Autonomic Nerve Endings Research Articles

Paraventricular Hypothalamic Nucleus Upregulates Intraocular Pressure Via Glutamatergic Neurons.

The neuroregulatory center of intraocular pressure (IOP) is located in the hypothalamus. An efferent neural pathway exists between the hypothalamic nuclei and the autonomic nerve endings in the anterior chamber of the eye. This study was designed to investigate whether the paraventricular hypothalamic nucleus (PVH) regulates IOP as the other nuclei do. Optogenetic manipulation of PVH neurons was used in this study. Light stimulation was applied via an optical fiber embedded over the PVH to activate projection neurons after AAV2/9-CaMKIIα-hChR2-mCherry was injected into the right PVH of C57BL/6J mice. The same methods were used to inhibit projection neurons after AAV2/9-CaMKIIα-eNpHR3.0-mCherry was injected into the bilateral PVH of C57BL/6J mice. AAV2/9-EF1α-DIO-hChR2-mCherry was injected into the right PVH of Vglut2-Cre mice to elucidate the effect of glutamatergic neuron-specific activation. IOP was measured before and after light manipulation. Associated nuclei activation was clarified by c-Fos immunohistochemical staining. Only mice with accurate viral expression and fiber embedding were included in the statistical analysis. Activation of projection neurons in the right PVH induced significant bilateral IOP elevation (n = 11, P < 0.001); the ipsilateral IOP increased more noticeably (n = 11, P < 0.05); Bilateral inhibition of PVH projection neurons did not significantly influence IOP (n = 5, P > 0.05). Specific activation of glutamatergic neurons among PVH projection neurons also induced IOP elevation in both eyes (n = 5, P < 0.001). The dorsomedial hypothalamic nucleus, ventromedial hypothalamic nucleus, locus coeruleus and basolateral amygdaloid nucleus responded to light stimulation of PVH in AAV-ChR2 mice. The PVH may play a role in IOP upregulation via glutamatergic neurons.

Morphological and neurochemical characterisation of anterogradely labelled spinal sensory and autonomic nerve endings in the mouse bladder.

The bladder is innervated by axons of sympathetic and parasympathetic efferent nerves, and by spinal afferent neurons. The objective was to characterise anatomically and immunohistochemically the terminal endings of sensory and autonomic motor nerve endings in wholemount preparations of the mouse bladder. We used both anterograde labelling of pelvic and hypogastric nerves ex vivo and anterograde labelling from lumbosacral dorsal root ganglia (DRG) in vivo in male and female mice. These were combined with immunohistochemistry for major markers of sensory, sympathetic and parasympathetic nerves. Selective labelling of spinal afferent endings following dextran biotin-labelling from DRGs in vivo showed no co-localisation of VAChT or TH in sensory terminals in the detrusor and suburothelial plexus. Biotinamide was applied ex vivo to nerve trunks arising in the pelvic ganglion and running towards the bladder. Among the filled axons, 38% of detrusor fibres and 47% of suburothelial axons were immunoreactive for calcitonin-gene related peptide (CGRP). Vesicular acetylcholine transporter (VAChT) immunoreactivity was present in 26% of both detrusor and suburothelial axons. For tyrosine hydroxylase (TH), the proportions were 15% and 17%, respectively. Three major morphological types of CGRP-immunoreactive nerve endings were distinguished in the bladder wall: simple, branching and complex. VAChT-immunoreactive parasympathetic axons had simple and branching endings; TH immunoreactive axons all had simple morphologies. Our findings revealed that different subtypes of sensory and autonomic nerve endings can be reliably identified by combining anterograde labelling ex vivo with specific immunohistochemical markers, although morphologically some of these types of endings were indistinguishable.

