Normal Ganglia Research Articles

AAV Provides an Alternative for Gene Therapy of the Peripheral Sensory Nervous System

Recombinant adeno-associated virus (rAAV) is a promising vector for applications in the central nervous system (CNS). Clinical phase I/II studies have demonstrated safety of rAAV-expressing therapeutic transgenes for a variety of benign (i.e., noncancer) CNS disorders, namely, Canavan disease,1 Parkinson’s disease,2,3,4 and Batten disease.5 Human studies were also recently reported targeting the retina.6,8,9,9 From the early stages of preclinical development, rAAV was appealing because it was found to be neurotrophic and capable of long-term activity without immune interference or toxicity when injected into the CNS.10

Aberrant synthesis of ATP synthase resulting from a novel deletion in mitochondrial DNA in an African patient with progressive external ophthalmoplegia

A young, adult, African male patient presented with progressive proximal muscle weakness, external ophthalmoplegia and ptosis, as well as cardiac conduction abnormalities resembling Kearns-Sayre syndrome (KSS). Magnetic resonance imaging (MRI) of the brain revealed normal basal ganglia but bilateral well-circumscribed lesions in the cerebellar peduncles. Enzyme deficiencies in oxidative phosphorylation (OXPHOS) complexes I, IV and V was measured in muscle tissue. Blue native polyacrylamide gel electrophoresis (BN-PAGE) confirmed decreased protein content and activity of these complexes and revealed the presence of two catalytically active complex V sub-complexes. Upon investigation by molecular genetics, the mitochondrial DNA (mtDNA) copy number was found to be elevated and a novel deletion of 3431bp was found in 80% of muscle mtDNA between positions 7115 and 10546, flanked by a 5bp direct repeat sequence. In addition, it could also be concluded that the absence of mtDNA-encoded ATPase6 and ATPase8 genes in this patient clearly resulted in aberrant synthesis of ATP synthase.

Anatomical changes in the primary visual cortex of the congenitally blind Crx−/− mouse

Mutations in the human cone-rod homeobox ( Crx) gene are associated with retinal dystrophies such as Leber Congenital Amaurosis (LCA), characterized by complete or near complete absence of vision from birth. The photoreceptors of Crx−/− mice lack outer segments, and therefore cannot capture light signals through rods and cones, thus resulting in a lack of normal retinal ganglion cell activity from birth. Using specific antibodies to subsets of neurons and markers of activity, we examined the impact of this absence of sensory input on the development of the primary visual cortex (V1) in early postnatal Crx−/− mice, before wiring of the visual system is complete, and in adulthood. We revealed that Crx−/− mice did not exhibit gross anatomical differences in V1; however, they exhibited significantly fewer calcium-binding protein (parvalbumin and calbindin-D28k) expressing interneurons, as well as reduced nonphosphorylated neurofilament expression in V1. These results reveal that the Crx mutation and lack of light stimulation through the photoreceptor pathway regulate the development and phenotype of different neuronal populations in V1 but not its general morphology. We conclude, therefore, that photoreceptor-mediated visual input during development is crucial for the normal postnatal development and maturation of subsets of cortical neurons.

The therapeutic concept of continuous dopaminergic stimulation (CDS) in the treatment of Parkinson's disease

Continuous dopaminergic stimulation is a therapeutic concept for the management of Parkinson's disease (PD) that proposes that continuous, as opposed to discontinuous or pulsatile, stimulation of striatal dopamine receptors will delay or prevent the onset of levodopa-related motor complications. This hypothesis has arisen from studies of the normal basal ganglia demonstrating that nigral dopaminergic neurons normally fire continuously and striatal dopamine levels are relatively constant. In MPTP monkeys, pulsatile stimulation of striatal dopamine receptors with short-acting agents is associated with the induction of molecular and physiologic changes in basal ganglia neurons and the development of motor complications. These are avoided when dopaminergic therapies are delivered in a more continuous manner. Studies in animal models support this hypothesis, demonstrating that long-acting dopamine agonists are associated with a decreased risk of motor complications in comparison to short-acting formulations of levodopa. Similarly, continuous infusion of dopamine agonists ropinirole and rodigotine reduces dyskinesia associated with intermittent doses of oral formulations of the same drug. The current challenge is to develop a long-acting formulation of levodopa that simulates the pharmacokinetic pattern seen with infusions of levodopa in attempt to provide comparable benefits with an oral levodopa treatment strategy.

