Acetylcholinesterase Staining Research Articles

Role of cholinergic innervation in biliary remnants of patients with biliary atresia.

Biliary innervation is considered important in regulating the function of bile ducts, whereas the role of innervation in the hepatobiliary system of patients with biliary atresia (BA) remains unknown. This current study aims to investigate the role of innervation in biliary remnants and analyze the relationship between the innervation and prognosis of BA after surgery. Eighty-seven patients with type III BA who underwent the Kasai procedure were consecutively enrolled from January 2017 to September 2020. Innervation and ductules in remnants were examined by pathologists. Liver function, onset of cholangitis, jaundice clearance, and survival with the native liver were recorded. Patients were followed up for 24 months. The relationship between innervation and prognosis was analyzed. In total, 67 patients had bile drainage postoperatively, and 21 biliary remnants contained neuronal plexuses where there was no neuron but nerve fiber bundles. Acetylcholinesterase staining was positive in all plexuses. In patients with bile drainage, those with plexuses had improved postoperative liver function, significantly better jaundice clearance 3 or 6 months postoperatively (50.0% vs. 19.1%, or 90.0% vs. 63.8%, respectively), fewer episodes of early cholangitis (10.0% vs. 34.0%), and better survival (80.0% vs. 61.7%) compared to those without. In addition, a larger area of plexuses was associated with a larger area of ductules (R2 = 0.786, p = 0.000), less frequent (p = 0.000) and later cholangitis onset (p = 0.012), and better jaundice clearance (p = 0.063). Increased cholinergic innervation in biliary remnants may help reduce the onset of cholangitis and lead to better and earlier jaundice clearance. Thus, it improves the postoperative prognosis of patients with BA.

Study on the Action of Chia Seed Oil in Rats’ Model of Autism

AbstractThis article mainly focuses on elucidating the effect of chia seed oil (CSO) on autistic traits in offspring rats induced with valproic acid (VPA). Postnatal maturity monitoring are conducted on postnatal day 1–15, and behavioral tests are implemented on postnatal day 49, subsequently, body weight, tail and body length, oxidative stress, the activity of acetylcholinesterase, histopathological analysis, and immunohistochemical staining in the hippocampal serotonin are investigated to confirm the rat model of autism and evaluate the effects of different doses of CSO. Offspring exposed to VPA prenatally exhibits autism‐like behavior, delayed eye‐opening, olfactory dysfunction, slow growth, poor sociability, high anxiety, and oxidative stress. In addition, neuronal degeneration and cytoplasmic vacuolation are found, and positive areas of hippocampal serotonin are aggrandized. However, treatment with CSO, exceptionally high doses of CSO, significantly improves the poor growth and behavioral disorder, enhanced the protective effect on neurodevelopment, and reduced oxidative stress.Practical application: It can be deduced that CSO has antioxidant and neuroprotective effects, which can provide good protection for autism, especially since high doses of CSO have a significant impact.

VP.05 Innervation defect: new pathomechanism of centronuclear myopathy?