Extracellular Diadenosine Tetraphosphate Suppresses Ectopic Proarrhythmicity in the Myocardial Tissue of the Pulmonary Veins in Adult but not in Neonatal Rats

Diadenosine tetraphosphate (Ap4A) belongs to a group of endogenous purine compounds that have recently been considered as new neurotransmitters or cotransmitters in the autonomic nervous system. It has been shown that Ap4A affects electrophysiology of a pacemaker and working myocardium and modulates adrenergic control of the heart in adult mammals. Nevertheless, the physiological role of Ap4A in the regulation of bioelectric properties in the pulmonary vein (PV) myocardium has not yet been investigated. It is well known that myocardial tissue in the wall of the PV acts as source of the ectopic proarrhythmic activity that underlies supraventricular arrhythmias like atrial fibrillation. The aim of the present study was to elucidate the effects Ap4A on bioelectrical properties and proarrhythmic ectopy in PVs in adult rats and at early postnatal ontogenesis. Thus, the resting potentials and the electrically evoked and spontaneous action potentials were recorded with the use of the standard microelectrode technique in multicellular isolated PV specimens from male Wistar rats at postnatal days 1-, 7-, 14-, and 21- and also from 60-day-old animals, which were considered as mature. The application of Ap4A caused significant reduction of action potential duration in PV specimens from rats of all ages. Ap4A also caused significant resting membrane potential hyperpolar-ization in quiescent PVs specimens from 14-, 21-, and 60-day-old rats. In addition, Ap4A caused complete and significant suppression of ectopic automaticity caused by preliminary noradrenaline administration in PV from 21- and 60-day-old rats, but Ap4A was unable to alter spontaneous intrinsic activity in PV from neo-nate (1-day-old) rats. The Ap4A-caused attenuation of noradrenaline-induced ectopy in PV was accompanied by substantial resting membrane potential hyperpolarization in all cases. Our results allow suggesting that the release of Ap4A as a cotransmitter from autonomic nerve endings can reduce proarrhythmic ectopy caused by sympathetic stimulation of the PV myocardium in vivo.

Plasma & urinary catecholamines & urinary vanillylmandelic acid levels in patients with generalized vitiligo.

Background & objectives:Vitiligo is an acquired skin disease characterized by depigmented areas of the skin. Increased release of catecholamines from autonomic nerve endings in microenvironment of melanocytes in affected skin might be involved in the aetiopathogenesis of vitiligo. Levels of catecholamines are considered as being related to onset or worsening of the disease. Therefore, in this study, the role of catecholamines was evaluated in mapping disease stability and outcome of vitiligo patients undergoing melanocyte transfer.Methods:In this study, circulatory and urinary levels of catecholamine (CA) and vanillylmandelic acid (VMA) were determined in 45 individuals (30 vitiligo patients and 15 healthy controls) using ELISA.Results:A significant increase for plasma and urinary catecholamines along with VMA was observed as compared to healthy controls. When the pre- and post-intervention levels were analyzed in responders and non-responders, respectively, only dopamine showed significant decline in urine, rest of the molecules in plasma as well as urine showed non-significant decline except VMA which showed insignificant increase.Interpretation & conclusions:Levels of plasma/urinary epinephrine, and plasma dopamine, could not be established as biomarkers for disease stability or successful outcome of autologous melanocyte transfer in generalized vitiligo patients. However, dopamine (urine) might be of help in determining the stability in patients with generalized vitiligo undergoing melanocyte transfer. Further studies need to be done on a large sample of patients to confirm our findings.

Da li terapija beta-blokatorima utiče na promene arterijskog krvnog pritiska u toku izvođenja spinalne anestezije? - beta-blokatori i regionalna anestezija

Introduction: Spinal anesthesia is one of the oldest, most useful and most commonly used techniques of regional anesthesia. The injection of local anesthetics into the subarachnoidal space creates a transient interruption of nerve impulses in the spinal nerve roots and paralysis of autonomic, sensitive and motor nerve endings. Sympathic blockade caused by spinal anesthesia leads to haemodynamic changes. Hypotension and bradycardia are the most common cardiovascular effects seen in sympathetic denervation. The goal of the work: Determine the effects of chronic therapy with beta-blockers on changes in arterial blood pressure during spinal anesthesia, since their effects are similar, and see the importance of hemodynamic monitoring (measurement of arterial pressure) in performing spinal anesthesia in this group of patients. Material and methods: After the approval by the Ethics Board of the University Clinical Center Banja Luka, a prospective, case-control study was conducted, which lasted from 01.06.2013. until 31.05.2016. at the surgical clinics of this institution. The study included 70 patients divided into two groups, aged 35-65 years. Group N1, a working group, consisted of 35 patients who, due to essential hypertension, used a beta-blocker, metoprolol, in chronic therapy. The second group was the control group, N2, and it consisted of 35 patients of the same age, ASA I status. Patients were subjected to spinal anesthesia for 'bloodless' inguinal hernia repair, bladder tumor operation, prostatic surgery, the urethra and lower extremities operations. Pregnant women, as well as patients with diabetes, renal, hepatic and heart diseases were not included in the work. Results: The results show that a decrease in arterial blood pressure (systolic, diastolic and mean) in the group of patients under beta-blocker therapy is significantly higher after performing spinal anesthesia. In the control group, systolic pressure decreased by about 11% after performing spinal anesthesia, while in the working group it decreased by 20%. Diastolic pressure after performing spinal anesthesia in the control group fell by 11.33%, which was a statistically significant decline. In the workgroup, this drop (from DP1 to DP2) was 20.4%.The mean arterial pressure after spinal anesthesia fell in the control group by 4%, while in the working group we recorded a decrease of MAP1-MAP2 of 20%. Conclusion: Patients on perioperative therapy with beta-blockers that underwent spinal anesthesia are hemodynamically unstable and require more efficient hemodynamic perioperative monitoring and intensive supervision.

