Myths and realities of the cardiac vagus.

There is continuing belief that cardiac parasympathetic postganglionic fibres are sparse or absent from the ventricles. This review of the literature shows that the supposition is a myth. Early studies considered that fine silver-stained fibres coursing amongst ventricle myocardial cells were most likely cardiac parasympathetic postganglionic fibres. The conclusions were later supported by acetyl cholinesterase staining using a method that appeared not to be associated with noradrenaline nerve fibres. The conclusion is critically examined in the light of several recent histological studies using the acetyl cholinesterase method and also a more definitive technique (CHAT), that suggest a widespread location of parasympathetic ganglia and a relatively dense parasympathetic innervation of ventricular muscle in a range of mammals including man. The many studies demonstrating acetylcholine release in the ventricle on vagal nerve stimulation and a high density of acetylcholine M2 receptors is in accord with this as are tests of ventricular performance from many physiological studies. Selective control of cardiac functions by anatomically segregated parasympathetic ganglia is discussed. It is argued that the influence of vagal stimulation on ventricular myocardial action potential refractory period, duration, force and rhythm is evidence that vagal fibres have close apposition to myocardial fibres. This is supported by clear evidence of accentuated antagonism between sympathetic activity and vagal activity in the ventricle and also by direct effects of vagal activity independent of sympathetic activity. The idea of differential control of atrial and ventricular physiology by vagal C and vagal B preganglionic fibres is examined as well as differences in chemical phenotypes and their function. The latter is reflected in medullary and supramedullary control. Reference is made to the importance of this knowledge to understanding the normal physiology of cardiac autonomic control and significance to pathology.

GABAergic control of micturition within the periaqueductal grey matter of the male rat.

In urethane-anaesthetised rats continuous infusion of saline into the bladder (6 ml h⁻¹) evoked periodic sharp rises in intravesicular pressure accompanied by rhythmic bursting of external urethral sphincter (EUS) EMG and expulsion of urine from the urethral meatus. Microinjection of the GABA agonist muscimol (250 pmol) into the caudal ventrolateral periaqueductal grey (PAG), but not at other sites in the PAG, either depressed reflex voiding frequency (-60%, n = 7) and tonic EUS EMG activity (-38%, n = 6) or completely inhibited voiding (four sites). Microinjection of the GABA antagonist bicuculline (BIC; 1 nmol) into the same region, to reduce ongoing GABA tone, increased reflex voiding frequency (+467%, n = 16) and tonic activity in the EUS (+56%, n = 7) whilst bursting activity in the EUS became desynchronised. Although muscimol failed to change reflex micturition when microinjected into the dorsal caudal PAG, microinjection of BIC at these sites evoked pronounced autonomic arousal and increased reflex voiding frequency (+237%, n = 34). The results demonstrate that the functional integrity of synapses in the caudal ventrolateral PAG is essential to permit micturition. Transmission through the region is normally regulated by a tonic GABAergic inhibitory influence. In contrast, the functional integrity of the dorsal caudal PAG is not essential for reflex micturition. However, micturition may be initiated from this region via projections to the caudal ventrolateral PAG, as part of the behavioural response to psychological threat or other stressful stimuli.

Palliative Lung Radiotherapy: Higher Dose Leads to Improved Survival?

AIMS: Choosing the optimal palliative lung radiotherapy regimen is challenging. Guidance from The Royal College of Radiologists recommends treatment stratification based on performance status, but evidence suggests that higher radiotherapy doses may be associated with survival benefits. The aim of this study was to investigate the effects of fractionation regimen and additional factors on the survival of palliative lung cancer radiotherapy patients. MATERIALS AND METHODS: A retrospective univariable (n = 925) and multivariable (n = 422) survival analysis of the prognostic significance of baseline patient characteristics and treatment prescription was carried out on patients with non-small cell and small cell lung cancer treated with palliative lung radiotherapy. The covariates investigated included: gender, age, performance status, histology, comorbidities, stage, tumour location, tumour side, smoking status, pack year history, primary radiotherapy technique and fractionation scheme. The overall mortality rate at 30 and 90 days of treatment was calculated. RESULTS: Univariable analysis revealed that performance status (P < 0.001), fractionation scheme (P < 0.001), comorbidities (P = 0.02), small cell histology (P = 0.02), 'lifelong never' smoking status (P = 0.01) and gender (P = 0.06) were associated with survival. Upon multivariable analysis, only better performance status (P = 0.01) and increased dose/fractionation regimens of up to 30 Gy/10 fractions (P < 0.001) were associated with increased survival. Eighty-five (9.2%) and 316 patients (34%) died within 30 and 90 days of treatment, respectively. CONCLUSION: In this retrospective single-centre analysis of palliative lung radiotherapy, increased total dose (up to and including 30 Gy/10 fractions) was associated with better survival regardless of performance status.

