Comparative specialization of intrinsic cardiac neurons in humans, mice, and pigs.

Intrinsic cardiac neurons (ICNs) play a crucial role in the proper functioning of the heart; yet a paucity of data pertaining to human ICNs exists. We took a multidisciplinary approach to complete a detailed cellular comparison of the structure and function of ICNs from mice, pigs, and humans. Immunohistochemistry of whole and sectioned ganglia, transmission electron microscopy, intracellular microelectrode recording and dye filling for quantitative morphometry were used to define the neurophysiology, histochemistry, and ultrastructure of these cells across species. The densely packed, smaller ICNs of mouse lacked dendrites, formed axosomatic connections, and had high synaptic efficacy constituting an obligatory synapse. At Pig ICNs, a convergence of subthreshold cholinergic inputs onto extensive dendritic arbors supported greater summation and integration of synaptic input. Human ICNs were tonically firing, with synaptic stimulation evoking large suprathreshold excitatory postsynaptic potentials like mouse, and subthreshold potentials like pig. Ultrastructural examination of synaptic terminals revealed conserved architecture, yet small clear vesicles (SCVs) were larger in pigs and humans. The presence and localization of ganglionic neuropeptides was distinct, with abundant VIP observed in human but not pig or mouse ganglia, and little SP or CGRP in pig ganglia. Action potential waveforms were similar, but human ICNs had larger after-hyperpolarizations. Intrinsic excitability differed; 93% of human cells were tonic, all pig neurons were phasic, and both phasic and tonic phenotypes were observed in mouse. In combination, this publicly accessible, multimodal atlas of ICNs from mice, pigs, and humans identifies similarities and differences in the evolution of ICNs.

A comprehensive study of the abdominal ganglia part 2: Aorticorenal ganglia.

Investigation into reports of pain treatment for abdominal cancer and abdominal pain syndromes revealed the lack of human studies on some of the abdominal sympathetic ganglia. Recent studies on renal artery denervation therapy as treatment for resistant hypertension has made the aorticorenal ganglia of particular importance. The aim of this study was to investigate the location, morphology, interconnections, and histological nature of aorticorenal ganglia. We dissected nine abdominal cavities and harvested 37 aorticorenal ganglia. Hematoxylin and Eosin, and Masson's staining techniques were used to study the histological structure. Additionally, ganglia harvested from five individuals were stained with immunohistochemical techniques to test for tyrosine hydroxylase activity. All aorticorenal ganglia were located in proximity to the renal artery, and the majority were close to the vessel origin. Identification of multiple aorticorenal ganglia was the norm, and ranged from 2 to 4 on the left and 1 to 3 on the right. While the pattern of aorticorenal ganglia seemed to be unique in each individual case, the interconnections between these and other ganglia were vast. The aorticorenal ganglia shared direct connections with the celiac, gonadal, inferior mesenteric, and first lumbar sympathetic trunk ganglion. Contributions from the greater, lesser, and least thoracic splanchnic nerves were also observed. While the results of our study may not have direct clinical implications in isolation, the vast number of interconnections with the other abdominal ganglia may cause complications in procedures such as celiac ganglion block. In addition, aorticorenal innervation interruption may lead to hypotension.

A comprehensive study of the abdominal ganglia part 1: Celiac, phrenic and superior mesenteric ganglia.

INTRODUCTION: Patients with pancreatic cancer, chronic pancreatitis and other abdominal pain syndromes may develop debilitating pain throughout the course of their illness with little to no relief by most conventional methods. While some form of relief is experienced by patients, not all benefit from these procedures and side effects, while transitory in most cases are severe and often not expected. Our aim was therefore to investigate the anatomy surrounding the abdominal sympathetic ganglia, the target for the invasive procedures in an attempt to understand the variations in results. MATERIALS AND METHODS: The abdominal cavities of nine individuals were dissected and the ganglia investigated, harvested and histologically and immunochemical stained. RESULTS: The phrenic ganglion was found inconsistently and more often in the left than the right. If present it was located in association with the inferior phrenic artery and often connected to the celiac ganglion. The celiac ganglion was located anterior to the diaphragmatic crus on both sides and specifically posteromedial to the suprarenal gland and superior to the renal artery on the left. On the right it was located posterior to the suprarenal gland and inferior vena cava also superior to the renal vessels. The superior mesenteric ganglion was only positively identified in one individual and was located on the left lateral aspect of the superior mesenteric artery. CONCLUSION: The blockade procedures for treatment of pain are developed to target the area around the celiac artery where the ganglion is commonly described to be located. However, based on our results of its location and interconnections the ganglion is not located in the targeted area.

