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Characterization of the neurons of the mouse hypogastric ganglion: morphology and electrophysiology.

The somata of mouse hypogastric ganglion cells injected with Lucifer yellow were ovoid in shape, lacked dendritic processes but gave rise to a single axonal process. Antidromic activation demonstrated that some of the cells contained in this ganglion innervated the vas deferens. The passive and active membrane properties of the ganglion cells were determined in current clamp experiments. Cells fired tetrodotoxin-sensitive action potentials in response to intracellularly-applied depolarizing current. In voltage clamp evidence was obtained for both a persistent inward calcium current at potentials between -30 and -40 mV and a transient calcium current evoked by step depolarizations to around -20 mV. In current clamp, however, cells did not fire calcium action potentials in the presence of tetrodotoxin. Three potassium currents, IM (blocked by 1 mM barium and by 30 microM bethanechol), IA (blocked by 2 mM 4-aminopyridine) and IK(Ca) fast (blocked by 100 microM cadmium and by 5 mM tetraethylammonium) were characterized in these neurons. In addition, IK(Ca) slow was observed in a small proportion of cells. Fast, all-or-nothing, excitatory synaptic potentials were recorded in response to single stimuli applied to the afferent fibres running to the ganglion. In most cells the excitatory synaptic potentials were suprathreshold for action potential initiation and were markedly reduced or abolished by 100 microM mecamylamine, 1 mM hexamethonium and following desensitization to 100 microM nicotine. Excitatory synaptic potentials arose from stimulation of a single presynaptic nerve process and are typical of strong synaptic inputs.

Action Potentials↗

Cystometric changes in alloxan diabetic rats: evidence for functional and structural correlates of diabetic autonomic neuropathy.

Autonomic neuropathy and urinary bladder function were compared in Sprague-Dawley rats with alloxan-diabetes of 3 months duration, rats fed sucrose for 8 weeks, and rats examined 8 weeks after pelvic nerve surgical axotomy; normal age-matched rats were used as controls. All experimental interventions induced bladder hypertrophy with increased bladder weight. In diabetic and sucrose-fed animals, water intake and urinary output increased. Cystometric recordings of normal rats in vivo showed rhythmic contractions (1.25 +/- 0.25 contr/min) with threshold volume for micturition reflex at 0.51 +/- 0.04 ml. In diabetic rats, bladder contractions were irregular and of lower frequency (0.60 +/- 0.04 contr/min), while threshold volume was significantly higher (1.00 +/- 0.11 ml). Bladder contractions were normal in sucrose-fed animals, though threshold volume was markedly augmented (1.27 +/- 0.19 ml). Pelvic nerve surgical ablation abolished micturition reflex. In bladder strips excised post-mortem, contractile response to field stimulation was reduced in diabetic rats compared to control and sucrose-fed animals. Morphological examination of pelvic and hypogastric nerves revealed abnormalities characteristic of diabetic neuropathy only in diabetic rats. These data suggest that in alloxan-induced diabetes the decrease in the rate of bladder contraction is the result of autonomic neuropathy; while bladder hypertrophy in sucrose-fed rats appears to be an organ adaptation to hyperdiuresis.

Alloxan↗

Neurons labelled after the application of tracer to the distal stump of the transected hypogastric nerve in the rat.