Expression of nitric oxide-containing structures in the rat carotid body

The carotid body (CB) is a major peripheral arterial chemoreceptor organ that evokes compensatory reflex responses so as to maintain gas homeostasis. It is dually innervated by sensory fibers from petrosal ganglion (PG) neurons, and autonomic fibers from postganglionic sympathetic neurons of the superior cervical ganglion (SCG) and parasympathetic vasomotor fibers of intrinsic ganglion cells in the CB. The presence of nitric oxide (NO), a putative gaseous neurotransmitter substance in a number of neuronal and non-neuronal structures, was examined in the CB, PG and SCG of the rat using nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry, nitric oxide synthase (NOS) immunohistochemistry and retrograde tracing. One week after injecting the retrograde tracer Fast Blue (FB) in the CB, we found that a subset of perikarya in the caudal portions of the PG and SCG were FB-labeled. Histochemistry and immunohistochemistry revealed that the majority of large- and medium-sized PG and SCG cells were NADPH-d positive and displayed a strong NOS immunostaining. We also observed that many varicose nerve fibers penetrating the CB and enveloping the glomus cells and blood vessels were NADPH-d reactive and expressed the constitutive isoforms of NOS, nNOS and eNOS. In addition, some autonomic microganglion cells embedded within, or located at the periphery of the CB, and not glomus or sustentacular cells were nNOS-immunopositive while CB microvasculature expressed eNOS. The present results suggest that NO is a transmitter in the autonomic nerve endings supplying the CB and is involved in efferent chemoreceptor inhibition by a dual mechanism.

Translocation and dissemination to target neurons of botulinum neurotoxin type B in the mouse intestinal wall.

Botulinum neurotoxins (BoNTs) are responsible for severe flaccid paralysis (botulism), which in most cases enter the organism via the digestive tract and then disseminate into the blood or lymph circulation to target autonomic and motor nerve endings. The passage way of BoNTs alone or in complex forms with associated nontoxic proteins through the epithelial barrier of the digestive tract still remains unclear. Here, we show using an in vivo model of mouse ligated intestinal loop that BoNT/B alone or the BoNT/B C-terminal domain of the heavy chain (HCcB), which interacts with cell surface receptors, translocates across the intestinal barrier. The BoNT/B or HCcB translocation through the intestinal barrier occurred via an endocytosis-dependent mechanism within 10-20 min, because Dynasore, a potent endocytosis inhibitor, significantly prevented BoNT/B as well as HCcB translocation. We also show that HCcB or BoNT/B specifically targets neuronal cells and neuronal extensions in the intestinal submucosa and musculosa expressing synaptotagmin, preferentially cholinergic neurons and to a lower extent other neuronal cell types, notably serotonergic neurons. Interestingly, rare intestinal epithelial cells accumulated HCcB suggesting that distinct cell types of the intestinal epithelium, still undefined, might mediate efficient translocation of BoNT/B.

Flibanserin for Low Sexual Desire in Women: A Molecule From Bench to Bed?

Flibanserin for Low Sexual Desire in Women: A Molecule From Bench to Bed?

