Deep brain stimulation of the nucleus basalis of Meynert in Alzheimer's dementia.

Cholinergic neurons of the medial forebrain are considered important contributors to brain plasticity and neuromodulation. A reduction of cholinergic innervation can lead to pathophysiological changes of neurotransmission and is observed in Alzheimer's disease. Here we report on six patients with mild to moderate Alzheimer's disease (AD) treated with bilateral low-frequency deep brain stimulation (DBS) of the nucleus basalis of Meynert (NBM). During a four-week double-blind sham-controlled phase and a subsequent 11-month follow-up open label period, clinical outcome was assessed by neuropsychological examination using the Alzheimer's Disease Assessment Scale-cognitive subscale as the primary outcome measure. Electroencephalography and [(18)F]-fluoro-desoxyglucose positron emission tomography were, besides others, secondary endpoints. On the basis of stable or improved primary outcome parameters twelve months after surgery, four of the six patients were considered responders. No severe or non-transitional side effects related to the stimulation were observed. Taking into account all limitations of a pilot study, we conclude that DBS of the NBM is both technically feasible and well tolerated.

Changes in Nutritional Status after Deep Brain Stimulation of the Nucleus Basalis of Meynert in Alzheimer's Disease--Results of a Phase I Study.

OBJECTIVE: The progression of Alzheimer's disease (AD) is associated with impaired nutritional status. New methods, such as deep brain stimulation (DBS), are currently being tested to decrease the progression of AD. DBS is an approved method in the treatment of Parkinson's disease, and its suitability for the treatment of AD patients is currently under experimental investigation. To evaluate the advantages and disadvantages of this new treatment, it is important to assess potential side effects of DBS regarding the nucleus basalis of Meynert; this new treatment is thought to positively affect cognition and might counteract the deterioration of nutritional status and progressive weight loss observed in AD. This study aims to assess the nutritional status of patients with AD before receiving DBS of the nucleus basalis of Meynert and after 1 year, and to analyze potential associations between changes in cognition and nutritional status. DESIGN: A 1-year phase I proof-of-concept study. SETTING: The Department of Psychiatry and Psychotherapy at the University of Cologne. PARTICIPANTS: We assessed a consecutive sample of patients with mild to moderate AD (n=6) who fulfilled the inclusion criteria and provided written informed consent. INTERVENTION: Bilateral low-frequency DBS of the nucleus basalis of Meynert. MEASUREMENTS: Nutritional status was assessed using a modified Mini Nutritional Assessment, bioelectrical impedance analysis, a completed 3-day food diary, and analysis of serum levels of vitamin B12 and folate. RESULTS: With a normal body mass index (BMI) at baseline (mean 23.75 kg/m²) and after 1 year (mean 24.59 kg/m²), all but one patient gained body weight during the period of the pilot study (mean 2.38 kg, 3.81% of body weight). This was reflected in a mainly stable or improved body composition, assessed by bioelectrical impedance analysis, in five of the six patients. Mean energy intake increased from 1534 kcal/day (min 1037, max 2370) at baseline to 1736 kcal/day (min 1010, max 2663) after 1 year, leading to the improved fulfillment of energy needs in four patients. The only nutritional factors that were associated with changes in cognition were vitamin B12 level at baseline (Spearman's rho = 0.943, p = 0.005) and changes in vitamin B12 level (Spearman's rho = -0.829, p = 0.042). CONCLUSION: Patients with AD that received DBS of the nucleus basalis of Meynert demonstrated a mainly stable nutritional status within a 1-year period. Whether DBS is causative regarding these observations must be investigated in additional studies.

The ionic mechanism of postsynaptic inhibition in motoneurones of the frog spinal cord.

