Hyperactivity and Hypermotivation Associated With Increased Striatal mGluR1 Signaling in a Shank2 Rat Model of Autism

Mutations in the SHANK family of genes have been consistently identified in genetic and genomic screens of autism spectrum disorder (ASD). The functional overlap of SHANK with several other ASD-associated genes suggests synaptic dysfunction as a convergent mechanism of pathophysiology in ASD. Although many ASD-related mutations result in alterations to synaptic function, the nature of those dysfunctions and the consequential behavioral manifestations are highly variable when expressed in genetic mouse models. To investigate the phylogenetic conservation of phenotypes resultant of Shank2 loss-of-function in a translationally relevant animal model, we generated and characterized a novel transgenic rat with a targeted mutation of the Shank2 gene, enabling an evaluation of gene-associated phenotypes, the elucidation of complex behavioral phenotypes, and the characterization of potential translational biomarkers. The Shank2 loss-of-function mutation resulted in a notable phenotype of hyperactivity encompassing hypermotivation, increased locomotion, and repetitive behaviors. Mutant rats also expressed deficits in social behavior throughout development and in the acquisition of operant tasks. The hyperactive phenotype was associated with an upregulation of mGluR1 expression, increased dendritic branching, and enhanced long-term depression (LTD) in the striatum but opposing morphological and cellular alterations in the hippocampus (HP). Administration of the mGluR1 antagonist JNJ16259685 selectively normalized the expression of striatally mediated repetitive behaviors and physiology but had no effect on social deficits. Finally, Shank2 mutant animals also exhibited alterations in electroencephalography (EEG) spectral power and event-related potentials, which may serve as translatable EEG biomarkers of synaptopathic alterations. Our results show a novel hypermotivation phenotype that is unique to the rat model of Shank2 dysfunction, in addition to the traditional hyperactive and repetitive behaviors observed in mouse models. The hypermotivated and hyperactive phenotype is associated with striatal dysfunction, which should be explored further as a targetable mechanism for impairment in ASD

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Synaptic pathways related to Shank3 and its interaction partner ProSAPiP1

Autism spectrum disorders (ASD) affect approximately 1% of the human population. The underlying mechanisms for disease formation and pathology have been addressed in many studies. Due to the high complexity of ASD, however, these mechanisms are still not completely understood. The three SHANK genes are frequently mutated in ASD patients with the highest contribution of SHANK3. Mutations in SHANK3 are causing several neuropsychiatric diseases including Phelan-McDermid syndrome, ASD and intellectual disability. Many studies investigated molecular alterations in Shank3 mutant mouse models. However, a comprehensive approach analysing an unbiased set of proteins has not yet been performed in Shank3 mutant mouse models. In this study, we established a new protocol to isolate the postsynaptic density (PSD) fraction from mouse brain tissue. Applying this protocol, we generated striatal and hippocampal PSD fractions of wild type and Shank3Δ11-/- mutant mice to compare the molecular setup of their PSD. Therefore, we performed ion-mobility enhanced data-independent label-free LC-MS/MS on the isolated PSD fractions to obtain an unbiased and comprehensive dataset of molecular changes in vivo in striatal and hippocampal PSD fractions lacking the major isoforms of Shank3. In the absence of these Shank3 isoforms, we identified brain region specific molecular alterations in the PSD, indicating disruptions in striatal glutamatergic synaptic transmission, while in the hippocampal PSD of Shank3Δ11-/- mutant mice mainly cytoskeleton-associated functions and processes were affected. Postsynaptic Homer1 was the only protein found significantly altered in both brain regions. We observed reductions in Homer1 in both brain regions, which are in line with previous observations in Shank3Δ11-/- and other Shank3 mutant mouse models. Our study is the first comprehensive analysis of brain region specific PSD proteomes of Shank3 mutant mice. We provide information on Shank3Δ11-/- mutant PSD composition, which will be important for further Shank3 based studies and pharmaceutical treatment experiments. Furthermore, we investigated functional aspects of ProSAP-interacting protein 1 (ProSAPiP1), a member of the Fezzin protein family and interaction partner of Shank3 and Spine-associated RapGAP (SPAR) with yet unknown function. We virally knocked down and overexpressed ProSAPiP1 to analyse functional consequences in primary hippocampal neurons. While general synapse numbers and the protein levels of main PSD scaffold proteins Shank3 and PSD95 were not altered, we observed that ProSAPiP1 regulates the postsynaptic protein levels of its interaction partner SPAR. In line with previously described consequences of alterations in postsynaptic SPAR protein levels, we observed an impact of ProSAPiP1 protein levels on dendritic spine maturation. Our results therefore suggest that ProSAPiP1 regulates SPAR levels to modulate dendritic spine maturity

