Cholinergic Structures Research Articles

Participation of Modulating Rhythms of the Decasecond Range in Intersystem Correlation Interactions in Early Rat Ontogeny.

On newborn non-narcotized 1-day-old (P1) and 16-day-old (P16) rats, a detailed analysis of intersystem somatovisceral interactions (ISI) mediated by decasecond (D1, 5-50 sec) range modulating rhythms was performed. Correlation interactions of the main body systems - cardiovascular, respiratory and somatomotor systems in norm and under conditions of changes in the level of cholinergic regulation were studied. Spectral correlation analysis was used to determine the participation of D1 range modulating rhythms in ISI. It was found that at P1, D1 range rhythms do not play a significant role in integrative processes. In P16 the activation of cholinergic structures, caused by the introduction of the acetylcholinesterase inhibitor physostigmine (eserine) leads to significant disturbances in the degree of correlation in the D1 range. Blockade of muscarinic and nicotinic cholinoreceptors does not alter the degree of correlation of systemic pairs in the slow-wave region (D1-low, 8-50 sec) of the D1 range. Under the influence on the cholinoreactive structures, the most significant changes in the degree of correlation in the ISI affect the somatorespiratory systemic pair. The results obtained indicate that the representation of the slow-wave components of D1 range modulating rhythms involved in the ISI increase during the first weeks of postnatal ontogeny. Changes in the level of cholinergic activation do not directly influence on ISI mediated by D1-low sub-band rhythms.

STRUCTURE OF INTRAMURAL NERVE ELEMENTS OF THE URINARY BLADDER IN PATIENTS WITH DIABETIC CYSTOPATHY

Using histological, and histochemical methods, morphological changes in the intramural nerve elements of the urinary bladder were studied in 6 patients suffering from diabetic cystopathy and who died from various complications of diabetes mellitus. The control group consisted of 4 practically healthy persons who died as a result of a car accident. For histological examination, tissue pieces of the nerve elements of the bladder were fixed with 12% formalin and impregnated according to the method of Bilshchovsky-Gross and Rasskazov. Adrenergic nerve structures were detected by incubation of sections in a 2% solution of glyoxylic acid, and cholinergic elements by the Karnovsky-Roots method. It was revealed that along with degenerative changes, reactive-compensatory processes also occur in the intramural nerve elements of the bladder in diabetic cystopathy. It was found that the activity of mediators in adrenergic and cholinergic structures decreases.

REPRODUCTIVE HORMONE LEVELS IN WOMEN WITH THALASSEMIA

Using histological, and histochemical methods, morphological changes in the intramural nerve elements of the urinary bladder were studied in 6 patients suffering from diabetic cystopathy and who died from various complications of diabetes mellitus. The control group consisted of 4 practically healthy persons who died as a result of a car accident. For histological examination, tissue pieces of the nerve elements of the bladder were fixed with 12% formalin and impregnated according to the method of Bilshchovsky-Gross and Rasskazov. Adrenergic nerve structures were detected by incubation of sections in a 2% solution of glyoxylic acid, and cholinergic elements by the Karnovsky-Roots method. It was revealed that along with degenerative changes, reactive-compensatory processes also occur in the intramural nerve elements of the bladder in diabetic cystopathy. It was found that the activity of mediators in adrenergic and cholinergic structures decreases.

ACETYLCHOLINE AND ITS SYNTHESIZING ENZYME CHOLINE ACETYLTRANSPHERASE IN THE ENTERIC NERVOUS SYSTEM

The enteric nervous system is the largest and most complex division of the peripheral nervous system. Located in the wall of the gastrointestinal tract, it is a regulatory and coordination unit of the nervous system. Neurochemical, pharmacological, and functional studies describe three main classes of neurons in the enteric nervous system– primary afferent, interneurons and motor neurons. These are grouped in ganglia, which are connected and form plexuses. Acetylcholine is a major neurotransmitter that plays a pivotal role in the enteric nervous system and several non-neuronal structures. Internal cholinergic neurons and vagus terminals in the enteric nervous system use acetylcholine as the main excitatory neurotransmitter, regulating motility and mucosal function in the digestive system. The enzyme choline acetyltransferase (ChAT) that catalyzes the synthesis of acetylcholine represents the most specific cholinergic marker. Recent markers used to visualize cholinergic structures are the splicing variants of ChAT mRNA that are transcribed from the ChAT gene. Different alternatively spliced ChAT mRNA variants are transcribed in many animal species, including humans. In the mouse, seven variants in the 5’-noncoding region and two variants that differ in their coding region are described. Morphological, genetic and molecular analysis of ChAT and its splicing variants, as the most reliable and frequently used marker for cholinergic structures, would contribute to a better understanding of the physiological and pathological states of cholinergic neurons in the enteric nervous system.