Electrophysiological characteristics of vasomotor preganglionic neurons and related neurons in the thoracic spinal cord of the rat: an intracellular study in vivo.

Sympathetic preganglionic neurons (SPN) represent the final central neurons in the sympathetic pathways which regulate vasomotor tone; they therefore play a pivotal role in the re-distribution of cardiac output to different vascular beds in response to environmental challenges. While the consensus view is that activity in these neurons is due mainly to supraspinal inputs, the possibility that some activity may be generated intrinsically and modified by synaptic inputs cannot be excluded. Therefore, in order to distinguish between these two possibilities, the electrophysiological properties of cardiovascular-like SPN in the upper thoracic spinal cord of the anesthetized rat were examined and their response to activation of vasodepressor inputs was investigated. Intracellular recordings were made from 22 antidromically identified SPN of which 17 displayed irregular, but maintained, spontaneous activity; no evidence of bursting behavior or pacemaker-like activity was observed. Stimulation of the aortic depressor nerve or a vasodepressor site within the nucleus tractus solitarius (NTS) resulted in a membrane hyperpolarization, decrease in cell input resistance and long-lasting cessation of neuronal firing in SPN including a sub-population which had cardiac-modulated patterns of activity patterns. Recordings were also undertaken from 80 non-antidromically-activated neurons located in the vicinity of SPN; 23% of which fired in phase with the cardiac cycle, with this peak of activity occurring before similar increases in cardiac-modulated SPN. Stimulation of vasodepressor regions of the NTS evoked a membrane hyperpolarization and decrease in cell input resistance in cardiac-modulated but not non-modulated interneurons. These studies show that activity patterns in SPN in vivo are determined principally by synaptic inputs. They also demonstrate that spinal interneurons which exhibit cardiac-modulated patterns of activity are postsynaptically inhibited following activation of baroreceptor pathways. However, the question as to whether these inhibitory pathways and/or disfacilitation of tonic excitatory drive underlies the baroreceptor-mediated inhibition of SPN remains to be determined.

A physiological role for nitric oxide in the centrally mediated sympathetic and somatomotor ejaculatory response in anesthetized male Wistar rats.

Neurones in the lumbosacral spinal cord are known to play a significant role in ejaculation. In these same areas neurones containing nitric oxide synthase (NOS) have been described. This raised the question as to whether there is a physiological role for nitrous oxide (NO) in the spinal cord in sexual behavior. We first established immunohistochemical localization of NOS positive neurones in the lumbosacral spinal cord. NOS positive neurones were found in several areas of the lumbosacral cord. Namely the intermediolateral column (IML) at L(1)-L(4) and sacral cord; the dorsal gray commissure above the central canal at L(1)-L(2); the ventral gray area of lamina X below the central canal at L(3)-L(4); the superficial laminae of the dorsal horn at all levels. Secondly, we examined the role of NO in the generation of synchronized bursting activity within the vas deferens nerve and associated penile muscles, typical of sexual responses in the male anesthetized rat. NO modulators were applied directly to the spinal cord at T(13)-L(4) via a catheter placed subdurally (intrathecal) and their effect on the genital responses evoked by systemic administration of p-chloroamphetamine (PCA) or apomorphine (apo) (both 1 mg/kg) was observed. All responses evoked by PCA (n=4) or apo (n=3) were abolished or reduced (n=1) during intrathecal NOS inhibition using N((G)) nitro-L-Arginine methyl ester (l-NAME, 200 mM, 20-microl). NOS inhibition using l-NAME was reversed with simultaneous intrathecal application of the NO substrate l-arginine (100 mM, 20-microl, n=3). The selective neuronal NOS inhibitor 1-(2-trifluoro-methylphenyl) imidazole (100 mM, 20-microl, TRIM) also abolished all responses evoked by PCA (n=3). There was evidence that the responses within the vas deferens nerve (VDN) after PCA or apo were enhanced following NO donation using sodium nitroprusside (SNP, 1 mM, 20-microl). Furthermore, a PCA-like response within the VDN was evoked following intrathecally applied l-glutamic acid (200 mM, 20-microl) in six of 26 animals and also by intrathecal SNP in two of eight animals. In conclusion the results suggest a significant excitatory role for NO in the bursting pattern of synchronized discharge generated in autonomic and somatic outflows from the lumbosacral cord by neurones governing ejaculation.

