Role of subplate neurosecretion in early cortical circuit formation

Lead Research Organisation: University of Oxford
Department Name: Physiology Anatomy and Genetics

Abstract

The developing brain is not simply a smaller version of the adult's, but one that has a completely different composition, connectivity, and logic of processing information. The neuronal circuits in the neonate are set up to interpret neural signals while the construction of the brain is still ongoing, with certain cell populations present only during such formation stages. These early cells and their functions are key components for the eventual maturation of the brain similarly to the dynamic transient scaffolds used for the construction of complex buildings.
We have a long-standing interest in studying these early-born cells and their involvement in both the normal and abnormal development of the brain. In particular, we focus on a transient compartment characterized by transient neuronal population and temporary synaptic connections called the subplate. Subplate cells are amongst the first born in the cerebral cortex, they initially represent a large cell group distributed in a zone between the site of neurogenesis and their final destination providing a platform for cortical development. However, these cells are only present during embryonic and early postnatal life and if the cortical development proceeds normally, they die at a particular stage.
The transient nature of these cells highlights their importance in the assembly of a fully functional brain as well as their susceptibility to damage. We investigate the consequences of prematurity or perinatal brain damage on these cells, and the consequences of ablating them during the developmental time window for neurosecretion.

With previous MRC grant funding during the past fourteen years, we characterised the early connections arriving to the cortex, gene expression patterns in subplate neurons at various developmental stages and also identified molecular markers for transient subplate neurons which now help us in the detailed study of these cells in animal models and in humans. This highlighted that many genes that are active in the subplate encode extracellular proteins. We recently described that subplate cells have the shape and intracellular machinery to secrete such proteins (Kondo et al., 2015). Protein secretation and vesicular release is downregulated following conditional knock-down of SNAP25 in subplate cells. Additionally, we demonstrated that one of these proteins - neuroserpin - is upregulated in a rat model of neonatal hypoxia-ischemia, as well as in normal brains at the peak of cell death. Combined with evidence from the adult stroke literature, this suggests that neuroserpin may play a neuroprotective role.
We wish to capitalise on our previous achievements by identifying the role that secretion from subplate cells might play in normal brain development and in perinatal damage, particularly by testing whether neuroserpin can reduce the injury following hypoxia ischemia in mice.

Our major objectives are:
1. Characterise the timing and expression levels of neuroserpin and other secretory proteins in mice during development and after hypoxia. Study neuroserpin expression (mRNA and protein) in human.
2. Prevent secretion from subplate cells by blocking regulated vesicle fusion or by removing subplate cells altogether from early postnatal stages and study the consequences this has on brain development.
3. Study the effects of hypoxia in mice after viral delivery of neuroserpin into the brain or in neuroserpin knock-out mice.

The outcome of this project will bring fundamental insights into the workings of brain development as well as its susceptibility and response to early brain injury. They will help to establish methods of ensuring healthy brain maturation by developing for the understanding necessary for effective treatments of brain damage after hypoxic ischemia and management of development in premature infants.

Technical Summary

Our long-term research programme is to study the mechanisms underlying the formation of transient early cortical circuits that are crucial for the development of subsequent more permanent adult neuronal networks. Our hypotheses for this grant are:
1. Subplate cells have a secretory function. This function is transient and it is necessary for the normal maturation of the brain including vascularisation and myelination and maybe neuronal cell survival.
2. Neuroserpin, a serin protease released from subplate neurons in the developing brain has a stage-specific function in cell survival of subplate and/or surrounding cells.
3. Neuroserpin has an additional endogenous function in preventing or reducing injury following neonatal hypoxia. This is based on our preliminary results demonstrating up regulation of neuroserpin following hypoxia in rats.
4. Increasing neuroserpin expression (via viral delivery) shall have a stage specific positive effect on the outcome (histopathological and behavioural) of neonatal hypoxia, whereas reduced neuroserpin levels (in neuroserpin KO mice) have a negative effect.
These hypotheses shall be tested in animal experiments, and some findings extended to human post-mortem tissue. We will characterise the cellular localisation of proteins encoded by genes for extracellular molecules in mouse and human including neuroserpin and CTGF. We will test the proportions of these proteins that are made in subplate cells and the proportions that are taken up by them. We will characterise the role of neurosecretion from subplate cells by using animal models with reduced membrane vesicle fusion in subplate or without subplate cells. For this, we will use subplate cell specific Cre expression in the period combined with Snap25-flox or STOP-flox diphtheria toxin A mice. Neuroserpin protein levels will be analysed in hypoxic-ischemic animal models and the effectiveness of increasing or decreasing neuroserpin expression before hypoxia shall be tested.

Planned Impact

Brain disorders are a major cause of human suffering and disability. In the developed world, there has been a gradual increase in the number of premature births and survival, but often with devastating disabilities. Additionally, a rise in the diagnoses of neurodevelopmental disorders such as autism has been documented. In developed countries, brain diseases impose a tremendous direct economic burden on public healthcare and, indirectly, an even higher burden on society as a whole through reduced ability to work and increased caring responsibilities of relatives. Advances in treatment and prevention of early-onset or developmental brain abnormalities will have a positive impact on an individual's well-being and their family, as well as reducing healthcare costs and improving economic prosperity of society. Our research will investigate new intervention and treatment strategies for neonatal hypoxia-ischemia and shed light on poorly described secretory functions of transient cell populations that may contribute to normal brain maturation.
The most common causes of brain disorders that have their onset at birth are related to hypoxic-ischemic brain damage and prematurity and current monitoring and medication of the diseased newborn brain is limited. Additionally, autism and schizophrenia are now generally considered to result from subtle developmental abnormalities of the brain. A complex interaction between the environment and susceptibility factors encoded in the genome drive a phenotype that only manifest after decades. Currently, we lack a basic understanding of the neurobiology of these disorders and our research intends to address some of those shortfalls. Both animal and human studies have shown that the immature brain is not just a small version of the adult brain - there are great differences between them. For example, an antiepileptic drug that is effective in the adult can in fact be harmful in the neonate. To be able to contemplate prevention, achieve early diagnosis and establish treatment, we need to increase our knowledge of the basic biology of the immature human brain, including non-classical neuronal functions like secretion of extracellular proteins, and this is where our proposed research programme is focused.