Acrylamide Neurotoxicity as a Possible Factor Responsible for Inflammation in the Cholinergic Nervous System.

Acrylamide (ACR) is a chemical compound that exhibits neurotoxic and genotoxic effects. It causes neurological symptoms such as tremors, general weakness, numbness, tingling in the limbs or ataxia. Numerous scientific studies show the effect of ACR on nerve endings and its close connection with the cholinergic system. The cholinergic system is part of the autonomic nervous system that regulates higher cortical functions related to memory, learning, concentration and attention. Within the cholinergic system, there are cholinergic neurons, anatomical cholinergic structures, the neurotransmitter acetylcholine (ACh) and cholinergic receptors. Some scientific reports suggest a negative effect of ACR on the cholinergic system and inflammatory reactions within the body. The aim of the study was to review the current state of knowledge on the influence of acrylamide on the cholinergic system and to evaluate its possible effect on inflammatory processes. The cholinergic anti-inflammatory pathway (CAP) is a neuroimmunomodulatory pathway that is located in the blood and mucous membranes. The role of CAP is to stop the inflammatory response in the appropriate moment. It prevents the synthesis and the release of pro-inflammatory cytokines and ultimately regulates the local and systemic immune response. The cellular molecular mechanism for inhibiting cytokine synthesis is attributed to acetylcholine (ACh), the major vagal neurotransmitter, and the α7 nicotinic receptor (α7nAChR) subunit is a key receptor for the cholinergic anti-inflammatory pathway. The combination of ACh with α7nAChR results in inhibition of the synthesis and release of pro-inflammatory cytokines. The blood AChE is able to terminate the stimulation of the cholinergic anti-inflammatory pathway due to splitting ACh. Accordingly, cytokine production is essential for pathogen protection and tissue repair, but over-release of cytokines can lead to systemic inflammation, organ failure, and death. Inflammatory responses are precisely regulated to effectively protect against harmful stimuli. The central nervous system dynamically interacts with the immune system, modulating inflammation through the humoral and nervous pathways. The stress-induced rise in acetylcholine (ACh) level acts to ease the inflammatory response and restore homeostasis. This signaling process ends when ACh is hydrolyzed by acetylcholinesterase (AChE). There are many scientific reports indicating the harmful effects of ACR on AChE. Most of them indicate that ACR reduces the concentration and activity of AChE. Due to the neurotoxic effect of acrylamide, which is related to the disturbance of the secretion of neurotransmitters, and its influence on the disturbance of acetylcholinesterase activity, it can be concluded that it disturbs the normal inflammatory response.

Cholinergic receptor binding in unimpaired older adults, mild cognitive impairment, and Alzheimer\u2019s disease dementia