Axons within the hypogastric nerve (HGN) provide sympathetic innervation to various tissues of the pelvic viscera (e.g. bladder, urethra, ureter, colon and sexual organs). Traditionally the HGN was considered to carry efferent impulses from the lumbar spinal cord to the periphery and afferent information from the periphery to the lumbar cord. In recent years however, there have been a number of reports of axons in the hypogastric nerve carrying impulses in directions opposite to these traditional ones. To further study this phenomenon fast blue dye was applied to the distal (or for comparison the proximal) stump of one transected hypogastric nerve and the locations of retrogradely labelled neurons determined in the spinal cord, the dorsal root ganglia (DRG), the sympathetic chain ganglia and the major pelvic ganglia (MPG). Labelled neurons were found in the spinal cord only for proximal stump dips. Labelled DRG neurons were mainly located in the L6-S1 ganglia when dye was applied to the distal stump, and mainly in the L1-L2 ganglia for proximal stump dips. The distribution profile of labelled sympathetic chain neurons was shifted caudally about one segment when the distal stump was dipped compared to the distribution obtained following a proximal stump dip. Labelled neurons were found contralateral to the dipped distal stump in all categories although in reduced numbers. More labelled neurons were found in male animals than in female animals. Fast blue in neurons in the DRGs and sympathetic chain labelled from the distal HGN reached these structures via the pelvic nerves. Labelled MPG neurons were found when the distal stump was dipped. Labelled MPG neurons were larger in the male than in the female. These fast blue labelled MPG neurons were also tested immunohistochemically for the presence of tyrosine hydroxylase (TH), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP). Labelled neurons which were TH+ were only found in the male; labelled neurons which were NPY+ or VIP+ were found in males and in females.

Animals↗

Hypogastric nerve section reveals a role for both afferent and efferent fibres in the feline continence process.

The role of the hypogastric afferent and efferent innervation in the process of urine storage during natural rate filling was examined in the pentobarbitone anaesthetized cat. The observed changes in bladder pressure following hypogastric nerve section demonstrated the presence of baseline sympatho-inhibitory and phasic sympatho-excitatory influences for a significant portion of each distension, which proceeded to a volume just over halfway through the continence process. A reduction in bladder wall compliance at the end of the distensions provided evidence for a net sympatho-inhibitory influence in the second half of the continence phase. Hypogastric nerve section also resulted in a reduction in the reflex increase in hypogastric efferent nerve activity, measured in-continuity. This suggested that hypogastric afferent fibres made a significant contribution to the regulation of sympathetic hypogastric nerve activity during natural rate filling.

Animals↗

Localisation of NADPH-diaphorase and acetylcholinesterase activities and of tyrosine hydroxylase and neuropeptide-Y immunoreactivity in neurons of the hypogastric ganglion of young adult and aged rats.

Neurons in the hypogastric (main pelvic) ganglia of 4- and 24-month-old male rats were investigated by enzyme histochemical methods for NADPH-diaphorase (NADPH-d) and acetylcholinesterase (AChE) activities and by immunofluorescence for tyrosine hydroxylase (TH) and neuropeptide Y (NPY) immunoreactivities. Systematic random sampling of standard sized areas of ganglion parenchyma revealed a content (per unit area) of 40.9 +/- 8.41 NADPH-d-positive neurons and 14.42 +/- 6.7 intensely AChE-positive neurons. In the aged rats the staining intensity was unchanged, but reductions in the numbers of cells stained for NADPH-d and AChE were not significantly different. Similar counts of TH- and NPY-immunoreactive neurons revealed values of 23.2 +/- 1.77 and 19.94 +/- 4.9, respectively, suggesting frequent co-localisation. The numbers of TH- and NPY-immunoreactive neurons were found to have decreased with age by 53% and 60%, respectively. Staining of consecutive sections revealed that those neurons which stained positively for NADPH-d did not show immunoreactivity for TH or NPY, and those neurons that were immunoreactive for TH or NPY did not contain intense NADPH-d staining. Occasional NPY-1R neurons were both TH- and NADPH-d-negative. These results suggest that NADPH-d staining is predominantly confined to the parasympathetic neuron population of the hypogastric ganglion and that it is the sympathetic neuron population alone that decreases in number with age.

Acetylcholinesterase↗

Arterial baroreflex inhibition by uterine distension in rats.