The impact of automatic retractors on the esophagus during anterior cervical surgery: an experimental in vivo study in a sheep model

Postoperative dysphagia is a well-recognized complication of the anterior surgical approach to the cervical spine. However, its incidence and etiology remain unknown. The aim of this study was to investigate the impact of automatic retractor use on the esophagus and to describe the related pathological changes that might occur during cervical spine surgery. A single-level cervical discectomy was performed via an anterior approach in 16 skeletally mature female sheep. Continuous retraction was applied with an automatic retractor system during surgery. The sheep model was chosen because of anatomical similarities to the human esophagus. The esophageal tract in every animal was examined using contrast radiographic examination. Eight animals were killed 3 days after the operation (Group 1). The remaining sheep were killed 4 weeks after the operation (Group 2). The esophagi were removed for histopathological study, which was performed using H & E and Masson trichrome staining. The changes in esophageal innervation were examined with nicotinamide adenine dinucleotide diphosphate-diaphorase histochemical staining. Only 1 animal (a Group 1 sheep) demonstrated any postoperative radiographic abnormality. In Group 1 sheep, histopathological study of the esophagi at the treated level revealed edema between the muscular fibers in the outer longitudinal and inner circular layers of the muscularis propria. At some points, obvious signs of vascular congestion, vascular damage, and inflammation were observed. In the Group 2 animals, there was mild-to-moderate fibrosis extending from the outer surface of the esophagus to the longitudinal layers of the muscularis propria in the area to which retraction had been applied. Enzyme-histochemical staining revealed the presence of normal myenteric plexus and ganglion cells, and nitrergic innervation in all parts of the esophagus wall. The results of this study demonstrate that direct pressure induced by the medial retractor blade on the esophagus wall leads to local injury. Postoperative dysphagia in human patients who have undergone anterior cervical spine surgery could be a clinical manifestation of this phenomenon.

Expression of endothelin system in neuroblastic tumors: close association of endothelin-1 and endothelin B receptor expression with differentiation of tumor cells

Although a critical role of the endothelin (ET) system in differentiation of neural crest cells has been reported, implication of the ET system in human neuroblastic tumors has not been fully elucidated. We immunohistochemically examined for localization of ET-1, ET-3, ET-A receptor (ET-A), and ET-B receptor (ET-B) in 24 ganglioneuromas, 8 ganglioneuroblastomas, 37 neuroblastomas, 14 normal sympathetic ganglia, and 10 fetal adrenal glands with regard to neuroblastic cell differentiation. Neuroblasts in fetal adrenal glands expressed ET-B (100%) alone. Immature ganglionic cells in sympathetic ganglia of fetus frequently expressed ET-1 (86%) and ET-B (100%), while ET-A was occasionally detected (28%). Ganglionic cells of mature adult ganglia consistently harbored ET-1 (100%) and, infrequently, ET-3 (21%) or ET-B (29%). Expression of ET-1 and ET-B was closely associated with tumor cell differentiation: the expression frequency of ET-1 or ET-B was 16% or 46% in neuroblastomas; 100% or 88% in ganglioneuroblastomas; and 96% or 92% in ganglioneuromas. In contrast, ET-3 and ET-A showed no association with tumor cell differentiation: the expression frequency of ET-3 or ET-A was 26% or 14% in neuroblastomas; 63% or 13% in ganglioneuroblastomas; and 29% or 21% in ganglioneuromas. In conclusion, ET-1 and ET-B are expressed with differentiation of neuroblastic tumors.

Stercoral perforation in a 17-year old

Stercoral perforation is a rare cause of perforation. This is the first reported case where a partial eating disorder (ED) is the primary causative differential. We present the case of a 17-year-old girl who presented to her local Emergency Department with a 24-h history of left-sided abdominal pain. She subsequently deteriorated and a computed tomography scan of her abdomen showed gross distension of the large bowel with a sigmoid perforation. She underwent total colectomy with end ileostomy. Histology reported stercoral perforation but normal bowel ganglia. While an inpatient she was reviewed by the Psychiatric team who were concerned she was suffering from a partial ED. This case highlights the importance of a multidisciplinary approach in optimally treating patients such as these. Aggressive medical management with involvement of a psychiatric team and dietetics addresses any underlying causative psychiatric issues and helps prevent recurrence.

