scholarly journals “Shepherd’s crook” neurons drive and synchronize the enhancing and suppressive mechanisms of the midbrain stimulus selection network

2018 ◽  
Vol 115 (32) ◽  
pp. E7615-E7623 ◽  
Author(s):  
Florencia Garrido-Charad ◽  
Tomas Vega-Zuniga ◽  
Cristián Gutiérrez-Ibáñez ◽  
Pedro Fernandez ◽  
Luciana López-Jury ◽  
...  
Keyword(s):  
Point Of View ◽  
Stimulus Selection ◽  
Common Input ◽  
Computational Point ◽  
Specific Subset ◽  
Neural Processes ◽  
Neural Tracing ◽  
Input Signals

The optic tectum (TeO), or superior colliculus, is a multisensory midbrain center that organizes spatially orienting responses to relevant stimuli. To define the stimulus with the highest priority at each moment, a network of reciprocal connections between the TeO and the isthmi promotes competition between concurrent tectal inputs. In the avian midbrain, the neurons mediating enhancement and suppression of tectal inputs are located in separate isthmic nuclei, facilitating the analysis of the neural processes that mediate competition. A specific subset of radial neurons in the intermediate tectal layers relay retinal inputs to the isthmi, but at present it is unclear whether separate neurons innervate individual nuclei or a single neural type sends a common input to several of them. In this study, we used in vitro neural tracing and cell-filling experiments in chickens to show that single neurons innervate, via axon collaterals, the three nuclei that comprise the isthmotectal network. This demonstrates that the input signals representing the strength of the incoming stimuli are simultaneously relayed to the mechanisms promoting both enhancement and suppression of the input signals. By performing in vivo recordings in anesthetized chicks, we also show that this common input generates synchrony between both antagonistic mechanisms, demonstrating that activity enhancement and suppression are closely coordinated. From a computational point of view, these results suggest that these tectal neurons constitute integrative nodes that combine inputs from different sources to drive in parallel several concurrent neural processes, each performing complementary functions within the network through different firing patterns and connectivity.

Digestion of soybean globulins, glycinin, α-conglycinin and β-conglycinin in the preruminant and the ruminant calf

1993 ◽  
Vol 73 (4) ◽  
pp. 891-905 ◽  
Author(s):  
H. M. Tukur ◽  
J. P. Lallès ◽  
C. Mathis ◽  
I. Caugant ◽  
R. Toullec
Keyword(s):  
Key Words ◽  
Soybean Protein ◽  
Point Of View ◽  
Protein Concentrate ◽  
Protein Digestion ◽  
Soybean Globulins ◽  
Milk Replacers ◽  
Meal Product

Two experiments involving either preruminant (exp. 1) or ruminant (exp. 2) fistulated calves were conducted to study the in vivo digestion of glycinin, α-conglycinin and β-conglycinin from soybean. It was incorporated as a flour (product A, antigenic in vitro) or a protein concentrate (product B, non antigenic in vitro) in milk replacers (exp. 1), or as a meal (product C, antigenic in vitro) in a weaning starter (exp. 2). ELISA detection of residual globulin immunoreactivity was determined on ileal digesta in exp. 1, and on duodenal, ileal and faecal digesta in exp. 2. Ileal flow of glycinin, α-conglycinin and β-conglycinin represented 10.3, 1.2 and 0.9% of corresponding globulin amounts ingested, in the case of product A (exp. 1). Immunoreactive α-conglycinin could only be detected in ileal digesta (1.3% of intake) of calves fed the diet containing product B. In exp. 2, immunoreactive globulins entered the duodenum in low amounts (below 1% of respective intake), especially after weaning. Accordingly, their flow at the ileum and in feces, although measurable, had no meaning from the nutritional point of view. Fermentation in the rumen of weaned calves appeared to be efficient in inactivating most of the potentially harmful dietary constituents. Key words: Soybean, protein, digestion, calf, weaning, glycinin

scholarly journals Immunity against cancer cells may promote their proliferation and metastasis