Sympathetic genital responses induced by p-chloroamphetamine in anaesthetized female rats.

In urethane-anesthetized female rats, a branch of the hypogastric nerve equivalent to the vas deferens nerve in males was shown anatomically and electrophysiologically to supply the uterine horns and we have consequently termed this the uterine nerve. Administration of p-chloroamphetamine i.v. elicited patterned bursting uterine nerve activity responses together with contractions of the uterine horns and musculature of the vaginal wall. These responses are qualitatively similar to ejaculatory responses observed following p-chloroamphetamine administration to anesthetized male rats and the urethrogenital reflex in females, suggesting they represent responses occurring during sexual processes. This response to p-chloroamphetamine was still present after complete transection of the spinal cord at T8. These data indicate that common neurophysiological and pharmacological mechanisms regulate genital reflexes at the lumbosacral spinal level in both the female and the male rat.

Activation by p-chloroamphetamine of the spinal ejaculatory pattern generator in anaesthetized male rats.

In urethane-anesthetized male rats, a branch of the hypogastric nerve was shown, anatomically and electrophysiologically, to supply the vas deferens. Recordings from this nerve revealed a low level of tonic activity, which was predominantly efferent motor activity. Administration of p-chloroamphetamine i.v., elicited a rhythmic burst of neuronal activity, coherent with rhythmic pressure increases in the vas deferens and contractions of the bulbospongiosus muscles, which together comprise ejaculation. This response to p-chloroamphetamine was still present after complete transection of the spinal cord at T8-T9. These data indicate that p-chloroamphetamine is capable of activating the spinal neuronal circuits that generate the pattern of autonomic and somatic responses similar to those of sexual climax. Furthermore based on the best documented action of p-chloroamphetamine, the results suggest that the excitability of the pattern generator is regulated by serotonergic, dopaminergic or noradrenergic receptors in the lumbosacral spinal cord. We conclude this animal model will enable robust studies of the pharmacology and physiology of central neural mechanisms involved in ejaculation and sexual climax.

Activation of D2-like receptors induces sympathetic climactic-like responses in male and female anaesthetised rats.

In anaesthetised male rats an intravenous (i.v.) injection of p-chloroamphetamine (PCA) produced a specific patterned bursting response in the sympathetic vas deferens nerve (VDN) that corresponds to ejaculation. In the present, study selective dopamine agonists and antagonists were used to investigate whether dopaminergic mechanisms influence the generation of this ejaculatory-related response. Administration of a mixed D(1/2) receptor agonist (0.1-1.0 mg kg(-1) apomorphine i.v.) also evoked the characteristic bursting pattern responses in the VDN. Similar, but fewer, burst pattern responses could also be evoked by a selective D(2/3) receptor agonist (0.1-2.0 mg kg(-1) piribedil). Responses to 1.0 mg kg(-1) apomorphine were blocked by pretreatments with either 0.5 mg kg(-1) remoxipride (D(2) receptor antagonist) or 0.5 mg kg(-1) nafadotride (D(3) receptor antagonist), suggesting that D(2)-like receptors were involved. Responses could not be evoked by i.v. injections of apomorphine (1.0 mg kg(-1)) in anaesthetised male rats with a midthoracic spinal section, indicating that activation of D(2)-like receptors at supraspinal sites leads to an increase in the excitability of the lumbosacral pattern generator for ejaculation. In anaesthetised female rats a similar patterned bursting response occurred in the uterine nerve (UN) in response to apomorphine (0.5-2.0 mg kg(-1) i.v.). Thus a common neural mechanism may regulate sexual climactic reflexes in both sexes.

The sympathetic innervation of the heart: Important new insights.