Evaluation of the Objectives of Teaching Practice in Athletic Associations for Physical Education Teaching Students Based on the General Framework of Athletic Associations and Other Extramural Factors

Durch Kultusministerium und BLSV wurde 1997 ein Vereinspraktikum ins Leben gerufen mit dem Ziel, bei Studierenden bereits in der Ausbildungsphase eine Sensibilisierung für die Zusammenarbeit zwischen Schule und Verein zu erreichen und gleichzeitig die berufliche Eignung durch Erprobung der sportpädagogischen Begabung zu testen. Zu überprüfen galt es, ob Studierende durch das Praktikum in die Lage versetzt werden, diese beiden angestrebten Zielsetzungen zu erfüllen. Dazu war es in einem ersten Schritt notwendig, Kriterien für die Zielerreichung zu bilden. Praktika haben bei Studierenden einen hohen Stellenwert. Untersuchungen haben ergeben, dass Praktika nicht nur positive, sondern auch negative Wirkungen mit sich bringen können. Um die positiven Wirkungen zu vergrößern, müssen Vorschriften für die Gestaltung und Begleitung der Praktika einfließen. Kooperationsprogramme erfüllen unterschiedliche Funktionen. Es werden ihnen unterstützende und innovative Funktionen für Schule und Verein zugeschrieben. Die kompensative Funktion wird aber auch kritisch bewertet. Um eine partnerschaftliche Programmentwicklung zu fördern, sind Kriterien wie z.B. Vertrauen und Überzeugung nötig. Auf dem Gebiet der Begabungsforschung dominieren derzeit die Person-Umwelt-Interaktionsmodelle. Danach ist Begabung ein nach Anlage und Umwelt determiniertes, dynamisches Produkt von persönlichkeitsbildenden Lernprozessen. Pädagogisch begabte Menschen treten dadurch in Erscheinung, dass sie intuitiv eine komplexe Sache und soziale Beziehungen zugleich erkennen und handhaben. Voraussetzung hierfür sind inhaltliche und soziale Fähigkeiten. Die Untersuchung sollte zeigen, wie das Praktikum auf der Grundlage von unterschiedlichen Rahmenbedingungen wirkt und ob dabei die festgelegten Ziele verwirklicht werden können. Zu diesem Zweck wurde im SS 03 an allen bayerischen Sportinstituten eine schriftliche Befragung durchgeführt, die zu folgenden Erkenntnissen führte: Ein Großteil der Studierenden ist bereits als aktives Mitglied im wettkampforientierten Vereinssport organisiert. Das Praktikum wird daher hauptsächlich im Heimatverein absolviert. Oft stimmen Sportarten im Praktikum mit der betriebenen Vereinssportart überein. Zum Themenbereich Kooperation zwischen Schule und Verein werden nur wenige Informationen und Einblicke gegeben. In den Unterrichtsversuchen sind die Praktikanten auf eigene Erfahrungen angewiesen. Betreuung von Vereinsseite erfolgt selten. Auf Grund dieser Rahmenbedingungen lässt sich die Wirkung mit Hilfe eines faktoranalytischen Verfahrens anhand von sechs Faktoren verdeutlichen. Wirkungen werden in den Bereichen soziale Selbsterfahrung, Wissen über Verein, Selbstbestätigung, Interesse an Vereinsarbeit, Perspektiven/Einblicke und Bestätigung der Berufswahl erzielt. Dabei werden die Wirkungen durch unterschiedliche vereinsinterne Bedingungen signifikant beeinflusst. Das Praktikum scheint ein geeignetes Mittel zum Test der eigenen Fähigkeiten und zur Bestätigung der Berufswahl zu sein. Allerdings zeichneten sich dabei keine erkennbaren Wirkungen hinsichtlich einer Sensibilisierung für die Zusammenarbeit zwischen Schule und Verein ab. Von der Grundidee stellen sich die beiden Ziele positiv dar, sind aber nur schwer miteinander zu vereinbaren. Um sinnvolle Erfahrungen hinsichtlich der sportpädagogischen Begabung zu gewährleisten, lässt sich das Praktikum nur eingeschränkt abwechslungsreich gestalten. Auf Grund der freien Gestaltungsmöglichkeit liefert es ein recht einseitiges Bild dieser Begabung. Der Beitrag des Praktikums zur Förderung der Kooperation Schule und Verein muss kritisch betrachtet werden. Studierende müssten bei Initiierung und Betreuung der Kooperationen das Vertrauen und die Überzeugung gewinnen, dass sie nicht zum Kompensieren all der Mängel dienen, die an Schul- wie Vereinssport derzeit erkennbar sind. Qualitativ hochwertige Kooperationen sind vorwiegend leistungssportlich orientiert. Breitensportliche Bereiche sind eher unterrepräsentiert. Studierende orientieren sich vorwiegend im leistungssportlichen Bereich. Die Vielschichtigkeit von Kooperationen wird durch diese vereinsexternen Bedingungen kaum erfüllt. Hier müsste eine Sensibilisierung durch explizite Steuerung für breitensportliche Belange geweckt werden. Die erhobenen Daten belegen den defizitären Informationsstand von Vereinen und Praktikanten und die mangelnde kritische Auseinandersetzung mit diesen Themen. Es entsteht der Eindruck, dass die Ergebnisse, die eine Sensibilisierung erkennen lassen, eine soziale Erwünschtheit widerspiegeln. Für eine bessere Umsetzung der