The ionic mechanism of postsynaptic inhibition in frog spinal motoneurones was studied with conventional and with ion-sensitive microelectrodes. In these neurones the inhibitory postsynaptic potential was depolarizing, its reversal potential being 15 mV less negative than the resting membrane potential. During the inhibitory postsynaptic potential the input resistance of the motoneurones was reduced to 20% of the resting value, indicating a strong increase of membrane conductance. The Cl- equilibrium potential calculated from intra- and extracellular Cl- activity measurements coincided with the reversal potential of the inhibitory postsynaptic potential to within a few millivolts. During repetitive inhibitory postsynaptic activity the intracellular Cl- activity decreased markedly, while the extracellular Cl- activity increased slightly. These changes of intra- and extracellular Cl- activities were no longer found after suppression of the inhibitory postsynaptic potential by strychnine. Blockade of an active, inward-going Cl- transport system in motoneurones by NH+4 led to a shift of the Cl- equilibrium potential and the reversal potential of the inhibitory postsynaptic potential towards the resting membrane potential. After prolonged action of NH+4, the Cl- equilibrium potential approached the membrane potential to within 5 mV, while the reversal potential of the inhibitory postsynaptic potential and resting membrane potential coincided. The difference between Cl- equilibrium potential and membrane potential after blockade of the Cl- pump is traced back to interfering intracellular ions, such as HCO-3 or SO42-, leading to an overestimation of intracellular Cl- activity and to the calculation of an erroneous Cl- equilibrium potential. Inhibitory amino acids like gamma-aminobutyrate or beta-alanine evoked depolarizations with reversal potentials similar to that of the inhibitory postsynaptic potential. These depolarizations were associated with a marked decrease of neuronal input resistance during inhibition. During the actions of these compounds a decrease of intracellular and a small increase of extracellular Cl- activity were found. The activities of other ions (K+, Ca2+ and Na+) did not change significantly, with the exception of extracellular K+ activity, which was slightly increased. Evidence is presented that the inhibitory postsynaptic potential, as well as the depolarizing action of inhibitory amino acids in motoneurones, is the result of an increase in membrane Cl- permeability and an efflux of Cl- from these cells, while other ions do not seem to be involved.

Epithelioid cells: membrane potential changes induced by substances influencing renin secretion.

Microelectrode recordings were performed in renin-containing epithelioid (JG) and vascular smooth muscle (VSM) cells of the afferent arteriole in the isolated hydronephrotic mouse kidney. Both cell types had a membrane potential of about -75 mV and exhibited small, spontaneous depolarizing transients, probably resulting from random transmitter release by sympathetic axon terminals. Substances depressing renin secretion, such as angiotensin II, arginine-vasopressin, and alpha 1-adrenergic agents reversibly depolarized both JG and VSM cells. On a molar basis, the action of angiotensin II was strongest. Stimulators of renin release, e.g. isoproterenol, histamine, and prostaglandin E2 did not influence the membrane potential of both cell types. VIP and NPY, possible co-transmitters of norepinephrine, as well as AP II, were also without effect. It is proposed that suppression of renin secretion from JG cells is mediated by depolarization and Ca2+ influx, whereas stimulation is triggered independently from membrane potential changes, e.g. by adenylate cyclase activation.

A piezotranslator with variable movement pattern: experiences with the penetration of very small cells.

Successful recording of intracellular potentials strongly depends on the quality of the impalement of the cells by microelectrodes. A substantial improvement of the penetration process could be obtained by using a piezotranslator which accurately controls the forward and backward movement of the electrode tip. The relevant movement amplitudes and velocities can be adjusted independently. The described piezotranslator was used in experiments with cultured cells of the glomerular mesangium of rat kidney, forming a flat monolayer 1-3 micrometers in height. Many successful impalements and long-term, stable recordings demonstrate the usefulness of the translator.

Atrial natriuretic factor stimulates renin release from the isolated rat kidney.

Intrarenal infusion of atrial natriuretic factor has been reported to increase renal blood flow and sodium excretion, and either to depress or not to affect renin release. We examined the effect of a synthetic atrial peptide, AP II, on renin release, renal perfusate flow and glomerular filtration rate in isolated rat kidneys, perfused at constant pressure with a synthetic Krebs-Henseleit hydroxyethyl-starch-albumin medium. Atrial peptide II, 0.3 to 30 nmol/l, produced a concentration-dependent increase in renin release. Renal perfusate flow increased initially but returned to pre-infusion levels during infusion. During vasodilatation by continuous infusion of hydralazine, AP II produced only minor changes of perfusate flow but higher increases of renin release than in controls, indicating that this effect is not secondary to vasodilatation. During suppression of prostaglandin synthesis by indomethacin, the effects of AP II were unaltered. Furthermore, AP II in concentrations up to 200 nmol/l did not alter resting membrane potential in juxtaglomerular cells.

External trial deep brain stimulation device for the application of desynchronizing stimulation techniques.