BackgroundCholinergic neurotransmitter system dysfunction contributes to cognitive impairment in Alzheimer’s disease and other syndromes. However, the specific cholinergic mechanisms and brain structures involved, time course of alterations, and relationships with specific cognitive deficits are not well understood.MethodsThis study included 102 older adults: 42 cognitively unimpaired (CU), 28 with mild cognitive impairment (MCI), and 32 with Alzheimer’s disease (AD) dementia. Each participant underwent a neuropsychological assessment. Regional brain α4β2 nicotinic cholinergic receptor binding (VT/fp) was measured using 2-[18F]fluoro-3-(2(S)azetidinylmethoxy)pyridine (2FA) and PET imaging. Voxel-wise analyses of group differences were performed. Relationships between receptor binding and cognition, age, and cholinesterase inhibitor medication use were assessed using binding values in six prespecified regions of interest.ResultsSPM analysis showed the group VT/fp binding differences in the bilateral entorhinal cortex, hippocampus, insula, anterior cingulate, thalamus, and basal ganglia (p < .05, FWE-corrected). Pairwise comparisons revealed lower binding in the AD group compared to the CU group in similar regions. Binding in the entorhinal cortex was lower in the MCI group than in the CU group; binding in the hippocampus was lower in the AD group than in the MCI group. AD participants taking cholinesterase inhibitor medication had lower 2FA binding in the bilateral hippocampus and thalamus compared to those not taking medication. In the CU group, age was negatively associated with 2FA binding in each region of interest (rs = − .33 to − .59, p < .05 for each, uncorrected). Attention, immediate recall, and delayed recall scores were inversely associated with 2FA binding in most regions across the full sample. In the combined group of CU and MCI participants, attention was inversely associated with 2FA binding in most regions, beyond the effect of hippocampal volume.ConclusionsNicotinic cholinergic receptor binding in specific limbic and subcortical regions is lower in MCI and further reduced in AD dementia, compared to CU older adults, and is related to cognitive deficits. Cognitive decline with age may be a consequence of reduced cholinergic receptor density or binding affinity that may also promote vulnerability to other Alzheimer’s processes. Contemporary modification of the “cholinergic deficit” of aging and AD may reveal opportunities to prevent or improve clinical symptoms.

Spatiotemporal mapping of sensory and motor innervation of the embryonic and postnatal mouse urinary bladder

The primary function of the urinary bladder is to store urine (continence) until a suitable time for voiding (micturition). These distinct processes are determined by the coordinated activation of sensory and motor components of the nervous system, which matures to enable voluntary control at the time of weaning. Our aim was to define the development and maturation of the nerve-organ interface of the mouse urinary bladder by mapping the organ and tissue distribution of major classes of autonomic (motor) and sensory axons. Innervation of the bladder was evident from E13 and progressed dorsoventrally. Increasing defasciculation of axon bundles to single axons within the muscle occurred through the prenatal period, and in several classes of axons underwent further maturation until P7. Urothelial innervation occurred more slowly than muscle innervation and showed a clear regional difference, from E18 the bladder neck having the highest density of urothelial nerves. These features of innervation were similar in males and females but varied in timing and tissue density between different axon classes. We also analysed the pelvic ganglion, the major source of motor axons that innervate the lower urinary tract and other pelvic organs. Cholinergic, nitrergic (subset of cholinergic) and noradrenergic neuronal cell bodies were present prior to visualization of these axon classes within the bladder. Examination of cholinergic structures within the pelvic ganglion indicated that connections from spinal preganglionic neurons to pelvic ganglion neurons were already present by E12, a time at which these autonomic ganglion neurons had not yet innervated the bladder. These putative preganglionic inputs increased in density prior to birth as axon terminal fields continued to expand within the bladder tissues. Our studies also revealed in numerous pelvic ganglion neurons an unexpected transient expression of calcitonin gene-related peptide, a peptide commonly used to visualise the peptidergic class of visceral sensory axons. Together, our outcomes enhance our understanding of neural regulatory elements in the lower urinary tract during development and provide a foundation for studies of plasticity and regenerative capacity in the adult system.

Changes Caused by Low Doses of Bisphenol A (BPA) in the Neuro-Chemistry of Nerves Located in the Porcine Heart.