CEREBELLAR MIGRATION DEFECTS IN AICARDI SYNDROME: AN EXTENSION OF THE NEUROPATHOLOGICAL SPECTRUM

The Aicardi syndrome is characterized by infantile spasms, corpus callosum agenesis, and chorioretinal lacunae and almost exclusively affects females (very rarely, 47, XXY males). The crucial genetic mishap likely occurs in the postzygotic stage, but the variable clinical phenotype among the ∼ 450 known cases has not been explained. No consistent mutations or deletions exist among patients. We encountered a baby girl with early onset infantile spasms. She had left-sided cleft lip/palate, costovertebral defects, scoliosis, callosal agenesis, and microphthalmia. She expired at the age of 3 months of respiratory infection. On autopsy she had thoracic hemivertebrae with rib defects, bilateral microphthalmia, microcornea, posterior colobomata, abnormalities of the retinal pigment epithelium, absence of normal ganglion cells in the retina, gross asymmetry of the brain with cerebral polymicrogyria, total callosal agenesis, cerebral subcortical and subependymal nodular heterotopias, cerebellar nodular heterotopias, and tegmental/basal unilateral brainstem hypoplasia. Cerebellar and retinal migration defects have not been described before in Aicardi syndrome and may have had a bearing on this patient's eventual outcome.

Error processing in normal aging and in basal ganglia disorders

Recently it has been shown that effects of aging and pathologically induced changes of basal ganglia structures may have quite similar effects on cognitive functions mediated by the medial prefrontal cortex. The question appears, if this pattern may be assignable to other cognitive functions that are mediated via the basal ganglia and medial prefrontal brain areas. Error processing is a component of executive functions that also depends on these areas and especially on the anterior cingulate cortex (ACC). Hence we ask, if error processing functions are differentially modulated by normal aging and basal ganglia diseases. Error processing mechanisms in these groups were investigated using a cognitive event-related potential (ERP), the error negativity. Enrolling an extended sample of young and elderly controls, as well as patients with Parkinson's and Huntington's disease, we show that modulations of error processing differ between aging, different basal ganglia diseases. Despite that the examined basal ganglia disorder groups (Parkinson's and Huntington's disease) differ in their age they show similar modulations in error processing, suggesting that aging effects are overridden by pathogenic effects. The study shows that it may be valuable to compare aging not only to different forms of basal ganglia disorders in order to gain knowledge about age- and disease-related mechanisms and the effects of these on cognitive functions. Diseases of the basal ganglia may impact error processing above and beyond the effects of normal aging. Although many aging, Parkinson's disease and Huntington's disease studies on error processing functions have already been published, this study ties together several related observations across all of these groups in one experiment.

Endogenous Polysialylated Neural Cell Adhesion Molecule Enhances the Survival of Retinal Ganglion Cells

During development, all retinal cells express polysialylated neural cell adhesion molecule (PSA-NCAM). PSA is localized only on glia in the adult retina, but as Müller glial processes ensheathe most retinal neurons, PSA remains in the extracellular environment of adult neurons. The authors sought to investigate the influence of endogenous PSA on the survival of neonatal as well as adult normal and injured retinal ganglion cells (RGCs). Endogenous retinal PSA was selectively degraded by application of endoneuraminidase. PSA presence and removal were confirmed by immunohistochemistry and levels were assessed by Western Blot analysis. Neonatal RGC survival after PSA removal was assessed in vitro in RGCs immunopanned from rat pups. Adult RGC survival was assessed in vivo in mice by investigating RGC densities after removal of PSA in normal retinas and after optic nerve transection. Virtually all neonatal RGCs express PSA-NCAM and survive well in vitro; however, removal of PSA resulted in a 42% loss of these cells 3 days after the treatment. Similarly, removal of PSA in the adult retina in vivo induced a loss of 25% of RGCs at 14 days, and significantly reduced RGC densities after optic nerve transection by an additional 27% (relative to injured retinas with a vehicle injection) at 7 days. Together, these findings demonstrate that endogenous PSA supports the survival of neonatal as well as injured and normal adult RGCs and provide the first functional evidence of a role for PSA in the adult retina.