Mutations in <i>DNM2, BIN1</i> and <i>MTM1</i> cause centronuclear myopathy (CNM). T-tubule malformation and its resultant defects in excitation-contraction coupling has been suggested to be the pathomechanism of this disease. Recently, in biopsied skeletal muscles from some CNM patients, we noticed the circumferential localization of acetylcholinesterase (AChE) on the sarcolemma on histochemical stain for AChE, which may well be due to the failure of AChE clustering in neuromuscular junction (NMJ). In support of our notion, NMJ anomalies were recently described in <i>Dnm2</i>-mutant animals. To check whether circumferential AChE expression on the sarcolemma is associated with CNM, and if so, to know whether NMJ is structurally and electrophysiologically abnormal in patients with CNM. We re-reviewed muscle pathology slides of the 40 patients with genetically-confirmed CNM (34 <i>DNM2, 1BIN1</i>, and 5 <i>MTM1</i>) and evaluated the localization of AChE. We also analyzed the morphology of NMJ in CNM by electron microscopy (EM). We collected the data of repetitive nerve stimulation (RNS) study and the effects of administration of AChE inhibitors (AChE-I) in CNM patients from the physicians. Circumferential AChE activity on the sarcolemma was observed in all patients with CNM. The frequency of the fibers with circumferential AChE was 100% in all CNM_DNM2, CNM_BIN1 and CNM_MTM1. On EM, postsynaptic structure of NMJ was abnormal in CNM_DNM2 muscles. Decremental response was seen on RNS in all three patients with CNM_DNM2 who received RNS, indicating the presence of myasthenic feature in CNM. One of two patients who received AChE-I showed improvement in muscle manual testing. Abnormal NMJ formation and a failure in AChE clustering indicate defects in NMJ transmission in CNM skeletal muscles. Decremental muscle action potential amplitudes and effective AChE-I suggest that myasthenic features in CNM contribute to muscle weakness in this disease. NMJ transmission defect due to postsynaptic structural abnormality is a new pathomechanism of CNM.

Effects of Smart Drugs on Cholinergic System and Non-Neuronal Acetylcholine in the Mouse Hippocampus: Histopathological Approach.

In recent years, people in the United States and other countries have been using smart drugs, called nootropic or cognitive enhancers, to improve concentration and memory learning skills. However, these drugs were originally prescribed for attention-deficit hyperactivity disorder and dementia, and their efficacy in healthy people has not yet been established. We focused on acetylcholine in the hippocampus, which is responsible for memory learning, and elucidate the long-term effects of smart drugs on the neural circuits. Smart drugs were administered orally in normal young mice for seven weeks. The hippocampus was sectioned and compared histologically by hematoxylin and eosin (HE) staining, immunohistochemistry for acetylcholine, and immunoelectron microscopy. There were no significant changes in acetylcholinesterase staining. However, in HE, we found perivascular edema, and choline acetyltransferase staining showed increased staining throughout the hippocampus and new signal induction in the perivascular area in the CA3, especially in the aniracetam and α-glyceryl phosphoryl choline group. Additionally, new muscarinic acetylcholine receptor signals were observed in the CA1 due to smart drug intake, suggesting that vasodilation might cause neuronal activation by increasing the influx of nutrients and oxygen. Moreover, these results suggest a possible new mechanism of acetylcholine-mediated neural circuit activation by smart drug intake.

Diagnostic accuracy of calretinin and acetylcholinesterase staining of rectal suction biopsies in Hirschsprung disease examined by unexperienced pathologists

Rectal suction biopsy (RSB) is a gold standard for diagnosing Hirschsprung disease (HD). Calretinin staining of RSB is increasingly used by experienced pathologists due to non-complex examination and comparable diagnostic accuracy with acetylcholinesterase (AChE). However, the diagnostic accuracy of calretinin examined by unexperienced pathologists remains to be elucidated. Therefore, we aim to compare diagnostic accuracy of calretinin with AChE on RSB for diagnosing HD when examined by unexperienced pathologists. We prospectively analyzed sections from RSB stained with AChE + HE and calretinin. Blinded examination was done by five unexperienced pathologists (pathology residents) and three experienced pathologists (senior pediatric gastro-enterology pathologists) assessing for the presence of HD. Cases for the study included ones proven to be HD on resection specimens and cases without HD. Diagnostic accuracy was determined calculating area under the curve (AUC), sensitivity, specificity, likelihood ratio, and posttest probability. Fleiss’ kappa analysis was performed to assess interobserver agreement between reviewers. Eleven of 18 included patients (61%) were diagnosed with HD. Comparing the diagnostic accuracy of unexperienced pathologists, calretinin versus AChE + HE showed sensitivity of 80.0% versus 74.5% and specificity of 100% versus 65.4%, AUC of 0.87 (0.78–0.96) versus 0.59 (0.45–0.72). Unexperienced pathologists showed substantial agreement with calretinin (kappa 0.72 [0.61–0.84]) and fair agreement with AChE + HE (kappa 0.34 [0.23–0.44]). We found calretinin having higher diagnostic accuracy in diagnosing HD compared to AChE + HE when examined by unexperienced pathologists. Therefore, we recommend to use calretinin as the standard technique for staining RSB in diagnosing HD.