Simple SummaryBisphenol A (BPA) is a substance commonly used in the plastics industry, which is a part of many everyday items. It may leach from plastics and penetrate food, water, soil and air. It is known that BPA negatively affects living organisms. It impairs the functions of the intestine, neurons, reproductive organs, endocrine glands and immune cells. Previous studies have also reported that BPA negatively influences the cardiovascular system, leading to heart arrhythmia, intensification of atherosclerosis, blood hypertension and increased risk of a heart attack. However, many aspects of the influence of BPA on the heart are still poorly understood. One of these aspects is the BPA impact on heart innervation. Therefore, this article aimed to investigate the influence of low doses of BPA on the number of nerves containing selected active substances taking part in neuronal stimuli conduction located in the porcine heart apex. The results indicate that even relatively low doses of BPA are not neutral to the cardiovascular system, because they affect the neurochemical characterization of nerves in the heart. These changes may underlie the negative effects of BPA on the heart.Bisphenol A (BPA) contained in plastics used in the production of various everyday objects may leach from these items and contaminate food, water and air. As an endocrine disruptor, BPA negatively affects many internal organs and systems. Exposure to BPA also contributes to heart and cardiovascular system dysfunction, but many aspects connected with this activity remain unknown. Therefore, this study aimed to investigate the impact of BPA in a dose of 0.05 mg/kg body weight/day (in many countries such a dose is regarded as a tolerable daily intake–TDI dose of BPA–completely safe for living organisms) on the neurochemical characterization of nerves located in the heart wall using the immunofluorescence technique. The obtained results indicate that BPA (even in such a relatively low dose) increases the number of nerves immunoreactive to neuropeptide Y, substance P and tyrosine hydroxylase (used here as a marker of sympathetic innervation). However, BPA did not change the number of nerves immunoreactive to vesicular acetylcholine transporter (used here as a marker of cholinergic structures). These observations suggest that changes in the heart innervation may be at the root of BPA-induced circulatory disturbances, as well as arrhythmogenic and/or proinflammatory effects of this endocrine disruptor. Moreover, changes in the neurochemical characterization of nerves in the heart wall may be the first sign of exposure to BPA.

Forensic characteristics of the cardio-toxic effect of organophosphorus compounds

Objective - to identify the morphological equivalents of the cardiotoxic action of organophosphate poisoning. It was studied 110 fatal organophosphate poisonings in toxicogenic and somatogenic stages. The study of the heart was a comprehensive in term of clinical, biochemical, histological, micromorphometric, histochemical and histoenzymological approaches. We have identified a complex of deep metabolic disorders and necrobiotic changes in the heart muscle and the level of cholinesterase activity inhibition in the cholinergic structures of the heart in the toxicogenic and somatogenic stages of fatal organophosphate poisoning.

Multimodal neuroimaging correlates of NREM and REM sleep: EEG spectral power in aging

AbstractBackgroundSleep disturbances have been associated with cognitive decline and Alzheimer’s disease (AD), with growing efforts to understand how these changes may relate to AD pathophysiology. However, we lack a comprehensive overview of the associations between objectively‐measured sleep oscillations and brain changes in aging so that efficient treatment strategies could be developed. Using an unbiased, fast, and powerful quantitative (spectral) analysis of sleep EEG rhythms, we assessed the relationships between both non‐rapid eye movement (NREM) and REM sleep oscillations, and amyloid burden, gray matter (GM) volume and perfusion in cognitively unimpaired older adults (CUOA).Method125 CUOA from the Age‐Well randomized controlled trial underwent a neuropsychological assessment, an ambulatory polysomnography recording and a structural MRI. A Florbetapir‐PET scan was also performed with early and late acquisitions to measure brain perfusion and amyloid burden respectively. Normalized EEG spectral power values were obtained using the ASEEGA® algorithm for delta (0.1‐4 Hz), theta (4‐8 Hz), alpha (8‐12 Hz) and beta (16‐50 Hz) EEG frequency bands, over the Cz‐Pz derivation (Berthomier et al., 2007). Voxel‐wise multiple regressions were then conducted between EEG spectral power values and neuroimaging data (i.e., GM volume, perfusion and amyloid burden), separately for NREM and REM sleep, controlling for age, gender, education, the apnea‐hypopnea index, sleep medication use, and the ApoE4 status.ResultLower NREM sleep delta power, reflecting a decrease in slow wave activity, was related to decreased GM volume and perfusion in fronto‐cingulate areas, but not to amyloid deposition. By contrast, decreased REM sleep theta power was associated with i) greater amyloid deposition in frontal, cingulate, parietal and temporal areas; ii) lower GM volume in frontal, temporal, occipital and insular cortices, and iii) increased perfusion in frontal and parietal regions.ConclusionBoth NREM and REM sleep alterations were associated with structural and functional changes in regions sensitive to aging and AD pathophysiology. Importantly, REM sleep changes were associated with greater amyloid deposition. This may be due to early changes in cholinergic structures, acetylcholine playing a neuroprotective role against amyloid. Thus older individuals with REM sleep alterations may be more at risk of belonging to the Alzheimer’s continuum.