2019 ◽  
Vol 117 (1) ◽  
pp. 426-431
Author(s):  
Chih-Wei Lin ◽  
Jia Xie ◽  
Ding Zhang ◽  
Kyung Ho Han ◽  
Geramie Grande ◽  
...  
Keyword(s):  
Immune Response ◽  
Growth Factor ◽  
Cell Growth ◽  
Cancer Patients ◽  
Growth Factor Receptor ◽  
Point Of View ◽  
Breast Cancer Cell Growth ◽  
Agonist Antibody

Herein we present a concept in cancer where an immune response is detrimental rather than helpful. In the cancer setting, the immune system is generally considered to be helpful in curtailing the initiation and progression of tumors. In this work we show that a patient’s immune response to their tumor can, in fact, either enhance or inhibit tumor cell growth. Two closely related autoantibodies to the growth factor receptor TrkB were isolated from cancer patients’ B cells. Although highly similar in sequence, one antibody was an agonist while the other was an antagonist. The agonist antibody was shown to increase breast cancer cell growth both in vitro and in vivo, whereas the antagonist antibody inhibited growth. From a mechanistic point of view, we showed that binding of the agonist antibody to the TrkB receptor was functional in that it initiated downstream signaling identical to its natural growth factor ligand, brain-derived neurotrophic factor (BDNF). Our study shows that individual autoantibodies may play a role in cancer patients.

scholarly journals Disruption of CCL21-Induced Chemotaxis In Vitro and In Vivo by M3, a Chemokine-Binding Protein Encoded by Murine Gammaherpesvirus 68

2003 ◽  
Vol 77 (1) ◽  
pp. 624-630 ◽  
Author(s):  
Kristian K. Jensen ◽  
Shu-Cheng Chen ◽  
R. William Hipkin ◽  
Maria T. Wiekowski ◽  
Martin A. Schwarz ◽  
...  
Keyword(s):  
Lymphoid Tissues ◽  
Transgenic Expression ◽  
Murine Gammaherpesvirus ◽  
Chemokine Signaling ◽  
Specific Subset ◽  
Gammaherpesvirus 68 ◽  
Murine Gammaherpesvirus 68 ◽  
Human Chemokines

ABSTRACT Chemokine-binding proteins represent a novel class of antichemokine agents encoded by poxviruses and herpesviruses. One such protein is encoded by the M3 gene present in the murine gammaherpesvirus 68 (MHV-68) genome. The M3 gene encodes a secreted 44-kDa protein that binds with high affinity to certain murine and human chemokines and has been shown to block chemokine signaling in vitro. However, there has been no direct evidence that M3 blocks chemokine activity in vivo, nor has the nature of M3-chemokine interaction been defined. To better understand the ability of M3 to block chemokine activity in vivo, we examined its interaction with a specific subset of chemokines expressed in lymphoid tissues, areas where gammaherpesviruses characteristically establish latency. Here we show that M3 blocks in vitro chemotaxis induced by CCL19 and CCL21, chemokines expressed constitutively in secondary lymphoid tissues. Moreover, we provide evidence that chemokine M3 binding exhibits positive cooperativity. In vivo, the expression of M3 in the pancreas of transgenic mice inhibits recruitment of lymphocytes induced by transgenic expression of CCL21 in this organ. The ability of M3 to block the biological activity of chemokines may represent an important strategy used by MHV-68 to evade immune detection and favor viral replication in the infected host.

scholarly journals Hemoglobin E in Northeast India: A review on its origin, distribution, migration and health implication

2016 ◽  
Vol 79 (3) ◽  
pp. 241-263 ◽  
Author(s):  
Mithun Sikdar
Keyword(s):  
Systematic Review ◽  
Northeast India ◽  
Point Of View ◽  
In Vivo Studies ◽  
Mt Dna ◽  
Hemoglobin E ◽  
Health Implication ◽  
Probable Origin