Cellular resolution anatomical and molecular atlases for prenatal human brains.

Increasing interest in studies of prenatal human brain development, particularly using new single‐cell genomics and anatomical technologies to create cell atlases, creates a strong need for accurate and detailed anatomical reference atlases. In this study, we present two cellular‐resolution digital anatomical atlases for prenatal human brain at postconceptional weeks (PCW) 15 and 21. Both atlases were annotated on sequential Nissl‐stained sections covering brain‐wide structures on the basis of combined analysis of cytoarchitecture, acetylcholinesterase staining, and an extensive marker gene expression dataset. This high information content dataset allowed reliable and accurate demarcation of developing cortical and subcortical structures and their subdivisions. Furthermore, using the anatomical atlases as a guide, spatial expression of 37 and 5 genes from the brains, respectively, at PCW 15 and 21 was annotated, illustrating reliable marker genes for many developing brain structures. Finally, the present study uncovered several novel developmental features, such as the lack of an outer subventricular zone in the hippocampal formation and entorhinal cortex, and the apparent extension of both cortical (excitatory) and subcortical (inhibitory) progenitors into the prenatal olfactory bulb. These comprehensive atlases provide useful tools for visualization, segmentation, targeting, imaging, and interpretation of brain structures of prenatal human brain, and for guiding and interpreting the next generation of cell census and connectome studies.

Limb Muscle Reinnervation with the Nerve-Muscle-Endplate Grafting Technique: An Anatomical Feasibility Study.

Background Peroneal nerve injuries results in tibialis anterior (TA) muscle paralysis. TA paralysis could cause “foot drop,” a disabling condition that can make walking difficult. As current treatment methods result in poor functional recovery, novel treatment approaches need to be studied. The aim of this study was to explore anatomical feasibility of limb reinnervation with our recently developed nerve-muscle-endplate grafting (NMEG) in the native motor zone (NMZ). Methods As the NMEG-NMZ technique involves in nerves and motor endplates (MEPs), the nerve supply patterns and locations of the MEP bands within the gastrocnemius (GM) and TA muscles of rats were investigated using Sihler's stain and whole-mount acetylcholinesterase (AChE) staining, respectively. Five adult rats underwent TA nerve transaction. The denervated TA was reinnervated by transferring an NMEG pedicle from the ipsilateral lateral GM. At the end of a 3-month recovery period, maximal muscle force was measured to document functional recovery. Results The results showed that the TA was innervated by the deep peroneal nerve. A single MEP band was located obliquely in the middle of the TA. The GM was composed of two neuromuscular compartments, lateral (GM-l) and medial (GM-m), each of which was innervated by a separate nerve branch derived from the tibial nerve and had a vertically positioned MEP band. The locations of MEP bands in the GM and TA muscles and nerve supply patterns demonstrated that an NMEG pedicle can be harvested from the GM-l and implanted into the NMZ within the TA muscle. The NMEG-NMZ pilot study showed that this technique resulted in optimal muscle force recovery. Conclusion NMEG-NMZ surgery is feasible for limb reinnervation. Specifically, the denervated TA caused by peroneal nerve injuries can be reinnervated with a NMEG from the GM-l.

Hirschsprung Disease.

Hirschsprung Disease.

Cholinergic preservation in the oldest-old prefrontal cortex.

