Facial Nucleus Research Articles

Diazoxide blocks or reduces microgliosis when applied prior or subsequent to motor neuron injury in mice

Diazoxide (DZX), an anti-hypertonic and anti-hypoglycemic drug, was shown to have anti-inflammatory effects in several injured cell types outside the central nervous system. In the brain, the neuroprotective potential of DZX is well described, however, its anticipated anti-inflammatory effect after acute injury has not been systematically analyzed. To disclose the anti-inflammatory effect of DZX in the central nervous system, an injury was induced in the hypoglossal and facial nuclei and in the oculomotor nucleus by unilateral axonal transection and unilateral target deprivation (enucleation), respectively. On the fourth day after surgery, microglial analysis was performed on tissue in which microglia were DAB-labeled and motoneurons were labeled with immunofluorescence. DZX treatment was given either prophylactically, starting 7 days prior to the injury and continuing until the animals were sacrificed, or postoperatively only, with daily intraperitoneal injections (1.25 mg/kg; in 10 mg/ml dimethyl sulfoxide in distilled water). Prophylactically + postoperatively applied DZX completely eliminated the microglial reaction in each motor nuclei. If DZX was applied only postoperatively, some microglial activation could be detected, but its magnitude was still significantly smaller than the non-DZX-treated controls. The effect of DZX could also be demonstrated through an extended period, as tested in the hypoglossal nucleus on day 7 after the operation. Neuronal counts, determined at day 4 after the operation in the hypoglossal nucleus, demonstrated no loss of motor neurons, however, an increased Feret's diameter of mitochondria could be measured, suggesting increased oxidative stress in the injured cells. The increase of mitochondrial Feret's diameter could also be prevented with DZX treatment.

CD4+ T cell expression of the IL-10 receptor is necessary for facial motoneuron survival after axotomy

BackgroundAfter peripheral nerve transection, facial motoneuron (FMN) survival depends on an intact CD4+ T cell population and a central source of interleukin-10 (IL-10). However, it has not been determined previously whether CD4+ T cells participate in the central neuroprotective IL-10 cascade after facial nerve axotomy (FNA).MethodsImmunohistochemical labeling of CD4+ T cells, pontine vasculature, and central microglia was used to determine whether CD4+ T cells cross the blood-brain barrier and enter the facial motor nucleus (FMNuc) after FNA. The importance of IL-10 signaling in CD4+ T cells was assessed by performing adoptive transfer of IL-10 receptor beta (IL-10RB)-deficient CD4+ T cells into immunodeficient mice prior to injury. Histology and qPCR were utilized to determine the impact of IL-10RB-deficient T cells on FMN survival and central gene expression after FNA. Flow cytometry was used to determine whether IL-10 signaling in T cells was necessary for their differentiation into neuroprotective subsets.ResultsCD4+ T cells were capable of crossing the blood-brain barrier and associating with reactive microglial nodules in the axotomized FMNuc. Full induction of central IL-10R gene expression after FNA was dependent on CD4+ T cells, regardless of their own IL-10R signaling capability. Surprisingly, CD4+ T cells lacking IL-10RB were incapable of mediating neuroprotection after axotomy and promoted increased central expression of genes associated with microglial activation, antigen presentation, T cell co-stimulation, and complement deposition. There was reduced differentiation of IL-10RB-deficient CD4+ T cells into regulatory CD4+ T cells in vitro.ConclusionsThese findings support the interdependence of IL-10- and CD4+ T cell-mediated mechanisms of neuroprotection after axotomy. CD4+ T cells may potentiate central responsiveness to IL-10, while IL-10 signaling within CD4+ T cells is necessary for their ability to rescue axotomized motoneuron survival. We propose that loss of IL-10 signaling in CD4+ T cells promotes non-neuroprotective autoimmunity after FNA.