Evidence for a Role of Connexin 43 in Trigeminal Pain Using RNA Interference In Vivo

The importance of glial cells in the generation and maintenance of neuropathic pain is becoming widely accepted. We examined the role of glial-specific gap junctions in nociception in the rat trigeminal ganglion in nerve-injured and -uninjured states. The connexin 43 (Cx43) gap-junction subunit was found to be confined to the satellite glial cells (SGCs) that tightly envelop primary sensory neurons in the trigeminal ganglion and we therefore used Cx43 RNA interference (RNAi) to alter gap-junction function in SGCs. Using behavioral evaluation, together with immunocytochemical and Western blot monitoring, we show that Cx43 increased in the trigeminal ganglion in rats with a chronic constriction injury (CCI) of the infraorbital nerve. Reducing Cx43 expression using RNAi in CCI rats reduced painlike behavior, whereas in non-CCI rats, reducing Cx43 expression increased painlike behavior. The degree of painlike behavior in CCI rats and intact, Cx43-silenced rats was similar. Our results support previous suggestions that increases in glial gap junctions after nerve injury increases nociceptive behavior but paradoxically the reduction of gap junctions in normal ganglia also increases nociceptive behavior, possibly a reflection of the multiple functions performed by glia.

S240 – Human Cochlear Implant Histopathology

Objectives 1) To exam the histopathology of multichannel cochlear implant temporal bones. 2) To evaluate the relationship of residual spiral ganglion cell counts to clinical hearing performance. Methods 8 temporal bones from 4 cochlear implant patients were examined histologically. Paired comparisons were made between implanted and nonimplanted temporal bones. Clinical performance data was obtained from patient charts. Results There were varying amounts of inflammation (fibrosis and ossification) in the basal turn of the cochlear in all implanted temporal bones. Trauma to the facial nerve at facial recess site was noticed in 1 case. Compared with nonimplanted ears, 2 implanted bones with less than 10-year duration of implantation had no significant changes of spiral ganglion cell population. One case with prolong implant duration (15 years) showed about 36% decrease of spiral ganglion cells at the implanted site. The case with best speech recognition (89% with CID sentence) had the highest residual spiral ganglion cells (30% of normal spiral ganglion cell population). 2 cases with poor clinical performance (< 10% with CID sentence) had the residual spiral ganglion cells at 11% and 22%. The case with moderate clinical performance (30% with CID sentence) had 14% of normal spiral ganglion cell population. Surviving dendrites varied from 5% to 30% among 4 cases with no relationship to clinical performance. Conclusions Our findings suggest prolonged implantation may affect spiral ganglion cell population. There is no reverse relationship between residual spiral ganglion cells in implanted temporal bones to clinical speech performance observed from our limited cases.

Comparative study of the distribution of the α‐subunits of voltage‐gated sodium channels in normal and axotomized rat dorsal root ganglion neurons

We compared the distribution of the alpha-subunit mRNAs of voltage-gated sodium channels Nav1.1-1.3 and Nav1.6-1.9 and a related channel, Nax, in histochemically identified neuronal subpopulations of the rat dorsal root ganglia (DRG). In the naïve DRG, the expression of Nav1.1 and Nav1.6 was restricted to A-fiber neurons, and they were preferentially expressed by TrkC neurons, suggesting that proprioceptive neurons possess these channels. Nav1.7, -1.8, and -1.9 mRNAs were more abundant in C-fiber neurons compared with A-fiber ones. Nax was evenly expressed in both populations. Although Nav1.8 and -1.9 were preferentially expressed by TrkA neurons, other alpha-subunits were expressed independently of TrkA expression. Actually, all IB4(+) neurons expressed both Nav1.8 and -1.9, and relatively limited subpopulations of IB4(+) neurons (3% and 12%, respectively) expressed Nav1.1 and/or Nav1.6. These findings provide useful information in interpreting the electrophysiological characteristics of some neuronal subpopulations of naïve DRG. After L5 spinal nerve ligation, Nav1.3 mRNA was up-regulated mainly in A-fiber neurons in the ipsilateral L5 DRG. Although previous studies demonstrated that nerve growth factor (NGF) and glial cell-derived neurotrophic factor (GDNF) reversed this up-regulation, the Nav1.3 induction was independent of either TrkA or GFRalpha1 expression, suggesting that the induction of Nav1.3 may be one of the common responses of axotomized DRG neurons without a direct relationship to NGF/GDNF supply.