Sex differences in the cardiac cholinergic response to adrenalin-induced myocardial necrosis and light desynchronosis

The aim was to identify sex differences in the cardiac response to cholinergic stimuli in the development of myocardial necrosis and light desynchronosis.Materials and methods. Experiments were conducted in 144 albino rats (72 males, 72 females). Myocardial necrosis was induced by adrenaline (0.5 mg/kg, intramuscularly).The intensity of bradycardia that occurred as a response to intravenous acetylcholine injection and electrical stimulation of vagus nerve was measured. The animals were divided into 4 groups: 1 – rats that were in a light balance (day/night); 2 – rats exposed to light deprivation for 10 days; 3 – rats exposed to permanent lighting for 10 days; 4 – animals exposed to permanent lighting for 10 days and injected with melatonin (5 mg/kg) intraperitoneally 1 hour before the necrosis modeling.Results. The development of myocardial necrosis when occurring in light deprivation was characterized by an increased responsiveness of the rat heart to cholinergic stimulation due to an increase in both cholinoreceptors sensitivity and release of acetylcholine from the vagus nerve terminals in females, and due to only an increased cholinoreceptors sensitivity in males. The development of myocardial necrosis in rats exposed to permanent lighting was characterized by significantly higher sensitivity of cholinergic structures of the heart than that in light balance, especially in females. Melatonin injection in the development of myocardial necrosis contributed to the heart cholinergic structures response to the stimulation, which was close to that observed in conditions of light balance, though it remained somewhat higher: due to the facilitation of acetylcholine release from vagus nerve in females, and it was combined with a higher sensitivity of cholinoreceptors in males.Conclusions. The development of myocardial necrosis in light desynchronosis (light deprivation or permanent lighting) causes an increased heart response to cholinergic stimulation in rats. Such effects are stronger, especially in females in conditions of permanent lighting. The injection of melatonin in rats with myocardial necrosis and permanent lighting results in an approximation (but not restoration) of the heart sensitivity to cholinergic stimuli parameters to those that are observed in light balance. However, the heart response to cholinergic stimulation remains higher, especially in males.

Volume Change in Frontal Cholinergic Structures After Traumatic Brain Injury and Cognitive Outcome.

The cholinergic nuclei in the basal forebrain innervate frontal cortical structures regulating attention. Our aim was to investigate if cognitive test results measuring attention relate to the longitudinal volume change of cholinergically innervated structures following traumatic brain injury (TBI). During the prospective, observational TBIcare project patients with all severities of TBI (n = 114) and controls with acute orthopedic injuries (n = 17) were recruited. Head MRI was obtained in both acute (mean 2 weeks post-injury) and late (mean 8 months) time points. T1-weighted 3D MR images were analyzed with an automatic segmentation method to evaluate longitudinal, structural brain volume change. The cognitive outcome was assessed with the Cambridge Neuropsychological Test Automated Battery (CANTAB). Analyses included 16 frontal cortical structures, of which four showed a significant correlation between post-traumatic volume change and the CANTAB test results. The strongest correlation was found between the volume loss of the supplementary motor cortex and motor screening task results (R-sq 0.16, p < 0.0001), where poorer test results correlated with greater atrophy. Of the measured sum structures, greater cortical gray matter atrophy rate showed a significant correlation with the poorer CANTAB test results. TBI caused volume loss of frontal cortical structures that are heavily innervated by cholinergic neurons is associated with neuropsychological test results measuring attention.

Cholinergic and adrenergic innervation of the pancreas in chinchilla (Chinchilla Laniger Molina).