Universal nature of cholinergic regulation demonstrated with nicotinic acetylcholine receptors.

Mammalian nicotinic acetylcholine receptors (nAChRs) were initially discovered as ligand-gated ion channels mediating fast synaptic transmission in the neuro-muscular junctions and autonomic ganglia. They were further found to be involved in a wide range of basic biological processes within the brain and in non-excitable tissues. The present review summarizes the data obtained in our laboratory during last two decades. Investigation of autonomic ganglia with the nAChR subunit-specific antibodies was followed by identification of nAChRs in B lymphocytes, discovery of mitochondrial nAChRs and their role in mitochondrial apoptosis pathway, and revealing the role of α7 nAChRs and α7-specific antibodies in neuroinflammation-related Alzheimer disease and COVID-19. The data obtained demonstrate the involvement of nAChRs in cell survival, proliferation, cell-to-cell communication and inflammatory reaction. Together with the ability of nAChRs to function in both ionotropic and metabotropic way, these data illustrate the universal nature of cholinergic regulation mediated by nAChRs.

Procedural and short-term results of electroanatomic-mapping-guided ganglionated plexus ablation by first-time operators: A multicenter study.

INTRODUCTION: Single-center observational studies have shown promising results with fragmented electrogram (FE)-guided ganglionated plexus (GP) ablation in patients with vagally mediated bradyarrhythmia (VMB). We aimed to compare the acute procedural characteristics during FE-guided GP ablation in patients with VMB performed by first-time operators and those of a single high-volume operator. METHODS AND RESULTS: This international multicenter cohort study included data collected over 2 years from 16 cardiac hospitals. The primary operators were classified according to their prior GP ablation experience: a single high-volume operator who had performed > 50 GP ablation procedures (Group 1), and operators performing their first GP ablation cases (Group 2). Acute procedural characteristics and syncope recurrence were compared between groups. Forty-seven consecutive patients with VMB who underwent FE-guided GP ablation were enrolled, n = 31 in Group 1 and n = 16 in Group 2. The mean number of ablation points in each GP was comparable between groups. The ratio of positive vagal response during ablation on the left superior GP was higher in Group 1 (90.3% vs. 62.5%, p = .022). Ablation of the right superior GP increased heart rate acutely without any vagal response in 45 (95.7%) cases. The procedure time was longer in group 2 (83.4 ± 21 vs. 118.0 ± 21 min, respectively, p < .001). Over a mean follow-up duration of 8.0 ± 3 months (range 2-24 months), none of the patients suffered from syncope. CONCLUSION: This multi-center pilot study shows for the first time the feasibility of FE-guided GP ablation across a large group of procedure-naïve operators.

Simpaticotomía bilateral video asistida por hiperhidrosis: análisis de 102 pacientes / Bilateral video-assisted thoracic sympathicotomy for hiperhidrosis: analysis of 102 patients