The incidence of finding abnormal protein aggregates characteristic of neurodegeneration, such as β-amyloid (Aβ), tau, α-synuclein, and phosphorylated-TDP-43 (pTDP-43), increases with age. While a small degree of cognitive impairment is associated with the aging brain, however, a subpopulation of the oldest-old, individuals 85 years of age and older, maintain cognition comparable to a much younger individual. The rostral prefrontal (rPFC) is a region implicated in executive function and working memory, two aspects of cognition determined to decline with age. The role of the rPFC in cognition is thought to be mediated by its high cholinergic innervation. This study aims to evaluate the incidence of abnormal protein aggregates in the oldest-old and evaluate cholinergic system preservation as a potential mechanism of resilience. Human formalin-fixed rPFC and hippocampus from male and female cognitively intact individuals over the age of 80 will be examined compared to sex- and age-matched Alzheimer's disease. Neuropathological load will be examined with immunohistochemical staining using specific antibodies for Aβ, tau, α-synuclein, and pTDP-43. The presence of cholinergic dysfunction will be evaluated using histochemical staining for butyrylcholinesterase and acetylcholinesterase, and immunohistochemical staining with a specific antibody for choline acetyltransferase. Preliminary results indicate that the oldest-old exhibit a high prevalence of Aβ and tau deposition but not α-synuclein or pTDP-43 in the rPFC. Additionally, abnormal intraneuronal Aβ and abnormal tau deposition in the white matter was observed in a subset of the oldest-old rPFC. Examination of the hippocampus while provide more insight into the presence of co-pathologies such as early pTDP-43 deposition in limbic predominant age-related TDP-43 encephalopathy (LATE). The results of this study will further our understanding of the prevalence of abnormal protein deposition in the oldest-old and the potential mechanisms of resilience which prevents clinical correlates.

Abstract 13827: The Effect of Tanshinone IIA on Megakaryopoiesis in Immune Vasculitis

Introduction and Objective: Tanshinone IIA is an active component of Danshen (Radix Salviae Miltiorrhiza Bge), which has been used to treat hypercoagulation-related diseases for centuries. However, its role in megakaryopoiesis remains unclear. Therefore, the present study aimed to examine the effect of Tanshinone IIA on megakaryocytes and platelets in immune vasculitis, and to identify the underlying mechanism(s). Methods: Albino rabbits, aged 3-4 weeks, were used in the present study. Immune vasculitis was established by intravenous injection of BSA (2.5 ml/kg). Tanshinone IIA (5 mg/kg/d) was given through intravenous injection as well. Blood sample was collected from the ear vein and bone marrow was harvested from the femur. Megakaryocytes and CFU-MK were counted with Wright’s and acetylcholine esterase staining, respectively. The coagulation activity and IL-1β level were determined by ELISA. Megakaryocytic cell line CHRF was used to examine the effect of Tanshinone IIA (1-30 μg/mL) on apoptosis; the assay was performed with Annexin V/PI, Caspase 3 and JC-1 using flow cytometry. Results: In rabbits with immune vasculitis, the platelet count, platelet aggregation and prothrombin fragment were significantly increased when compared to their counterparts. Megakaryocytes and CFU-MK were also significantly increased in immune vasculitis. In addition, the level of IL-1β was significantly higher in rabbits with immune vasculitis, which was positively correlated with platelet count and platelet aggregation. Tanshinone IIA (5 mg/kg/d, 7d) significantly reduced the number of megakaryocytes, CFU-MK and platelets, and decreased the coagulation activity. It also significantly lowered the IL-1β level. In megakaryocytic cell line CHRF, Tanshinone IIA (1-30 μg/mL) induced dose-dependent apoptosis. Conclusion: The present study shows that not only platelets, but also megakaryocytes are involved in the pathogenesis of immune vasculitis. IL-1β contributes to the thrombocytosis by inducing megakaryopoiesis. Tanshinone IIA exerts anti-platelet, anti-inflammation and anti-megakaryocyte effects in immune vasculitis; the underlying mechanism may be related to its inhibitory effect on IL-1β and the proapoptotic effect on megakaryocytes.