In the past decade deep brain stimulation (DBS)-the application of electrical stimulation to specific target structures via implanted depth electrodes-has become the standard treatment for medically refractory Parkinson's disease and essential tremor. These diseases are characterized by pathological synchronized neuronal activity in particular brain areas. We present an external trial DBS device capable of administering effectively desynchronizing stimulation techniques developed with methods from nonlinear dynamics and statistical physics according to a model-based approach. These techniques exploit either stochastic phase resetting principles or complex delayed-feedback mechanisms. We explain how these methods are implemented into a safe and user-friendly device.

Intracellular ion activities and equilibrium potentials in motoneurones and glia cells of the frog spinal cord.

Intra-and extracellular ion activities were measured with ion sensitive microelectrodes in motoneurones and glia cells of the spinal cord of the frog. These data were corrected for cross sensitivities of the ion exchangers to intracellular interfering ions, and equilibrium potentials for K +, Na +, Ca2 + and C1- (EK, ENa, ECa and EC1) were calculated. In motoneurones with membrane potentials exceeding -60 mV the following mean equilibrium potentials were determined. ENa = + 29.4mV, EK = -87.9 mV, ECa = + 52.6 mV, EC1 = -34.1 mV. The corresponding values for glia cells were: ENa = + 40.5 mV, EK = -84.0 mV, ECa = + 35.7 mV, EC1 = -59.7 mV. The intracellular ionic milieu is probably disturbed by the impalement of the cells. This transiently decreases the intracellular K + and increases intracellular Na +. These effects were estimated and their origin is discussed. The results of the experiments suggest a non-passive transmembrane distribution of K +, Na + and Ca2 + in motoneurones and glia cells, a non-passive transmembrane distribution of C1- in motoneurones, and a passive transmembrane distribution of C1- in glia cells.

The ionic mechanism of the excitatory action of glutamate upon the membranes of motoneurones of the frog.

Simultaneous intra- and extracellular recordings with K+, Na+, Ca2+, and Cl- sensitive microelectrodes were performed in motoneurones of the spinal cord of the frog during depolarizations mediated by glutamate (GLUT) and by experimentally increased extracellular K+. Depolarization resulting from increased K+ activity (alpha K+) in the bathing solution evoked a decrease of intracellular Na+ activity (alpha Na+i); a transient increase of alpha Na+i accompanied by a decrease of alpha Na+e was observed during the depolarization induced by GLUT. Both modes of depolarization led to an increase of alpha Cl-i and a concomitant decrease of alpha Cl-e. An experimental increase of alpha K+e led to a threshold dependent increase of alpha Ca2+i by at least one order of magnitude and to an equally threshold dependent strong decrease of alpha Ca2+e. The threshold of these changes of alpha Ca2+ was at a membrane potential of -25 mV. During a depolarization of half the amplitude induced by GLUT a comparable increase of alpha Ca2+i and a smaller decrease of alpha Ca2+e were observed. The GLUT mediated changes of alpha Ca2+ were not threshold dependent and occurred synchronously with the onset of depolarization. A transient decrease of alpha K+i and a parallel strong increase of alpha K+e occurred during the GLUT induced depolarization. Depolarization evoked by an experimental increase of alpha K+e led to an increase of alpha K+i. The observed changes in the ionic composition of the intra- and extracellular fluids indicate that GLUT evokes an increase in membrane permeability to Na+ and Ca2+ and a subsequent influx of these ions into motoneurones, while the inward shift of Cl- and the outward shift of K+ are presumably passive. A voltage dependent Ca2+ influx is triggered at -25 mV membrane potential.

On the postsynaptic action of glutamate in frog spinal motoneurons.

In the isolated frog spinal cord depolarization of motoneurons (MNs) induced by glutamate (GLUT) was not accompanied by measurable changes of neuronal input resistance when chemical synaptic transmission was blocked by Mn2+ or Mg2+. The GLUT depolarization was, however, paralleled by a considerable increase in K+ in the extracellular space. To clarify, whether the GLUT depolarization was exclusively due to a reduction of the transmembrane K+ gradient or whether ion conductances not detectable by measurements of neuronal input resistance were involved, membrane potential (MP) was plotted semilogarithmically versus extracellular K+ activity (aKe+). During experimental elevation of aKe+ the function delta MP/dec. delta aKe+ was found to agree fairly with the Nernst equation. The slope of this function was much steeper during GLUT superfusion, indicating an influx of positive ions. The elevation of aKe+ during the GLUT action can mimic postsynaptic effects by release of transmitter from presynaptic terminals synapsing with the recorded cell. In vivo preparations do not allow blockade of chemical synaptic transmission. Therefore, it is impossible to decide, whether the recorded cell is depolarized either postsynaptically by GLUT or by K+ release from surrounding GLUT sensitive cells. As an experimental proof of the postsynaptic GLUT action is not feasible in such preparations, the ubiquitous action of GLUT in the CNS may have been overestimated.