Arterial baroreflexes are inhibited during activation of some viscero-sensory receptors. The present study was designed to determine whether stimulation of mechanoreceptors of the uterus also inhibits the baroreflex bradycardia (BB). The aortic depressor nerve (ADN) was electrically stimulated to elicit BB in chloralose/urethane-anesthetized, succinylcholine-immobilized, and artificially ventilated rats. Hydraulic distension of the uterus with warm 0.9% NaCl solution was found to suppress BB with a threshold intra-uterine pressure of 25-100 mmHg. The inhibition was variably affected by successive transections of the hypogastric nerve (HgN) and the pelvic nerve (PN). In 6 rats out of the 16 tested, preceding transection of either HgN or PN (HgN, 4; PN, 2) abolished the inhibition, whereas in 8 rats the inhibition was not affected by preceding transection of either nerve but was abolished by subsequent transection of the other nerve (PN, 3; HgN, 5). In 2 rats, preceding HgN or PN section reduced the inhibition and subsequent PN or HgN section completed the withdrawal of the inhibition. Recording study confirmed that both the HgN and PN contain afferent fibers, signalling mechanoceptive information arising in the uterine wall. Electrical stimulation of the HgN as well as the PN remarkably suppressed the BB. In conclusion, uterine distension suppresses the BB and this inhibition is mediated by afferent fibers running in the HgN and PN with variable contributions among animals. The reflex suppression of BB due to uterine sensory activation may contribute to cardiovascular regulation during parturition.

Animals↗

Gender, bladder distension and hypogastric nerve activity in the cat.

In the pentobarbitone anaesthetised cat, postganglionic activity was recorded from the whole hypogastric nerve and from hypogastric filaments destined for the bladder wall during bladder distension. A gender difference was observed in whole hypogastric nerve recordings, cardiac-related activity obscuring other components in females with intact internal reproductive organs. In contrast, nerve activity in the vesical hypogastric filaments was dominated by bladder distension-related activity in both sexes. These observations suggest that bladder distension-related activity, which has a continence promoting action on the bladder detrusor muscle, is destined for the bladder wall, while the cardiac-related activity is largely destined for the uterine and vaginal vasculature.

Animals↗

Changes in neuronal contribution to contractile responses of vas deferens of young and adult guinea pigs.

The response characteristics of vas deferens to electrical hypogastric nerve stimulation at various frequencies was studied in guinea pigs of 2 to 15 weeks old. In 2-week-old guinea pigs the stimulation induced monophasic contraction, some of which remained after blocking alpha 1-adrenoceptor and desensitizing P2-purinoceptors with prazosin and alpha, beta-methylene ATP, respectively. In guinea pigs of 10 to 15 weeks old stimulation induced biphasic contraction, which was almost completely inhibited by both blockers. These results suggest that some unknown component other than ATP and norepinephrine is involved in the transmission at 2 weeks, and that its relative significance changes during development.

Adenosine Triphosphate↗

Sympathetic and parasympathetic regulation of the uterine blood flow and contraction in the rat.

The effects of electrical stimulation of hypogastric sympathetic and pelvic parasympathetic nerves on uterine blood flow and contraction in anesthetized female non-pregnant normal cycling rats were examined. Electrical stimulation of the efferent pelvic nerve with supramaximal intensity induced marked increase of uterine blood flow accompanied by uterine contraction. On the other hand, the stimulation of efferent hypogastric nerve caused decrease of uterine blood flow accompanied by uterine contraction. These responses could only be elicited with stimulus intensity above the threshold for unmyelinated C fibers in both the hypogastric and pelvic nerves. Intravenous administration of atropine (0.5 mg/kg) totally blocked the response of uterine contraction elicited by pelvic and hypogastric nerve stimulation and also the increase of blood flow induced by pelvic nerve stimulation. Intravenous administration of phenoxybenzamine (0.5 mg/kg) blocked the decreased response of uterine blood flow induced by hypogastric nerve stimulation. It was concluded that uterine blood flow and contraction were regulated by both the parasympathetic and sympathetic nerves, but in different manners; blood flow is regulated reciprocally (1) by parasympathetic vasodilators mainly via activation of muscarinic cholinergic receptors, and (2) by sympathetic vasoconstrictors via activation of alpha-adrenergic receptors; contraction is produced by activation of both parasympathetic and sympathetic nerves via muscarinic cholinergic receptors.

Adrenergic alpha-Antagonists↗

Neural mechanisms accounting for the increase in blood pressure and heart rate during vagino-cervical stimulation.