Resumen La hiperhidrosis es un trastorno caracterizado por la producción excesiva de sudor por las glándulas sudoríparas ecrinas que influye negativamente en las actividades sociales, laborales y fundamentalmente en la calidad de vida. Se divide en primaria o secundaria. La primaria es una enfermedad benigna caracterizada por una excesiva sudoración con mayor frecuencia en palmas, plantas, axilas y cara. Su incidencia es del 1% de la población y su causa es desconocida. La mayoría de los tratamientos médicos no logran un buen control sintomático y frecuentemente son transitorios. La simpaticotomía torácica bilateral videoasistida se ha vuelto el tratamiento de elección en pacientes muy sintomáticos. En el período de 1998 a 2018 se realizaron 174 simpaticotomias bilaterales videoasistidas por hiperhidrosis primaria, de las cuales 102 cumplieron los criterios de inclusión. Se excluyeron a 72 pacientes. El 20.5% fueron hombres y el 79.5% mujeres con una edad media de 29.22 años. En cuanto a la localización fue palmoplantar axilar en un 50.9%, axilar en un 23.5%, palmoplantar en un 10.7%, palmar en un 7.8%, palmoaxilar 6.8% y facial 5.8%. Los pacientes con sudoración palmar presentaron 94.9% de mejoría, axilar 84.51%, plantar 46.25% y facial 84% respectivamente. El post operatorio arrojó una media de internación de 1.1 días. Como efecto no deseado, se presentó sudoración compensatoria en 53 casos y complicaciones postoperatorias en 18 casos. Concluimos que es una técnica segura, que resuelve de manera significativa la sudoración, mejorando la calidad de vida.

Abstract Hyperhidrosis is a disorder consisting of excessive sweating through the different body sweat glands, which produces a negative impact socially and in work-related activities in those that suffer this condition. There are primary and secondary forms. The primary form is a benign condition with excessive sweating mainly in palms, soles of feet, axillae and face. It affects a 1% of the population, and its cause is unknown. Most medical treatments are unsuccessful, and at best, transitory. In patients who are very troubled by the condition, videoassisted bilateral thoracic sympathicotomy has become the elective treatment. In the period ranging from 1998 to 2018, 174 procedures were undertaken for primary hyperhidrosis, of which 102 satisfied the inclusion criteria. 72 patients were excluded. A 20.5% were males, and 79.5% were females, with an average age of 29.22 years at surgery. As to localization of sweating, a 50.9% was palmar-plantar-axillary, 23.5% axillary, 10.7% palmarplantar, 7.8% palmar, 6.8% palmar-axillary, and a 5.8% facial. Those patients with palmar sweating showed a 94.9% improvement, those with axillary sweating a 88.51%, with plantar a 46.25% and those with facial sweating a 84% improvement. The average admission time was 1.1 days. As an undesired effect, compensatory sweating occurred in 53 cases and postoperative complications in 18 cases. We conclude this is a safe technique, that diminishes sweating significantly, improving patient's quality of life.

[Bilateral video-assisted thoracic sympathicotomy for hiperhidrosis: analysis of 102 patients]. / Simpaticotomía bilateral video asistida por hiperhidrosis: análisis de 102 pacientes.

Hyperhidrosis is a disorder consisting of excessive sweating through the different body sweat glands, which produces a negative impact socially and in work-related activities in those that suffer this condition. There are primary and secondary forms. The primary form is a benign condition with excessive sweating mainly in palms, soles of feet, axillae and face. It affects a 1% of the population, and its cause is unknown. Most medical treatments are unsuccessful, and at best, transitory. In patients who are very troubled by the condition, videoassisted bilateral thoracic sympathicotomy has become the elective treatment. In the period ranging from 1998 to 2018, 174 procedures were undertaken for primary hyperhidrosis, of which 102 satisfied the inclusion criteria. 72 patients were excluded. A 20.5% were males, and 79.5% were females, with an average age of 29.22 years at surgery. As to localization of sweating, a 50.9% was palmar-plantar-axillary, 23.5% axillary, 10.7% palmarplantar, 7.8% palmar, 6.8% palmar-axillary, and a 5.8% facial. Those patients with palmar sweating showed a 94.9% improvement, those with axillary sweating a 88.51%, with plantar a 46.25% and those with facial sweating a 84% improvement. The average admission time was 1.1 days. As an undesired effect, compensatory sweating occurred in 53 cases and postoperative complications in 18 cases. We conclude this is a safe technique, that diminishes sweating significantly, improving patient's quality of life.