AbstractA systematic review of the studies on hemoglobin E in Northeast India has been carried out to understand the magnitude of research undertaken on this aspect during the last seven decades. Owing to the high prevalence of hemoglobin E in this part of India different authors have studied this hemoglobin from different perspectives and found conflicting results. However a systematic review of such studies is lacking from a holistic point of view. Most of the epidemiological, in vitro as well as in vivo studies show signatures of selection with this hemoglobin locus. However, how this polymorphism is maintained at different rates at different geographical region is still a matter of contention. This review will fill the gap from all perspectives starting from the frequency distribution of hemoglobin E and its spread in different parts of Northeast India, its relationship with malaria hypothesis, the population migration, population affinity and most importantly the health implication arising out of it. A probable origin of hemoglobin E among an Austroasiatic population of Northeast India has been postulated with the help of advance molecular anthropological knowledge like the deep rooted markers of mt DNA and Y-chromosome haplotypes.

scholarly journals Soluble Urokinase Receptors in Focal Segmental Glomerulosclerosis: A Review on the Scientific Point of View

2016 ◽  
Vol 2016 ◽  
pp. 1-14 ◽  
Author(s):  
Andreas Kronbichler ◽  
Moin A. Saleem ◽  
Björn Meijers ◽  
Jae Il Shin
Keyword(s):  
Focal Segmental Glomerulosclerosis ◽  
Point Of View ◽  
Soluble Form ◽  
Glomerular Diseases ◽  
Segmental Glomerulosclerosis ◽  
Type Plasminogen Activator ◽  
Urokinase Type Plasminogen Activator ◽  
End Stage

Focal segmental glomerulosclerosis (FSGS) is one of the primary glomerular disorders in both children and adults which can progress to end-stage renal failure. Although there are genetic and secondary causes, circulating factors have also been regarded as an important factor in the pathogenesis of FSGS, because about 40% of the patients with FSGS have recurrence after renal transplantation. Soluble urokinase-type plasminogen activator receptor (suPAR) is a soluble form of uPAR, which is a membrane-bound protein linked to GPI in various immunologically active cells, including podocytes. It has recently been suggested as a potential circulating factor in FSGS by in vitro podocyte experiments, in vivo mice models, and human studies. However, there have also been controversies on this issue, because subsequent studies showed conflicting results. suPAR levels were also increased in patients with other glomerular diseases and were inversely correlated with estimated glomerular filtration rate. Nevertheless, there has been no balanced review on this issue. In this review, we compare the conflicting data on the involvement of suPAR in the pathogenesis of FSGS and shed light on interpretation by taking into account many points and the potential variables and confounders influencing serum suPAR levels.

scholarly journals S. mansoni SmKI-1 Kunitz-domain: Leucine point mutation at P1 site generates enhanced neutrophil elastase inhibitory activity

2021 ◽  
Vol 15 (1) ◽  
pp. e0009007
Author(s):  
Fábio Mambelli ◽  
Bruno P. O. Santos ◽  
Suellen B. Morais ◽  
Enrico G. T. Gimenez ◽  
Duana C. dos S. Astoni ◽  
...  
Keyword(s):  
Inhibitory Activity ◽  
Neutrophil Elastase ◽  
Point Mutations ◽  
Point Of View ◽  
Mutant Proteins ◽  
Inflammatory Parameters ◽  
C Terminus ◽  
Kunitz Domain