Early and effective causal treatment of brain developmental abnormalities has the real potential to have a huge impact on the life of the individual, family and society in general. Preterm and term infants who need early interventions and treatment will be the primary beneficiaries of our research and families of these infants will benefit from an increased understanding and better clinical care, possibly reduced caring burden.
Clinicians responsible for the care of infants with hypoxic brain injury will also greatly benefit from this research as it has the potential for informing best clinical practice and new clinical trials. We will have the greatest impact in this field by regular exchanges with practicing clinicians and by providing training and information to future practicioners.
Our laboratory provides a wider benefit to society and the economy by training future generations of scientists and medics, and frequently hosting international student exchanges and visits (China, Germany, US, Canada, Brazil, Japan), which promotes overall economic prosperity by greater international understanding and cooperation. Our public engagement work also brings benefits by increasing understanding of animal experimentation. In addition, reaching the wider public can lead to greater understanding of brain abnormalities and disabilities which, consequently,brings a positive impact on equality and inclusion, especially amongst children.
Lastly, our image data can be of interest to artists and primary school teachers trying to engage children in science through art and collage techniques using scientific materials. AHS's images of fluorescent brains have previously been used for this purpose.

Publications

10 25 50
 
Title Art of Anatomy 
Description This is a paper on the link between anatomy and art: This paper arose from exhibitions in Oxford and Dublin and comprises three experiments which look at the relationship between anatomy and art. In the first experiment, a passport photograph, photographic portrait and portrait in oils, all of the same sitter, show how artistic input transforms anatomy from a mere likeness into works of art. In the second, the reverse is true, as computer techniques render idealized old master images anatomically accurate. The third experiment addresses the biomechanical consequences of anatomical variation and shows that vehicular design is based on mean body shapes, and so it is the average, rather than the idealized, form that is safer in a collision. 
Type Of Art Creative Writing 
Year Produced 2019 
Impact This paper arose from part of The Art of Anatomy exhibition in the Kendrew Barn, St John's College Oxford which was part of the Anatomical Society Summer Meeting, 23-25 July 2018 (https://www.youtube.com/watch?v=GY1YHiYwomw). The use of anatomical distortion can aid in creating a work of art. Swanlike necks add grace to paintings, and may go undetected by the casual viewer. Marked distortions can add to the fantasy element, or idealisation, in a painting. When normal anatomy is applied, this is lost, and what remains is a mediocre academy nude, rather than great art. Car safety has radically improved in recent years by the inclusion of features based on 'average' body shapes. However, this is a onesizefitsall approach. For those of us who are outliers, a bespoke approach is required to improve car safety. In art, anatomy may be transformed in the creation of a great work of art, and its accurate restoration can reverse the process. However, where safety is paramount, such accuracy is essential in both biomechanics and medicine. 
URL https://www.youtube.com/watch?v=GY1YHiYwomw
 
Title Cajal's Interactions with Sherrington and the Croonian Lecture 
Description Publication on history of science in The Anatomical Record: Sherrington was a major proponent of the neuron doctrine and he was inspired by Santiago Ramón y Cajal's theory of dynamic polarization of nerve cells (Ley de la polarización dinámica de la célula nerviosa). Sherrington coined the term "synapse" to name the Cajal description of interneuronal contact and he gave the term, for functional nerve endings, as "Boutons terminaux," still used today. These two giants of neuroscience met only once, but they had a lifelong friendship. It was Sherrington who wrote Cajal's Obituary for the Royal Society. We review here some of the scientific exchanges between Cajal and Sherrington, with particular attention to 1894, when the two neuroscientist met in London during Cajal's visit to deliver the Croonian Lecture to the Royal Society. We shall examine not only the scientific exchanges but also their friendship, which was immediate and strong. Anat Rec, 2019. 
Type Of Art Creative Writing 
Year Produced 2019 
Impact The paper emphasises the need for European and international research collaborations: Sherrington and Cajal met only once, but they had a lifelong friendship. Cajal and Sherrington shared several views on the organization of the nervous system, contiguity, importance of the synapse, and convergence and divergence of neural connections, although they devoted different importance to inhibition. The close friendship between Sherrington and Cajal lasted throughout their lives. It was Sherrington who wrote Cajal's Obituary for the Royal Society (Sherrington, 1935). 
URL https://anatomypubs.onlinelibrary.wiley.com/doi/full/10.1002/ar.24189
 
