase (AChE) characterized by their sedimentation coefficients (5S, 7.5S, 11.5S, and 20S), are found in chick ciliary ganglion. After transection of the preganglionic nerve (denervation), total AChE activity in the ganglion dropped by 35% in 2 days. By then, 11.5s and 20s forms had diminished by 60 and 75% respectively, where as 7.5s remained practically unchanged. Since presynaptic structures disappeared 2 days after denervation, we inferred that at most 35% of total ganglion AChE was presynaptic: 11.5s and 20s might be mainly presynaptic and 7.5S, postsynaptic. At later time intervals. total AChE continued to decline up to day 5, possibly as a result of orthograde transynaptic regulation of the enzyme activity. After transection of postganglionic nerves (axotomy), total ganglion activity showed little change; 11.5s and 20s decreased by 40 and 6076, respectively, in 5 days, but these drops were compensated for by an early increase in 7 5S, which started the day after axotomy. After simultaneous transection of both pre- and postganglionic nerves (double section), total ganglion AChE dropped rapidly by 35% in 1 day and remained at that level up to 21 days. The 11.5S diminished rapidly by 60% in 1 day. The early increase of the 7.5s form induced by axotomy alone did not occur. Since the effect resulting from double section was not the equivalent of the cumulative effects observed after denervation and axotomy, respectively, the level of AChE forms in the ganglion may be regulated by reciprocal interaction of pre- and postsynaptic elements. After denervation and double section but not after axotomy alone, the contralateral non-operated ganglion exhibited a fall in the 20s form. This suggests that a transynaptic effect is exerted on AChE by the contralateral preganglionic neuron. Taken together, these results indicate that the various AChE molecular forms in chick ciliary ganglion are preferentially but not exclusively distributed as follows: the pre- and postganglionic axons contain mainly the 11.5S form, whereas nerve endings and synaptic structures are enriched in 20S, and ganglion cell bodies, in 7.5s.     

Decentralisation of neurones in the pelvic ganglion of the guinea-pig: Reinnervation by adrenergic nerves

An electron-microscopic study has been made of adrenergic and cholinergic nerve fibres and synapses in the pelvic ganglion of the guinea-pig at intervals of up to 60 days following section of the hypogastric and pelvic nerves. Transection of the hypogastric nerves led to degeneration of 80-90% of the cholinergic nerve profiles and synapses in the ganglion. The small number of adrenergic nerves and synapses did not change, but 30-60 days after section, this number increased 8-10 times. Transection of the pelvic nerves led to degeneration of about 15% of the cholinergic nerve terminals, but no change in adrenergic terminals. After transection of both hypogastric and pelvic nerves, only about 1% of cholinergic nerves survived, but after 30-60 days, the number of adrenergic nerves increased 8-10 times. It is concluded that following cholinergic nerve degeneration in the ganglion, adrenergic nerves, probably originating as collateral sprouts from postganglionic neurones and granule-containing cells, can replace them to some extent.     

The localization of the beta-subtype of protein kinase C (PKC-beta) in rat sympathetic neurons

The localization of PKC-beta was studied in rat sympathetic neurons using a polyclonal antibody specific for the beta 1- and beta 2-subspecies. The tissues studied included the superior cervical (SCG) and hypogastric (HGG) ganglia and the target tissues of the SCG and HGG neurons: the submandibular gland, iris, prostate and vas deferens. PKC-beta-LI was found in nerve fibers in both ganglia. A proportion of the fibers in the SCG disappeared after decentralization, suggesting that the fibers were of both pre- and postganglionic origin. The somata of the HGG and SCG neurons expressed varying amounts of PKC-beta-LI, the majority of SCG neurons being labelled only after colchicine treatment. In all target tissues there were PKC-beta-immunoreactive nerve fibers in bundles, but the most peripheral branches of the fibers were negatively labelled. The results show that PKC-beta-LI is widely present in sympathetic postganglionic neurons with mainly quantitative differences. The lack of PKC-beta in the most peripheral branches of nerve fibers might be a general feature of sympathetic postganglionic neurons, suggesting that the participation of PKC-beta in neurotransmitter release and in other functions in nerve terminals in sympathetic adrenergic neurons is unlikely.     