The Schistosoma mansoni SmKI-1 protein is composed of two domains: a Kunitz-type serine protease inhibitor motif (KD) and a C-terminus domain with no similarity outside the genera. Our previous work has demonstrated that KD plays an essential role in neutrophil elastase (NE) binding blockage, in neutrophil influx and as a potential anti-inflammatory molecule. In order to enhance NE blocking capacity, we analyzed the KD sequence from a structure-function point of view and designed specific point mutations in order to enhance NE affinity. We substituted the P1 site residue at the reactive site for a leucine (termed RL-KD), given its central role for KD’s inhibition to NE. We have also substituted a glutamic acid that strongly interacts with the P1 residue for an alanine, to help KD to be buried on NE S1 site (termed EA-KD). KD and the mutant proteins were evaluated in silico by molecular docking to human NE, expressed in Escherichia coli and tested towards its NE inhibitory activity. Both mutated proteins presented enhanced NE inhibitory activity in vitro and RL-KD presented the best performance. We further tested RL-KD in vivo in an experimental model of monosodium urate (MSU)-induced acute arthritis. RL-KD showed reduced numbers of total cells and neutrophils in the mouse knee cavity when compared to KD. Nevertheless, both RL-KD and KD reduced mice hypernociception in a similar fashion. In summary, our results demonstrated that both mutated proteins showed enhanced NE inhibitory activity in vitro. However, RL-KD had a prominent effect in diminishing inflammatory parameters in vivo.

scholarly journals New Insight to Overcome Tumor Resistance: An Overview from Cellular to Clinical Therapies

Life ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 1131
Author(s):  
Giulia Mitola ◽  
Paolo Falvo ◽  
Francesco Bertolini
Keyword(s):  
Residual Disease ◽  
Drug Effects ◽  
Point Of View ◽  
Tumor Resistance ◽  
Cancer Treatments ◽  
Secondary Tumors ◽  
Molecular Features ◽  
Preclinical And Clinical Studies

Disease relapse caused by drug resistance still represents a major clinical hurdle in cancer treatments. Tumor cells may take advantage of different intracellular and genetic systems attenuating the drug effects. Resistant cells or minimal residual disease (MRD) cells have strong clinical relevance, as they might give rise to secondary tumors when the therapy is concluded. Thus, MRDs are crucial therapeutic targets in order to prevent tumor relapse. Therefore, several groups aim at understanding how MRDs are orginated, characterizing their molecular features, and eradicating them. In this review, we will describe MRD from a genetic, evolutionary, and molecular point of view. Moreover, we will focus on the new in vitro, in vivo, preclinical, and clinical studies that aim at eradicating tumor resistance.

scholarly journals Μελέτη της έκφρασης των κοινών οδών των γονιδίων in vitro και in vivo στους όγκους του κεντρικού νευρικού συστήματος για την ανεύρεση αιματολογικών δεικτών

2013 ◽  
Author(s):  
Γεώργιος Λάμπρου
Keyword(s):  
Gene Structure ◽  
Statistical Physics ◽  
Point Of View ◽  
Henri Poincaré