Title The Art of Anatomy Exhibition - St John's College, Oxford (July 2018) 
Description Art of Anatomy Exhibition, The Barn, June-July, 2018 Professor Clive Lee (Royal College of Surgeons in Ireland & Royal Hibernian Academy of Arts) Dr Sarah Simbler (Tutor of Anatomy at the Ruskin School of Art, University of Oxford) Dr Tian S. Liang (Early Career Leverhulme Fellow, St John's College) Professor Zoltán Molnár (Tutor of Human Anatomy, St John's College, Oxford) This exhibition will explore the artistic representation of anatomy during different historic periods and in different parts of the world. It will coincide with the Summer Meeting of the Anatomical Society and the Editor of Journal of Anatomy has invited papers from the anatomists and artists involved (1). Anatomy Transformed Professor Clive Lee is Professor of Anatomy at the Royal College of Surgeons in Ireland and the Royal Hibernian Academy of Arts, Visiting Professor of Biomechanics at Trinity College Dublin and has applied to be a Visiting Researcher at St John's College during Trinity Term 2018. In medicine and engineering, anatomy is applied, but in art it is transformed. Transformation by distortion is examined by comparing masterpieces, such as Ingres' La Grande Odalisque, estimated to have 3 extra lumbar vertebrae, with anatomically accurate computer images generated by Oscar winning engineer, Anil Kokaram (2,3). The balance between accuracy and artistry is explored in a triptych from passport photo to photographic portrait, by Amelia Stein RHA, to a portrait in oils by Mick O'Dea, President of the Royal Hibernian Academy of Arts www.rhagallery.ie The biomechanics of beauty will explore the practicalities of aesthetic ideals from an engineering standpoint. Finally, Anatomists, Engineers & Artists is a video installation showing the creation of an on-line 3D surface anatomy course: www.rcsi.ie/surfaceanatomy Science, History, Art Dr Sarah Simblet is a fine artist, writer, broadcaster and anatomist, Tutor in Anatomy at the Ruskin School of Art (BFA) and she teaches Short Courses in Anatomy, Botany and Drawing on the Summer School Programme (4). She is author of Anatomy for the Artist (2001), The Drawing Book (2005), Botany for the Artist (2010) and co-author of The New Sylva (2014). Sarah is dedicated to sharing and encouraging visual intelligence in others through her drawings, teaching and broadcasting worldwide. www.rsa.ox.ac.uk/people/sarah-simblet Anatomising the Body: Art, Science and the Humanities in Modern China Dr Tian S. Liang is a Leverhulme Fellow at St John's College and co-editor of Hong Ling: A Retrospective. Taipei: Soka Art (2016). She specialises in modern and contemporary East Asian art and explores understandings of human anatomy and its relations to artistic representations of the human body in China between 1851 and 1949 (5). By placing the knowledge of human anatomy at the centre of a broader nexus of artistic, medical, educational, publishing and humanitarian contexts, this project will provide the first history of the ways in which the 'body' was scientifically assessed and artistically represented across different visual media - painting, sculpture, print, woodcut and photography. https://www.leverhulme.ac.uk/awards-made/awards-focus/anatomising-body-art-science-and-humanities-modern-china-1851%E2%80%931949 Cerebral Art Professor Zoltan Molnar is Tutor of Human Anatomy at St John's College Oxford, Professor of Developmental Neuroscience at Department of Physiology, Anatomy and Genetics. His main research interest is in the development of the cerebral cortex. He has been running and organising the Neuroanatomy Practicals since 2000 at the University of Oxford. He can see art in the structure of the brain, in its histology, images. He has a huge collection of microscopic images. For some examples see (6) and for journal covers selected from Molnár Laboratories illustration see: https://www.dpag.ox.ac.uk/research/molnar-group/publications-front-covers/journal-covers 
Type Of Art Artistic/Creative Exhibition 
Year Produced 2018 
Impact This exhibition will explore the artistic representation of anatomy during different historic periods and in different parts of the world. It will coincide with the Summer Meeting of the Anatomical Society and the Editor of Journal of Anatomy has invited papers from the anatomists and artists involved. 
URL https://www.sjc.ox.ac.uk/college-life/art/
 
Title The Oxford Science and Ideas Festival - The consciousness field 
Description https://www.dpag.ox.ac.uk/news/the-oxford-science-and-ideas-festival http://www.theconsciousnessfield.com/index.html Once again researchers from across our Oxford Neuroscience Community pulled all the stops out for the Oxford Science and Ideas Festival. Extensive programmes of activities were run by the Nuffield Department of Clinical Neurosciences and the Department of Experimental Psychology. In addition, the Department of Physiology, Anatomy and Genetics' Zoltan Molnar in collaboration with Colin Blakemore and St John's College put on a holographic presentation 'The Consciousness Field' created by artist Maria Lopez. 
Type Of Art Artistic/Creative Exhibition 
Year Produced 2018 
Impact The exhibition was very well attended and the general public had the chance to ask questions from researchers and artists. http://www.theconsciousnessfield.com/index.html 
URL https://www.dpag.ox.ac.uk/news/the-oxford-science-and-ideas-festival
 
Description Senior Founding Member of the University of Oxford Cortex Club
Geographic Reach Local/Municipal/Regional 
Policy Influence Type Influenced training of practitioners or researchers
Impact The Cortex Club connects researchers at the University of Oxford with world-leading neuroscientists through a unique educational forum dealing with cutting-edge topics and significant challenges in neuroscience. Our events range from small intense debates with up-and-coming scientists to large discussion sessions led by internationally prominent speakers, followed by the opportunity to ask them questions over drinks. News release on the establishment of the cortex club: https://www.dpag.ox.ac.uk/news/cortex-club-celebrates-10-years-of-student-run-talks-and-discussions-for-oxfords-neuroscience-community
URL http://cortexclub.com
 
Description St John's College Research Centre Project Grant
Amount £90,000 (GBP)
Organisation University of Oxford 
Department St John's College Oxford
Sector Academic/University
Country United Kingdom
Start 01/2017 
End 01/2019
 
Description The Academy of Medical Sciences Newton Advanced Fellowship
Amount £111,000 (GBP)
Organisation Academy of Medical Sciences (AMS) 
Department Newton Advanced Fellowship
Sector Academic/University
Country United Kingdom
Start 03/2017 
End 02/2020
 
Title Allen Brain Research Institute - Non Human Primate Developmental Transcriptome Database 
Description The Allen Brain Institute contacted us to advise them on the collection and analysis of the developmental transcriptomic atlas in macaque cerebral cortex. The approach is similar that we developed previously - Hoerder-Suabediessen et al., 2009 - PMID: 23401504 and 2013 - PMID: 23401504; Oeschger et al., 2013 - PMID: 21862448; Belgard et al., 2010 - PMID: 21867878 
Type Of Material Biological samples 
Year Produced 2014 
Provided To Others? Yes  
Impact Our work that was supported by MRC lead to establishing methodologies and approaches that now used by the Allen Brain Institute. We were invited to advise the recent project on Macaque Brain Development Transcriptome where we shall lead the analysis of the subplate layer. This is based on - Hoerder-Suabediessen et al., 2009 - PMID: 23401504 and 2013 - PMID: 23401504; Oeschger et al., 2013 - PMID: 21862448; Belgard et al., 2010 - PMID: 21867878 
URL http://www.alleninstitute.org/science/public_resources/nhp_atlas.html
 