MORPHOLOGY AND FUNCTION OF THE AUTONOMOUS NERVOUS SYSTEM

The autonomic nervous system consists of the sympathetic and parasympathetic divisions. The central part is represented by supra-segmental and segmental centres. Parasympathetic segmental centres in the brain are accessory nucleus of the oculomotor nerves, superior salivary nucleus of the facial nerve, inferior salivary nucleus of the glossopharyngeal nerve and dorsal nucleus of the vagus nerve. In the spinal cord, these are the intermediate lateral nuclei. Sympathetic segmental centres in the brain are absent, and in the spinal cord, intermediate-lateral nuclei are located in the lateral horns in the eighth cervical, all thoracic and 1-2 lumbar spinal segments. The peripheral part of the autonomic nervous system is represented by pre-nodal and post-nodal branches, paravertebral, prevertebral and terminal nodes and plexuses. The intramural part of the autonomic nervous system lies in the larger part of a wide and narrow-loop net and represented with a large number of nerve cells different by their shapes and sizes and clustered as intramural nodes, or individual nerve cells included along the net loops. The autonomic plexuses of the abdominal cavity are topographically divided into celiac, superior and inferior mesenteric, abdominal aortic, mesenteric, superior and inferior hypogastric region.

Sex‐ and Time‐Dependent Differences in Glutamate Receptor Subunit (GLUN1) in the Rat Rostral Ventrolateral Medulla

The cardioprotective effects of ovarian hormones are evident by the fact that women have a lower risk of cardiovascular disease compared to men, at least until menopause, at which time the relative risks reverse. Several studies have also demonstrated the importance of ovarian hormones in the regulation of sympathetic outflow, potentially via actions in the rostral ventrolateral medulla (RVLM), a brain region critical to sympathetic outflow and blood pressure. However, the time course and mechanisms of sex‐dependent changes in excitation of RVLM neurons have not been fully characterized. Therefore, the purpose of this study was to compare glutamate receptor (GluN1) subunit expression in the RVLM of male and female rats, focusing specifically on ages of development in female rats which precede the onset of the estrous cycle (4 wks old) versus when the estrous cycle is fully established (8 wks old). We hypothesized that eight week‐old female rats would exhibit reduced GluN1 compared to four week‐old females. In contrast, we also predicted that eight week‐old male rats would have higher GluN1 compared to four week‐old males. To test our hypothesis, we obtained brain punches from RVLM subregions of male and female Sprague‐Dawley rats (n=3 ea; 4 and 8 wks old) based on their anatomical location relative to the facial nucleus (FN). Punches were processed for western blotting of GluN1 subunit in four subregions: FN‐480; FN‐240; FN+240 and FN+480. At FN‐480, GluN1 was lower in eight week‐compared to four week‐old female rats (0.67 ± 0.11 vs. 1.11 ± 0.08 protein/GAPDH, respectively). In contrast, at FN‐480, eight week‐old male rats exhibited increased NR1 expression compared to four week old males (1.05 ± 0.06 vs. 0.65 ± 0.10; protein/GAPDH, respectively p<0.05). These results suggest that lower levels of ovarian hormones in prepubescent females may increase the excitability of RVLM neurons and lead to greater sympathoexcitation, which then decrease with the onset of the reproductive cycle. The absence of ovarian hormones in males (perhaps coupled with a lack of regular exercise) may exacerbate RVLM excitability, leading to greater sympathoexcitation in sedentary males. Collectively, these data suggest that ovarian hormones exert cardioprotective effects via actions in the RVLM, reducing sympathetic outflow and contributing to a lower incidence of cardiovascular disease in women of reproductive age.Support or Funding InformationHL096787‐08; AHA25810010