Τα νεοπλασματικά νοσήματα έχουν πάρει πλέον τη μορφή επιδημίας. Η αιτιολογία της ογκογένεσηςπαραμένει στο μεγαλύτερο μέρος της άγνωστη. Έχουν διατυπωθεί αρκετές θεωρίες για τουςμηχανισμούς ογκογένεσης αλλά καμία μέχρι σήμερα δεν έχει δώσει μια ικανοποιητική εξήγηση.Ειδικότερα όταν αναφερόμαστε στις παιδικές νεοπλασίες η κατανόηση του φαινομένου λαμβάνει πιοιδιαίτερες διαστάσεις λόγω της ευαισθησίας του πληγμένου πληθυσμού. Η πλειοψηφία των παιδικώννεοπλασιών αφορά στις αιματολογικές κακοήθειες, όπως είναι η Οξεία Λεμφοβλαστική Λευχαιμία(ΟΛΛ). Στην περίπτωση της ΟΛΛ έχει γίνει τεράστια πρόοδος στη θεραπευτική προσέγγισή τηςαφού τα επίπεδα επιβίωσης αγγίζουν το 75%-80%. Το αποτέλεσμα είναι εκπληκτικό αν σκεφθείκανείς ότι 40 έτη πριν η λευχαιμία ήταν θανατηφόρα στο 90% των περιπτώσεων. Παρά την πρόοδοπου έχει επιτευχθεί υπάρχει ακόμα ένα 10-20% των ασθενών, που υποτροπιάζει. Τα αίτια τωνυποτροπών ακόμα δεν έχουν εξηγηθεί ή κατανοηθεί πλήρως.Η περίπτωση του Κεντρικού Νευρικού Συστήματος (ΚΝΣ) είναι αρκετά πιο περίπλοκη. Ταποσοστά επιβίωσης είναι αρκετά χαμηλά και το προσδόκιμο επιβίωσης δεν ξεπερνάει τα δύο με τρίαέτη. Τα νεοπλάσματα του ΚΝΣ έχουν ιδιαίτερο ενδιαφέρον και βαρύτητα για δύο λόγους. Ο πρώτοςλόγος είναι ότι τα κύτταρα του εγκεφάλου δεν πολλαπλασιάζονται, ή τουλάχιστον δεν επιτελούν αυτήτη διαδικασία στο βαθμό που γίνεται στα υπόλοιπα κύτταρα του σώματος και ο δεύτερος λόγος είναιότι η λειτουργία του εγκεφάλου αποτελεί από μόνη της μια πρόκληση, που ακόμα δεν έχει απαντηθείή κατανοηθεί.Το κύριο βάρος στην έρευνα έχει δοθεί στην κατανόηση των μηχανισμών της ογκογένεσης, τηνταυτοποίηση προγνωστικών παραγόντων και εν γένει την ταξινόμηση και διαφοροποίηση τωννεοπλασιών με τη βοήθεια μοριακών χαρακτηριστικών. Στο σημείο αυτό πρέπει να θυμίσουμε ότι τοκύριο δόγμα της έρευνας ήταν η διατύπωση μιας υπόθεσης και η μετέπειτα διερεύνησή της. Με τηνεμφάνιση των νέων πειραματικών τεχνολογιών, όπως οι μικροσυστοιχίες, η έρευνα άρχισε να κινείταιπρος την κατεύθυνση της διερεύνησης με βάση την ανακάλυψη. Δηλαδή, πρώτα παρατηρούμε καιμετά διερευνούμε. Κατά την προσωπική μας άποψη η παρατήρηση μεγάλου όγκου δεδομένων δεναποκλείει την πρότερη διατύπωση υποθέσεως αλλά αντιθέτως της δίνει και μεγαλύτερη βαρύτητα.Η εφαρμογή αυτών των τεχνολογιών φάνηκε ιδιαίτερα χρήσιμη σε εφαρμογές, όπως η ταυτοποίησητύπων νεοπλασμάτων με καλύτερα αποτελέσματα από τις κλασσικές μεθόδους που μπορούσαν ναεξετάσουν μόνο μερικούς παράγοντες ανά βιολογικό φαινόμενο. Από την άλλη όμως, οι νέεςτεχνολογίες δεν μας βοήθησαν αρκετά στο να κατανοήσουμε το μηχανισμό ασθενειών, όπως οινεοπλασίες, αλλά δημιούργησαν περισσότερα ερωτήματα. Πιθανότατα αυτό να οφείλεται στογεγονός ότι τα βιολογικά φαινόμενα είναι εξαιρετικά περίπλοκα και η κατανόησή τους περνάει μέσααπό άλλες οδούς. Η εφαρμογή των νέων τεχνολογιών στη βιολογία και βιοϊατρική οδήγησε στη δημιουργία μιας νέαςπροσέγγισης στη θεραπευτική που ονομάστηκε ασθενο-κεντρική προσέγγιση. Η προσέγγιση αυτήστοχεύει στο να ανιχνεύσει ειδικούς παράγοντες που διαφοροποιούν, όχι πλέον τη νόσο αλλά τονασθενή. Στο σημείο αυτό