Effects of carbamazepine on spinal cord ischemia

Prophylactic treatment with carbamazepine has been shown to reduce the cerebral damage and neurologic deficit in ischemic conditions. A randomized controlled study based on a rabbit model was designed to study the effect of carbamazepine on a spinal cord ischemic reperfusion injury. Thirty New Zealand rabbits were randomly assigned to 1 of the 2 groups (n = 15 per group): group I (control group) and group II (carbamazepine group). Spinal cord ischemia was induced by infrarenal aortic crossclamp for 25 minutes in both groups. Functional evaluation with the Tarlov score during a 2-day observation period and histopathologic assessment of the lumbar spinal cord were performed. Changes in spinal cord morphology were observed with hematoxylin-eosin staining and electron microscopy. Gray matter damage was assessed on the basis of the number of normal neurons in the ventral horn. Diffuse destruction of gray matter with moderate to severe vacuolization and essentially no normal ganglion cells was observed in the spinal cord of rabbits in the control group, whereas specimens of rabbits assigned to the carbamazepine group showed ganglion cells with normal nuclei and cytoplasm (P < .0001). Neurologic impairment was significantly attenuated in the carbamazepine group compared with the Tarlov scores of the control group (P < .0001 at day 2). Carbamazepine may protect the spinal cord from ischemic reperfusion injury that is associated with ameliorated neurologic and histopathologic results.

Timing the initiation of treatment in Parkinson’s disease

It is estimated that the dopaminergic neuronal loss that causes the characteristic motor features of Parkinson’s disease (PD) has reached 50–70% by diagnosis and it is progressive. Clinical, imaging and...

PDGF- and Insulin/IGF-1–Specific Distinct Modes of Class IAPI 3-Kinase Activation in Normal Rat Retinas and RGC-5 Retinal Ganglion Cells

To compare PDGF- and insulin/IGF-1-induced class I(A) PI 3-kinase/Akt survival signaling in normal retinas and retinal ganglion cells (RGCs). Normal rat retinas and RGC-5 cells were used for (1) immunohistochemical and immunoblot studies to detect PDGF receptor (PDGFR) subtypes and (2) immunoprecipitation, immunoblot, and in vitro lipid kinase assays to determine basal and PDGF-induced class I(A) PI 3-kinase/Akt survival signaling, in comparison with insulin or IGF-1 responses. Furthermore, RGC-5 cells were exposed to broad-spectrum (LY294002) or p110 isoform-selective (PI-103) PI 3-kinase inhibitors (versus Akt inhibitor) to assess the consequent effects on Akt phosphorylation, caspase-3/PARP cleavage, apoptotic phenotype, and cell viability, as a function of serum trophic factors. PDGFR-alpha and -beta immunoreactivity was observed in rat retinal Müller cells and in the RGC layer and blood vessels, respectively. In addition, PDGFR-alpha and -beta protein expression was observed in RGC-5 cells. Both retinas and RGC-5 cells exhibited a similar pattern of subunit-specific basal class I(A) PI 3-kinase activity, which was stimulated in a temporal and signal-specific manner by PDGF and insulin/IGF-1. Furthermore, RGC-5 cells showed PDGFR-alpha/beta tyrosine phosphorylation that induced the p85alpha regulatory subunit to activate p110alpha/beta-associated class I(A) PI 3-kinase, which in turn enhanced Akt phosphorylation. Exposure of serum-deprived RGC-5 cells to PI 3-kinase or Akt inhibitors increased susceptibility to apoptotic phenotype as revealed by caspase-3 and PARP cleavage. The present findings provide direct evidence of two distinct modes of retinal class I(A) PI 3-kinase activation that occurs in response to PDGF receptor and insulin/IGF-1 receptor stimulation. PDGF-induced PI 3-kinase/PIP3/Akt axis may provide new therapeutic approaches to ameliorate cell death in diabetic retinopathy and other retinal neurodegenerations.

