Isolation of neural stem and oligodendrocyte progenitor cells from the brain of live rats

Summary Postnatal brain neural stem and progenitor cells (NSPCs) cluster in anatomically inaccessible stem cell niches, such as the subependymal zone (SEZ). Here, we describe a method for the isolation of NSPCs from live animals, which we term “milking.” The intracerebroventricular injection of a release cocktail, containing neuraminidase, integrin-β1-blocking antibody, and fibroblast growth factor 2, induces the controlled flow of NSPCs in the cerebrospinal fluid, where they are collected via liquid biopsies. Isolated cells retain key in vivo self-renewal properties and their cell-type profile reflects the cell composition of their source area, while the function of the niche is sustained even 8 months post-milking. By changing the target area more caudally, we also isolate oligodendrocyte progenitor cells (OPCs) from the corpus callosum. This novel approach for sampling NSPCs and OPCs paves the way for performing longitudinal studies in experimental animals, for more in vivo relevant cell culture assays, and for future clinical neuro-regenerative applications

ΔΙΕΡΕΥΝΗΣΗ ΤΩΝ ΕΠΙΜΟΡΦΩΤΙΚΩΝ ΑΝΑΓΚΩΝ ΤΩΝ ΕΚΠΑΙΔΕΥΤΙΚΩΝ ΠΡΩΤΟΒΑΘΜΙΑΣ ΚΑΙ ΔΕΥΤΕΡΟΒΑΘΜΙΑΣ ΕΚΠΑΙΔΕΥΣΗΣ ΣΤΗ ΔΙΔΑΣΚΑΛΙΑ ΤΗΣ ΕΛΛΗΝΙΚΗΣ ΩΣ ΔΕΥΤΕΡΗΣ/ΞΕΝΗΣ ΓΛΩΣΣΑΣ

Περίληψη Οι σύγχρονες κοινωνίες, και κατ’ επέκταση και η Ελλάδα, χαρακτηρίζονται από ανομοιογένεια, όσον αφορά στους πληθυσμούς που κατοικούν σε αυτές. Οι νέες πολυπολιτισμικές συνθήκες στην ελληνική εκπαιδευτική πραγματικότητα δημιουργούν την ανάγκη ενός πλήρως καταρτισμένου εκπαιδευτικού, ο οποίος θα αναλάβει να διδάξει τη νέα ελληνική ως δεύτερη/ξένη γλώσσα. Η επιμόρφωση, λοιπόν, αποτελεί ένα σημαντικό εργαλείο προκειμένου να οπλιστεί με νέες γνώσεις και δεξιότητες, δίχως να παραβλέπει τις εκάστοτε ανάγκες και προσδοκίες που παρουσιάζονται σε μια κοινωνία που αλλάζει διαρκώς. Η παρούσα έρευνα διεξήχθη το 2018-2019 με σκοπό τη διερεύνηση των επιμορφωτικών αναγκών των εκπαιδευτικών Πρωτοβάθμιας και Δευτεροβάθμιας Εκπαίδευσης που διδάσκουν την ελληνική γλώσσας ως δεύτερη/ξένη γλώσσα.. Η έρευνα ήταν ποσοτική και εργαλείο συλλογής των δεδομένων ήταν το ερωτηματολόγιο, που χορηγήθηκε σε δείγμα 70 εκπαιδευτικών Πρωτοβάθμιας και Δευτεροβάθμιας Εκπαίδευσης. Το πρώτο μέρος του ερωτηματολογίου περιελάμβανε τα δημογραφικά στοιχεία των συμμετεχόντων, ενώ το δεύτερο εξέταζε τις ανάγκες επιμόρφωσης ανάλογα με τη φάση της διδασκαλίας (προετοιμασία-σχεδίαση διδασκαλίας, περιεχόμενο διδασκαλίας, υλοποίηση διδασκαλίας, εκπαιδευτικό υλικό διδασκαλίας και αποτελεσματικότητα διδασκαλίας). Σύμφωνα με τα αποτελέσματα της έρευνας, οι περισσότεροι εκπαιδευτικοί θεωρούν ότι χρειάζονται να επιμορφωθούν πάρα πολύ στον προσδιορισμό των εκπαιδευτικών αναγκών, των ιδιαίτερων ενδιαφέροντών και χαρακτηριστικών των μαθητών τους. Επίσης, κρίνουν πολύ αναγκαία την επιμόρφωσή τους στην αξιοποίηση των Τ.Π.Ε και την επιλογή κατάλληλων εκπαιδευτικών τεχνικών και στο ρόλο τους τους στις απαιτήσεις της σύγχρονης κοινωνίας. Οι εκπαιδευτικοί ζητούν να επιμορφωθούν πάρα πολύ στις τεχνικές διαμόρφωσης κατάλληλου ομαδοσυνεργατικού κλίματος και πολύ στη διαμόρφωση ισότιμων σχέσεων συνεργασίας και επικοινωνίας με τους μαθητές τους. Όσον αφορά το εκπαιδευτικό υλικό διδασκαλίας, εκφράζουν έντονα την επιμόρφωσή τους και πάνω σε αυτό το αντικείμενο.Abstract Modern societies, included Greece, are characterized by heterogeneity in the populations that inhabit them. The new multicultural conditions in the Greek educational reality create the need for a fully trained educator who will undertake to teach Greek as a second / foreign language. Training is therefore an important tool for arming new knowledge and skills, without overlooking the needs and aspirations of a constantly changing society. This survey was conducted in 2018-2019 to investigate the training needs of Primary and Secondary Education teachers who teach the Greek language as second / foreign language. The survey was quantitative and the data collection tool was the questionnaire, which was given in a sample 70 Primary and Secondary Education Teachers. The first part of the questionnaire included the demographics of the participants, while the second examined the training needs according to the teaching phase (teaching-preparation, teaching content, teaching, teaching material and teaching effectiveness). According to the results of the survey, most teachers feel that they need to be highly trained in identifying the educational needs, the particular interests and characteristics of their pupils. They also find it very necessary to train them in the use of ICTs and the choice of appropriate educational techniques and their role in the demands of modern society. Teachers are asking for a great deal of training in the techniques of shaping an appropriate co-operative climate, and much in equating cooperative and communication relationships with their students. As far as the teaching material is concerned, they strongly express their training on this subject as well