Cholinergic and adrenergic innervation of the pancreas in chinchilla (Chinchilla Laniger Molina) was examined in this study. The pancreas is both an exocrine and endocrine gland with autonomic and sensory innervation presented by the numerous nerve fibers and small agglomerations of nerve cells. Investigations were performed on 16 adult chinchillas of both sexes. The material was collected immediately after death of the animals. Histochemical methods: AChE and SPG were used, in addition to routine technique of single and double immunohistochemical (IHC) staining using whole mount specimens and freezing sections with a thickness of 8 to 12 μm. In the immunofluorescence staining, primary antibodies directed against markers used to identify cholinergic - ChAT and VAChT, and adrenergic - DbH and TH neurons. Secondary antibodies were coupled to Alexa Fluor 488 and Alexa Fluor 555 fluorophores. Histochemical studies (AChE) revealed that chinchilla pancreatic cholinergic innervation consisted of ganglionic neurocytes and numerous nerve fibers. These structures are located in the parenchyma of the exocrine part of the organ in close proximity to blood vessels and are present within the walls of the pancreatic ducts and interstitial connective tissue. A delicate fiber network around the Langerhans islets was also observed. The most numerous cholinergic structures were found in the head and tail, and the least numbers were found in the body of the pancreas. The SPG method revealed that adrenergic fibers form a network in the adventitia of blood vessels, and individual fibers run throughout the pancreatic parenchyma. Moreover, adrenergic nerve fibers were observed around the ganglionic neurocytes. This innervation was similar in all parts of the investigated organ. IHC investigations allowed observations of both the cholinergic and adrenergic activities of autonomic nerve structures. Additionally, using ChAT/DbH double staining, colocalization of these substances was observed in the fibers of the pancreatic parenchyma that passed through the cholinergic ganglia. Colocalization of VAChT and TH was found in nerve fibers of the exocrine part, in the walls of blood vessels, and in individual nerve cells. Colocalization of ChAT/DbH and VAChT/TH was observed in the single nerve cells and in the small (2-3 cell) ganglia. ChAT- and DbH-immunopositive nerve fibers were found in the area of the islets of Langerhans. The results indicate a more intense cholinergic innervation of the chinchilla's pancreas, which is represented by both ganglia and nerve fibers, while adrenergic structures are mainly represented by fibers and only single neurocytes. This arrangement of the investigated structures in this species may imply a major role for hormonal control of exocrine secretion in rodents.

Иммуногистохимическое исследование иннервации надпочечника крысы

The aim of the study was to study the innervation of the cortical part and medulla of the rat adrenal gland using neuroimmunohistochemical markers. Original data on the innervation of the rat Wistar (n = 8) adrenal cortex and medulla were obtained using immunohistochemical detection of the PGP 9.5 protein, synaptophysin, tyrosine hydroxylase, peripherin and serotonin. It was established that all zones of the cortex (glomerular, bundle, and reticular) are innervated by nerve fibers of various neurotransmitter identity. In the cortex, the innervation of the glomerular zone is most pronounced. A thick terminal plexus of varicose axons around islets of endocrinocytes, arterioles and nerve cells was detected in this zone. Cholinergic nerve structures (plexus of preterminal nerve fibers and the terminal synaptic network, which consists of varicose parasympathetic axons located around groups of neuroendocrine cells and sinusoidal capillaries) prevail in the medulla. Parasympathetic and sympathetic intramural ganglia were found in the medulla. Chromaffin cells immunoreactive to various catecholamines and serotonin were found in the adrenal medulla. It was assumed that different sources of sympathetic innervation of the organ are present. These are the medulla's own neurons and sympathetic trunk neurons. Such innervation is important for the regulation of local blood circulation.The results obtained in this study can be used both in scientific research and in the diagnosis of adrenal diseases.

A Novel Antiserum Against a Predicted Human Peripheral Choline Acetyltransferase (hpChAT) for Labeling Neuronal Structures in Human Colon.

Choline acetyltransferase (ChAT), the enzyme synthesizing acetylcholine (ACh), has an exon-skipping splice variant which is expressed preferentially in the peripheral nervous system (PNS) and thus termed peripheral ChAT (pChAT). A rabbit antiserum previously produced against rat pChAT (rpChAT) has been used for immunohistochemistry (IHC) to study peripheral cholinergic structures in various animals. The present study was undertaken to develop a specific antiserum against a predicted human pChAT (hpChAT) protein. A novel mouse antiserum has been successfully raised against a unique 14-amino acid sequence of hpChAT protein. Our Western blot using this antiserum (termed here anti-hpChAT serum) on human colon extracts revealed only a single band of 47 kDa, matching the deduced size of hpChAT protein. By IHC, the antiserum gave intense staining in many neuronal cells and fibers of human colon but not brain, and such a pattern of staining seemed identical with that reported in colon of various animals using anti-rpChAT serum. In the antibody-absorption test, hpChAT-immunoreactive staining in human colon was completely blocked by using the antiserum pre-absorbed with the antigen peptide. Double immunofluorescence in human colon moreover indicated that structures stained with anti-hpChAT were also stained with anti-rpChAT, and vice versa. hpChAT antiserum allowed the identification of cell types, as Dogiel type cells in intramural plexuses, and fiber innervation of colon muscles and mucosae. The present results demonstrate the specificity and reliability of the hpChAT antiserum as a novel tool for immunohistochemical studies in human colon, opening venues to map cholinergic innervation in other human PNS tissues.