Zielsetzungen sind organisatorische Veränderungen überlegenswert.In 1997, the German Ministry for Culture and the BLSV (Bavarian State Sports Association) introduced compulsory teaching practice in athletic associations for physical education teaching students. The aim was to sensitize students to the working environment involving both schools and athletic associations while still in the training phase and to simultaneously check the students’ suitability for the profession by putting their aptitude for teaching PE to the test. It was necessary to assess whether this teaching practice puts students in a position to fulfil both the above objectives. In order to make such an assessment, the first step had to be to determine criteria which signalled the achievement of the objectives. Working practice and internships are highly significant for students. Investigations have shown that such practice periods can have not only positive, but also negative effects. Regulations concerning the structure and supervision of teaching practice must therefore be created to enhance the positive effects. Cooperation programmes fulfil various functions; ostensibly of a supportive and innovative nature for schools and athletic associations. The compensative role, however, is also subject to critical assessment. In order to foster the development of partner programmes, criteria such as trust and conviction must be fulfilled. Currently dominant in the field of aptitude research is the ‚individual-environment-interaction model’. According to this model, aptitude is a dynamic product of personality-forming learning processes determined by disposition and environment. People with an aptitude for teaching appear to intuitively and simultaneously recognize and deal with complex matters and social relationships. The prerequisites for this are textual and social abilities. The investigation should show the effects of teaching practice based on various general conditions and whether the determined objectives can be realized. To this end, a written survey was conducted at all Bavarian sports institutes in the summer semester 2003 which resulted in the following findings: The majority of students are already active members of competition-orientated athletic associations. Teaching practice is therefore primarily undertaken at the students’ home-town athletic associations. The types of sports involved in the teaching practice often coincide with those run by the association. Little information is given and few insights are gained concerning the topic of cooperation between schools and associations. During teaching practice lessons, the students have to rely on their own experiences. Supervision is rarely supplied by the association. Due to this general framework, the effects can be clarified with the aid of a factor analytic procedure using six factors. Effects are achieved in the fields of social self-awareness, knowledge about the association, self-affirmation, interest in the association’s work, perspectives/insights and affirmation of the choice of profession. These effects are significantly influenced by various intramural conditions. Teaching practice appears to be a suitable method for students to test their own abilities and to affirm their vocation. Admittedly, recognizable effects regarding sensitization to cooperation between schools and associations were not apparent. Both objectives are fundamentally positive, yet difficult to combine. In order to guarantee meaningful experiences with regard to PE teaching aptitude, teaching practice can only be structured with limited diversity. Due to the freedom of structure, the picture created of a student’s talent is rather one-sided. The contribution of teaching practice to promoting cooperation between schools and associations has to be critically examined. When cooperation programmes are initiated and supervised, students should gain the trust and conviction that they do not serve to compensate all the deficits currently apparent in school sports and athletic associations. Qualitative cooperation is primarily competitive-sport oriented. Amateur areas are rather underrepresented. Students orient themselves primarily in the field of competitive sport. The diversity of cooperation is hardly fulfilled due to these extramural conditions. Sensitization should be inspired here through explicit supervision of amateur sport matters. The submitted data prove the deficient state of information in athletic associations and students undertaking teaching practice and the lack of critical discussion about these topics. The overriding impression is that the results which reveal sensitization actually reflect social desirability. Organizational changes are worth considering in order to better realize the objectives