The rise in blood pressure and heart rate produced by the mechanical stimulation of the uterine cervix (VS) was examined after adrenalectomy, after pelvic or hypogastric neurectomies or after spinal cord transection in anesthetized rats. Neither adrenalectomy, nor hypogastric neurectomy prevented the rise in heart rate and blood pressure produced by VS. After the spinal cord transection at T6 level, VS was still able to produce the rise in blood pressure. However, the rise in blood pressure was significantly lower than that produced in the same animals before the transection. No changes in heart rate were produced by VS after spinal cord transection. This result can be explained because this level of transection prevents the reach of the afferent inflow to the superior cervical ganglia. Pelvic neurectomy abolished completely the effects of VS on blood pressure and heart rate. Low intensity (1-2 times the threshold) electrical stimulation of the pelvic nerve produced a rise in blood pressure. Even though heart rate increased during electrical stimulation, the change in heart rate was not statistically different from the pre-stimulation value. These results suggest that the changes in blood pressure and heart rate produced by VS represent a neuronal reflex response mediated by the pelvic nerve. The fact that the effects of VS on blood pressure persist in spinal cord-transected animals suggests that the reflex is integrated at the spinal level. However, the cardiovascular responses to VS were significantly lower than before transection, suggesting that supraspinal centers are also involved in the reflex.

Adrenalectomy↗

Biochemical and morphological effects of castration on the postorganizational development of the hypogastric ganglion.

The biochemical and morphological development of the sympathetic hypogastric ganglion (HG) was examined subsequent to postnatal castration at 10-11 days of age. Previous studies suggested that tyrosine hydroxylase (T-OH) activity, an index of noradrenergic maturation, and choline acetyltransferase (ChAT) activity, a marker for preganglionic terminal formation, were dependent on gonadal hormones during normal ontogeny. In the present studies, morphometric analyses of the HG revealed that the cross-sectional area of the cell soma and nucleus were significantly reduced following postnatal castration at day 10. Conversely, castration produced no change in the number of HG neurons. In addition, postnatal castration prevents the development of postsynaptic T-OH activity to a greater extent than ganglionic protein resulting in a significant loss of T-OH specific activity. In contrast, presynaptic ChAT activity was reduced in parallel with ganglionic protein, thus ChAT specific activity was unchanged. Testosterone replacement therapy, even in groups where treatment was delayed for up to 2 weeks after castration, completely reversed deficits in both T-OH and ChAT activities. These studies suggest that altered development of ganglion protein subsequent to postnatal castration is related to decreases in the size of neurons and not to the loss of neurons. The lack of cell loss also suggest that decreased levels of postsynaptic T-OH activity results from a loss of enzyme activity per cell and the decreased levels of ChAT activity probably represent fewer presynaptic terminals per neuron. In addition, delayed testosterone replacement subsequent to castration was effective in restoring enzyme activities suggesting an 'activational' not 'organizational' role for testosterone after postnatal day 10.

Animals↗

Colocalization of vasoactive intestinal polypeptide and nitric oxide in penis-innervating neurons in the major pelvic ganglion of the rat.

By using a combination of fluorescent retrograde labeling and double immunofluorescence histochemistry, the present study revealed in the rat that the vast majority of major pelvic ganglion neurons innervating penile erectile tissue contained both vasoactive intestinal polypeptide (VIP) and nitric oxide synthase. The result suggests that VIP and nitric oxide may exert a cooperative action in penile erection.

Amino Acid Oxidoreductases↗

Reactions of rat sympathetic neurons to ethanol exposure are age-dependent.

Age-differences in the sensitivity of peripheral sympathetic neurons to chronic ethanol exposure and ethanol withdrawal were studied in male Wistar rats aged 4 months, 12 months, or 24 to 25 months. The superior cervical ganglia (SCG) of the young (4 months) and the 2-year-old rats responded to a 12-day or 4-week ethanol exposure with significantly increased catecholamine turnover, while the ganglia of the middle-aged rats (12 months) showed only a minor increase in the intensity of catecholamine fluorescence and tyrosine hydroxylase immunoreactivity. Extensive neuronal vacuolation was found in the 4 months ethanol-exposed SCG, probably as a reaction of a subpopulation of neurons to increased stimulation. Ethanol-induced neuronal loss was most prominent in the SCG of the oldest age group. Contrary to the marked changes in SCG functional and morphometric parameters, the pelvic sympathetic neurons in the hypogastric ganglion showed no significant changes after ethanol exposure. The pattern of ethanol-induced morphological alterations found in the present study did not provide unambiquous support for either the "accelerated aging" or the "increased vulnerability" concept regarding ethanol-aging interactions in the nervous system.