Autonomic control of the heart has a significant influence over development of life threatening arrhythmias that can lead to sudden cardiac death. Sympathetic activity is known to be upregulated during these conditions and hence the sympathetic nerves present a target for treatment. However, a better understanding of the anatomy and physiology of cardiac sympathetic nerves is required for the progression of clinical interventions. This review explores the organization of the cardiac sympathetic nerves, from the preganglionic origin to the postganglionic innervations, and provides an overview of literature surrounding anti-arrhythmic therapies including thoracic sympathectomy and dorsal spinal cord stimulation. Several features of the innervation are clear. The cardiac nerves differentially supply the nodal and myocardial tissue of the heart and are dependent on activity generated in spinal neurones in the upper thoracic cord which project to synapse with ganglion cells in the stellate complex on each side. Networks of spinal interneurones determine the pattern of activity. Groups of spinal neurones selectively target specific regions of the heart but whether they exhibit a functional selectivity has still to be elucidated. Electrical or ischemic signals can lead to remodeling of nerves in the heart or ganglia. Surgical and electrical methods are proving to be clinically beneficial in reducing atrial and ventricular arrhythmias, heart failure and severe cardiac pain. This is a rapidly developing area and we need more basic understanding of how these methods work to ensure safety and reduction of side effects.

Pharmacological control of altitude sickness.

Acute mountain sickness has long been recognized as a potentially life-threatening condition afflicting otherwise healthy individuals who ascend rapidly to high altitude, where the partial pressure of oxygen in the air is reduced. The symptoms of acute mountain sickness are probably a consequence of disturbances in fluid balance brought about by severe tissue hypoxia. Acute mountain sickness can be prevented by an adequately slow ascent, which is the best method, but for those with limited time there are several drug therapies that provide a relatively good protection. John Coote describes these treatments and their possible mechanisms of pharmacological action.

Effect of electrical vs. chemical deep brain stimulation at midbrain sites on micturition in anaesthetized rats.

AIM: To understand how deep brain stimulation of the midbrain influences control of the urinary bladder. METHODS: In urethane-anaesthetized male rats, saline was infused continuously into the bladder to evoke cycles of filling and voiding. The effect of electrical (0.1-2.0 ms pulses, 5-180 Hz, 0.5-2.5 V) compared to chemical stimulation (microinjection of D,L-homocysteic acid, 50 nL 0.1 M solution) at the same midbrain sites was tested. RESULTS: Electrical stimulation of the periaqueductal grey matter and surrounding midbrain disrupted normal coordinated voiding activity in detrusor and sphincters muscles and suppressed urine output. The effect occurred within seconds was reversible and not secondary to cardiorespiratory changes. Bladder compliance remained unchanged. Chemical stimulation over the same area using microinjection of D,L-homocysteic acid (DLH) to preferentially activate somatodendritic receptors decreased the frequency of micturition but did not disrupt the coordinated pattern of voiding. In contrast, chemical stimulation within the caudal ventrolateral periaqueductal grey, in the area where critical synapses in the micturition reflex pathway are located, increased the frequency of micturition. CONCLUSION: Electrical deep brain stimulation within the midbrain can inhibit reflex micturition. We suggest that the applied stimulus entrained activity in the neural circuitry locally, thereby imposing an unphysiological pattern of activity. In a way similar to the use of electrical signals to 'jam' radio transmission, this may prevent a synchronized pattern of efferent activity being transmitted to the spinal outflows to orchestrate a coordinated voiding response. Further experiments to record neuronal firing in the midbrain during the deep brain stimulation will be necessary to test this hypothesis.

Nitric oxide and cardiac autonomic control in humans.