Clinical comparison of botulinum toxin in motor and autonomic disorders: Similarities and differences

Botulinum toxin is a well established, highly effective and safe treatment option for movement disorders and autonomic diseases with excellent long term results. There is increasing evidence that the beneficial effect in both motor and autonomic indication is based on a complex mode of botulinum toxin action modulating efferent as well as afferent nerve fiber activity. In particular, this has been shown for the treatment of dystonia, spasticity and overactive bladder. A unique observation is that botulinum toxin has a markedly longer duration of action in autonomic than in motor disorders for which the reason remains unclear. Although botulinum toxin type B seems to have an initially higher affinity to autonomic nerve endings there is currently no clear evidence that type B is superior to type A in autonomic disorders. The risk of antibody formation probably does not depend on the target tissue injected and seems to be similar for movement disorders and autonomic indications. More research is needed to better understand similarities and differences of treatment outcome in motor and autonomic disorders.

Imaging in Parkinson's disease: time to look below the neck.

This scientific commentary refers to ‘Imaging acetylcholinesterase density in peripheral organs in Parkinson’s disease with 11C-donepezil PET’ by Gjerloff et al. (10.1093/brain/awu369). The possibility of assessing disease progression in Parkinson’s disease was revolutionized by the ability to image dopamine synthesis in vivo , first described by Garnett and colleagues more than 30 years ago (Garnett et al. , 1983). Since that landmark paper, there have been hundreds of publications presenting the results of molecular imaging using either SPECT or PET tracers for the synthesis, synaptic packaging and/or reuptake of dopamine. While the relationship between changes in dopamine function assessed by these techniques and clinical deterioration is imperfect and may be confounded by compensatory changes and the effects of medications, it is nonetheless possible to obtain an independent measure of disease progression and indeed to detect pre-motor dopaminergic dysfunction in subjects at high risk of future Parkinson’s disease (Nandhagopal et al. , 2008; Iranzo et al. , 2011). However, in recent years it has been proposed that changes in the dopamine system (and motor impairment) may not become manifest until relatively late in the disease course. By contrast, the autonomic nervous system shows early involvement; indeed it has been suggested that the deposits of aberrantly folded α-synuclein associated with Parkinson’s disease may originate in autonomic nerve endings of the gastrointestinal tract and be retrogradely transmitted to the caudal brainstem via the vagus nerve (Braak et al. , 2003) (Fig. 1). Several studies have used imaging to …

Cajal-like interstitial cells as a novel target in detrusor overactivity treatment: true or myth?

IntroductionThe Cajal–like intestitial cells (ICCs) act as a pacemaker and are responsible for generating smooth muscle activity in the gastrointestinal tract (GI). Interstitial cells that resemble ICCs in the GI have been identified in the urinary bladder.Materials and methodsThis review is based on a systemic literature research. The medline/pubmed, scopus, embase, and Web of Science databases were browsed in order to identify original and review articles, as well as editorials relating to cajal–like cells, urinary bladder, detrusor overactivity, overactive bladder, glivec, etc. The controlled vocabulary of the Medical Subject Headings (MeSH) database was used to ensure the sensitivity of the searches. 40 papers met the criteria and were used for this review.ResultsCajal cells lie in close proximity to the muscle cells, autonomic nerve endings, and urothelial cells. There is increasing evidence that ICCs play role in urinary tract dysfunction development (e.g. detrusor overactivity, primary obstructive megaureter, congenital ureteropelvic junction obstruction, etc.). ICCs may be responsible for generating electrical potentials and induction of detrusor muscle contractions. Novel pathomechanisms of detrusor overactivity development have been postulated, as follows: 1) the disturbance of spontaneous contractility caused by altered signal transduction of ICCs between nerves and detrusor muscle cells, and 2). the alteration in signal transduction between urothelium and afferent nerve endings via suburothelial ICCs. The c–kit receptor is not only a detection marker of these cells, but may also play a crucial role in the control of bladder function.ConclusionsCajal cells in urinary bladder suggest that the c–kit receptor may provide a novel target for treating detrusor overactivity. The review presents the current knowledge of ICCs, its role in urinary bladder function, and potential novel therapeutic strategy.