Toxic effects of subacute exposure to acrylamide on motor endplates of the gastrocnemius in rats

Acrylamide (ACR) is a recognized toxin that is known to induce neurotoxicity in humans and experimental animals. This study aimed to investigate the toxic effects of subacute exposure of the motor endplate (MEP) of the gastrocnemius in rats to ACR. All rats were randomly divided into control, 9, 18, and 36 mg/kg ACR groups, and ACR was administered by gastric gavage for 21 days. The behavioral tests were performed weekly. On the 22nd day, the wet weight of the gastrocnemius was measured. The changes in muscle fiber structure, nerve endings, and MEP in the gastrocnemius were examined by hematoxylin–eosin (HE) and gold chloride staining. Acetylcholinesterase (AChE) content in the gastrocnemius was detected by AChE staining. The expression of AChE and calcitonin gene-related peptide was detected by immunohistochemistry and western blot. Rats exposed to ACR showed a significant increase in gait scores and hind limb splay distance compared with the control group, and the wet weight of the gastrocnemius was reduced, HE staining showed that the muscle fiber structure of the gastrocnemius became thin and the arrangement was dense with nuclear aggregation, gold chloride staining showed that nerve branches decreased and became thin, nerve fibers became short and light, the number of MEPs was decreased, the staining became light, and the structure was not clear. AChE staining showed that the number of MEPs was significantly reduced after exposure to ACR, the shape became small, and the AChE content decreased in a dose-dependent manner. Immunohistochemistry and western blot analysis results of the expression levels of AChE and CGRP showed a decreasing trend as compared to the control group with increasing ACR exposure dose. The reduction in protein levels may be the mechanism by which ACR has a toxic effect on the MEP in the gastrocnemius of rats.

Modulation of prefrontal cortex function by basal forebrain cholinergic and GABAergic neurons at the behavioral and molecular level.

The present research aimed to study the effects of selective immunolesions of cholinergic or gamma-aminobutyric acid (GABA)ergic neurons in the nucleus basalis magnocellularis (NBM) on memory function as well as cholinergic activity and the level of expression of glutamatergic [NR2B subunit of N-methyl-D-aspartate(NMDA)] receptors in the medial prefrontal cortex (mPFC) and hippocampus of behaviorally characterized rats. In behavioral experiments, working memory was assessed by a spatial alternation testing procedure in a plus-maze, and acquisition and retention of spatial memory was evaluated in a Morris water maze. The rats were divided into three groups: the NBM cholinergic, GABAergic immunolesioned groups and the normal control group. Cholin acetyltransferase or parvalbumin staining of the NBM and acetylcholinesterase staining of the mPFC and hippocampal sections were performed to visualize the effects of immunotoxins. The electrophoresis and immunoblotting were run to evaluate the effect of NBM lesions on the amount of the NR2B subunit of NMDA receptors. The results indicate that the immunolesion of cholinergic NBM neurons impair spatial working memory, as well as long-term spatial memory which is accompanied by significant changes in glutamatergic (the NR2B subunit of NMDA receptor) and cholinergic markers in the mPFC, whereas immunolesion of GABAergic NBM neurons does not affect long-term spatial memory, it does though cause the impairment of working memory with a reduction of the NMDA NR2B receptor signaling in the mPFC. The present results demonstrate that the cholinergic and GABAergic NBM cell groups play diverse and complementary roles and are integrated in distinct NBM-mPFC networks that may play different roles in mPFC memory function.

Examination of cortically projecting cholinergic neurons following exercise and environmental intervention in a rodent model of fetal alcohol spectrum disorders.