Title In Utero Electroporation Methods in the Study of Cerebral Cortical Development 
Description Electroporation or electropermeabilization is a method that uses electric pulses to deliver molecules into cells and tissues. The in utero electroporation method has enabled the field to administer plasmids to these neural progenitors, allowing temporal and cell type-specific control for the manipulation of gene expression. My laboratory shared our methodological expertise with the field in a book chapter: Protocol Prenatal and Postnatal Determinants of Development; Volume 109 of the series Neuromethods pp 21-39 In Utero Electroporation Methods in the Study of Cerebral Cortical Development Authors Isabel Martínez-Garay, Fernando García-Moreno, Navneet Vasistha, Andre Marques-Smith, Zoltan Molnar Publication date 2016 Journal Prenatal and Postnatal Determinants of Development Pages 21-39 Publisher Springer New York 
Type Of Material Technology assay or reagent 
Year Produced 2015 
Provided To Others? Yes  
Impact Research in the field of cortical development has benefited from technical advances in recent years, and tools are now available to label, monitor, and modulate cohorts of cerebral cortical neurons using in vivo approaches. Substantial populations of cerebral cortical neurons are generated in a specific sequence by the radial glia progenitors that line the ventricular surface during development. These radial progenitors self-renew and generate intermediate progenitors or neurons in a precisely choreographed fashion. in utero electroporation has become a central technique in the study of key aspects of neural development, such as progenitor proliferation, neurogenesis, neuronal migration, and circuit formation. This method has also facilitated the exploitation of cell lineage and optogenetic techniques in various species from chick to gyrencephalic higher mammals. This chapter provides a description of the method and gives some examples for its utility in the study of cerebral cortical development and evolution. 
URL http://link.springer.com/protocol/10.1007/978-1-4939-3014-2_2
 
Title Low-cost printed tissues that address the global catastrophe of brain disease 
Description Low-cost printed tissues that address the global catastrophe of brain disease Hagan Bayley (Oxford Chemistry: Division of Mathematical, Physical and Life Sciences) Francis Szele (Oxford Physiology, Anatomy and Genetics: Division of Medical Sciences) Zoltan Molnar (Oxford Physiology, Anatomy and Genetics: Division of Medical Sciences) Brain damage through injury or disease has devastating social and economic consequences that are expanding exponentially in impact. We will initiate a radical approach in which the brain is repaired with 3D-printed neural tissue derived from stem cells. Ultimately, we aim to produce a low-cost, "democratized" technology that addresses an indisputable global catastrophe. We will address the emerging global catastrophe of brain disease by a radical new approach to 3D cell patterning. We will repair the brain by using anatomically correct implants produced from patients' own tissues with stem cell technology. Advances in diagnosis, which allow the early identification of brain disorders, will provide windows of opportunity for effective intervention. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2017 
Provided To Others? Yes  
Impact Graham AD, Olof SN, Burke MJ, Armstrong JPK, Mikhailova EA, Nicholson JG, Box SJ, Szele FG, Perriman AW, Bayley H: High-Resolution Patterned Cellular Constructs by Droplet-Based 3D Printing. Sci Rep 2017, 7:7004. We are currently preparing a manuscript on printed cortical neurons. 
URL http://bayley.chem.ox.ac.uk
 
Title A Transcriptomic Atlas of Mouse Neocortical Layers - supplementary website for Belgard, et al. (2011) 
Description A transcriptomic atlas of mouse neocortical layers. Belgard TG, Marques AC, Oliver PL, Abaan HO, Sirey TM, Hoerder-Suabedissen A, García-Moreno F, Molnár Z, Margulies EH, Ponting CP. Neuron. 2011 Aug 25;71(4):605-16. We sequenced transcriptomes from layers 1-6b of different areas (primary and secondary) of the adult (postnatal day 56) mouse somatosensory cortex to understand the transcriptional levels and functional repertoires of coding and noncoding loci for cells constituting these layers. A total of 5,835 protein-coding genes and 66 noncoding RNA loci are differentially expressed ("patterned") across the layers, on the basis of a machine-learning model (naive Bayes) approach. Layers 2-6b are each associated with specific functional and disease annotations that provide insights into their biological roles. This new resource (http://genserv.anat.ox.ac.uk/layers) greatly extends currently available resources, such as the Allen Mouse Brain Atlas and microarray data sets, by providing quantitative expression levels, by being genome-wide, by including novel loci, and by identifying candidate alternatively spliced transcripts that are differentially expressed across layers. 
Type Of Material Database/Collection of data 
Year Produced 2011 
Provided To Others? Yes  
Impact This is the first website that describe the layer-specific transcriptome of the adult mouse cortex using deep sequencing method. This enables the study of splice variants and non-coding transcripts. 
URL http://genserv.anat.ox.ac.uk/layers
 
Title Online database of subplate gene expression at https://molnar.dpag.ox.ac.uk/subplate/. 
Description Here we present gene expression evidence for distinct roles of the mouse subplate across development as well as unique molecular markers to extend the repertoire of subplate labels. Performing systematic comparisons between different ages (embryonic days 15 and 18, postnatal day 8, and adult), we reveal the dynamic and constant features of the markers labeling subplate cells during embryonic and early postnatal development and in the adult. This can be visualized using the online database of subplate gene expression at https://molnar.dpag.ox.ac.uk/subplate/. We also identify embryonic similarities in gene expression between the ventricular zones, intermediate zone, and subplate, and distinct postnatal similarities between subplate, layer 5, and layers 2/3. 
Type Of Material Database/Collection of data 
Year Produced 2013 
Provided To Others? Yes  
Impact Our database (https://molnar.dpag.ox.ac.uk/subplate/) help the general research community to find suitable markers to reveal subplate, the early generated, largely transient cell population. 
URL https://molnar.dpag.ox.ac.uk/subplate/
 