The Lethal Toxin from Australian Funnel-Web Spiders Is Encoded by an Intronless Gene

Australian funnel-web spiders are generally considered the most dangerous spiders in the world, with envenomations from the Sydney funnel-web spider Atrax robustus resulting in at least 14 human fatalities prior to the introduction of an effective anti-venom in 1980. The clinical envenomation syndrome resulting from bites by Australian funnel-web spiders is due to a single 42-residue peptide known as δ-hexatoxin. This peptide delays the inactivation of voltage-gated sodium channels, which results in spontaneous repetitive firing and prolongation of action potentials, thereby causing massive neurotransmitter release from both somatic and autonomic nerve endings. Here we show that δ-hexatoxin from the Australian funnel-web spider Hadronyche versuta is produced from an intronless gene that encodes a prepropeptide that is post-translationally processed to yield the mature toxin. A limited sampling of genes encoding unrelated venom peptides from this spider indicated that they are all intronless. Thus, in distinct contrast to cone snails and scorpions, whose toxin genes contain introns, spiders may have developed a quite different genetic strategy for evolving their venom peptidome.

Impaired Autonomic Regulation of Resistance Arteries in Mice With Low Vascular Endothelial Growth Factor or Upon Vascular Endothelial Growth Factor Trap Delivery

Control of peripheral resistance arteries by autonomic nerves is essential for the regulation of blood flow. The signals responsible for the maintenance of vascular neuroeffector mechanisms in the adult, however, remain largely unknown. Here, we report that VEGF( partial differential/ partial differential) mice with low vascular endothelial growth factor (VEGF) levels suffer defects in the regulation of resistance arteries. These defects are due to dysfunction and structural remodeling of the neuroeffector junction, the equivalent of a synapse between autonomic nerve endings and vascular smooth muscle cells, and to an impaired contractile smooth muscle cell phenotype. Notably, short-term delivery of a VEGF inhibitor to healthy mice also resulted in functional and structural defects of neuroeffector junctions. These findings uncover a novel role for VEGF in the maintenance of arterial neuroeffector function and may help us better understand how VEGF inhibitors cause vascular regulation defects in cancer patients.

Characterization of ouabain‐induced noradrenaline and acetylcholine release from in situ cardiac autonomic nerve endings

Although ouabain modulates autonomic nerve ending function, it is uncertain whether ouabain-induced releasing mechanism differs between in vivo sympathetic and parasympathetic nerve endings. Using cardiac dialysis, we examined how ouabain induces neurotransmitter release from autonomic nerve ending. Dialysis probe was implanted in left ventricle, and dialysate noradrenaline (NA) or acetylcholine (ACh) levels in the anaesthetized cats were measured as indices of neurotransmitter release from post-ganglionic autonomic nerve endings. Locally applied ouabain (100 microm) increased in dialysate NA or ACh levels. The ouabain-induced increases in NA levels remained unaffected by cardiac sympathetic denervation and tetrodotoxin (Na+ channel blocker, TTX), but the ouabain-induced increases in ACh levels were attenuated by TTX. The ouabain-induced increases in NA levels were suppressed by pretreatment with desipramine (NA transport blocker) and augmented by reserpine (vesicle NA transport blocker). In contrast, the ouabain-induced increases in ACh levels remained unaffected by pretreatment with hemicholinium-3 (choline transport blocker) but suppressed by vesamicol (vesicle ACh transport blocker). The ouabain-induced increases in NA levels were suppressed by pretreatment with omega-conotoxin GVIA (N-type Ca2+ channel blocker), verapamil (L-type Ca2+ channel blocker) and TMB-8 (intracellular Ca2+ antagonist). The ouabain-induced increases in ACh levels were suppressed by pretreatment with omega-conotoxin MVIIC (P/Q-type Ca2+ channel blocker), and TMB-8. Ouabain-induced NA release is attributable to the mechanisms of regional exocytosis and/or carrier-mediated outward transport of NA, from stored NA vesicle and/or axoplasma, respectively, while the ouabain-induced ACh release is attributable to the mechanism of exocytosis, which is triggered by regional depolarization. At both sympathetic and parasympathetic nerve endings, the regional exocytosis is because of opening of calcium channels and intracellular calcium mobilization.