Up to 1 in 5 infants in the United States are exposed to alcohol prenatally, resulting in neurodevelopmental deficits categorized as fetal alcohol spectrum disorders (FASD). Choline supplementation ameliorates some deficits, suggesting that alcohol exposure (AE) perturbs cholinergic neurotransmission and development. Behavioral interventions, which upregulate cholinergic neurotransmission, rescue cognitive deficits in rodent models of FASD. We investigated the impacts of two interventions (either wheel-running (WR) or "super intervention," WR plus exposure to a complex environment) on cholinergic neuronal morphology in the nucleus basalis of Meynert (NBM), the source of cortical cholinergic input, and prefrontal cortex (PFC) in a rodent model of FASD. One third of the total 47 male pups received intragastric intubation of ethanol in milk substitute during postnatal days (PD) 4-9. Another third served as sham-intubated procedural controls while the final third served as suckle controls. Rats from each group were exposed to either intervention during PD 30-72. Choline acetyltransferase (ChAT+ ) and acetylcholinesterase staining were used to quantify cholinergic neuron number, soma volume, and axon number. Our data indicate a main effect of postnatal treatment on ChAT+ neuron number in NBM in adulthood. Post hoc analysis demonstrates that ChAT+ neuron number is reduced in AE compared to suckle control rodents (p < .01). We examined the cytoarchitectonics of cholinergic neurons in NBM and PFC in adulthood following early postnatal AE and two interventions. We show that AE reduces ChAT+ neuron number in NBM, and this is not mitigated by either intervention.

Distribution of acetylcholinesterase in the hippocampal formation of the Atlantic white-sided dolphin (Lagenorhynchus acutus).

The cetacean hippocampal formation has been noted to be one of the smallest relative to brain size of all mammals studied. This region, comprised of the dentate gyrus, hippocampus proper, subiculum, presubiculum, parasubiculum and the entorhinal cortex, is important in learning, memory, and navigation. There have been a number of studies detailing the distribution of acetylcholinesterase (AChE) in the hippocampal formation of terrestrial mammals with the goal of gaining a greater understanding of some aspects of the cholinergic innervation to this region, as well as its parcellation. The present study was undertaken to describe the organization, cytoarchitecture, and distribution of AChE in the hippocampal formation of the Atlantic white-sided dolphin (AWSD) with the view to understand similarities and differences between this aquatic mammal and terrestrial mammals. Nissl-staining demonstrated cytoarchitecture of the hippocampal formation in the AWSD comparable to that reported in other cetaceans. In addition, the AWSD had a rich pattern of AChE staining that distinctly varied between regions and laminae. A number of differences in the distribution of AChE staining in areas comparable to those of terrestrial species reported suggested possible alterations in connectivity of this region. Overall, however, AChE-staining suggested that cholinergic innervation, neural pathways and function of the hippocampal formation of the AWSD is conserved, similar to other mammals.

Acetylcholinesterase staining for the pathological diagnosis of Hirschsprung's disease.

Hirschsprung's disease (HD) is a congenital disease manifesting various degrees of functional bowel obstruction caused by the absence of enteric ganglion cells, which are usually absent in the colonic segment of the HD patient. Because the aganglionic segment of HD always includes the rectum, pathological diagnosis can be made using a rectal sample. HD should be diagnosed as early as possible because serious complications, such as acute enterocolitis or toxic megacolon, can develop without a definitive diagnosis and appropriate treatment. In the mid-1900s, HD was diagnosed by HE staining of specimens obtained by full-thickness biopsy. Since then, the combination of rectal mucosal biopsy and rubeanic acid-amplificated AChE staining has been brought about by the following milestones: the discovery that the submucosal plexus and the intermuscular plexus had the same level of nerve migration; the findings of research on acetylcholine (ACh) and acetylcholinesterase (AChE) in the intestinal tract; and the establishment of a rubeanic acid amplification method. Consequently, the diagnostic rate of HD improved dramatically in the 1980s. This review outlines the history of diagnostic methods for HD, the roles of ACh and AChE in the intestine, and the method of AChE staining.

Histochemical Staining of Acetylcholinesterase in Carnolian Honeybee (Apis mellifera carnica) Brain after Chronic Exposure to Organophosphate Diazinon

Abstract Organophosphate insecticides are known to inhibit the activity of enzyme acetylcholinesterase. They affect olfactory learning and memory formation in honeybees. These insecticides cause mushroom body inactivation in honeybees, but their influence on other brain regions involved in olfactory perception and memory is unknown. The goal of this study was to study the effects of organophosphate insecticide diazinon on carnolian honeybee (Apis mellifera carnica) acetylcholinesterase activity in the olfactory brain regions of antennal lobe, mushroom body and lateral procerebrum (lateral horn). The lamina, medulla and lobula of optic lobes were also analyzed. The level of acetylcholinesterase activity was visualized using the histochemical staining method. Densitometric analysis of histochemical signals indicated that diazinon inhibited acetylcholinesterase activity only in the lip of calyces of mushroom body, but not in other analyzed olfactory regions, antennal lobe and lateral procerebrum. The visual brain system optic lobes were also unaffected. This is in accordance with the literature reporting that mushroom body is the main brain center for olfactory learning and memory formation in honeybees.