Cardiac autonomic control is of prognostic significance in cardiac disease, yet the control mechanisms of this system remain poorly defined. Animal data suggest that nitric oxide (NO) modulates cardiac autonomic control. We investigated the influence of NO on the baroreflex control of heart rate in healthy human subjects. In 26 healthy male volunteers (mean age, 23+/-5 years), we measured heart rate variability and baroreflex sensitivity during inhibition of endogenous NO production with N(G)-monomethyl-L-arginine (L-NMMA) (3 mg/kg per hour) and during exogenous NO donation with sodium nitroprusside (1 to 3 mg/h). Increases from baseline (Delta) in high-frequency (HF) indexes of heart rate variability were smaller with L-NMMA in comparison to an equipressor dose of the control vasoconstrictor phenylephrine (12 to 42 microg/kg per hour): Deltaroot mean square of successive RR interval differences (DeltaRMSSD)=23+/-32 versus 51+/-48 ms (P<0.002); Deltapercentage of successive RR interval differences >50 ms (DeltapNN50)=5+/-15% versus 14+/-12% (P<0.05); and DeltaHF normalized power=-2+/-7 versus 9+/-8 normalized units (P<0.01), respectively. Relative preservation of these indexes was observed during unloading of the baroreflex with sodium nitroprusside compared with a matched fall in blood pressure produced by a control vasodilator, hydralazine (9 to 18 mg/h): DeltaRMSSD=-8+/-8 versus -24+/-15 ms (P<0.001); DeltapNN50=-6+/-11% versus -15+/-19% (P<0.01); DeltaHF normalized power=-7+/-13 versus -13+/-11 normalized units (P<0.05), respectively. The change in cross-spectral alpha-index calculated as the square root of the ratio of RR interval power to systolic spectral power in the HF band (although not alpha-index calculated in the same way for the low-frequency bands or baroreflex sensitivity assessed by the phenylephrine bolus method) was attenuated with L-NMMA compared with phenylephrine (Delta=4+/-8 versus 14+/-15 ms/mm Hg, respectively; P<0.02) and with sodium nitroprusside compared with hydralazine (Delta=-7+/-6 and -9+/-7 ms/mm Hg, respectively; P<0.05). In conclusion, these data demonstrate that NO augments cardiac vagal control in humans.

Modulation of cardiac autonomic control in humans by angiotensin II.

Angiotensin II (Ang II) exerts an inhibitory action on vagal activity in animals and may also facilitate sympathetic activity. The object of this study was to compare autonomic activity resulting from equivalent steady-state baroreflex activation during intravenous Ang II infusion with that resulting from a control infusion of phenylephrine. Eight healthy subjects aged 22 to 34 years were studied in a single-blind, randomized, prospective crossover study. Autonomic activity was determined by computer analysis of RR interval variability in the time and frequency domains. Despite equal experimental hypertension with Ang II and phenylephrine infusion, at peak infusion rates the mean RR interval was significantly shorter with Ang II (983 +/- 179 milliseconds; mean +/- SD) than with phenylephrine (1265 +/- 187 milliseconds, P < .01). The variability of RR intervals was not significantly different, but the variability (median interquartile difference) of RR interval successive differences was significantly lower with Ang II (66 milliseconds) than with phenylephrine (104 milliseconds, P < .02). Power spectral analysis revealed the power of the 0.25-Hz component in normalized units to be significantly smaller during Ang II infusion (20.5 +/- 12.7 U) than during phenylephrine (38.2 +/- 14.7 U, P < .05), whereas the power of the 0.1-Hz component was significantly greater during Ang II infusion (67.8 +/- 17.1 U) than phenylephrine (38.8 +/- 20.3 U, P < .05). Measures of vagal modulation of heart rate were significantly attenuated, and sympathetic modulation appeared to be increased during Ang II infusion compared with control phenylephrine infusions. These observations may underlie reports of increased vagal activity during angiotensin-converting enzyme inhibitor therapy.

Evidence that sympathetic preganglionic neurones are arranged in target-specific columns in the thoracic spinal cord of the rat.

It is recognised that selective activation of different target-specific sympathetic preganglionic neurones forms the basis of many autonomic responses. The anatomical basis for this could be the spatial arrangement of these neurones in the spinal cord nuclei. The present study tested this possibility in the rat by determining the location in single animals of three distinct groups of sympathetic preganglionic neurones, one group projecting to the superior cervical ganglion, another to the stellate ganglion and one to the adrenal medulla. Sympathetic preganglionic neurones to each of these targets were simultaneously labeled with fluorescent dyes, either Fluorogold, Fast Blue, or Diamidino Yellow. The numbers and general morphology of the neurones were similar to previous descriptions, and they were distributed in four subnuclei, the nucleus intermediolateralis pars principalis, the nucleus intermediolateralis pars funiculus, the nucleus intercalatus spinalis, and the nucleus intercalatus spinalis pars paraependymalis. It was shown that all three groups of neurones were represented in the more medial sympathetic nuclei, but in the nuclei at the lateral border of the intermediate grey matter each one of the three groups of neurones occupied a discrete location. Adrenal medullary sympathetic preganglionic neurones occupied a lateral aspect, the superior cervical ganglion sympathetic preganglionic neurones a medial aspect, and the stellate ganglion sympathetic preganglionic neurones a space between. Some sympathetic preganglionic neurones were double labeled after dye injections into the superior cervical and stellate ganglion thus indicating that they projected to both ganglia.(ABSTRACT TRUNCATED AT 250 WORDS)