P68. A double-blind, sham-controlled phase-1b study of combined STN and NBM-DBS in PD and mild to moderate Parkinson’s dementia (DEMPARK-DBS)

Introduction Subthalamic nucleus (STN)-DBS has proven to be a safe and long-term effective treatment for advanced Parkinson’s disease (PD). One of the contraindications for DBS is marginal or definite cognitive decline, which excludes a relevant amount of PD patients otherwise eligible for this effective therapy. There is growing evidence from case or series studies of DBS in cholinergic brain structures to stabilize or improve cognitive function in dementia. We aim to provide proof of safety of combined STN and Nucleus Basalis Meynert (NBM) stimulation in treating advanced PD having mild to moderate dementia. Design/Method In a prospective, monocentric study with double-blinded, randomized delayed-activation of NMB-stimulation 12 patients with idiopathic PD (Hoehn and Yahr stage ⩾2 in medication off, aged 45–75, disease duration ⩾5 years, improve of PD symptoms by ⩾30% in a levodopa challenge) can be included when the following criteria are met: medical refractory motor fluctuations or tremor with eligibility for bilateral STN-DBS but presence of mild to moderate Parkinson’s disease dementia (PDD) as defined by the MDS consensus guidelines (Mini–Mental State Examination score: 10–24). Eligible patients will be implanted with a rechargeable DBS System (Boston Scientific Vercise TM) with electrodes placed bilaterally in each STN and NBM. All 4 electrodes will be connected via a splitter to a single pacemaker. To analyze sole effects of DBS surgery on motor state and cognition, STN-DBS will be activated after 4 ± 1 weeks in all subjects. After 12 weeks of active STN-DBS a further motor-functional and cognitive assessment will be performed before randomization to either 24-weeks of sham- or instant active NBM-DBS. Subjects randomized to NBM-sham will have a delayed activation of NBM-DBS after the 24-weeks-sham-period. Subsequent assessment will be performed for all subjects after 24 and 48 weeks of active NBM stimulation. Outcome parameters and assessments SAE and safety parameters; UPDRS, Clinical Dyskinesia Rating Scale; ADCS-CGIC & -ADL, Zarit-Burden Interview, SF-36; BDI-II, Starkstein Apathy Scale, PDQ-39, EQ-5D; Neuropsychiatric Inventory, ADAS-Cog, Verbal Fluency (RWT), Wisconsin Card Sorting test, Trail Making test, Stroop test (Victoria vs.), Symbol Digit Modalities test, Brief Test of Attention. Current state Up to now 4 patients could be enrolled and were already implanted. First results of STN-stimulation will be presented at the congress. Conclusion Our study will determine safety of additional NBM stimulation in PD patients with PDD and may give first insight if it can slow or improve cognitive decline. We therefore hope to encourage other centers to refer eligible patients.

Heart Rhythm Disturbances Caused by Injection of Cholinesterase Inhibitor Physostigmine to Rats during the Early Ontogeny.

Parameters of cardiac activity after administration of the cholinesterase inhibitor physostigmine were analyzed in newborn rats and on day 16 of postnatal development. The type of cardiovascular response to acetylcholine excess in newborns and 16-day-old rats were similar, but they significantly differed by the magnitude, which suggests that that maturation of cholinergic structures involved in the regulation of cardiac activity is completed during the early postnatal ontogeny.