Congenital hyperinsulinism associated with Hirschsprung\u2019s disease\u2014a report of an extremely rare case

BackgroundCongenital hyperinsulinism (CH) is a rare disease, characterized by severe hypoglycemia induced by inappropriate insulin secretion from pancreatic beta-cells in neonate and infant. Hirschsprung’s disease (HD) is also a rare disease in which infants show severe bowel movement disorder. We herein report an extremely rare case of combined CH and HD.Case presentationThe patient was a full-term male infant who showed poor feeding, vomiting, and hypotonia with lethargy on the day of birth. He was transferred to tertiary hospital after a laboratory analysis revealed hyperinsulinemic hypoglycemia. The patient showed remarkable abdominal distension without meconium defecation. An abdominal X-ray showed marked dilatation of the large bowel. He was diagnosed with CH (nesidioblastosis) associated with suspected HD. He was initially treated with an intravenous infusion of high-dose glucose with the intermittent injection of glucagon. This was successfully followed by treatment with diazoxide and octreotide (a somatostatin analog). At 8 months of age, HD was confirmed by the acetylcholinesterase staining of a rectal mucosal biopsy specimen, and a transanal pull-through operation was performed to treat HD. At 14 months of age, subtotal pancreatectomy was performed for the treatment of focal CH located in the pancreatic body. His postoperative course over the past 12 years has been uneventful without any neurologic or bowel movement disorders.ConclusionsAlthough it is extremely rare for CH to be associated with HD, associated HD should be considered when a patient with CH presents severe constipation.

Cholinergic and inflammatory phenotypes in transgenic tau mouse models of Alzheimer's disease and frontotemporal lobar degeneration.

An early and sizeable loss of basal forebrain cholinergic neurons is a well-characterized feature associated with measurable deficits in spatial learning and cognitive impairment in patients with Alzheimer’s disease. In addition, pro-inflammatory glial cells such as astrocytes and microglia may play a key role in the neurodegenerative cascade of Alzheimer’s disease and tauopathies. We recently presented two mouse models: Line 1, expressing the truncated tau fragment identified as the core of the Alzheimer’s paired helical filament, and Line 66, expressing full-length human tau carrying a double mutation (P301S and G335D). Line 1 mice have a pathology that is akin to Alzheimer’s, whilst Line 66 resembles frontotemporal lobar degeneration. However, their cholinergic and inflammatory phenotypes remain elusive. We performed histological evaluation of choline acetyltransferase, acetylcholinesterase, p75 neurotrophin receptor, microglial ionized calcium binding adaptor molecule 1 and astrocytic glial fibrillary acidic protein in the basal forebrain, hippocampus and cortex of these models. A significant lowering of choline acetyltransferase-positive neurons and p75-positive neurons in the basal forebrain of Line 1 at 3, 6 and 9 months was observed in two independent studies, alongside a significant decrease in acetylcholinesterase staining in the cortex and hippocampus. The reductions in choline acetyltransferase positivity varied between 30% and 50% at an age when Line 1 mice show spatial learning impairments. Furthermore, an increase in microglial ionized calcium binding adaptor molecule 1 staining was observed in the basal forebrain, hippocampus and entorhinal cortex of Line 1 at 6 months. Line 66 mice displayed an intact cholinergic basal forebrain, and no difference in p75-positive neurons at 3 or 9 months. In addition, Line 66 exhibited significant microglial ionized calcium binding adaptor molecule 1 increase in the basal forebrain and hippocampus, suggesting a prominent neuroinflammatory profile. Increased concentrations of microglial interleukin-1β and astrocytic complement 3 were also seen in the hippocampus of both Line 1 and Line 66. The cholinergic deficit in Line 1 mice confirms the Alzheimer’s disease-like phenotype in Line 1 mice, whilst Line 66 revealed no measurable change in total cholinergic expression, a phenotypic trait of frontotemporal lobar degeneration. These two transgenic lines are therefore suitable for discriminating mechanistic underpinnings between the Alzheimer’s and frontotemporal lobar degeneration-like phenotypes of these mice.