A New Mouse Strain to Study Motoneurons in Pompe Disease

Pompe disease is a lysosomal storage disorder resulting from a deficiency in the lysosomal enzyme acid α‐glucosidase (GAA). The most severe form of the disease is associated with complete or near complete absence of functional GAA which results in cardiorespiratory failure and death within the first year of life. The only approved treatment for Pompe disease is human recombinant GAA (enzyme replacement therapy (ERT)) which improves overall survival rates but the recombinant protein does not reach the central nervous system. Respiratory impairment in Pompe disease has historically been attributed to muscle weakness but an accumulation of studies of Pompe patients and animal models indicate that motoneuron pathology is functionally relevant. In an effort to better understand the role of motoneurons in Pompe disease, we used breeding strategies to create a new mouse model with motoneuron histopathology features seen in Pompe disease while also allowing easy visualization of motoneurons via a fluorescent tag. Chat‐cre mice (Jackson) were crossed with Gaa−/− mice (Taconic). The resulting heterozygous strain were crossed with eYFP+/+ mice (courtesy Dr. Brian Harfe). Further breeding resulted in homozygous mice which were Chat‐cre+/+/Gaa−/−/eYFP+/+. These mice were then crossed to a Flox‐Gaa+/+ (Taconic) to create a heterozygous strain (Chat‐cre+/−/Gaa+/−/eYFP+/−/Flox‐Gaa+/−). Further inbreeding produced mice that were Chat‐cre+/+/Gaa+/−/eYFP+/+/Flox‐Gaa+/+. The intent of this final cross was to produce mice in which alpha motoneurons express eYFP but do not express GAA. Histological evaluation of the brainstem and spinal cord of these mice showed robust eYFP expression in the brainstem including the hypoglossal motor nucleus, nucleus ambiguous, and facial motor nucleus. In the cervical spinal cord, alpha motoneurons throughout the ventral horn also had robust fluorescence. This included putative phrenic motoneurons as determined based on location and size. The robust eYFP expression was restricted to motoneurons which indicates that motoneurons are Gaa−/− (based on the breeding scheme). Ongoing studies are evaluating GAA protein expression in peripheral tissues, glycogen accumulation in motoneurons, and also brainstem and spinal cord motoneuron loss over the lifespan. In addition, the eYFP signal in motoneurons will permit rigorous evaluation of motoneuron transduction efficacy in gene therapy experiments.Support or Funding Information2R01HD052682‐11A1 (DDF/BJB)

Prevention of denervated muscle atrophy with accelerated nerve-regeneration by babysitter procedure in rat facial nerve paralysis model.

The "babysitter" procedure is a reconstruction technique for facial nerve complete paralysis and uses the movement source from the healthy facial nerve with a cross-nerve graft. First, an end-to-side neurorrhaphy is performed between the affected facial nerve trunk and hypoglossal nerve for continuously delivering stimuli to the mimetic muscles for preventing the atrophy of mimetic muscles. Despite favorable clinical results, histological and physiological mechanisms remain unknown. This study attempted to establish a model for the "babysitter" procedure and find its efficacy in rats with facial nerve complete paralysis. A total of 16 Lewis rats were used and divided into 2 groups; cross nerve graft (n = 8) and babysitter groups (n = 8). The facial nerve trunk was transected in both groups. Babysitter group underwent a two-stage procedure. Cross nerve graft group underwent only the transfer of nerve graft from the healthy side to affected side. The animals were assessed physiologically by compound muscle action potential (CMAP), and the regenerated nerve tissues were evaluated histopathologically at 13 weeks after surgery. Facial nucleus stained with retrograde tracers proved the re-innervation of affected facial muscle by the babysitter procedure. In CMAP, the amplitude of babysitter group was significantly higher than that of the cross-facial nerve graft group (p < .05). Histological examination found a significant difference in myelin g-ratio between two groups (p < .05). This study investigated the "babysitter" procedure for rat facial nerve palsy. Babysitter procedure shortened the denervation period without mimic muscle atrophy.

Interference with SRF expression in skeletal muscles reduces peripheral nerve regeneration in mice

Traumatic injury of peripheral nerves typically also damages nerve surrounding tissue including muscles. Hence, molecular and cellular interactions of neighboring damaged tissues might be decisive for successful axonal regeneration of injured nerves. So far, the contribution of muscles and muscle-derived molecules to peripheral nerve regeneration has only poorly been studied. Herein, we conditionally ablated SRF (serum response factor), an important myofiber transcription factor, in skeletal muscles of mice. Subsequently, the impact of this myofiber-restricted SRF deletion on peripheral nerve regeneration, i.e. facial nerve injury was analyzed. Quantification of facial nerve regeneration by retrograde tracer transport, inspection of neuromuscular junctions (NMJs) and recovery of whisker movement revealed reduced axonal regeneration upon muscle specific Srf deletion. In contrast, responses in brainstem facial motor neuron cell bodies such as regeneration-associated gene (RAG) induction of Atf3, synaptic stripping and neuroinflammation were not overly affected by SRF deficiency. Mechanistically, SRF in myofibers appears to stimulate nerve regeneration through regulation of muscular satellite cell (SC) proliferation. In summary, our data suggest a role of muscle cells and SRF expression within muscles for regeneration of injured peripheral nerves.

Sources and lesion-induced changes of VEGF expression in brainstem motoneurons.

Motoneurons of the oculomotor system show lesser vulnerability to neurodegeneration compared to other cranial motoneurons, as seen in amyotrophic lateral sclerosis (ALS). The overexpression of vascular endothelial growth factor (VEGF) is involved in motoneuronal protection. As previously shown, motoneurons innervating extraocular muscles present a higher amount of VEGF and its receptor Flk-1 compared to facial or hypoglossal motoneurons. Therefore, we aimed to study the possible sources of VEGF to brainstem motoneurons, such as glial cells and target muscles. We also studied the regulation of VEGF in response to axotomy in ocular, facial, and hypoglossal motor nuclei. Basal VEGF expression in astrocytes and microglial cells of the cranial motor nuclei was low. Although the presence of VEGF in the different target muscles for brainstem motoneurons was similar, the presynaptic element of the ocular neuromuscular junction showed higher amounts of Flk-1, which could result in greater efficiency in the capture of the factor by oculomotor neurons. Seven days after axotomy, a clear glial reaction was observed in all the brainstem nuclei, but the levels of the neurotrophic factor remained low in glial cells. Only the injured motoneurons of the oculomotor system showed an increase in VEGF and Flk-1, but such an increase was not detected in axotomized facial or hypoglossal motoneurons. Taken together, our findings suggest that the ocular motoneurons themselves upregulate VEGF expression in response to lesion. In conclusion, the low VEGF expression observed in glial cells suggests that these cells are not the main source of VEGF for brainstem motoneurons. Therefore, the higher VEGF expression observed in motoneurons innervating extraocular muscles is likely due either to the fact that this factor is more avidly taken up from the target muscles, in basal conditions, or is produced by these motoneurons themselves, and acts in an autocrine manner after axotomy.

Generation of a ChATCre mouse line without the early onset hearing loss typical of the C57BL/6J strain

The development of knockin mice with Cre recombinase expressed under the control of the promoter for choline acetyltransferase (ChAT) has allowed experimental manipulation of cholinergic circuits. However, currently available ChATCre mouse lines are on the C57BL/6J strain background, which shows early onset age-related hearing loss attributed to the Cdh23753A mutation (a.k.a., the ahl mutation). To develop ChATCre mice without accelerated hearing loss, we backcrossed ChATIRES-Cre mice with CBA/CaJ mice that have normal hearing. We used genotyping to obtain mice homozygous for ChATIRES-Cre and the wild-type allele at the Cdh23 locus (ChATCre,Cdh23WT). In the new line, auditory brainstem response thresholds were ∼20 dB lower than those in 9 month old ChATIRES-Cre mice at all frequencies tested (4–31.5 kHz). These thresholds were stable throughout the period of testing (3–12 months of age).We then bred ChATCre,Cdh23WT animals with Ai14 reporter mice to confirm the expression pattern of ChATCre. In these mice, tdTomato-labeled cells were observed in all brainstem regions known to contain cholinergic cells. We then stained the tissue with a neuron-specific marker, NeuN, to determine whether Cre expression was limited to neurons. Across several brainstem nuclei (pontomesencephalic tegmentum, motor trigeminal and facial nuclei), 100% of the tdTomato-labeled cells were double-labeled with anti-NeuN (n = 1896 cells), indicating Cre-recombinase was limited to neurons. Almost all of these cells (1867/1896 = 98.5%) also stained with antibodies against ChAT, indicating that reporter label was expressed almost exclusively in cholinergic neurons. Finally, an average 88.7% of the ChAT+ cells in these nuclei were labeled with tdTomato, indicating that the Cre is expressed in a large proportion of the cholinergic cells in these nuclei.We conclude that the backcrossed ChATCre,Cdh23WT mouse line has normal hearing and expresses Cre recombinase almost exclusively in cholinergic neurons. This ChATCre,Cdh23WT mouse line may provide an opportunity to manipulate cholinergic circuits without the confound of accelerated hearing loss associated with the C57BL/6J background. Furthermore, comparison with lines that do show early hearing loss may provide insight into possible cholinergic roles in age-related hearing loss.

MiR-206 Reduces the Severity of Motor Neuron Degeneration in the Facial Nuclei of the Brainstem in a Mouse Model of SMA.

Spinal muscular atrophy (SMA) is a severe neuromuscular disease affecting infants caused by alterations of the survival motor neuron gene, which results in progressive degeneration of motor neurons (MNs). Although an effective treatment for SMA patients has been recently developed, the molecular pathway involved in selective MN degeneration has not been yet elucidated. In particular, miR-206 has been demonstrated to play a relevant role in the regeneration of neuromuscular junction in several MN diseases, and particularly it is upregulated in the quadriceps, tibialis anterior, spinal cord, and serum of SMA mice. In the present paper, we demonstrated that miR-206 was transiently upregulated also in the brainstem of the mouse model of SMA, SMAΔ7, in the early phase of the disease paralleling MN degeneration and was down-regulated in the late symptomatic phase. To prevent this downregulation, we intracerebroventricularly injected miR-206 in SMA pups, demonstrating that miR-206 reduced the severity of SMA pathology, slowing down disease progression, increasing survival rate, and improving behavioral performance of mice. Interestingly, exogenous miRNA-206-induced upregulation caused a reduction of the predicted target sodium calcium exchanger isoform 2, NCX2, one of the mainregulators of intracellular [Ca2+] and [Na+]. Therefore, we hypothesized that miR-206 might exert part of its neuroprotective effect modulating NCX2 expression in SMA disease.

Active expiratory oscillator regulates nasofacial and oral motor activities in rats.

What is the central question of this study? Does the parafacial respiratory group (pFRG), which mediates active expiration, recruit nasofacial and oral motoneurons to coordinate motor activities that engage muscles controlling airways in rats during active expiration. What is the main finding and its importance? Hypercapnia/acidosis or pFRG activation evoked active expiration and stimulated the motoneurons and nerves responsible for the control of nasofacial and oral airways patency simultaneously. Bilateral pFRG inhibition abolished active expiration and the simultaneous nasofacial and oral motor activities induced by hypercapnia/acidosis. The pFRG is more than a rhythmic oscillator for expiratory pump muscles: it also coordinates nasofacial and oral motor commands that engage muscles controlling airways. Active expiration is mediated by an expiratory oscillator located in the parafacial respiratory group (pFRG). Active expiration requires more than contracting expiratory muscles as multiple cranial nerves are recruited to stabilize the naso- and oropharyngeal airways. We tested the hypothesis that activation of the pFRG recruits facial and trigeminal motoneurons to coordinate nasofacial and oral motor activities that engage muscles controlling airways in rats during active expiration. Using a combination of electrophysiological and pharmacological approaches, we identified brainstem circuits that phase-lock active expiration, nasofacial and oral motor outputs in an in situ preparation of rat. We found that either high chemical drive (hypercapnia/acidosis) or unilateral excitation (glutamate microinjection) of the pFRG evoked active expiration and stimulated motoneurons (facial and trigeminal) and motor nerves responsible for the control of nasofacial (buccal and zygomatic branches of the facial nerve) and oral (mylohyoid nerve) motor outputs simultaneously. Bilateral pharmacological inhibition (GABAergic and glycinergic receptor activation) of the pFRG abolished active expiration and the simultaneous nasofacial and oral motor activities induced by hypercapnia/acidosis. We conclude that the pFRG provides the excitatory drive to phase-lock rhythmic nasofacial and oral motor circuits during active expiration in rats. Therefore, the pFRG is more than a rhythmic oscillator for expiratory pump muscles: it also coordinates nasofacial and oral motor commands that engage muscles controlling airways in rats during active expiration.