Location of bodies of peripheral (sympathetic) afferent neurons participating in peripheral reflexes of the small intestine

Acute electrophysiological experiments on cats have shown that after preliminary decentralization of the solar plexus, accompanied by degeneration of spinal and vagal afferent and preganglionic efferent fibers in its postganglionic (mesenteric) nerves, only slow activity of the C-afferents is recorded in the peripheral segments of the mesenteric nerves instead of activity of the A-, B-, and C-fibers in the control (before degeneration). Activity of the C-afferents is intensified with the appearance of spontaneous contractions of the small intestine and also after gentle stretching of the corresponding segment of the intestine by inflation of a rubber balloon. After preliminary division of the mesenteric nerves, accompanied by degeneration of the postganglionic fibers in their peripheral segments, activity of C-afferents only also was observed, but it was much weaker than in the first series of experiments. After preliminary decentralization of the solar plexus and division of the mesenteric nerves application of a single electrical stimulus to the central part of one of the divided mesenteric nerves evokes a reflex electrical response in the other mesenteric nerves which disappears after treatment of the ganglia of the solar plexus with azamethonium bromide and also after electrical stimulation of the mesenteric nerves at 10–20 Hz. However, after decentralization only, this response was much weaker than after division of the mesenteric nerves. It is concluded that these peripheral responses of the intestinal nerves are due to excitation of two types of peripheral afferent neurons: the bodies of some lie in the small intestine and their long axons (C-afferents) run to the ganglia of the solar plexus; the bodies of the others lie in the ganglia of the solar plexus and their long axons (also C-afferents) run to the intestine, where they terminate in its receptors.Institute of Physiology, Academy of Sciences of the Belorussian SSR, Minsk. Translated from Neirofiziologiya, Vol. 6, No. 2, pp. 175–185, March–April, 1974.     

Projection pathways,co-existence of peptides and synaptic organization of nerve fibers in the inferior mesenteric ganglion of the guinea-pig

Summary The presence of immunoreactive enkephalin, dynorphin, vasoactive intestinal polypeptide, cholecystokinin, substance P and neuropeptide Y in nerve fibers that project to the guinea-pig inferior mesenteric ganglion was analysed, after different denervation and ligation procedures. A quantitative analysis demonstrates that enkephalin- and substance P fibers reach the ganglion mainly via lumbar splanchnic and partly via intermesenteric nerves. Dynorphin-, vasoactive intestinal polypeptide- and cholecystokinin fibers reach the ganglion mainly via colonic and partly via hypogastric or intermesenteric nerves. Neuropeptide Y fibers enter via intermesenteric, lumbar splanchnic and hypogastric nerves and pass through the ganglion. Analysis of serial 0.5 m sections tends to confirm co-existence: of dynorphin, vasoactive intestinal polypeptide and cholecystokinin in fibers projecting from the colon; of dynorphin with substance P in the lumbar splanchnic nerves; and of neuropeptide Y with substance P in the hypogastric and colonic fibers. Synaptic contacts, predominantly axodendritic, onto the ganglion cells from enkephalin-, vasoactive intestinal polypeptide-, and substance P-containing terminals were revealed by electron microscopy. Enkephalin-immunoreactive axon varicosities are filled with small, clear vesicles with a few large, cored vesicles and form asymmetric synapses; dynorphin-, vasoactive intestinal polypeptide- and cholecystokinin-immunoreactive axon varicosities are rich in large, dense-cored vesicles and form symmetric synapses.     

Visceral afferent pathways relaying synaptically in the caudal mesenteric sympathetic ganglion

The caudal mesenteric sympathetic ganglion of cats was isolated and perfused, and responses of the preganglionic trunks of the ganglion to electrical stimulation of the central end of the hypogastric nerve were studied. Stimulation of the nerve with single square pulses gives rise to early and late responses. Early responses appear after various latent periods and are the result of excitation of transit fibers of groups A, B, and C, whereas the appearance of late responses is associated with the synaptic transmission of excitation in the sympathetic ganglion from afferent sympathetic neurons at the first level (from the pelvic organs to the caudal ganglion) to afferent sympathetic neurons of the second level (from the caudal ganglion and above). Early responses are not blocked, but late responses are blocked by perfusion of the ganglion with azamethonium bromide and magnesium salts, and also by tetanization of the hypogastric nerve at 10–50 Hz. Other facts indicating the synaptic relaying of visceral sympatho-sympathetic afferent pathways in the ganglion are also described.Institute of Physiology, Academy of Sciences of the Belorussian SSR, Minsk. Translated from Neirofiziologiya, Vol. 2, No. 5, pp. 507–514, September–October, 1970.     

EFFECTS OF SURGICAL DECENTRALIZATION AND NERVE GROWTH FACTOR ON THE MATURATION OF ADRENERGIC NEURONS IN A MOUSE SYMPATHETIC GANGLION

Abstract— The increase in tyrosine hydroxylase activity in mouse superior cervical ganglion during postnatal development was prevented by administration of the protein synthesis inhibitor cycloheximide. Surgical section of the preganglionic nerves in 4-day-old mice prevented the normal increases in tyrosine hydroxylase and monoamine oxidase activity in the ganglion during development. Surgical decentralization also prevented the developmental increases in ganglion size and cell numbers. The preganglionic fibres thus appear to exert a general regulatory effect on the growth and biochemical maturation of postganglionic adrenergic neurons in sympathetic ganglia. Administration of nerve growth factor to young mice failed to eliminate the differences in ganglion size, cell numbers and tyrosine hydroxylase activity between normally innervated and decentralized ganglia. Nerve growth factor, however, caused an increase in all these parameters in both control and decentralized ganglia–the magnitude of these increases being greatest in the control ganglia. Administration of carbachol and physostigmine to neonatal mice did not influence the normal development of tyrosine hydroxylase activity in the superior cervical ganglion.     

