scholarly journals Bioorthogonal Labeling Reveals Different Expression of Glycans in Mouse Hippocampal Neuron Cultures during Their Development

Molecules ◽  
2020 ◽  
Vol 25 (4) ◽  
pp. 795 ◽  
Author(s):  
Diana Soares da Costa ◽  
João C. Sousa ◽  
Sandro Dá Mesquita ◽  
Nevena I. Petkova-Yankova ◽  
Fernanda Marques ◽  
...  
Keyword(s):  
Cell Fate ◽  
Hippocampal Neurons ◽  
Temporal Distribution ◽  
Physiological Conditions ◽  
Tissue Sections ◽  
Maturation In Vitro ◽  
Bioorthogonal Labeling ◽  
The Central Nervous System ◽  
Live Mouse

The expression of different glycans at the cell surface dictates cell interactions with their environment and other cells, being crucial for the cell fate. The development of the central nervous system is associated with tremendous changes in the cell glycome that is tightly regulated. Herein, we have employed bioorthogonal Cu-free click chemistry to image temporal distribution of different glycans in live mouse hippocampal neurons during their maturation in vitro. We show development-dependent glycan patterns with increased fucose and decreased mannose expression at the end of the maturation process. We also demonstrate that this approach is biocompatible and does not affect glycan transport although it relies on an administration of modified glycans. The applicability of this strategy to tissue sections unlocks new opportunities to study the glycan dynamics under more complex physiological conditions.

F-spondin and mindin: two structurally and functionally related genes expressed in the hippocampus that promote outgrowth of embryonic hippocampal neurons

Development ◽  
1999 ◽  
Vol 126 (16) ◽  
pp. 3637-3648 ◽  
Author(s):  
Y. Feinstein ◽  
V. Borrell ◽  
C. Garcia ◽  
T. Burstyn-Cohen ◽  
V. Tzarfaty ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Type I ◽  
Neural Connections ◽  
Putative Receptor ◽  
Rodent Brain ◽  
C Terminus ◽  
The Central Nervous System ◽  
Overlapping Domains ◽  
System F

Extracellular matrix (ECM) proteins play an important role in early cortical development, specifically in the formation of neural connections and in controlling the cyto-architecture of the central nervous system. F-spondin and Mindin are a family of matrix-attached adhesion molecules that share structural similarities and overlapping domains of expression. Genes for both proteins contain a thrombospondin type I repeat(s) at the C terminus and an FS1-FS2 (spondin) domain. Both the vertebrate F-spondin and the zebrafish mindins are expressed on the embryonic floor plate. In the current study we have cloned the rat homologue of mindin and studied its expression and activity together with F-spondin in the developing rodent brain. The two genes are abundantly expressed in the developing hippocampus. In vitro studies indicate that both F-spondin and Mindin promote adhesion and outgrowth of hippocampal embryonic neurons. We have also demonstrated that the two proteins bind to a putative receptor(s) expressed on both hippocampal and sensory neurons.

Imaging Molecular Targeting In Living Neural Cultures

1999 ◽  
Vol 5 (S2) ◽  
pp. 1228-1229
Author(s):  
Christopher S. Wallace ◽  
Michael A. Silverman ◽  
Michelle A. Burack ◽  
Janis E. Lochner ◽  
Richard G. Allen ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Hippocampal Neurons ◽  
Protein Targeting ◽  
Fluorescent Protein ◽  
Complex Structure ◽  
Time Lapse ◽  
The Central Nervous System

Recent technical advances in the ability to attach an endogenously fluorescent protein sequence—i.e., green fluorescent protein or GFP and its derivatives--to any protein of experimental interest promises to mark a new era of progress in the study of protein targeting. Bringing these new tools to bear on neurons of the central nervous system has been challenging, however, because they have a very complex structure and are relatively difficult to transfect because they are post-mitotic.We use two cell culture approaches to characterize protein trafficking within neurons of the central nervous system in vitro. The first is a dissociated culture of hippocampal neurons from embryonic (El8) rats which is especially suited to analysis by conventional light microscopy because these neurons are grown on glass coverslips at low density. Neurons cultured in this way develop a morphology comparable to that seen in vivo and permit the establishment of axons and dendrites to be analyzed by time-lapse microscopy.