πρέπει να αναφέρουμε ότι η παρούσα εργασία κινείται στον αντίθετο άξονααπό την επικρατούσα τάση. Η φιλοσοφία της παρούσης εργασίας αφορά στην κατανόηση τωνβιολογικών φαινομένων και στη διατύπωση γενικών νόμων ή θεωριών που να περιγράφουν ταβιολογικά φαινόμενα. Κατά συνέπεια αν αυτό το προεκτείνουμε στην καρκινική οντογένεση θαλέγαμε, ότι διερευνήσαμε την πιθανότητα ύπαρξης κοινών οδών και μηχανισμών στην καρκινικήβιολογία. Η ιδέα πίσω από τη φιλοσοφία προκύπτει από το γεγονός (και πράγματι αυτό είναι έναγεγονός) ο καρκινικός φαινότυπος είναι παρόμοιος είτε μιλάμε για όγκους του ΚΝΣ είτε του ήπατοςείτε του αίματος. Τέτοια παρόμοια χαρακτηριστικά είναι η μη ελεγχόμενη πολλαπλασιαστικήδυναμική, η απώλεια των μηχανισμών της απόπτωσης, η απώλεια της ιδιότητας της αναστολής λόγωμεμβρανικής αλληλεπίδρασης και άλλα. Αυτή η κοινή μηχανική πιθανότατα να προκύπτει απόκοινούς μοριακούς μηχανισμούς.Για το σκοπό αυτό χρησιμοποιήσαμε τέσσερα εργαλεία. Το πρώτο είναι ο πειραματισμός, τοδεύτερο ήταν η θεωρία συστημάτων, το τρίτο ήταν η χαοτική μηχανική ήτοι τα μη-γραμμικάσυστήματα και το τέταρτο ήταν η φυσική βιολογία. Δεν πιστεύουμε ότι υπάρχει καλύτερος τρόποςνα περιγράψουμε την παρούσα προσέγγιση παρά μέσα από τα λόγια του μαθηματικού του 19ου αιώνα, Henri Poincaré:“…life is a relationship among molecules and not a property of any molecule…”“…Science is built up of facts, as a house is with stones. But a collection of facts is no more a science, than a heapof stones is a house…”Επίσης έχει τεράστιο ενδιαφέρον να παραθέσουμε τα λόγια ενός φυσικού του 20ου αιώνα του ErwinSchrödinger του οποίου οι εργασίες μας επηρέασαν κατά τη μελέτη των παρόντων φαινομένων, “…howcan we, from the point of view of statistical physics, reconcile the facts that the gene structure seems to involve only acomparatively small number of atoms and that nevertheless it displays a most regular and lawful activity withadurability or permanence that borders upon them miraculous?...the evidence that two features, similar inappearance, are based on the same principle, is always precarious as long as the principle itself is unknown.”Για να συνοψίσουμε, η παρούσα εργασία επικεντρώθηκε σε μια βασική ερώτηση: ποιες είναι οιαλλαγές και μηχανισμοί που λαμβάνουν χώρα κατά τον καρκινικό πολλαπλασιασμό πριν τηδιάγνωση; Για να επιχειρήσουμε να δώσουμε μια απάντηση σε αυτό το ερώτημα χρησιμοποιήσαμετέσσερις άξονες διερεύνησης. Ο πρώτος άξονας αφορούσε στη δημιουργία ενός in vitro,πολλαπλασιαστικού μοντέλου με στόχο να εξετάσουμε την πορεία των καρκινικών κυττάρων από τηστιγμή της εμφάνισής τους μέχρι τη στιγμή λίγο πριν τη διάγνωση. Ο δεύτερος άξονας αφορούσεστη διερεύνηση κρίσιμων σηματοδοτικών οδών κατά τον κυτταρικό πολλαπλασιασμό με τη χρήση αναστολέων και/ή φαρμάκων γνωστών για τη δράση τους. Ο τρίτος άξονας αφορούσε στη γονιδιακήέκφραση και ρύθμιση κατά τα στάδια του πολλαπλασιασμού και τέλος ο τέταρτος άξονας αφορούσεστη διερεύνηση των αλλαγών στο εξωκυττάριο περιβάλλον και ειδικότερα στις αλλαγές στιςφυσικοχημικές/ηλεκτροχημικές ιδιότητες των περιβαλλόντων βιολογικών υγρών.