Cardiac NGF and GDNF expression during Trypanosoma cruzi infection in rats

In rats, autonomic nerve endings are damaged during Trypanosoma cruzi-induced myocarditis. Gradual recovery occurs after the acute phase. The present work shows the cardiac levels of glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF), and their cellular sources during T. cruzi infection in rats. Atrial and ventricular NGF levels (ELISA) increased significantly at day 20 post inoculation, the time-point of maximal sympathetic denervation. ELISA failed to show significant increase of cardiac GDNF levels. However immunohistochemistry showed a significant increase of anti-GDNF gold particles over atrial granules at day 20. Light microscopy showed stronger NGF immunostaining in atrial cardiomyocytes and several blood capillaries. In situ hybridization showed NGF and GDNF mRNAs in atrial and ventricular myocytes of both infected and uninfected animals. Endothelial cells exhibited NGF mRNA and protein only in infected rats. No evidence of neurotrophic factor expression by the infiltrating mononuclear cells was found. This is the first report on neurotrophic factor expression during T. cruzi infection. Our findings indicate an important role for NGF in the regenerative phenomena subsequent to a myocarditis able to damage sympathetic nerve endings, with preservation of preterminals and nerve trunks. GDNF could have a minor or a more transient participation.

Water Channels and Zymogen Granules in Salivary Glands

Salivary secretion occurs in response to stimulation by neurotransmitters released from autonomic nerve endings. The molecular mechanisms underlying the secretion of water, a main component of saliva, from salivary glands are not known; the plasma membrane is a major barrier to water transport. A 28-kDa integral membrane protein, distributed in highly water-permeable tissues, was identified as a water channel protein, aquaporin (AQP). Thirteen AQPs (AQP0 - AQP12) have been identified in mammals. AQP5 is localized in lipid rafts under unstimulated conditions and translocates to the apical plasma membrane in rat parotid glands upon stimulation by muscarinic agonists. The importance of increases in intracellular calcium concentration [Ca(2+)](i) and the nitric oxide synthase and protein kinase G signaling pathway in the translocation of AQP5 is reviewed in section I. Signals generated by the activation of Ca(2+) mobilizing receptors simultaneously trigger and regulate exocytosis. Zymogen granule exocytosis occurs under the control of essential process, stimulus-secretion coupling, in salivary glands. Ca(2+) signaling is a principal signal in both protein and water secretion from salivary glands induced by cholinergic stimulation. On the other hand, the cyclic adenosine monophosphate (cAMP)/cAMP-dependent protein kinase system has a major role in zymogen granule exocytosis without significant increases in [Ca(2+)](i). In section II, the mechanisms underlying the control of salivary protein secretion and its dysfunction are reviewed.

Phenytoin as a novel anti-vitiligo weapon

Vitiligo is a psychologically devastating clinical conundrum which affects approximately 1% of the general population. The exact cause of the illness is an enigma, but several hypotheses about its pathogenesis are advanced.The autoimmune hypothesis proposes an autoimmune attack against melanocytes. Although anti-melanocyte autoantibodies have been demonstrated in vitiligo, recent research casts doubt on their pathogenic role and instead supports the involvement of cell-mediated autoimmune response in the pathobiology of this disorder, which is characterized by increase of suppressor T-cells and decrease of the helper/suppressor ratio in association with the presence of type-1 cytokine secreting cytotoxic T cells in the vicinity of disappearing melanocytes.The neural hypothesis proposes that increased release of norepinephrine, a melanocytotoxin, from the autonomic nerve endings in the microenvironment of melanocytes injures these cells. Moreover, norepinephrine induces the catecholamine degrading enzyme monoamine oxidase (MAO), which favors the formation of toxic levels of hydrogen peroxide in the vicinity of melanocytes.Another theory suggests that abnormal permeability of melanosome membrane, which normally prevents the diffusion of toxic melanin precursors into the cytoplasm, may cause melanocyte damage.Phenytoin, the widely-used anticonvulsant, has been employed both topically and systemically in the treatment of some dermatological disorders. The drug has been shown to significantly suppress mitogen-induced activation of lymphocytes and cytotoxic T lymphocyte activity and to polarize the immune response toward the type-2 pathway. It also significantly decreases suppressor T cells and increases the helper/suppressor ratio.At high concentrations, the drug inhibits the release of norepinephrine and the activity of MAO. Moreover, phenytoin is suggested to interact with membrane lipids, which may promote stabilization of the membranes.The hydantoin moiety of phenytoin exerts a direct stimulatory action on melanocytes; facial hyperpigmentation is a recognized side effect of orally administered phenytoin.Altogether, the above evidence suggests that phenytoin could be therapeutically effective against vitiligo. As phenytoin stimulates collagen production and inhibits its breakdown, its concomitant use with topical steroids could prevent steroid-induced skin atrophy while potentiating the anti-vitiligo effect of these agents.