The mesangial cell culture: a tool for the study of the electrophysiological and pharmacological properties of the glomerular mesangial cell.

Cultured rat glomerular mesangial cells (MC) were evaluated as a tool for reliable electrophysiological measurements as well as for fluorimetric determinations of intracellular Ca++. They had a resting potential similar to that observed in cultured vascular smooth muscle cells (VSMCs), in VSMCs of mouse kidney arterioles, or in glomerular--presumably mesangial--cells of kidney slices. The comparison with the other cell types was carried out in order to look for features distinguishing them from these cells, e.g., active and passive electrical membrane properties or electrical membrane responses to vasoactive pharmacological agents. In MCs, as well as in the other cell types, the average membrane potential was approx. -50 mV. The vasoconstrictor peptides angiotensin II (ANG II) and arginine-vasopressin (AVP) caused depolarizations that could be blocked by the respective specific inhibitors of these compounds. The agonist-induced depolarizations have to be attributed, at least in part, to a Ca++ inward current. Norepinephrine, if any, had only a weak action upon MCs, whereas isoproterenol either did not influence the membrane potential or hyperpolarized the cells. Other substances tested, which had no influences upon the membrane potential, were neuropeptide Y and atriopeptin 3. As to their resting electrical properties and their responses to pharmacological agents, cultured mesangial cells did not differ from glomerular, i.e., most probably mesangial, cells in the kidney slice. The difference between mesangial cells and VSMCs consists in their reaction to noradrenaline. Whereas VSMCs respond with a marked depolarization, the noradrenaline effect upon MCs in culture and in the kidney slice is either absent or very weak. Repeated passage of the cells (more than six passages) led to a gradual loss of their responsiveness to the agonists, indicating reduced receptor expression which may be interpreted as dedifferentiation. This held for both cultured MCs and VSMCs. Fluorimetric measurements using the Ca++-specific indicators quin-2 and fura-2 were performed with a purpose-developed, ultrasensitive photon-counting microspectrofluorimeter. Individual MCs as well as isolated glomeruli responded to the vasoconstrictors ANG II and AVP with an increase in Ca++-dependent fluorescence indicating that these agents indeed depolarize the cells partly via a Ca++ influx and increase cytosolic free Ca++.(ABSTRACT TRUNCATED AT 400 WORDS)

Junctional transmission in renin-containing and smooth muscle cells of the afferent arteriole.

Intracellular recordings were done in renin-containing juxtaglomerular (JG) and vascular smooth muscle (VSM) cells of the mouse kidney afferent arteriole. Both cell types exhibited a membrane potential around -75 mV and spontaneous depolarizing transients resembling spontaneous excitatory junction potentials (SEJPs) in the arterioles of other organs. The amplitude distribution of these randomly occurring transients was skewed in both cell types with a modal value of 1.2-1.9 mV. Activation of presumably postjunctional alpha 1-, P2-, ANG II- and AVP-receptors depolarized JG and VSM cells. Application of the P1-purinoceptor agonist 2-chloroadenosine strongly increased frequency and amplitude of the SEJP-like events, whereas these transients were abolished by the P1-purinoceptor antagonist 8-phenyltheophylline, both substances presumably acting on prejunctional receptors. The SEJP-like events were completely depressed by reserpine treatment, but not abolished by alpha 1-, alpha 2-, and P2-antagonists. At present, it cannot be decided, whether norepinephrine is the sole transmitter in the afferent arteriole, acting on specialized junctional adrenoceptors with the P2-purinoceptors being irrelevant for junctional transmission, or whether both substances are co-transmitters. Except norepinephrine and ATP, all other transmitter candidates tested were ruled out for various reasons.

Improvement of freeze-fracture autoradiography for localization of soluble substances in tissue samples.

Freeze-fracture autoradiography is accepted as an adequate technique for localization studies of soluble substances at the electron microscopical level. The method, however, involves many critical preparation steps, among them a protective carbon coating of the developed nuclear emulsion adhering to the replica. We demonstrate here that this additional carbon coating may be omitted. This simplification leads to a significant improvement of the sample yield as compared with the previously described procedures.

Intracellular recordings from renin-positive cells of the afferent glomerular arteriole.