Increased risk of developing schizophrenia in animals exposed to cigarette smoke during the gestational period

Cigarette smoking during the prenatal period has been investigated as a causative factor of obstetric abnormalities, which lead to cognitive and behavioural changes associated with schizophrenia. The aim of this study was to investigate behaviour and AChE activity in brain structures in adult rats exposed to cigarette smoke during the prenatal period. Pregnant rats were divided into non-PCSE (non-prenatal cigarette smoke exposure) and PCSE (prenatal cigarette smoke exposure) groups. On post-natal day 60, the rats received saline or ketamine for 7days and were subjected to behavioural tasks. In the locomotor activity task, the non-PCSE+ketamine and PCSE+ketamine groups exhibited increased locomotor activity compared with the saline group. In the social interaction task, the non-PCSE+ketamine and PCSE+ketamine groups exhibited an increased latency compared with the control groups. However, the PCSE+ketamine group exhibited a decreased latency compared with the non-PCSE+ketamine group, which indicates that the cigarette exposure appeared to decrease, the social deficits generated by ketamine. In the inhibitory avoidance task, the non-PCSE+ketamine, PCSE, and PCSE+ketamine groups exhibited impairments in working memory, short-term memory, and long-term memory. In the pre-pulse inhibition (PPI) test, cigarette smoke associated with ketamine resulted in impaired PPI in 3 pre-pulse (PP) intensity groups compared with the control groups. In the biochemical analysis, the AChE activity in brain structures increased in the ketamine groups; however, the PCSE+ketamine group exhibited an exacerbated effect in all brain structures. The present study indicates that exposure to cigarette smoke during the prenatal period may affect behaviour and cerebral cholinergic structures during adulthood.

CHARACTERISTIC OF BRAIN ELECTROBIOLOGICAL ACTIVITY IN ELDERLY WOMEN WITH DIFFERENT LEVEL OF COGNITIVE DISORDERS

The paper shows the study results of a brain electrobiological activity in elderly women with various level of cognitive impairment. Importance of early diagnostic of cognitive disorders and applicability of methods of functional brain imaging in revealing disorders in higher cortical functions are shown in the paper. To assess cognitive functions we applied express-method of evaluating of cognitive functions during normal aging. According to the test results three groups were formed: 1 - women without cognitive disorders, 2 -with mild cognitive impairment, and 3 - with moderate cognitive impairment. The EEG was registered for all participants by means of 128-channel system GES-300. Absolute spectral power for all bands was calculated. According to the spectral analysis it has been revealed that higher power of the slow-wave rhythms is characteristic of groups with disorders of cognitive functions. Prevalence of a delta rhythm in the general EEG has been also stated. These changes are perhaps caused by dysfunction of the activating cholinergic systems and cortical structures, and also both dyscirculatory and structural changes of brain matter.

Localization of choline acetyltransferase and tyrosine hydroxylase immunoreactivities in the superior colliculus of the microbat, Rhinolophus ferrumequinum.

The purpose of this study was to determine whether the superior colliculus (SC) of the microbat has the same neurochemical makeup as that of other mammals. We examined the organization of choline acetyltransferase (ChAT)- and tyrosine hydroxylase-immunoreactive (TH-IR) fibers/cells using standard immunohistochemistry with antibodies against ChAT and TH. ChAT-IR fibers observed in the superficial layers were denser than those in the deeper layers, and these fibers were classified into two types: small varicose fibers and large varicose fibers. ChAT-IR cells were predominantly located in the superficial layers with diverse morphologies. Among the well-known sources of cholinergic fibers in the mammalian SC, pedunculopontine tegmental nucleus (PPTN) and laterodorsal tegmental nucleus (LDTN) contained strongly labeled ChAT-IR cells, while no cholinergic structures were found in the parabigeminal nucleus (PBG) in the microbat brain. TH-immunoreactivity was found within fibers but not within cells. The density of TH-IR fibers was high in the zonal layer, moderate in the superficial gray and optic layers, and low in the deeper layers. Well-labeled TH-IR cells were also observed within area 13 and the locus coeruleus, known as the sources of catecholaminergic fibers in other mammalian SC. Although there are some cytoarchitectural variations among species, our results clearly showed elaborately organized ChAT-IR and TH-IR fibers/cells in the microbat SC. Our findings will contribute significantly to the understanding of actively constructed microbat visual systems.