The central site of the sympatho-inhibitory action of 5-hydroxytryptamine in the cat.

The aim of this study was to determine the site in the CNS at which 5-hydroxytryptamine (5-HT) inhibits efferent sympathetic nerve activity in the cat. 5-Hydroxytryptamine (3 and 10 micrograms/kg), given into the lateral cerebral ventricle, produced immediate non dose related increases in mean blood pressure (MBP), heart rate (HR) and renal nerve activity (RNA). Larger doses (30 and 100 micrograms/kg i.c.v.) produced gradual decreases in blood pressure, heart rate and renal nerve activity, which did not occur when access of the drug to the fourth ventricle was prevented. Administration of 5-HT (10 and 30 micrograms/kg) into the fourth ventricle produced only decreases in blood pressure, heart rate and renal nerve activity after 15-40 min, which were accompanied by decreases in cardiac output and renal vascular resistance, but little or no change in total peripheral resistance. Application of 5-HT onto the ventral surface of the medulla, into the subarachnoid space at various levels along the spinal cord or into various parts of the nucleus tractus solitarius produced no effect on blood pressure heart rate or renal nerve activity. However, application of a cotton wool pledget soaked in a 5-HT solution (3 mg/ml) over the entire obex/NTS region produced immediate decreases in blood pressure, heart rate and renal nerve activity. These studies suggest that the sympatho-inhibitory effect of 5-HT is due to an action at a site near the caudal end of the dorsal surface of the medulla.

Identification of an efferent projection from the paraventricular nucleus of the hypothalamus terminating close to spinally projecting rostral ventrolateral medullary neurons.

The paraventricular nucleus of the hypothalamus is increasingly being viewed as an important site for cardiovascular integration because of its connections to regions in the brain and spinal cord which are known to be important in cardiovascular control. Like the vasomotor neurons of the rostral ventrolateral medulla, descending axons from paraventricular neurons can be identified that form synapses on sympathetic preganglionic neurons in the thoracic spinal cord. The purpose of this study was to determine whether paraventricular axons project to the rostral ventrolateral medulla and whether they are closely apposed to reticulospinal neurons in this region. Descending paraventricular axons were labelled with biotin dextran amine, while rostral ventrolateral medullary neurons were retrogradely labelled from the spinal cord with wheatgerm agglutinin conjugated to horseradish peroxidase. This revealed, within the rostral ventrolateral medulla, paraventricular axon and terminal varicosities closely apposed to and apparently contiguous with retrogradely labelled spinally projecting neurons. Thus our study at the light microscopical level has shown the potential for the paraventricular nucleus to directly influence rostral ventrolateral reticulospinal neurons. We suggest these connections, if confirmed by electron microscopy, could be one means by which activation of paraventricular neurons elicits alterations in blood pressure.

Identification of branching paraventricular neurons of the hypothalamus that project to the rostroventrolateral medulla and spinal cord.