Expression of gLTP in Sympathetic Ganglia from Stress-hypertensive Rats: Molecular Evidence

We previously reported behavioral and electrophysiological evidence indicating that superior cervical ganglia (SCG) from rats that developed hypertension as a result of chronic psychosocial stress expressed ganglionic long-term potentiation (gLTP) in vivo. In the present study, we present additional supportive evidence by measuring changes in protein levels of essential signaling molecules in ganglia from chronically stressed rats. We compared protein levels of essential, LTP-related signaling molecules in ganglia isolated from chronic stress-hypertensive rats, known to have expressed gLTP, with those of the same molecules in normal ganglia 1h after eliciting gLTP by high frequency stimulation (HFS) in vitro. Immunoblot analysis showed a significant increase in the levels of phosphorylated CaMKII, total CaMKII, nitric oxide synthase (NOS-1), and calmodulin in SCG from both chronically stressed rats and from normal rat ganglia in which gLTP was expressed by HFS in vitro. Additionally, there was a parallel reduction in calcineurin protein levels in ganglia from both groups. The present results confirm that ganglia from stressed rats have expressed gLTP in vivo and that synaptic plasticity in sympathetic ganglia may involve a molecular cascade largely similar to that of LTP in the hippocampal CA1 region.

Expression of Pancreatitis Associated Proteins in Urothelium and Urinary Afferent Neurons Following Cyclophosphamide Induced Cystitis

We examined the expression profile of the members of the pancreatitis associated proteins/regenerating gene family in the bladder and in the primary afferent neurons of dorsal root ganglia using an animal model of cystitis. We examined the expression of pancreatitis-associated protein-I and pancreatitis-associated protein-III in the bladder and the dorsal root ganglia of female rats 4 hours, 48 hours or 10 days after cyclophosphamide (Sigma) injection using immunohistochemistry and reverse transcriptase-polymerase chain reaction. No pancreatitis-associated protein-III immunoreactivity was identified in control bladders but prominent expression was observed in the urothelium of animals with chronic cystitis. Cells expressing pancreatitis-associated protein-I were seen in the dorsal root ganglia but not in the bladder. In normal dorsal root ganglia pancreatitis-associated protein-I was expressed in a minor population of small diameter neurons (2.4%) that were also positive for isolectin-B4. However, by 10 days following the onset of cystitis the number of pancreatitis-associated protein-I positive neurons was increased (7.6%) and pancreatitis-associated protein-I immunoreactivity was further observed in a slightly larger group of neurons and tyrosine kinase A positive small neurons. The current results suggest that pancreatitis-associated protein-III is associated with bladder inflammation and they implicate pancreatitis-associated protein-I in the abnormal sensation in cystitis.

MRI findings of the normal and diseased trigeminal nerve ganglion and branches: a pictorial review

The trigeminal nerve is the largest cranial nerve with both sensory and motor functions. Magnetic resonance imaging (MRI) is an excellent technique to evaluate the trigeminal nerve. We report the MRI findings and illustrate the normal and diseased trigeminal nerve at Meckel's cave, the cavernous sinus, the skull base foramina, the pterygopalatine fossa, and the peripheral branches.

Clinicopathological characteristics of neck ganglioneuroma

Ganglioneuroma of the head and neck is rare. The clinical, histopathological and immunohistochemical characteristics of 6 cases of cervical ganglioneuroma were analyzed. The average age of the patients in this study was 32.8 years (6-62 years). The tumors grew slowly and the patients were asymptomatic. Grossly, they were well encapsulated. Under the microscope, the tumors consisted of primarily Schwann cells, tangled masses of neurites in bundles, and variably-distributed large ganglion cells. The ganglion cells showed positive immunohistochemical reactivity to neuronspecific enolase, neurofilament, chromogranin A, and synaptophysin but negative for S100, the same as in the controlled normal sublingual ganglion cells. All the tumors were treated with surgical excision. There was no recurrence and metastasis during a follow-up time of 3-5 years.