Title The History of Medical Sciences University of Oxford Project 
Description The History of Medical Sciences Project, University of Oxford. Our project aims to preserve and bring to life the rich history of science through the followings streams: 1. Seminars, given by experts in their field, on the history of important discoveries and the individuals behind them 2. Digitisation and display of historical microscope slides used for research and teaching and associated letters, notes and other documents, 360° photography of historical models and instruments 
Type Of Material Database/Collection of data 
Year Produced 2014 
Provided To Others? Yes  
Impact Pur historic repository helped to preserve unique letters, histological preparations, scientific instruments and models. The historic repository attracted several international scholars to further study the life and work of outstanding Oxford research academics, including Sherrington, Eccles, Fitzgerald. 
URL https://history.medsci.ox.ac.uk/
 
Title Web resource for adult forebrain expression in mouse and chicken at http://genserv.anat.ox.ac.uk/brainevo. 
Description We explored which genes are actively transcribed in the regions of controversial ancestry in a representative bird (chicken) and mammal (mouse) at adult stages. We conducted four analyses comparing the expression patterns of their 5,130 most highly expressed one-to-one orthologous genes that considered global patterns of expression specificity, strong gene markers, and coexpression networks. we provide a Web resource for adult forebrain expression in mouse and chicken at http://genserv.anat.ox.ac.uk/brainevo. 
Type Of Material Database/Collection of data 
Year Produced 2013 
Provided To Others? Yes  
Impact This dataset not only will enable insights into telencephalic gene expression evolution, but also can facilitate future functional investigations. For example, the cross-species marker lists for striatum and hippocampus can readily be expanded using the advanced search feature on the website. 
URL http://genserv.anat.ox.ac.uk/brainevo
 
Description Collaboration with Dr Istvan Adorjan (Semmelweis Medical University, Budapest Hungary) 
Organisation Semmelweiss University
Country Hungary 
Sector Academic/University 
PI Contribution My laboratory supervised the initial work of Dr Istvan Adorjan who is analysing the neuroserpin expression in developing human cortex (obtained from the MRC-Wellcome Human Brain Bank - Newcastle and Institute of Child Health, London) and also examining neuroserpin expression in human pathological specimen (obtained in collaboration with King's College, London).
Collaborator Contribution Dr Istvan Adorjan is analysing the neuroserpin expression in developing human cortex (obtained from the MRC-Wellcome Human Brain Bank - Newcastle and Institute of Child Health, London) and also examining neuroserpin expression in human pathological specimen (obtained in collaboration with King's College, London).
Impact We are planning to present the results of this collaboration on the Anatomical Society Summer Meeting in July 2018 (http://www.anatsocmeeting.co.uk) followed with publications.
Start Year 2016
 
Description Collaboration with Dr Lei Shi (Jinan University, PR of China) Roles of subplate-secreted factors in cortical development 
Organisation Jinan University
Country China 
Sector Academic/University 
PI Contribution I hosted Dr Lei Shi from Jinan University, People Republic of China for her sabbatical in 2015-2016. During her sabbatical we collaborated on projects examining the ER stress and neurosecretion from subplate neurons during normal development and during perinatal hypoxia-ischaemia. We continued this collaboration after her return and we succesfully applied for a The Academy of Medical Sciences Newton Advanced Fellowship. The details of your award are as follows: Applicant: Dr Lei Shi, Title: Roles of subplate-secreted factors in cortical development Overseas institution: Jinan University Co-applicant: Professor Zoltán Molnár UK host institution: University of Oxford Amount: £111000
Collaborator Contribution Dr Lei Shi is an expert in the in vitro analysis of protein secretion and trafficking. She is the main driver to move our anatomical observations to molecular and cellular level. Our previous observations were descriptive: Shinichi Kondo, Hannah Al-Hasani, Anna Hoerder-Suabedissen, Wei Zhi Wang, and Zoltán Molnár (2015) Secretory function in subplate neurons during cortical development. Front Neurosci. 9:100. With the help of Dr Lei Shi we now generated constructs to be able to monitor protein secretion and trafficking with great resolution in vitro.
Impact We contributed with a review to Frontiers in Neuroscience: Title: Neonatal Hypoxic-Ischaemic Brain Injury: Mechanisms, Models, and Therapeutic Challenges Authors: Lancelot Jamie Millar, Lei Shi, Anna Hoerder-Suabedissen, Zoltan Molnar Manuscript ID: 239973 Edited by: Daniela Tropea
Start Year 2015
 
Description Collaboration with Prof Xiaoqun Wang, Institute of Biophysics, Chinese Academy of Science, Beijing, China 
Organisation Chinese Academy of Sciences
Department Institute of Biophysics
Country China 
Sector Academic/University 
PI Contribution Professor Xiaoqun Wang (Principle of Investigator at the Institute of Biophysics, State Key laboratory of Brain and Cognitive Science, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Science, Beijing 100101, China) approached our laboratory to collaborate on cerebral cortical development in ferret and non-human primates and human using single cell transcriptomic methodologies. I sponsored his Royal Society Newton Fellowship application and regularly advise him on research as his UK mentor.
Collaborator Contribution Professor Xiaoqun Wang (Principle of Investigator, Institute of Biophysics, State Key laboratory of Brain and Cognitive Science, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Science, Beijing 100101, China) is conducting single cell sequencing experiments and analysis in various species (mouse, ferret, macaque monkey and human) and we collaborate on the analysis of the subplate relevant data.
Impact So far we had presentations on meetings (https://corticaldevelopment.wordpress.com/; http://www.fens.org/Meetings/The-Brain-Conferences/Cortex-Evolution-and-Development/) and Prof Xiaoqun Wang shall participate on the Oxford Meeting of the Anatomical Society that is specifically on human brain development that I shall host in Oxford (http://www.anatsocmeeting.co.uk).
Start Year 2015
 