The Shank3 interaction partner ProSAPiP1 regulates postsynaptic SPAR levels and the maturation of dendritic spines in hippocampal neurons

The postsynaptic density or PSD is a submembranous compartment containing a wide array of proteins that contribute to both morphology and function of excitatory glutamatergic synapses. In this study, we have analyzed functional aspects of the Fezzin ProSAPiP1, an interaction partner of the well-known PSD proteins Shank3 and SPAR. Using lentiviral-mediated overexpression and knockdown of ProSAPiP1, we found that this protein is dispensable for the formation of both pre- and postsynaptic specializations per se. We further show that ProSAPiP1 regulates SPAR levels at the PSD and the maturation of dendritic spines. In line with previous findings on the ProSAPiP1 homologue PSD-Zip70, we conclude that Fezzins essentially contribute to the maturation of excitatory spine synapses

Selective Localization of Shanks to VGLUT1-Positive Excitatory Synapses in the Mouse Hippocampus

Members of the Shank family of multidomain proteins (Shank1, Shank2, and Shank3) are core components of the postsynaptic density (PSD) of excitatory synapses. At synaptic sites Shanks serve as scaffolding molecules that cluster neurotransmitter receptors as well as cell adhesion molecules attaching them to the actin cytoskeleton. In this study we investigated the synapse specific localization of Shank1-3 and focused on well-defined synaptic contacts within the hippocampal formation. We found that all three family members are present only at VGLUT1-positive synapses, which is particularly visible at mossy fiber contacts. No costaining was found at VGLUT2-positive contacts indicating that the molecular organization of VGLUT2-associated PSDs diverges from classical VGLUT1-positive excitatory contacts in the hippocampus. In light of SHANK mutations in neuropsychiatric disorders, this study indicates which glutamatergic networks within the hippocampus will be primarily affected by shankopathies

Table_1_Hyperactivity and Hypermotivation Associated With Increased Striatal mGluR1 Signaling in a Shank2 Rat Model of Autism.xlsx