La hiperhidrosis es un trastorno caracterizado por la producción excesiva de sudor por las glándulas sudoríparas ecrinas que influye negativamente en las actividades sociales, laborales y fundamentalmente en la calidad de vida. Se divide en primaria o secundaria. La primaria es una enfermedad benigna caracterizada por una excesiva sudoración con mayor frecuencia en palmas, plantas, axilas y cara. Su incidencia es del 1% de la población y su causa es desconocida. La mayoría de los tratamientos médicos no logran un buen control sintomático y frecuentemente son transitorios. La simpaticotomía torácica bilateral videoasistida se ha vuelto el tratamiento de elección en pacientes muy sintomáticos. En el período de 1998 a 2018 se realizaron 174 impaticotomias bilaterales videoasistidas por hiperhidrosis primaria, de las cuales 102 cumplieron los criterios de inclusión. Se excluyeron a 72 pacientes. El 20.5% fueron hombres y el 79.5% mujeres con una edad media de 29.22 años. En cuanto a la localización fue palmoplantar axilar en un 50.9%, axilar en un 23.5%, palmoplantar en un 10.7%, palmar en un 7.8%, palmoaxilar 6.8% y facial 5.8%. Los pacientes con sudoración palmar presentaron 94.9% de mejoría, axilar 84.51%, plantar 46.25% y facial 84% respectivamente. El post operatorio arrojó una media de internación de 1.1 días. Como efecto no deseado, se presentó sudoración compensatoria en 53 casos y complicaciones postoperatorias en 18 casos. Concluimos que es una técnica segura, que resuelve de manera significativa la sudoración, mejorando la calidad de vida.

New basic insights on the potential of a chitosan-based medical device for improving functional recovery after radical prostatectomy.

OBJECTIVES: To evaluate: (i) the neuro-regenerative potential of chitosan membrane (CS-Me) on acutely axotomised autonomic neurones in vitro; (ii) to exclude the possibility that a pro-regenerative biomaterial could interfere with the proliferation activity of prostate cancer cell lines; (iii) to provide an in vivo proof of the biocompatibility and regeneration promoting effect of CS-Me in a standardised rat model of peripheral nerve injury and repair; (iv) finally, to evaluate the tissue reaction induced by the degrading material; as previous studies have shown promising effects of CS-Me for protection of the neurovascular bundles for potency recovery in patients that undergo nerve-sparing radical prostatectomy (RP). MATERIALS AND METHODS: Addressing aim (i), the neuro-regenerative potential, organotypic cultures derived from primary sympathetic ganglia were cultured on CS-Me over 3 days and neurite extension and axonal sprouting were evaluated. Addressing aim (ii), effects of CS on cancer cells, different human prostate cancer cell lines (PC3, DU-145, LN-Cap) were seeded on CS-coated plates or cultured in the presence of CS-Me dissolution products. Addressing aims (iii) and (iv), functional recovery of peripheral nerve fibres and tissue reaction with the biomaterial, CS-Me and CS nerve guides were used to repair a median nerve injury in the rat. Functional recovery was evaluated during the post-recovery time by the behavioural grasping test. RESULTS: CS-Me significantly stimulated axon elongation from autonomic ganglia in comparison to control conditions in organotypic three-dimensional cultures. CS coating, as well as the dissolution products of CS-Me, led to a significantly lower proliferation rate of prostate cancer cell lines in vitro. Tissue reaction towards CS-Me and standard CS nerve guides was similar in the rat median nerve model, as was the outcome of nerve fibre regeneration and functional recovery. CONCLUSION: The results of this study provide the first experimental evidence in support of the clinical safety of CS-Me and of their postulated effectiveness for improving functional recovery after RP. The presented results are coherent in demonstrating that acutely axotomised autonomic neurones show increased neurite outgrowth on CS-Me substrate, whilst the same substrate reduces prostate cancer cell line proliferation in vitro. Furthermore, CS-Me do not demonstrate any disadvantage for peripheral nerve repair in a standard animal model.

[Age-related expression of calcium-binding proteins in autonomic ganglionic neurons].

Calbindin 28 kDa (CB), calretinin (CR) and parvalbumin (PB) are belonged to calcium-binding proteins which are widely distributed in the nervous system and selectively expressed in certain population of neurons. These proteins are expressed not only in the central nervous system, but also in the autonomic ganglia. CB and PB are found in the sympathetic ganglia of rodents, CB and CR are found in metasympathetic intramural ganglia. Their functions are poor understood but one can suggest their important role in regulation of the Ca2+ level in the cell. Сalcium-binding proteins are also play an important role in the development of autonomic neurons. There is an increasing of the percentage of CB and CR in the metasympathetic intramural ganglia of small intestine in the early postnatal development, whereas in sympathetic ganglia the percentage of CB is decreased. Possibly, the functional meaning of such changes can be explained by the role of calcium currents in the development of neurons and the synaptic transmission.

Estimating sympathetic tone by recording subcutaneous nerve activity in ambulatory dogs.

INTRODUCTION: We tested the hypothesis that subcutaneous nerve activity (SCNA) of the thorax correlates with the stellate ganglion nerve activity (SGNA) and can be used to estimate the sympathetic tone. METHODS AND RESULTS: We implanted radio transmitters in 11 ambulatory dogs to record left SGNA, left thoracic vagal nerve activity (VNA), and left thoracic SCNA, including 3 with simultaneous video monitoring and nerve recording. Two additional dogs were studied under general anesthesia with apamin injected into the right stellate ganglion while the right SGNA and the right SCNA were recorded. There was a significant positive correlation between integrated SGNA (iSGNA) and integrated SCNA (iSCNA) in the first 7 ambulatory dogs, with correlation coefficient of 0.70 (95% confidence interval [CI] 0.61-0.84, P < 0.05 for each dog). Tachycardia episodes (heart rate exceeding 150 bpm for ≥3 seconds) were invariably preceded by SGNA and SCNA. There was circadian variation of both SCNA and SGNA. Crosstalk was ruled out because SGNA, VNA, and SCNA bursts had different timing and activation patterns. In an eighth dog, closely spaced bipolar subcutaneous electrodes also recorded SCNA, but with reduced signal to noise ratio. Video monitoring in additional 3 dogs showed that movement was not a cause of high frequency SCNA. The right SGNA correlated strongly with right SCNA and heart rate in 2 anesthetized dogs after apamin injection into the right stellate ganglion. CONCLUSIONS: SCNA recorded by bipolar subcutaneous electrodes correlates with the SGNA and can be used to estimate the sympathetic tone.

A histopathological study of bovine ganglia.

One hundred and sixty-eight ganglia from 54 cattle aged 10 days to 10 years were examined microscopically. Samples from six autonomic ganglia and one sensory ganglion were represented. Thirteen animals were clinically normal and 41 were submitted for post-mortem examination. Neuronal vacuolation, spheroid formation, lipofuscin accumulation and central chromatolysis were observed sporadically and were of varying magnitude. Neuronal vacuolation and spheroid formation were not age-related changes, while lipofuscin accumulation was more common in older animals and central chromatolysis was more common in younger cattle. Non-suppurative inflammation and neuronophagia were also common findings (23 out of 54 animals, 42.6%) in autonomic ganglia that did not contain herpesvirus DNA as determined by polymerase chain reaction. Renaut bodies, features of peripheral nerves, were most commonly noted in the vagus. None of the histopathological findings were related to any particular disease in which loss of autonomic nervous system function might be expected. Furthermore, all changes were as common in clinically normal animals as in animals with disease.

Objective evaluation of plantar hyperhidrosis after sympathectomy

OBJECTIVE: The aim of the present study was to prospectively, randomly, blindly, and objectively investigate how surgery affects plantar sudoresis in patients with palmar and plantar hyperhidrosis over a one-year period using a sudorometer (VapoMeter). METHODS: From February 2007 to May 2009, 40 consecutive patients with combined palmar hyperhidrosis and plantar hyperhidrosis underwent video-assisted thoracic sympathectomy at the T3 or T4 ganglion level (15 women and 25 men, with a mean age of 25 years). RESULTS: Immediately after the operation and during the one-year follow-up, all of the patients were free from palmar hyperhidrosis episodes. Compensatory hyperhidrosis of varying degrees was observed in 35 (87.5%) patients after one year. Only two (2.5%) patients suffered from severe compensatory hyperhidrosis. There was a large initial improvement in plantar hyperhidrosis in 46.25% of the cases, followed by a progressive regression of that improvement, such that only 30% continued to show this improvement after one year. The proportion of patients whose condition worsened increased progressively (from 21.25% to 47.50%), and the proportion of stable patients decreased (32.5% to 22.50%). This was not related to resection level; however, a lower intensity of plantar hyperhidrosis prior to sympathectomy correlated with worse evolution. CONCLUSION: Patients with palmar hyperhidrosis and plantar hyperhidrosis who underwent video-assisted thoracic sympathectomy to treat their palmar hyperhidrosis exhibited good initial improvement in plantar hyperhidrosis, which then decreased to lesser degrees of improvement over a one-year period following the surgery. For this reason, video-assisted thoracic sympathectomy should not be performed when only plantar hyperhidrosis is present.

Neurogenesis in the adult peripheral nervous system.

Most researchers believe that neurogenesis in mature mammals is restricted only to the subgranular zone of the dentate gyrus and the subventricular zone of the lateral ventricle in the central nervous system. In the peripheral nervous system, neurogenesis is thought to be active only during prenatal development, with the exception of the olfactory neuroepithelium. However, sensory ganglia in the adult peripheral nervous system have been reported to contain precursor cells that can proliferate in vitro and be induced to differentiate into neurons. The occurrence of insult-induced neurogenesis, which has been reported by several investigators in the brain, is limited to a few recent reports for the peripheral nervous system. These reports suggest that damage to the adult nervous system induces mechanisms similar to those that control the generation of new neurons during prenatal development. Understanding conditions under which neurogenesis can be induced in physiologically non-neurogenic regions in adults is one of the major challenges for developing therapeutic strategies to repair neurological damage. However, the induced neurogenesis in the peripheral nervous system is still largely unexplored. This review presents the history of research on adult neurogenesis in the peripheral nervous system, which dates back more than 100 years and reveals the evidence on the under estimated potential for generation of new neurons in the adult peripheral nervous system.

Neurotensin receptor 1 immunoreactivity in the peripheral ganglia and carotid body.

In the present study we investigated, through immunohistochemistry, the presence and location of neurotensin receptor 1 (NTR1) in the peripheral ganglia and carotid body of 16 humans and 5 rats. In both humans and rats, NTR1 immunostained ganglion cells were found in superior cervical ganglia (57.4±11.6% and 72.4±11.4%, respectively, p<0.05), enteric ganglia (51.9±10.4% and 64.6±6.1%, p<0.05), sensory ganglia (69.2±10.7% and 73.0±13.1%, p>0.05) and parasympathetic ganglia (52.1±14.1% and 59.4±14.0%, p>0.05), supporting a modulatory role for NT in these ganglia. Positivity was also detected in 45.6±9.2% and 50.8±6.8% of human and rat type I glomic cells, respectively, whereas type II cells were negative. Our findings suggest that NT produced by type I cells acts in an autocrine or paracrine way on the same cell type, playing a modulatory role on chemoception.

Effect of Fluoxetine on Calcium or Potassium Channels in the Neuron of Rat Major Pelvic Ganglia / 대한비뇨기과학회지

PURPOSE: The major pelvic ganglia (MPG) provide the majority of the innervations to the lower urinary tract. The pelvic ganglia are unique autonomic ganglia that contain both sympathetic and parasympathetic neurons. It has been known that the low-threshold voltage-gated (T-type) Ca2 channels are only expressed only in the sympathetic neurons, whereas these channels are absent in parasympathetic neurons. In the present study, we examined the effect of fluoxetine, a world-wide used antidepressant, on the voltage-dependent Ca2 and K currents in the adrenergic neurons of the MPG. MATERIALS AND METHODS: The effect of fluoxetine on the voltage-dependent Ca2 and K currents in the adrenergic neurons of the MPG were examined using the whole-cell patch-clamp technique. RESULTS: Fluoxetine inhibited the voltage-activated Ca2 currents in the adrenergic neurons of the MPG. Both high-threshold (HVA) and low- threshold (LVA, T-type) Ca2 currents were inhibited by fluoxetine with an IC50 of 5.3 and 10.8microM, respectively. Fluoxetine also decreased the both the peak amplitude and the plateau of the outward K currents. The inhibition of the peak K currents by fluoxetine was concentration- dependent with an IC50 of 3.2microM. The inhibitions of the Ca2 and K currents were quickly reversible upon washout of the fluoxetine. CONCLUSIONS: These results provide evidence for the direct inhibition of the voltage dependant Ca2 and K currents by fluoxetine and these inhibitory effects could modify the synaptic transmission in adrenergic neurons of the MPG.