scholarly journals Cytokinin Signaling and De Novo Shoot Organogenesis

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 265
Author(s):  
Katarzyna Hnatuszko-Konka ◽  
Aneta Gerszberg ◽  
Izabela Weremczuk-Jeżyna ◽  
Izabela Grzegorczyk-Karolak
Keyword(s):  
Cell Fate ◽  
Molecular Interactions ◽  
De Novo ◽  
Shoot Organogenesis ◽  
Control Mechanisms ◽  
Cytokinin Signaling ◽  
Tissue Sections ◽  
Transcriptional Reprogramming ◽  
Genetic Events

The ability to restore or replace injured tissues can be undoubtedly named among the most spectacular achievements of plant organisms. One of such regeneration pathways is organogenesis, the formation of individual organs from nonmeristematic tissue sections. The process can be triggered in vitro by incubation on medium supplemented with phytohormones. Cytokinins are a class of phytohormones demonstrating pleiotropic effects and a powerful network of molecular interactions. The present study reviews existing knowledge on the possible sequence of molecular and genetic events behind de novo shoot organogenesis initiated by cytokinins. Overall, the review aims to collect reactions encompassed by cytokinin primary responses, starting from phytohormone perception by the dedicated receptors, to transcriptional reprogramming of cell fate by the last module of multistep-phosphorelays. It also includes a brief reminder of other control mechanisms, such as epigenetic reprogramming.

scholarly journals Forever Young: How to Control the Elongation, Differentiation, and Proliferation of Cells Using Nanotechnology

2009 ◽  
Vol 18 (9) ◽  
pp. 1047-1058 ◽  
Author(s):  
R. G. Ellis-Behnke ◽  
Y. X. Liang ◽  
J. Guo ◽  
D. K. C. Tay ◽  
G. E. Schneider ◽  
...  
Keyword(s):  
Cell Activity ◽  
Local Environment ◽  
Nanofiber Scaffold ◽  
Self Assembling ◽  
Maturation In Vitro ◽  
The Matrix ◽  
The Central Nervous System ◽  
Differentiation And Proliferation

Within the emerging field of stem cells there is a need for an environment that can regulate cell activity, to slow down differentiation or proliferation, in vitro or in vivo while remaining invisible to the immune system. By creating a nanoenvironment surrounding PC12 cells, Schwann cells, and neural precursor cells (NPCs), we were able to control the proliferation, elongation, differentiation, and maturation in vitro. We extended the method, using self-assembling nanofiber scaffold (SAPNS), to living animals with implants in the brain and spinal cord. Here we show that when cells are placed in a defined system we can delay their proliferation, differentiation, and maturation depending on the density of the cell population, density of the matrix, and the local environment. A combination of SAPNS and young cells can be implanted into the central nervous system (CNS), eliminating the need for immunosuppressants.

scholarly journals Cultured Embryonic Hippocampal Neurons Deficient in Glucocorticoid (GC) Receptor: A Novel Model for Studying Nongenomic Effects of GC in the Neural System

Endocrinology ◽  
2005 ◽  
Vol 146 (9) ◽  
pp. 4036-4041 ◽  
Author(s):  
Lin Xiao ◽  
Aiqun Qi ◽  
Yizhang Chen
Keyword(s):  
Hippocampal Neurons ◽  
Neural System ◽  
Neural Cells ◽  
Rt Pcr ◽  
Nongenomic Action ◽  
The Central Nervous System ◽  
And Function ◽  
Nongenomic Effects ◽  
Novel Model

Abstract Glucocorticoid (GC) acts through both genomic and nongenomic mechanisms. It affects the structure and function of the central nervous system, especially the hippocampus. Here we report an in vitro culture system that can yield embryonic hippocampal neurons deficient in the expression of GC receptor as demonstrated by immunoblotting, immunocytochemistry, and RT-PCR. Owing to this unique feature, those neuron preparations can serve as an ideal model for studying the nongenomic actions of GC on neural cells. In this study, we found that the Erk1/2, c-Jun N-terminal kinase (JNK), and p38 MAPKs were activated in these neurons by BSA-conjugated corticosterone within 15 min of treatment. This activation was not blocked by RU38486, spironolactone, or cycloheximide. Therefore, it is concluded that the activation of MAPKs observed here was due to the nongenomic action of GC. Furthermore, a 24-h incubation with corticosterone at concentrations ranged from 10−11–10−5m did not have an effect on the viability of GC receptor-deficient neurons.

scholarly journals Amot regulates neuronal dendritic tree through Yap1

2018 ◽  
Author(s):  
Katarzyna O. Rojek ◽  
Joanna Krzemień ◽  
Hubert Doleżyczek ◽  
Paweł M. