Britta Fricke

Epithelial Na+ channels and stomatin are expressed in rat trigeminal mechanosensory neurons

Abstract. Caenorhabditis elegans MEC-4 and MEC-10 are subunits of the degenerin/epithelial Na+ channel (DEG/ENaC) ion channel superfamily thought to be associated with MEC-2 (a stomatin-like protein) in a mechanotransducing molecular complex in specialized touch sensory neurons. A key question is whether analogous molecular complexes in higher organisms transduce mechanical signals. To address this question, we selected mechanoreceptors of the rat vibrissal follicle-sinus complex in the mystacial pad and the trigeminal ganglia for an immunocytochemical and molecular biological study. RT-PCR of poly(A+) mRNA of rat trigeminal ganglia indicated that α-, β-, and γ-ENaC and stomatin mRNA are expressed in rat trigeminal ganglia. Using immunocytochemistry, we found that α-, β-, and γ-ENaC subunits and stomatin are localized in the perikarya of the trigeminal neurons and in a minor fraction of their termination site in the vibrissal follicle-sinus complex, where longitudinal lanceolate endings are immunopositive. We conclude that α-, β-, and γ-ENaC subunits as well as the candidate interacting protein stomatin are coexpressed in a mammalian mechanoreceptor, a location consistent with a possible role in mechanotransduction.

Homozygous mutations in caveolin-3 cause a severe form of rippling muscle disease

Heterozygous missense mutations in the caveolin‐3 gene (CAV3) cause different muscle disorders. Most patients with CAV3 alterations present with rippling muscle disease (RMD) characterized by signs of increased muscle irritability without muscle weakness. In some patients, CAV3 mutations underlie the progressive limb‐girdle muscular dystrophy type 1C (LGMD1C). Here, we report two unrelated patients with novel homozygous mutations (L86P and A92T) in CAV3. Both presented with a more severe clinical phenotype than usually seen in RMD. Immunohistochemical and immunoblot analyses of muscle biopsies showed a strong reduction of caveolin‐3 in both homozygous RMD patients similar to the findings in heterozygous RMD. Electron microscopy studies showed a nearly complete absence of caveolae in the sarcolemma in all RMD patients analyzed. Additional plasma membrane irregularities (small plasmalemmal discontinuities, subsarcolemmal vacuoles, abnormal papillary projections) were more pronounced in homozygous than in heterozygous RMD patients. A stronger activation of nitric oxide synthase was observed in both homozygous patients compared with heterozygous RMD. Like in LGMD1C, dysferlin immunoreactivity is reduced in RMD but more pronounced in homozygous as compared with heterozygous RMD. Thus, we further extend the phenotypic variability of muscle caveolinopathies by identification of a severe form of RMD associated with homozygous CAV3 mutations. Ann Neurol 2003

Four new cases of stomatin-deficient hereditary stomatocytosis syndrome: association of the stomatin-deficient cryohydrocytosis variant with neurological dysfunction

This report concerns congenitally Na+–K+ leaky red cells of the ‘hereditary stomatocytosis’ class. Three new isolated cases and one new pedigree are described, and one previously reported case is expanded. In all cases, Western blotting of red cell membranes revealed a deficiency in the 32 kDa membrane protein, stomatin. All showed pronounced cation leaks at 37°C with markedly abnormal intracellular Na+ and K+ concentrations, like all other such stomatin‐deficient cases. Consistent with recent findings in two previously described British pedigrees, immunocytochemistry demonstrated that the deficiency of stomatin was not complete. On typical blood films, some red cells showed positive stomatin immunoreactivity, while most were negative, although in one case only a minority were negative. All platelets and neutrophils were stomatin positive. The cases differed markedly between themselves with regard to the temperature dependence of the passive leak to K+. Three showed a simple monotonic temperature dependence, while two showed a minimum at around 20–25°C, such that the cells were extremely leaky at 0°C, giving the phenotype known as ‘cryohydrocytosis’. These patients are the only two known cases of stomatin‐deficient cryohydrocytosis. Both showed a congenital syndrome of mental retardation, seizures, cataracts and massive hepatosplenomegaly, probably defining a new haemato‐neurological syndrome.

Salivary cortisol and psychological mechanisms in patients with acute versus chronic low back pain

This study was designed to explore whether the basal adrenocortical activity is related with pain-related coping, nonverbal pain behavior, depressive mood, and fatigue in patients with acute and chronic nonspecific low back pain. 19 patients with acute low back pain (ALBP) and 24 with chronic low back pain (CLBP) participated in the study. The adrenocortical activity was assessed through the cortisol awakening response. All participants provided five saliva samples (0, 15, 30, 45, and 60min after waking) on two consecutive days off work. Pain-related coping [fear-avoidance coping (FAC) and endurance coping (EC)], nonverbal pain behavior (NPB), depressive mood, and fatigue were assessed through questionnaires. Among ALPB patients, EC was negatively associated with the cortisol release, whereas fatigue was positively associated with it. Among CLBP patients, FAC, NPB, depressive mood, and fatigue were negatively associated with the cortisol awakening response, whereas EC tended to be positively associated with it. The results indicate that pain-related coping strategies which are expected to be successful appear to lower the adrenocortical activity among ALBP patients, whereas affective distress may enhance the level of cortisol in this group. Among CLBP patients, long-term maladaptive coping strategies might contribute to hypocortisolism.

Topography and immunocytochemical characterization of nerve fibers in the leptomeningeal compartments of the rat. A light- and electron-microscopical study

Abstract.The localization of peptidergic, catecholaminergic, and nitroxidergic nerve fibers in the ventral leptomeningeal connective tissue compartment was studied in whole-mount preparations and serial semithin and ultrathin sections. For immunocytochemistry, whole-mount preparations of the leptomeninges and ventral brain slices with the meninges were incubated as free-floating specimens with primary antibodies against protein gene product 9.5 (PGP 9.5), substance P (SP), calcitonin gene-related peptide (CGRP), dopamine beta-hydroxylase (DβH), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), and nitric oxide synthase (NOS) using the avidin-biotin-peroxidase method. Based on the regional differences of the connective tissue organization, the leptomeninx is subdivided into the pial, trabecular, and adventitial leptomeninx. The antibody PGP 9.5 stains all unmyelinated nerve fibers in the leptomeninx. Although the highest density of nerve fibers occurs in the adventitial leptomeninx, nerve fibers, and terminals are additionally present in the trabecular and pial leptomeninx. DβH-, NPY-, VIP- and NOS-immunoreactive (IR) nerve fibers occur exclusively in the adventitial leptomeninx forming neuromuscular junctions. CGRP- and SP-IR nerve fibers are localized in all three leptomeningeal compartments where they terminate close to the subarachnoid space (type 1) or within the connective tissue (type 2). Due to their morphological and immunocytochemical characterization a possible chemo-, mechano- or nociceptive function is discussed in the context of pathophysiological aspects.

Vibrography During Tumor Neurosurgery

The aim of this study was to determine whether elastography, a sonographically based real‐time strain imaging method for registering the elastic properties of tissue, can be used in brain tumor surgery.

Stomatin is mis-trafficked in the erythrocytes of overhydrated hereditary stomatocytosis, and is absent from normal primitive yolk sac-derived erythrocytes

The 32 kD lipid‐raft‐associated membrane protein ‘stomatin’ is deficient from the erythrocyte membrane in the Na+–K+ leaky haemolytic anaemia, overhydrated hereditary stomatocytosis (OHSt). To date, no mutation in the gene coding for this protein has so far been found in OHSt. In this study, we have analysed the distribution of stomatin in both cultured erythroid cells from OHSt patients and in normal embryological and fetal erythroid development. In erythroid cell cultures from OHSt patients, stomatin‐immunoreactivity (stomatin‐IR) was present in progenitor cells but remained restricted to the area of the multivesicular complexes and the nucleus in the developing cells and was not seen in the plasma membrane. This could be consistent with the idea that stomatin is an innocent passenger in a more fundamental trafficking abnormality. In normal embryonic development, we found that, in extraembryonic (yolk sac) erythropoiesis, neither the nucleated red cells nor their enucleated mature derivatives displayed any stomatin‐IR. In contrast, all haemangiopoietic progenitor cells of intraembryonic haematopoiesis, starting with the mesodermal precursors in the aorta–gonad–mesonephros region, exhibited strong stomatin‐IR. The significance of this observation on these poorly understood cells is currently unclear.

Topography and distribution of nerve fibers in the posterior longitudinal ligament of the rat: an immunocytochemical and electron-microscopical study.

The distribution and immunocytochemical characterization of nerve fibers and their terminals in the posterior longitudinal ligament of the rat lumbar vertebral column was studied in whole-mount preparations and serial semithin and ultrathin sections. Differences in the localization, distribution pattern and density of peptidergic and catecholaminergic nerve fibers were found in the vertebral and intervertebral regions of the posterior longitudinal ligament. For immunocytochemistry, free floating specimens were incubated with primary antibodies against protein gene product 9.5, substance P, calcitonin gene-related peptide, dopamine-beta-hydroxylase, vasoactive intestinal polypeptide and neuropeptide Y together with the avidin-biotin-peroxidase method. In whole-mount preparations, the neural marker protein gene product 9.5 is immunostained in all unmyelinated nerve fibers in the posterior longitudinal ligament, thus giving a panoramic view of the nerve fiber plexus. The most striking nerve fiber plexus is localized in the intervertebral region. In this region, the posterior longitudinal ligament is rich in capillaries that form a dense plexus within its ventral part and extend to the outer layer of the annulus fibrosus. The peptidergic and catecholaminergic innervation of the posterior longitudinal ligament is discussed in the context of pain syndromes related to the vertebral column and degenerative lumbar spine diseases.

Individualized ventriculostomy in hydrocephalus: An intravital anatomical study

BACKGROUND Ventriculostomy is a common neuroendoscopic operation but one with disastrous complications in rare cases. AIMS The aim of this study was to perform an intravital analysis of the configuration at the floor of the third ventricle as a possible basis for selection of the ventriculostomy site. MATERIALS AND METHODS The study population consisted of 32 patients who underwent ventriculostomy for the treatment of hydrocephalus. Perforation of the floor of the third ventricle was carried out on an individual basis following evaluation of the anatomic situation. Video material and magnetic resonance images (MRI) were analyzed. RESULTS A classification system including three major groups was developed using the inner distance of the mamillary bodies as the key criterion. It was defined as narrow for values between 0 and 1 mm (observed range: 0-0.5 mm), medium for values between 1.1 and 3.4 mm (range 1.1-3.4 mm) and large for values greater than 3.4 mm (range: 3.8-6.9 mm). Statistical analysis of MR and video measurements revealed a good correlation. The ventriculostomy site was rostral of the mamillary bodies in 23 of the patients (n=27) and slightly occipital in four. The ventriculostomy site was located more to the left in 22 patients and more to the right in five. CONCLUSION As a conclusion the ventriculostomy site has to be chosen in each case following a careful review of all available information. A classification system for the anatomical variations as well as the exact size and site of ventriculostomy should be introduced.

Consideration of ergonomic aspects in the development of a new endoscopic navigation system

During the development of new navigation systems, the ergonomic aspects of the accompanying software are seldom taken into account. The VN system is a navigation system working with real and previously stored neuroendoscopic images. The latter enable the module to execute virtual back movement, e.g. in case of bleeding. Several other modules are offered. The aim of the study was the ergonomic analysis of different modules and their learning curves in cadaveric heads. The endoscope was navigated by the neurosurgeon while another person operated the computer. A total of 128 experiments were performed with two software versions. When the landmark tracking module was used, a real learning curve could be observed. By contrast, testing the measurement module did not produce a learning curve. A significant reduction of the time required by the three modules investigated could be observed with software version 2. The module for virtual back movement works best with a minimum time of 20 s for image storage. During machine-human interactions ergonomic software use is important, especially if operative procedures are performed.