Characterization of Acetylcholine-induced Currents in Male Rat Pelvic Ganglion Neurons

The pelvic ganglia provide autonomic innervations to the various urogenital organs, such as the urinary bladder, prostate, and penis. It is well established that both sympathetic and parasympathetic synaptic transmissions in autonomic ganglia are mediated mainly by acetylcholine (ACh). Until now, however, the properties of ACh-induced currents and its receptors in pelvic ganglia have not clearly been elucidated. In the present study, biophysical characteristics and molecular nature of nicotinic acetylcholine receptors (nAChRs) were studied in sympathetic and parasympathetic major pelvic ganglion (MPG) neurons. MPG neurons isolated from male rat were enzymatically dissociated, and ionic currents were recorded by using the whole cell variant patch clamp technique. Total RNA from MPG neuron was prepared, and RT-PCR analysis was performed with specific primers for subunits of nAChRs. ACh dose-dependently elicited fast inward currents in both sympathetic and parasympathetic MPG neurons (EC50; 41.4microliterM and 64.0microliterM, respectively). ACh-induced currents showed a strong inward rectification with a reversal potential near 0 mV in current-voltage relationship. Pharmacologically, mecamylamine as a selective antagonist for alpha3beta4 nAChR potently inhibited the ACh-induced currents in sympathetic and parasympathetic neurons (IC50; 0.53micrometer and 0.22micrometer, respectively). Conversely, alpha- bungarotoxin, alpha-methyllycaconitine, and dihydro-beta-erythroidine, which are known as potent and sensitive blockers for alpha7 or alpha4beta2 nAChRs, below micromolar concentrations showed negligible effect. RT-PCR analysis revealed that alpha3 and beta4 subunits were predominantly expressed in MPG neurons. We suggest that MPG neurons have nAChRs containing alpha3 and beta4 subunits, and that their activation induces fast inward currents, possibly mediating the excitatory synaptic transmission in pelvic autonomic ganglia.

The Actions of Adenosine on Voltage-dependent K+ Currents in Neurons of Male Rat Major Pelvic Ganglia / 대한비뇨기과학회지

PURPOSE: The major pelvic ganglia (MPG) function as a relay center for autonomic pathways to the urogenital organs, such as the urinary bladder, vas deference, and penis. It is well known that adenosine acts as an important neuromodulator in various neuronal tissues. Several studies have suggested that some of these actions are coupled with potassium conductances. However, the exact mechanisms are unclear. Therefore, the roles of adenosine on the various potassium channels, in MPG neurons, were investigated. MATERIALS AND METHODS: Single neurons of the MPGs, located on the lateral surfaces of the prostate gland, from male rats were enzymatically dissociated. Ionic currents were recorded using the whole-cell variant patch-clamp technique. RESULTS: Two types of voltage-dependent outward potassium channels were isolated in the MPG neurons using whole-cell voltage protocols. One was the transient outward potassium current (type A-current, IA), the other was the delayed rectifier potassium current (IKDR). The IA and IKDR were recorded in both adrenergic and nonadrenergic neurons, which were distinguished by the existence of T-type calcium currents. Both the adrenergic and nonadrenergic neurons had the same kind of outward potassium currents. Application of adenosine (10(-4)M) increased the IA reversibly. N-cyclopentyladenosine (CPA, 10(-5)M), an A1 selective agonist, produced the same effect. However, the delayed rectifier components were not affected by the adenosine or CPA. The effects of adenosine and CPA on the IA were mostly prevented by pretreatment with DPCPX, an A1 selective antagonist. CONCLUSIONS: Adenosine increased the IA only, via the selective activation of A1 adenosine receptors. The augmentation of A-currents by adenosine may reduce neuronal firings, and then contribute to regulation of neuronal excitability in male rat MPG neurons.