scholarly journals A Modeling Study Suggesting a Possible Pharmacological Target to Mitigate the Effects of Ethanol on Reward-Related Dopaminergic Signaling

2008 ◽  
Vol 99 (5) ◽  
pp. 2703-2707 ◽  
Author(s):  
Michele Migliore ◽  
Claudio Cannia ◽  
Carmen C. Canavier
Keyword(s):  
Dopaminergic Neurons ◽  
Temporal Structure ◽  
Ionic Current ◽  
Brain Regions ◽  
Point Of View ◽  
Dopamine Level ◽  
Addictive Behaviors ◽  
Brain Functions

Midbrain dopaminergic neurons are involved in several critical brain functions controlling goal-directed behaviors, reinforcing/reward processes, and motivation. Their dysfunctions alter dopamine release and contribute to a vast range of neural disorders, from Parkinson's disease to schizophrenia and addictive behaviors. These neurons have thus been a natural target of pharmacological treatments trying to ameliorate the consequences of several neuropathologies. From this point of view, a clear experimental link has been recently established between the increase in the pacemaker frequency of dopaminergic neurons in vitro after acute ethanol application and a particular ionic current ( Ih). The functional consequences in vivo, however, are not clear and they are very difficult to explore experimentally. Here we use a realistic computational model of dopaminergic neurons in vivo to suggest that ethanol, through its effects on Ih, modifies the temporal structure of the spiking activity. The model predicts that the dopamine level may increase much more during bursting than during pacemaking activity, especially in those brain regions with a slow dopamine clearance rate. The results suggest that a selective pharmacological remedy could thus be devised against the rewarding effects of ethanol that are postulated to mediate alcohol abuse and addiction, targeting the specific HCN genes expressed in dopaminergic neurons.

scholarly journals Identification of reference genes for RT-qPCR data normalisation in aging studies

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Lourdes González-Bermúdez ◽  
Teresa Anglada ◽  
Anna Genescà ◽  
Marta Martín ◽  
Mariona Terradas
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Quantitative Polymerase Chain Reaction ◽  
Expression Levels ◽  
Qpcr Data ◽  
Age Related ◽  
Specific Subset ◽  
Aging Studies

Abstract Aging is associated with changes in gene expression levels that affect cellular functions and predispose to age-related diseases. The use of candidate genes whose expression remains stable during aging is required to correctly address the age-associated variations in expression levels. Reverse transcription quantitative-polymerase chain reaction (RT-qPCR) has become a powerful approach for sensitive gene expression analysis. Reliable RT-qPCR assays rely on the normalisation of the results to stable reference genes. Taken these data together, here we evaluated the expression stability of eight frequently used reference genes in three aging models: oncogene-induced senescence (OIS), in vitro and in vivo aging. Using NormFinder and geNorm algorithms, we identified that the most stable reference gene pairs were PUM1 and TBP in OIS, GUSB and PUM1 for in vitro aging and GUSB and OAZ1 for in vivo aging. To validate these candidates, we used them to normalise the expression data of CDKN1A, APOD and TFRC genes, whose expression is known to be affected during OIS, in vitro and in vivo aging. This study demonstrates that accurate normalisation of RT-qPCR data is crucial in aging research and provides a specific subset of stable reference genes for future aging studies.

Sign in / Sign up

Export Citation Format

Share Document