Description Investigation of the effect of genetic perturbation of VIP+ interneurons during and beyond postnatal development 
Organisation University of Oxford
Department Department of Physiology, Anatomy and Genetics
Country United Kingdom 
Sector Academic/University 
PI Contribution Investigation of the changes in local and long range connectivity in early postnatal development after genetic perturbation of VIP+ interneurons using laser scanning photostimulation and an optogenetic viral approach.
Collaborator Contribution Investigation of the effect of genetic perturbation of VIP+ interneurons in adulthood during a sensory discrimination task.
Impact Early stages, no relevant outcomes at present
Start Year 2018
 
Description Brain Awareness week themed activity at local Woodcraft Elfins group 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Other audiences
Results and Impact The children aged 6-10 were encouraged to discuss their understanding of "senses", and we played some games relying specifically on vision or hearing. Then the children were shown a model of the human brain and we identified different brain structures, and some functional areas of the cortex. We then discussed that sensory areas of the cerebral cortex are found in roughly the same place in all mammals studied, but that the size of each area differs markedly between species depending upon their particular use of a sense organ. The children were then encouraged to "design" their own fantasy animal and the senses this animal would rely on, and then to create a 3D brain model out of play-doh. We then discussed that the brain contains cells, showed them some images of neurons (ranging from Cajal, to images of brains acquired as part of this MRC grant), and encouraged them to replicate the different cell shapes as pipe-cleaner models of neurons.
Year(s) Of Engagement Activity 2018
 
Description Brain Diaries Exhibition - Museum of Natural History, University of Oxford 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Extract from the introduction from the exhibition website: Baby, child, teenager, adult
Throughout your life, your brain undergoes extraordinary changes.
Fascinating physical developments in your brain unlock each new chapter as you grow. And with billions of neurons and trillions of connections, it responds to your experiences to make you the person you are.
What does the latest research tell us about how
our brains work, from the minute we are conceived to the moment we die?
Explore the
Year(s) Of Engagement Activity 2016
URL http://www.oum.ox.ac.uk/braindiaries/
 
Description Evolution of the Brain Masterclass - Museum of Natural History, Oxford 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Using the Museum's collections neuroscientist Professor Zoltán Molnár and his laboratory members (https://www.dpag.ox.ac.uk/research/molnar-group) and his medical students from St John's College will lead you through the amazing development and adaptations of the vertebrate brain.
Year(s) Of Engagement Activity 2017,2018
URL https://www.facebook.com/pg/molnarlab/photos/?tab=album&album_id=730283630497690
 
Description Interview to BBC on a publication on brain fording in Nature Physics by Tuomas Tallinen, a soft matter physicist at the University of Jyväskylä in Finland and a co-authors. 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Interview to BBC on a publication on brain fording in Nature Physics by Tuomas Tallinen, a soft matter physicist at the University of Jyväskylä in Finland and a co-authors.
I discussed research with Dr Jonathan Webb, Science reporter, BBC News and my comments were used in the Science and Environment BBC Website: http://www.bbc.co.uk/news/science-environment-35438294
Year(s) Of Engagement Activity 2016
URL http://www.bbc.co.uk/news/science-environment-35438294
 
Description Interview to the OX Magazine on Brain Awareness Week 2017 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact From the 10th March the Museum of Natural History are opening an exhibition and series of events on the mysteries of neurological development - Brain Diaries. I was approached by the OX Magazine to give an interview and discuss the highlights of this exhibition and also my own research. This was produced for the general public in Oxfordshire.
The interview can be accessed on: http://oxhc.co.uk/Brain-Diaries-In-conversation-with-Professor-Zoltan-Molnar.asp
Year(s) Of Engagement Activity 2016
URL http://oxhc.co.uk/Brain-Diaries-In-conversation-with-Professor-Zoltan-Molnar.asp
 
Description Lecture for the Oxford Medical Lecture Club on 28th January 2018 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Oxford Medical Lecture Club is for the Retired Clinical Professionals of University of Oxford. The members organise the meetings and invite lecturers. My lecture was on the History of Neuroscience in Oxford (with special attention to Willis and Sherrington). https://www.facebook.com/molnarlab/posts/768407353351984 The members of the group were encouraged to contribute to the History of Medical Sciences Project website with their memories and conserve historic letters, manuscripts with general importance.
Year(s) Of Engagement Activity 2018
URL https://www.facebook.com/molnarlab/posts/768407353351984
 
Description Molnar Laboratory FaceBook Page with over 200 followers 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact The Molnar Laboratory has its own Facebook page: https://www.facebook.com/molnarlab/
There are over 200 followers. The FB page advertises seminars, lectures, conferences and keeps in touch with alumni of the laboratory.
Year(s) Of Engagement Activity 2015,2016,2017,2018,2019
URL https://www.facebook.com/molnarlab/
 
Description Molnar Laboratory Public Facebook Page 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact The Molnar Laboratory's Facebook Page (https://www.facebook.com/molnarlab/) announces relevant seminars, publications, news releases to the audience that follows our activities (around 200 individuals at the moment).
Year(s) Of Engagement Activity 2015,2016,2017,2018
URL https://www.facebook.com/molnarlab/
 
Description News Release on: Unexpected origin of the subplate neurons (Pedraza et al., PNAS 2014) 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Our interview helped to release the following news in english and in Spanish (http://www.madrimasd.org/informacionidi/noticias/noticia.asp?id=60258?igen=notiweb&dia_suplemento=martes
and in:
http://www.abc.es/ciencia/20140429/abci-nuevos-hallazgos-corteza-cerebral-201404281708.html

A collaborative study between the groups of Professor Juan De Carlos (Instituto Cajal, C.S.I.C., Madrid, Spain) and Professor Zoltán Molnár (Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford UK) has provided evidence for a new source of the earliest generated cerebral cortical cells, the subplate. In their recent publication in the journal Proceeding of the National Academy of Science USA (Pedraza et al., 2014) they describe that a considerable proportion of these early generated cells (known as the subplate) originate from an unusual and unexpected source, in the anterior and medial part of the telencephalon, outside the cerebral cortex itself. Previously, it had been generally accepted that they originate locally from the germinal zone of the cortical neuroepithelium.