Conducting pathways of the superior cervical sympathetic ganglion of the cat

Individual nerves of the superior cervical sympathetic ganglion were stimulated in acute experiments on cats, and action potentials (AP) were recorded from other nerves of the ganglion in order to clarify whether or not there is transmission of excitation through the ganglion from one nerve to another and to establish whether this transmission is continuous or synaptic. The method of intracellular recording from neurons of the ganglion was also used. It is established that stimulation of the cervical sympathetic nerve evokes AP in all of the peripheral nerves of the ganglion, a circumstance that is the result of synaptic transmission of excitation. There is no transmission of excitation in the reverse direction or between any of the 12 peripheral nerves of the ganglion (including the four branches of the internal carotid nerve). Orthodromic excitation is recorded intracellularly from neurons of the ganglion during stimulation of the cervical sympathetic nerve, and antidromic excitation is recorded during stimulation of a peripheral nerve (the internal carotid nerve). It follows that the pathways through the ganglion which conduct excitation from the cervical sympathetic nerve into all of the remaining nerves of the ganglion are synaptic. Analysis of EPSP latent periods indicated that preganglionic fibers that differ sharply with respect to threshold and conduction rate (groups S₂ and S₄) converge on one and the same neurons of the ganglion.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 2, No. 2, pp. 216–224, March–April, 1970.     

The origin of VIP-containing nerves in the urinary bladder of rat

The innervation of the urinary bladder is known to include a considerable number of nerves containing vasoactive intestinal polypeptide (VIP). The origin of such nerves in the bladder of rat was investigated in this study using the methods of immunocytochemistry and radioimmunoassay combined with surgical sectioning of the hypogastric and/or pelvic nerves to the bladder. Eight days after pelvic nerve sectioning proximal to the main pelvic ganglion, VIP-immunoreactive nerves and VIP content were markedly increased from the level in the sham-operated rat bladder. Sectioning of hypogastric or both nerve pathways led to a less significant increase. It was therefore postulated that the majority of VIP-immunoreactive nerves originate from ganglia located either close to the bladder or within the bladder wall. It is interesting that in these experiments the VIP content of the bladder nerves is inversely related to the changes in motility that would be expected to result from the nerve sections.     

Synaptic transmission in thoracic autonomic ganglia of the dog

Afferent stimulation of one canine thoracic cardiopulmonary nerve can generate compound action potentials in another ipsilateral cardiopulmonary nerve. These compound action potentials persist after acute decentralization of the middle cervical ganglion, indicating that they result from neural activity in the middle cervical ganglion and thoracic nerves. Changing the frequency of stimulation can alter the compound action potentials, suggesting that temporal facilitation or inhibition occurs in this middle cervical ganglion preparation. The compound action potentials can be modified by stimulation of sympathetic preganglionic fibers and by hexamethonium, atropine, phentolamine, propranolol, and (or) manganese. It thus appears that afferent cardiopulmonary nerves can activate efferent cardiopulmonary nerves via synaptic mechanisms in the stellate and middle cervical ganglia. It also appears that these mechanisms involve adrenergic and cholinergic receptors and are influenced by preganglionic sympathetic fibers arising from the cord.     

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.     

Blockade of reflex venous capacitance responses in liver and spleen by hexamethonium, atropine, and surgical section.

Since hexamethonium and surgical section have been used to prevent reflex splanchnic capacitance responses, we examined the effectiveness of these procedures in blocking responses to direct stimulation of preganglionic fibres in the splanchnic nerves. Liver blood volume was measured by plethysmography and splenic blood volume by weighing in cats anesthetized by pentobarbital. The cats were adrenalectomized to prevent adrenal catecholamine secretion in response to splanchnic nerve stimulation. Hexamethonium (10 and 20 mg/kg) alone or atropine (1 mg/kg) alone caused only a small variable block of the responses to preganglionic nerve stimulation. A combination of the two drugs essentially produced a complete block of the liver capacitance response, but a significant response still persisted in the spleen. Surgical section of the postganglionic nerve bundles around the hepatic and splenic arteries completely abolished the responses to preganglionic stimulation. It is concluded that a relatively complete block of reflex splanchnic capacitance responses requires either a combination of hexamethonium and atropine or surgical section of the postganglionic nerves.