<p>Mutations in the SHANK family of genes have been consistently identified in genetic and genomic screens of autism spectrum disorder (ASD). The functional overlap of SHANK with several other ASD-associated genes suggests synaptic dysfunction as a convergent mechanism of pathophysiology in ASD. Although many ASD-related mutations result in alterations to synaptic function, the nature of those dysfunctions and the consequential behavioral manifestations are highly variable when expressed in genetic mouse models. To investigate the phylogenetic conservation of phenotypes resultant of Shank2 loss-of-function in a translationally relevant animal model, we generated and characterized a novel transgenic rat with a targeted mutation of the Shank2 gene, enabling an evaluation of gene-associated phenotypes, the elucidation of complex behavioral phenotypes, and the characterization of potential translational biomarkers. The Shank2 loss-of-function mutation resulted in a notable phenotype of hyperactivity encompassing hypermotivation, increased locomotion, and repetitive behaviors. Mutant rats also expressed deficits in social behavior throughout development and in the acquisition of operant tasks. The hyperactive phenotype was associated with an upregulation of mGluR1 expression, increased dendritic branching, and enhanced long-term depression (LTD) in the striatum but opposing morphological and cellular alterations in the hippocampus (HP). Administration of the mGluR1 antagonist JNJ16259685 selectively normalized the expression of striatally mediated repetitive behaviors and physiology but had no effect on social deficits. Finally, Shank2 mutant animals also exhibited alterations in electroencephalography (EEG) spectral power and event-related potentials, which may serve as translatable EEG biomarkers of synaptopathic alterations. Our results show a novel hypermotivation phenotype that is unique to the rat model of Shank2 dysfunction, in addition to the traditional hyperactive and repetitive behaviors observed in mouse models. The hypermotivated and hyperactive phenotype is associated with striatal dysfunction, which should be explored further as a targetable mechanism for impairment in ASD.</p

Image_2_Hyperactivity and Hypermotivation Associated With Increased Striatal mGluR1 Signaling in a Shank2 Rat Model of Autism.TIF

<p>Mutations in the SHANK family of genes have been consistently identified in genetic and genomic screens of autism spectrum disorder (ASD). The functional overlap of SHANK with several other ASD-associated genes suggests synaptic dysfunction as a convergent mechanism of pathophysiology in ASD. Although many ASD-related mutations result in alterations to synaptic function, the nature of those dysfunctions and the consequential behavioral manifestations are highly variable when expressed in genetic mouse models. To investigate the phylogenetic conservation of phenotypes resultant of Shank2 loss-of-function in a translationally relevant animal model, we generated and characterized a novel transgenic rat with a targeted mutation of the Shank2 gene, enabling an evaluation of gene-associated phenotypes, the elucidation of complex behavioral phenotypes, and the characterization of potential translational biomarkers. The Shank2 loss-of-function mutation resulted in a notable phenotype of hyperactivity encompassing hypermotivation, increased locomotion, and repetitive behaviors. Mutant rats also expressed deficits in social behavior throughout development and in the acquisition of operant tasks. The hyperactive phenotype was associated with an upregulation of mGluR1 expression, increased dendritic branching, and enhanced long-term depression (LTD) in the striatum but opposing morphological and cellular alterations in the hippocampus (HP). Administration of the mGluR1 antagonist JNJ16259685 selectively normalized the expression of striatally mediated repetitive behaviors and physiology but had no effect on social deficits. Finally, Shank2 mutant animals also exhibited alterations in electroencephalography (EEG) spectral power and event-related potentials, which may serve as translatable EEG biomarkers of synaptopathic alterations. Our results show a novel hypermotivation phenotype that is unique to the rat model of Shank2 dysfunction, in addition to the traditional hyperactive and repetitive behaviors observed in mouse models. The hypermotivated and hyperactive phenotype is associated with striatal dysfunction, which should be explored further as a targetable mechanism for impairment in ASD.</p