These subplate cells in the embryonic cerebral cortex are particularly interesting for clinicians concerned with brain developmental abnormalities, since they provide a transient scaffold for the developing cerebral cortex and assist in the development of cortical connectivity, effectively providing the foundation of the adult structure. Indeed, the first synaptic contact in the cerebral cortex is established between subplate cells and the first thalamic fibers that reach the neuroepithelium. Disruption or damage to these early cells could be the source of flaws that lead to cerebral cortical developmental disorders, such as cerebral palsy, schizophrenia and autism. The discoveries that these scaffold cells arrive after a longer and more arduous journey might be significant to understand developmental disorders, because the process of their migration could be vulnerable and could be the cause of some cortical developmental abnormalities. If the scaffold is not assembled in the right time at the right place, then other aspects of the building work of making a brain can suffer which may only manifest in later life. This makes the discovery of this unusual origin and migration of the subplate neurons important for the developmental neurobiology and clinical neurology community.

The team in Madrid, led by Professor De Carlos noticed that some subplate cells originate from an unusual anterior and medial location (rostral medial telencephalic wall) and migrate tangentially through long routes to the cerebral cortex. The team at Oxford studied a selective subgroup of subplate cells expressing Lpar1 (Edg2) which had similar distribution and characteristics to the labeled cells found by De Carlos and his team (Molecular diversity of early-born subplate neurons. Hoerder-Suabedissen A, Molnár Z. Cereb. Cortex. 2013 Jun;23(6):1473-83). When the two teams compared their results on a meeting they decided to use cell-tracker to label rostral medial telencephalic wall in the mouse strain with fluorescently labeled Lpar1-positive cells. These experiments showed a very good match and clearly demonstrated that indeed this subset of labeled subplate cells originated from this unexpected and unusual anterior-medial source in mice.

The layer of subplate cells is in the Cerebral Cortex of all mammals, but their size varies between species, being wider in humans and primates. This suggests that subplate cells are not a vestige of early neuronal structures, but a fundamental structure involved in early stage of cortical development. For this reason it will be important to study this source also in humans and hence, it will be imperative to study abnormalities of this particular subplate cell population in human cognitive disorders. It has been previously found that many of the schizophrenia and autism associated genes were expressed transiently in subplate during development (Expression profiling of mouse subplate reveals a dynamic gene network and disease association with autism and schizophrenia. Hoerder-Suabedissen et al., Proc Natl Acad Sci U S A. 2013 Feb 26;110(9):3555-60). Studying the way that nerves develop and connect this network can reveal where things can go wrong and trigger diseases such as the mentioned cerebral palsy, autism or schizophrenia.

This work was supported by the Spanish Ministerio de Ciencia e Innovación and by the Medical Research Council of the United Kingdom. Please visit the Proceedings of the National Academy of Sciences website for the full article (Pedraza et al., 2014)

After the publication of these news releases, I received more approaches from Spain (from students and postdocs).
Year(s) Of Engagement Activity 2013,2014
URL http://www.dpag.ox.ac.uk/news/unexpected-origin-of-the-subplate-scaffold-cells-of-our-developing-bra...
 
Description News release on: Autism and Schizophrenia May Develop During Short Period of Time in Infancy 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact News release was helped by our work and our interview:

http://healthland.time.com/2013/02/12/autism-and-schizophrenia-may-develop-during-short-period-of-time-in-infancy/
http://www.sciencedaily.com/releases/2013/02/130211162223.htm
http://www.genengnews.com/gen-news-highlights/autism-and-schizophrenia-genes-show-early-fleeting-activation/81247981/
http://www.enn.com/health/article/45586
http://scicasts.com/gene/2029-functional-genomics/5417-study-shows-autism-and-schizophrenia-genes-only-active-in-developing-brains/
http://medicalxpress.com/news/2013-02-genes-autism-schizophrenia-brains.html
http://www.myscience.org/news/2013/genes_for_autism_and_schizophrenia_only_active_in_developing_brains-2013-imperial
http://www.sify.com/news/genes-for-autism-and-schizophrenia-only-active-in-developing-brains-news-international-ncmp5rcadic.html

Genes connected to the two disorders may only be active for a brief window of time.
The study, while in mice, could provide clues about how the developmental disorders develop. Focusing on a region of the brain known as the subplate, where the first nerve cells develop, researchers at the University of Oxford, King's College London and Imperial College London found that genes linked to autism and schizophrenia were only active in these regions during early stages of brain development.

Neurons in the subplate region form the foundation for the network of neural connections that eventually crisscross the brain. Studying the way that nerves develop and join this network can reveal where the growth can go wrong and trigger diseases such as autism or schizophrenia.

I regularly advise journalists on publications related to autism and schizophrenia and on general issues on brain development.
These news releases gave us public exposure.
Year(s) Of Engagement Activity 2013,2014
URL http://healthland.time.com/2013/02/12/autism-and-schizophrenia-may-develop-during-short-period-of-ti...
 
Description Oxford - McGill Mini Symposium 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research and patient groups
Results and Impact https://www.mcgill.ca/brain/international-partners/oxford-university/oxford-neuroscience-news

On 14th January 2014 a mini Symposium organised by Andrew King and Zoltán Molnár served as the first joint cluster meeting between the systems and cellular neuroscience groups of the DPAG. The meeting covered a wide range of topics focussed around "Mammalian cerebral cortical circuits".