A case of refractory generalized atonic seizure and hemifacial spasm with the possible causative pontocerebellar lesion

The patient was a 35-year-old woman. At the age of 1, she had undergone resection and radiation therapy for neoplastic lesions in the pons. She had a history of gelastic seizures when she was in elementary school, and brief lapses of the neck and truncal muscular tone and convulsions on the left face occurred at the age of 23. After a generalized sharp wave in the ictal electroencephalogram and electromyogram recording, left orbicularis oris muscle contraction was observed followed by sudden cervical extensor atonia. Seizure propagation was noted in the cerebral cortex, left facial nerve nucleus, and brainstem reticular formation. In a simultaneous electroencephalography with functional MRI, the blood oxygen level-dependent effect related to generalized sharp waves was observed in the vicinity of brainstem lesions in addition to a decrease in bilateral frontal and parietal lobes signals, as detected in generalized seizures. These findings suggest that the lesion could be a part of the epilepsy network. Although most epileptic seizures are derived from the cerebral cortex, it is important to note that brainstem lesions are involved in seizures in the patient presented in this study.

Etv1 Controls the Establishment of Non-overlapping Motor Innervation of Neighboring Facial Muscles during Development

SUMMARYThe somatotopic motor-neuron projections onto their cognate target muscles are essential for coordinated movement, but how that occurs for facial motor circuits, which have critical roles in respiratory and interactive behaviors, is poorly understood. We report extensive molecular heterogeneity in developing facial motor neurons in the mouse and identify markers of subnuclei and the motor pools innervating specific facial muscles. Facial subnuclei differentiate during migration to the ventral hindbrain, where neurons with progressively later birth dates—and evolutionarily more recent functions—settle in more-lateral positions. One subpopulation marker, ETV1, determines both positional and target muscle identity for neurons of the dorsolateral (DL) subnucleus. In Etv1 mutants, many markers of DL differentiation are lost, and individual motor pools project indifferently to their own and neighboring muscle targets. The resulting aberrant activation patterns are reminiscent of the facial synkinesis observed in humans after facial nerve injury.

Sensorimotor integration in the whisker somatosensory brain stem trigeminal loop.

The rodent's vibrissal system is a useful model system for studying sensorimotor integration in perception. This integration determines the way in which sensory information is acquired by sensory organs and the motor commands that control them. The initial instance of sensorimotor integration in the whisker somatosensory system is implemented in the brain stem loop and may be essential to the way rodents explore and sense their environment. To examine the nature of these sensorimotor interactions, we recorded from lightly anesthetized rats in vivo and brain stem slices in vitro and isolated specific parts of this loop. We found that motor feedback to the vibrissal pad serves as a dynamic gain controller that controls the response of first-order sensory neurons by increasing and decreasing sensitivity to lower and higher tactile stimulus magnitudes, respectively. This delicate mechanism is mediated through tactile stimulus magnitude-dependent motor feedback. Conversely, tactile inputs affect the motor whisking output through influences on the rhythmic whisking circuitry, thus changing whisking kinetics. Similarly, tactile influences also modify the whisking amplitude through synaptic and intrinsic neuronal interaction in the facial nucleus, resulting in facilitation or suppression of whisking amplitude. These results point to the vast range of mechanisms underlying sensorimotor integration in the brain stem loop.NEW & NOTEWORTHY Sensorimotor integration is a process in which sensory and motor information is combined to control the flow of sensory information, as well as to adjust the motor system output. We found in the rodent's whisker somatosensory system mutual influences between tactile inputs and motor output, in which motor neurons control the flow of sensory information depending on their magnitude. Conversely, sensory information can control the magnitude and kinetics of whisker movement.