The paraventricular nucleus of the hypothalamus has efferent connections to autonomic nuclei known to ultimately regulate cardiovascular function. Studies have revealed projections to the sympathetic preganglionic neurons of the spinal cord and presympathetic motor neurons of the rostral ventrolateral medulla. This study set out to establish whether the same neurons in the paraventricular nucleus innervate both these regions. In rats the fluorescent neuroanatomical tracers FluoroGold, Fast Blue or Dextran tetramethyl rhodamine were injected into either the rostral ventrolateral medulla or T2 region of the spinal cord. After a suitable survival period (five to seven days) three populations of neurons could be identified in the paraventricular nucleus, double-labelled neurons and single-labelled neurons resulting from the injections into the spinal cord or injections into the rostral ventrolateral medulla. The neurons were of similar size regardless of the dye content. Most neurons were found in the parvocellular subdivision of the mid rostral paraventricular nucleus. The number of labelled neurons decreased in the caudal sections. This study provides an anatomical basis for three means of influence that the paraventricular nucleus can have on sympathetic activity; a hierarchical in series projection via the rostral ventrolateral medulla; a projection running in parallel with this but bypassing the rostroventrolateral medulla; and a branching population innervating neurons in both the rostral ventrolateral medulla and spinal cord. The paraventricular nucleus of the hypothalamus is an important brain area concerned with maintaining cardiovascular homeostasis. This anatomical study has not only provided confirmatory evidence that direct projections arising from the paraventricular hypothalamic nucleus do project to the rostral ventrolateral medulla and spinal cord, regions known to influence cardiovascular regulation. The study has identified a branching projection originating in the paraventricular nucleus of the hypothalamus that projects to both the rostral ventrolateral medulla and the spinal cord. Thus the paraventricular nucleus of the hypothalamus has three pathways in which to influence cardiovascular homeostasis.

Rostroventrolateral medulla neurons preferentially project to target-specified sympathetic preganglionic neurons.

The rostroventrolateral medulla is a key site for the regulation of vasomotor tone. Sympatho-excitatory neurons project from this region to contact sympathetic preganglionic neurons located in the intermediolateral nucleus of the thoracic and lumbat spinal cord. Functional studies show that stimulation of specific sites in the ventral medulla lead to selective activation of different vascular effectors. The present study was designed to determine the anatomical basis for this selectivity in vasomotor control. Anterograde and retrograde tracing methods were utilized to determine if the descending rostral ventrolateral projection is topographically organized such that neurons in particular locations within the nucleus project preferentially and contact a specific group of sympathetic preganglionic neurons. For this purpose spinally-projecting neurons at 15 sites from three separate rostrocaudal locations within the rostroventrolateral medulla in nine rats were anterogradely labelled with biotin dextran amine. The spinal cord was examined for axon terminals having close apposition to two groups of sympathetic preganglionic neurons, those projecting to the superior cervical ganglion and those to the adrenal medulla which were retrogradely labelled with cholera B chain-conjugated horseradish peroxidase. Areas of close apposition between retrogradely-labelled dendrites, cell bodies and anterogradely-labelled axons were found. Axons descending from the more rostral part of the rostroventrolateral medulla produced the highest density of close appositions to sympathetic preganglionic neurons in both target-specific populations. Caudal rostroventrolateral medulla injection sites gave rise to a less dense distribution of axons and terminals around the spinal sympathetic nuclei. This study has demonstrated that spinally-projecting neurons in the rostroventrolateral medulla are both topographically and viscerotopically organized. It is suggested that such an arrangement provides the means for selective and differential control of autonomic effectors and in particular those involved in cardiovascular regulation.

Paraventricular nucleus activation of renal sympathetic neurones is synaptically depressed by nitric oxide and glycine acting at a spinal level.

A high density of nitric oxide synthesising enzyme is present in sympathetic preganglionic neurones in the spinal cord. It has been shown that nitric oxide is released as a consequence of synaptic activity. In the present study in anaesthetised rats we determined if nitric oxide acted as a retrograde messenger molecule to modulate the excitatory effects on the renal sympathetic spinal network elicited by paraventricular nucleus stimulation. Neurones in the latter nucleus were stimulated by microinjecting DLH and drugs were applied to the spinal cord via an intrathecal catheter with the tip positioned at T9-T10. Intrathecal application of the nitric oxide donors, sodium nitroprusside or [3-(2-hydroxy-1-methyl-2-nitrosohydrazino)-N-methyl-1-propanamine] significantly increased tonic activity in the renal sympathetic nerve. In contrast synaptic activity evoked by intrathecal glutamate or by paraventricular nucleus stimulation was enhanced by preventing nitric oxide generation with intrathecal N(G)-monomethyl-L-arginine monoacetate (L-NMMA) a nitric oxide synthase inhibitor. Enhancement of synaptically induced renal nerve activity was also observed following intrathecal glycine receptor inhibitor strychnine. Strychnine was without effect when it was given after L-NMMA. It was concluded that paraventricular nucleus excitation of renal sympathetic neurones is subject to inhibitory modulation by released nitric oxide and it is suggested the latter acts via glycine interneurones.