The joint cluster meeting brought together groups who are experts in the field of cerebral cortical circuit formation and plasticity in mammals. Their expertise extends from neurogenesis and neuronal migration to the establishment and plasticity of connections. These groups use various techniques: laser-scanning photo stimulation, 2-photon microscopy, confocal microscopy, electrophysiology and behavioural testing. Their aim is to understand how mammalian cortical neurons control cortical development and plasticity and how they are integrated into functional cortical circuitry responsible for some of the most sophisticated sensory and motor processing.

The cluster meeting was linked to the visit of Professor Anne Mc Kinney, Vice Chair of McGill's Brain Institute, who delivered a keynote lecture on "Dendritic spines, autism and epilepsy". Her laboratory has made a great contribution to the insights into synapse maintenance and plasticity in the mammalian cerebral cortex and hippocampus.

The symposium gave an excellent opportunity to continue to build links between McGill and Oxford Neuroscience.
Year(s) Of Engagement Activity 2013,2014
URL https://www.mcgill.ca/brain/international-partners/oxford-university/oxford-neuroscience-news
 
Description Professor Molnar has a Twitter Account with Science Focus 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Professor Molnar has a Twitter Account (https://twitter.com/ZoltanMolnar64) that has outreach function. Seminars, lectures and conferences are advertised. It also draws attention to recent publications and presentations. There are over 300 followers.
Year(s) Of Engagement Activity 2015,2016,2017,2018,2019,2020
URL https://twitter.com/ZoltanMolnar64
 
Description Professor Zoltán Molnár features in BBC programme 'How the NHS Changed our World' 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact https://www.dpag.ox.ac.uk/news/professor-zoltan-molnar-features-in-bbc-programme-how-the-nhs-changed-our-world

The BBC recently visited Zoltán Molnár, Professor of Neuroscience here in DPAG, to discuss the research undertaken by Sir Charles Scott Sherrington, during his time in the Laboratory of Physiology, as part of a new series on 'How the NHS Changed our World'.

Sherrintgon kept hundreds of of microscope slides in a specially constructed Histological Box, which has been preserved and is now kept by Zoltán here in the Department.

In the interview, Zoltán sheds light on how Sherrington and his colleagues would shed their skin for the sake of scientific research. Sherrington's notes reveal how popular self-experimentation was in those days.

The interview can be viewed here and you can also watch the whole episode on iPlayer.
Year(s) Of Engagement Activity 2018
URL https://www.dpag.ox.ac.uk/news/professor-zoltan-molnar-features-in-bbc-programme-how-the-nhs-changed...
 
Description The 2015 Brain Awareness Week Lecture at Museum of the History of Science, Oxford 19th March 2015 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact The 2015 Brain Awareness Week Lecture at Museum of the History of Science, Oxford 19th March 2015.
This lecture was part of the Brain Awareness Week activities at Oxford.
The lecture was entitled: "Neuroscience in Oxford: Four Centuries of Discovery" and was aimed at the general public. The lecture was recorded and now available on: https://history.medsci.ox.ac.uk/seminars/history-of-medical-sciences-seminar-series/prof-zoltan-molnar-neuroscience-in-oxford-four-centuries-of-discovery/
Year(s) Of Engagement Activity 2015
URL https://history.medsci.ox.ac.uk/seminars/history-of-medical-sciences-seminar-series/prof-zoltan-moln...
 
Description The 2015 Brain Awareness Week Lecture at Museum of the History of Science, Oxford 19th March 2015 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact The lecture was hosted by the History of Science Museum and it was entitled: "Neuroscience in Oxford: Four Centuries of Discovery".
The lecture theatre was completely full and the lecture was recorded and posted to: https://history.medsci.ox.ac.uk/seminars/history-of-medical-sciences-seminar-series/prof-zoltan-molnar-neuroscience-in-oxford-four-centuries-of-discovery/
Year(s) Of Engagement Activity 2015
URL https://history.medsci.ox.ac.uk/seminars/history-of-medical-sciences-seminar-series/prof-zoltan-moln...
 
Description Zoltan Molnar organised two workshops on the neurobiology of taste and smell (St John's College and in Department of Physiology, Anatomy and Genetics) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact The Workshop started with a scientific preseantation on taste and smell by Professor Laura Lopez-Mascaraque. The talk was aimed to the general public. Then the attendees (30-40 on both occasions) had the opportunity to determine the number of their taste buds after staining the tip of their tongue with food colourant and then exploring the small fungiform papillae within a small selected area with magnifying glasses. Non-tasters have less then 8 buds, average tasters have 8-13 buds, whereas super-tasters have more than 18 buds in a unit area.

The participants also tested their olfactory threshold by exploring their sensitivity to capture the odours at different concentrations. Olfactory threshold of a given compound is the minimum odour concentration that is perceived by the human sense of smell. This allows the detection of the different sensitivity degree to capture the odours at different concentrations. Mirror image molecules (known as stereoisomers) have very different smells. The best-known stereoisomers that evoke very different aromas are D-carvone and L-carvone, which smell like caraway and spearmint, respectively.

The olfactory discrimination tests proved to be the most challenging tasks of the day. The participants were given 12 different small boxes with different natural aromas. Based on their previous experience they had to name these odours. Participants noted that they "knew what the aromas were" but that they just could not name them. Once the correct responses were announced, the participants could retrospectively identify the aromas of lemon, chocolate, rose etc.

The participants also tested their olfactory memory by recognising the hidden aromas in different small boxes, but this test was made easier using small images of three possibilities. This screening test of olfaction is reliable in recognising people with normal olfaction and can distinguish them from people with hyposmia or anosmia. A person with a score of 7 or 8 has a 99.74% probability of having a normal olfaction whereas patients with subjective hyposmia or anosmia score between 0 and 6. The workshop finished with wine tasting demonstrations by the current captain of the Oxford University Blind Tasting Society (Domen Presern and Janice Wang). https://www.dpag.ox.ac.uk/news/worlds-of-sense; https://www.sjc.ox.ac.uk/discover/news/smell-research
Year(s) Of Engagement Activity 2018
URL https://www.dpag.ox.ac.uk/news/worlds-of-sense