Aging↗

Effects of stimulation on the multiquantal spontaneous synaptic potentials in guinea pig hypogastric ganglia.

Loss frequency (0.5-2 Hz) stimulation of the preganglionic nerve produced calcium ion (Ca2+)-dependent increases in the frequency of spontaneous synaptic potentials recorded from guinea-pig hypogastric ganglion cells. The increased frequency was accompanied by a marked increase in the proportion of multiquantal spontaneous potentials and this was also dependent on extracellular Ca2+. The latter effect was interpreted as an increase in the degree of bursting within the spontaneous release process, supporting the hypothesis that the bursting is related to the cytoplasmic Ca2+ concentration.

Animals↗

Developmental changes of the hypogastric ganglion associated with the differentiation of the reproductive tracts in the mouse.

Prenatal exposure of male or female mice to diethylstilbestrol (DES) or testosterone propionate (TP) from days 9 to 16 of gestation produces pseudohermaphrodites with dual reproductive tracts. In these animals, the hypogastric ganglion which is thought to innervate the internal genitalia was examined at birth. The number of ganglion cells was significantly larger in prenatally DES-exposed male and TP-exposed female mice than that of controls. These results suggest that the number of hypogastric ganglion cells is dependent on the number of the target organs for innervation.

Animals↗

alpha-Bungarotoxin and P15 toxin binding site in mammalian sympathetic ganglia.

alpha-Bungarotoxin (alpha-Bgtx) demonstrates a specific and saturable binding to rat superior cervical ganglion, yet it does not block ganglionic response mediated by nicotinic receptors in the guinea pig vas deferens-hypogastric nerve preparation. P15 toxin, a probe for ganglionic nicotinic receptor, prevents the binding of alpha-Bgtx to rat ganglia, and alpha-Bgtx prevents the ganglioplegic action of hexamethonium. Hexamethonium does not block the binding of alpha-Bgtx and P15 to rat ganglia. It is concluded that alpha-Bgtx and P15 bind to ganglionic nicotinic receptor at a common site which is different from that of cholinergic agonists and antagonists.

Animals↗

Neural inputs from the uterus to the paraventricular magnocellular neurons in the rat.

Extracellular action potentials were recorded from antidromically identified, tonically firing cells in the hypothalamic paraventricular nucleus (PVN) of ovariectomized, estrogen-treated female rats under urethane anesthesia. Genital or somatic sensory stimuli, or electrical stimulation of the nerves innervating the pelvis were applied. Uterine horn or vaginal distension each excited 33% of the neurons tested. Probing of the cervix had no effect. Hindpaw pinch produced excitation in 39% and inhibition in 11% of the neurons tested. Non-noxious somatic stimuli had no effect. Stimulation of the uterine afferent nerves, the hypogastric and pelvic nerves, excited 55% and 30% of the neurons tested, respectively. Stimulation of a somatic nerve of the hindleg, the sciatic nerve, activated 80% of the neurons tested. These results indicate that specific sensory afferents arrive at the PVN from the uterus; in addition, somatic afferents converge in this hypothalamic nucleus.

Afferent Pathways↗

Responses of hypogastric nerve afferent fibers to uterine distension in estrous or metestrous rats.

The response of afferent fibers in the distal end of the cut hypogastric nerve to distension of the uterus to various pressure levels was investigated in anesthetized virgin female rats on two different days of the estrous cycle (estrus or metestrus). Whereas relatively intense levels of uterine distension produced an increase in afferent activity in a pressure-dependent manner in both stages of the estrous cycle, the minimal pressure necessary to activate the fibers was significantly less on the day of estrus. These results suggest that hypogastric nerve afferent fibers are more likely to respond to potentially damaging uterine stimuli during the time when such responsiveness would be most important for reproductive success.

Action Potentials↗