Mitotic activity, modulation of DNA processing, and purinergic signalling in the adult rat auditory brainstem following sensory deafferentation.

A complex scenario of cellular network reorganization is caused by unilateral sensory deafferentation (USD) in the adult rat central auditory system. We asked whether this plasticity response involves mitosis. Immunohistochemistry was applied to brainstem sections for the detection and localization of mitotic markers Ki67 and PCNA, the growth-associated protein Gap43 and purine receptor P2X4. Fluorescent double staining was done for Ki67:PCNA and for both of them with HuC/HuD (neurons), S100 (astrocytes), Iba1 (microglia) and P2X4. Inquiring 1-7days after USD, we found Ki67 expression to be changed in cellular profiles of cochlear nucleus (CN) with a significant increase in number by 1-3days, followed by reset to control level within 1week. USD-induced mitosis exclusively occurred in microglia and was absent elsewhere in the auditory brainstem. PCNA staining of small cellular profiles increased similarly but remained elevated. PCNA staining intensity also changed in CN, superior olive and inferior colliculus in neuronal nuclei, suggesting shifts in DNA processing. No apoptotic cell death was detected in any region of the adult auditory brainstem after USD. A comparison of anterograde and retrograde effects of nerve damage revealed proliferating microglia expressing P2X4 receptors in CN upon USD, but not in the facial nucleus after facial nerve transection. In conclusion, the deafferentation model studied here permits insight into the capacity of the adult mammalian brain to invoke mitosis among glia cells, adjustment of gene processing in neurons and purinergic signalling between them, jointly accounting for a multilayered neuro- and glioplastic response.

Pontine cavernous malformation: microsurgery evading the floor of the fourth ventricle.

This video demonstrates resection of a left pontine cavernous malformation that is abutting the floor of the fourth ventricle (f4V). Even though accessing the lesion through the f4V seems to be reasonable, we used a lateral supracerebellar approach through the middle cerebellar peduncle to preserve especially the abducens and facial nuclei. After total resection the patient was neurologically intact at the 3-month follow-up. Postoperative MRI revealed 3.5-mm pontine tissue between the cavity and f4V that appeared to be absent in preoperative MRI. Approaching pontine lesions through the f4V is not the first choice. In our opinion, the philosophy of safe entry zones is a concept to be reassessed.The video can be found here: https://youtu.be/1Jh6giZc-48.

MULTIPLE SCLEROSIS PRESENTING AS ISOLATED PERIPHERAL FACIAL NERVE PALSY

Isolated facial nerve palsy is commonly seen with Bell’s palsy. However, it can be associated with a central lesion at the level of the ipsilateral facial nucleus or the facial nerve at the pons. Multiple sclerosis is a chronic autoimmune inflammatory disease characterized by axonal degeneration and demyelination of the central nervous system. Isolated cranial neuropathies are rarely seen with multiple sclerosis. The patient being reported is a 20-year-old female who developed isolated facial nerve palsy, which was initially treated as Bell’s palsy. However, she was found to have multiple sclerosis on MRI brain. Isolated facial nerve palsy due to multiple sclerosis is a rare scenario and can often get misdiagnosed and treated as Bell’s palsy.

Synchrotron radiation-based FTIR spectro-microscopy of the brainstem of the hSOD1 G93A rat model of amyotrophic lateral sclerosis.