Intracellular recordings were made in juxtaglomerular granulated (JG) cells and in vascular smooth muscle (VSM) cells in afferent arterioles of hydronephrotic mouse kidneys. Both cell types did not differ in their passive and active electrical membrane properties; membrane potential was about -58 mV, input resistance exceeded 400 M omega, and JG as well as VSM cells showed spontaneous depolarizations resembling excitatory junction potentials and active responses observed in smooth muscle cells of other blood vessels in various species. These depolarizations, attributed to spontaneous transmitter release from adrenergic terminals, were extremely polymorphous and quite frequent. Epinephrine, norepinephrine, phenylephrine, arginine vasopressin, and angiotensin II depolarized JG and VSM cells, but isoproterenol and orciprenaline had no effect. A hyperpolarizing action of catecholamines was never observed. It is suggested that, in this in vitro preparation, isoproterenol increases renin secretion by a mechanism independent of membrane potential changes. Depolarizations mediated by alpha-mimetic agents, arginine vasopressin, and angiotensin II, as well as by the junctional activity may inhibit renin secretion by an increased calcium influx into JG cells.

Coexistence of renin and cathepsin B in epithelioid cell secretory granules.

Mature juxtaglomerular epithelioid cell secretory granules of the rat exhibit both renin- and cathepsin B-like immunoreactivity. On the basis of the coexistence with renin at a pH which, according to previous experiments, is probably in the range of that in lysosomes, cathepsin B is suggested to be involved in the activation of renin prior to secretion.

Ultrastructural changes associated with renin secretion from the juxtaglomerular apparatus of mice.

Thin sections and freeze-fracture replicas were used to investigate the ultrastructural changes associated with renin secretion from the juxtaglomerular part of the afferent arteriole of male mice. Adrenalectomized animals in which renin secretion was stimulated by furosemide application and bleeding were also studied. Exocytosis of mature electron-dense granules was found in all experimental groups. Before extrusion, the region of granule facing the cell membrane changed, with vesicular and/or stacked membrane-like profiles and a small local protrusion of the granule membrane appearance of. Concomitantly, punctuate sites of fusion between the cell and granule membranes were observed. Later, unaltered amorphous, and altered membrane-like granule content was released from omega-shaped cavities into the extracellular space. In stimulated animals the alteration and extrusion of several closely apposed granules was reminiscent of compound exocytosis. Coated pits were frequently seen, suggesting specific retrieval of the former granule membrane. The collapsing silhouette of a depleted granule very rarely took the form of a saccule whose narrow membrane-bounded neck was continuous with the extracellular space. Observed were two additional events by which active and inactive renin may be released. Small electron-lucent vacuoles of undetermined origin fused with the cell membrane and, in stimulated kidneys, some epithelioid cell processes disintegrated. However, the interpretation of the related ultrastructural phenomena was uncertain.

Cathepsin D coexists with renin in the secretory granules of juxtaglomerular epithelioid cells.

Mature juxtaglomerular epithelioid cell secretory granules of the rat exhibit both renin- and cathepsin D-like immunoreactivity. On the basis of the coexistence with renin at a pH which, according to previous experiments, is probably in the range of that in lysosomes, cathepsin D is suggested to be involved in the regulation of the granular renin stores available for secretion.

Influence of pulsatile perfusion upon renin release from the isolated perfused rat kidney.

It is well established that renin release from the juxtaglomerular epithelioid cells in the media of the afferent arteriole strongly depends on the mean renal perfusion pressure, whereas a possible influence of the pulsation of blood pressure on renin release has only occasionally been investigated, and the results are contradictory. Such an influence on renin release cannot be excluded because pulsation is known to modulate arterial baroreceptors and vascular tone in some resistance vessels. In the isolated perfused rat kidney, we found a pulsation amplitude-dependent inhibition of renin release that could be blocked either by vasodilatation or by calcium channel blockade. The inhibition occurred at perfusion pressures between 85 and 125 mm Hg. The underlying pulsation pressure-sensitive mechanism has to be ascribed integrating properties, because a constant-flow pressure rise to the "systolic" value of pulsatile perfusion resulted in virtually the same inhibition of renin release. Moreover, a reduced urine flow during pulsatile perfusion provides evidence for preglomerular constriction under these conditions. It is concluded that, besides pathological changes of renal perfusion pressure, variations of the pulse amplitudes, e.g. resulting from renal artery stenosis or atherosclerosis, may also influence renin release and contribute to renovascular hypertension.