Being a Neural Stem Cell: A Matter of Character But Defined by the Microenvironment

The cells that build the nervous system, either this is a small network of ganglia or a complicated primate brain, are called neural stem and progenitor cells. Even though the very primitive and the very recent neural stem cells (NSCs) share common basic characteristics that are hard-wired within their character, such as the expression of transcription factors of the SoxB family, their capacity to give rise to extremely different neural tissues depends significantly on instructions from the microenvironment. In this chapter we explore the nature of the NSC microenvironment, looking through evolution, embryonic development, maturity and even disease. Experimental work undertaken over the last 20 years has revealed exciting insight into the NSC microcosmos. NSCs are very capable in producing their own extracellular matrix and in regulating their behaviour in an autocrine and paracrine manner. Nevertheless, accumulating evidence indicates an important role for the vasculature, especially within the NSC niches of the postnatal brain; while novel results reveal direct links between the metabolic state of the organism and the function of NSCs

In silico screening of GMQ-like compounds reveals guanabenz and sephin1 as new allosteric modulators of acid-sensing ion channel 3.

Acid-sensing ion channels (ASICs) are voltage-independent cation channels that detect decreases in extracellular pH. Dysregulation of ASICs underpins a number of pathologies. Of particular interest is ASIC3, which is recognised as a key sensor of acid-induced pain and is important in the establishment of pain arising from inflammatory conditions, such as rheumatoid arthritis. Thus, the identification of new ASIC3 modulators and the mechanistic understanding of how these compounds modulate ASIC3 could be important for the development of new strategies to counteract the detrimental effects of dysregulated ASIC3 activity in inflammation. Here, we report the identification of novel ASIC3 modulators based on the ASIC3 agonist, 2-guanidine-4-methylquinazoline (GMQ). Through a GMQ-guided in silico screening of Food and Drug administration (FDA)-approved drugs, 5 compounds were selected and tested for their modulation of rat ASIC3 (rASIC3) using whole-cell patch-clamp electrophysiology. Of the chosen drugs, guanabenz (GBZ), an α2-adrenoceptor agonist, produced similar effects to GMQ on rASIC3, activating the channel at physiological pH (pH 7.4) and potentiating its response to mild acidic (pH 7) stimuli. Sephin1, a GBZ derivative that lacks α2-adrenoceptor activity, has been proposed to act as a selective inhibitor of a regulatory subunit of the stress-induced protein phosphatase 1 (PPP1R15A) with promising therapeutic potential for the treatment of multiple sclerosis. However, we found that like GBZ, sephin1 activates rASIC3 at pH 7.4 and potentiates its response to acidic stimulation (pH 7), i.e. sephin1 is a novel modulator of rASIC3. Furthermore, docking experiments showed that, like GMQ, GBZ and sephin1 likely interact with the nonproton ligand sensor domain of rASIC3. Overall, these data demonstrate the utility of computational analysis for identifying novel ASIC3 modulators, which can be validated with electrophysiological analysis and may lead to the development of better compounds for targeting ASIC3 in the treatment of inflammatory conditions.This work was supported by Versus Arthritis Research Grants (RG20930 and RG21973; GC and EStJS), BBSRC grant (BB/R006210/1; JRFH and EStJS), BBSRC-funded studentships (LAP and JCG, BB/M011194/1) and Gates Cambridge Trust (SC)

Isolation of neural stem and oligodendrocyte progenitor cells from the brain of live rats.

Postnatal brain neural stem and progenitor cells (NSPCs) cluster in anatomically inaccessible stem cell niches, such as the subependymal zone (SEZ). Here, we describe a method for the isolation of NSPCs from live animals, which we term "milking." The intracerebroventricular injection of a release cocktail, containing neuraminidase, integrin-β1-blocking antibody, and fibroblast growth factor 2, induces the controlled flow of NSPCs in the cerebrospinal fluid, where they are collected via liquid biopsies. Isolated cells retain key in vivo self-renewal properties and their cell-type profile reflects the cell composition of their source area, while the function of the niche is sustained even 8 months post-milking. By changing the target area more caudally, we also isolate oligodendrocyte progenitor cells (OPCs) from the corpus callosum. This novel approach for sampling NSPCs and OPCs paves the way for performing longitudinal studies in experimental animals, for more in vivo relevant cell culture assays, and for future clinical neuro-regenerative applications