Hepatic stellate cells do not derive from the neural crest

Hepatic stellate cells (HSC) have been hypothesised to derive from the neural crest, based on their expression of multiple neural/neuroendocrine features and their contacts with autonomic nerve endings. We studied the emergence of HSC in the liver during embryonic development in a transgenic mouse line expressing yellow fluorescent protein (YFP) in all neural crest cells and their derivatives. Cellular YFP expression in these mice was compared with desmin expression between embryonic day (E) 11.5 and adulthood. YFP was abundantly expressed in neural crest cells delaminating from the neural tube and in all known neural crest-derived structures and cell populations. In particular, YFP expressing cells perfectly mimicked the spatial and temporal pattern of enteric nervous system development from neural crest cells migrating from the postotic region. Cells within the adrenal medulla were also YFP positive. Analysis of the liver showed that desmin-expressing, stellate-shaped, perisinusoidally located HSC were evident from E11.5 onwards. However, no detectable YFP expression was seen in the developing liver or in HSC, from E11.5 until adulthood. These findings suggest HSC do not descend from the neural crest, and therefore may derive from the septum transversum mesenchyme, from endoderm or from the mesothelial liver capsule.

Characterization of a Ca2+-activated nonselective cation channel during dedifferentiation of cultured rat ventricular cardiomyocytes.

Cardiac hypertrophy is associated with electrical activity modifications, including sustained depolarization, that lead to a propensity for arrhythmias. The ionic currents underlying the sustained depolarization are not well defined. Similar modifications were reported on adult rat cardiomyocytes in primary culture undergoing dedifferentiation. Using the single-channel measurements on these cells, we identified the appearance of a Ca2+-activated nonselective cation channel (NSCCa) during the dedifferentiation process. In excised inside-out patches the channel presented a linear I/V relationship with a conductance of 26.5 pS. It was equally selective for Na+ and K+ and impermeable to Cl- and Ca2+ ions. The open probability increased with depolarization and with rise in intracellular calcium concentration. The channel activity was reduced by intracellular ATP and suppressed by flufenamic acid. Channel detection increased after incubation with a purinergic receptor agonist (ATPgS) or a PKC activator (PMA). Furthermore, occurrence of the channel developed during the culture. Absent at one day in vitro (d.i.v.), channel activity was present in 5, 46, 27 and 19% of patches after 4, 7, 14 and 21 d.i.v., respectively. We suggest that the channel may be associated with pro-arrhythmic signaling, in particular during the release of transmitters from autonomic nerve endings in the hypertrophied hearts.

Stimulation of Ca(2+) influx through ATP receptors on rat brain synaptosomes: identification of functional P2X(7) receptor subtypes.

1. ATP receptors of the P2X class have previously been identified on autonomic nerve endings and on a limited population of CNS neurons. 2. In the present study P2X receptors on mammalian cortical synaptosomes have been identified by a variety of functional and biochemical studies. In choline buffer ATP analogues caused concentration/time dependent Ca(2+) influx. Relative to the effects caused by ATP, benzoylbenzoyl ATP (BzATP) was about seven times more active than ATP while 2-me-S-ATP and ATPgammaS were much less active. alpha,beta-me- ATP and beta,gamma-me-ATP were virtually inactive. In sucrose buffer, relative to choline buffer, the activity of BzATP was more than doubled while activity in sodium buffer was reduced. Moreover, the P2X antagonists PPADS or Brilliant Blue G both significantly attenuated influx. These observations suggest the presence of P2X receptors on synaptosomes which subserve Ca(2+) influx. This activity profile of the ATP analogues and the response to blocking agents are characteristic of responses of P2X(7) receptors. 3. Influx was unaffected by the VSCC inhibitors omega-CTx-MVIIC and (-) 202 - 791, indicating that ATP induced Ca(2+) influx occurred primarily through P2X receptors. 4. P2X(7) receptor protein was identified by Western blotting and immunohistochemical staining. Purified preparations were devoid of significant concentrations of GFAP or the microglial marker OX-42 but contained greatly enriched amounts of syntaxin and SNAP 25. 5. The various pharmacological and biochemical studies were all consistent with the presence of functional P2X(7) receptors.