Pathological mechanisms in amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disease, are still poorly understood. One subset of familial ALS cases is caused by mutations in the metallo-enzyme copper-zinc superoxide dismutase (SOD1), increasing the susceptibility of the SOD1 protein to form insoluble intracellular aggregates. Here, we employed synchrotron radiation-based Fourier transform infrared spectroscopy and microscopy to investigate brainstem cross-sections from the transgenic hSOD1 G93A rat model of ALS that overexpresses human-mutated SOD1. We compared the biomacromolecular organic composition in brainstem tissue cross-sections of ALS rats and their non-transgenic littermates (NTg). We demonstrate that the proteins and especially their antiparallel β-sheet structure significantly differed in all three regions: the facial nucleus (FN), the gigantocellular reticular nucleus (GRN) and the trigeminal motor nucleus (TMN) in the brainstem tissue of ALS rats. The protein levels varied between different brainstem areas, with the highest concentration observed in the region of the FN in the brainstem tissue of NTg animals. Furthermore, the concentration of lipids and esters was significantly decreased in the TMN and FN of ALS animals. A similar pattern was detected for choline and phosphate assigned to nucleic acids with the highest concentrations in the FN of NTg animals. The spectroscopic analysis showed significant differences in phosphates, amide and lipid structure in the FN of NTg animals in comparison with the same area of ALS rats. These results show that the hG93A SOD1 mutation causes metabolic cellular changes and point to a link between bioorganic composition and hallmarks of protein aggregation.

Hindbrain neurovascular anatomy of adult goldfish (Carassius auratus).

The goldfish hindbrain develops from a segmented (rhombomeric) neuroepithelial scaffold, similar to other vertebrates. Motor, reticular and other neuronal groups develop in specific segmental locations within this rhombomeric framework. Teleosts are unique in possessing a segmental series of unpaired, midline central arteries that extend from the basilar artery and penetrate the pial midline of each hindbrain rhombomere (r). This study demonstrates that the rhombencephalic arterial supply of the brainstem forms in relation to the neural segments they supply. Midline central arteries penetrate the pial floor plate and branch within the neuroepithelium near the ventricular surface to form vascular trees that extend back towards the pial surface. This intramural branching pattern has not been described in any other vertebrate, with blood flow in a ventriculo-pial direction, vastly different than the pial-ventricular blood flow observed in most other vertebrates. Each central arterial stem penetrates the pial midline and ascends through the floor plate, giving off short transverse paramedian branches that extend a short distance into the adjoining basal plate to supply ventromedial areas of the brainstem, including direct supply of reticulospinal neurons. Robust r3 and r8 central arteries are significantly larger and form a more interconnected network than any of the remaining hindbrain vascular stems. The r3 arterial stem has extensive vascular branching, including specific vessels that supply the cerebellum, trigeminal motor nucleus located in r2/3 and facial motoneurons found in r6/7. Results suggest that some blood vessels may be predetermined to supply specific neuronal populations, even traveling outside of their original neurovascular territories in order to supply migrated neurons.

Tridimensional mapping of Phox2b expressing neurons in the brainstem of adult Macaca fascicularis and identification of the retrotrapezoid nucleus

Chemosensitivity is a key mechanism for the regulation of breathing in vertebrates. The retrotrapezoid nucleus is a crucial hub for respiratory chemoreception within the brainstem. It integrates chemosensory information that are both peripheral from the carotid bodies (via the nucleus of the solitary tract) and central through the direct sensing of extracellular protons. To date, the location of a genetically defined RTN has only been ascertained in rodents. We first demonstrated that Phox2b, a key determinant for the development of the visceral nervous system and branchiomotor nuclei in the brainstem including the RTN, had a similar distribution in the brainstem of adult macaques compared to adult rats. Second, based on previous description of a specific molecular signature for the RTN in rats, and on an innovative technique for duplex in situ hybridization, we identified parafacial neurons which coexpressed Phox2b and ppGal mRNAs. They were located ventrally to the nucleus of the facial nerve and extended from the caudal part of the nucleus of the superior olive to the rostral tip of the inferior olive. Using the previously described blockface technique, deformations were corrected to allow the proper alignment and stacking of digitized sections, hence providing for the first time a 3D reconstruction of the macaque brainstem, Phox2b distribution and the primate retrotrapezoid nucleus. This description should help bridging the gap between rodents and humans for the description of key respiratory structures in the brainstem.