Постнатальное развитие комплекса заднелатеральных ядер дорзального таламуса

Using histochemical stain for acetylcholinesterase (AChE), the lateral posterior pulvinar complex (LP-P) was analyzed in kittens aged 0–123 postnatal days and in adult cats. In the lateral nucleus of the complex (LPl) of kittens aged 4–21 days, a transient clustered AChE staining of neuropil was revealed. The average size of AChE clusters varied between 200 and 300 mkm; the inter-cluster distance varied between 430 and 600 mkm. Optical contrast of clusters increased rapidly from the 4th to the 14th postnatal day and, thereafter, rapidly decreased up to the 21st-28th postnatal day. Since AChE is directly linked to neuronal differentiation and development and AChE clusters are located in the same area with neurons responsible for thalamocortical connections, the transient clusters of AChE expression could be related to the development of visual structures.

Herbal complex 'Buyang Huanwu Tang' improves motor endplate function of denervated-dependent skeletal muscle atrophy in rat.

Denervated-dependent skeletal muscle atrophy is a disease induced by skeletal muscle associated peripheral neuro-disconnection. Its specific molecular mechanisms remain unknown. The treating for denervated-dependent skeletal muscle atrophy is applied with an herbal complex Buyang Huanwu Tang used in traditional Chinese medicine and subjected to the established denervated-dependent skeletal muscle atrophy in rat models, and the therapeutic effects and associated mechanisms were evaluated in the pathogenesis of denervated-dependent skeletal muscle atrophy. Denervated-dependent skeletal muscle atrophy in rats was established and randomly divided into eight groups, including Normal control, Model, Positive control, Model + Buyang Huanwu Tang, Model + astragalus extracts, Model + Buyang Huanwu Tang-astragalus, Buyang Huanwu Tang + LY294002, and astragalus extract + LY294002 group. Hematoxylin-eosin staining and quantitative RT-PCR (qRT-PCR) assay were used to examine the inflammatory response of muscle tissues. Quantitative RT-PCR and Western blotting assay were utilized to analyze mRNA and protein expression. Immunohistochemistry assay was used to detect molecule expression in anterior cervical muscle tissues. Motor endplate activity was examined using the wholemount acetylcholinesterase staining method. The wet mass ratio of anterior cervical muscle was measured. The results indicated that Buyang Huanwu Tang treatment significantly alleviated inflammatory response, enhanced acetylcholinesterase activity, and motor endplate functions, and promoted wet mass of anterior cervical muscle compared to denervated-dependent skeletal muscle atrophy rat models (P < 0.05). Buyang Huanwu Tang regulated molecules of PI3K/PKB/GSK3β/FOXO1 signaling pathway. Buyang Huanwu Tang significantly reduced muscle atrophy F-box protein, MuFR-1, Bax and caspase 9 expression, significantly enhanced Bcl-2 expression, and remarkably increased element-binding protein and vascular endothelial growth factor levels, compared to Model group (P < 0.05). Buyang Huanwu Tang suppressed caspase 9 and caspase 3 activity and associated apoptosis. Moreover, PI3K specific blocker, LY294002, significantly inhibited the effects of Buyang Huanwu Tang on the above molecule expression (P < 0.05). In conclusion, Buyang Huanwu Tang improved motor endplate functions of denervated-dependent skeletal muscle atrophy rat model through suppressing mitochondria-mediated apoptosis and activating PI3K/PKB/FOXO1 signaling pathway.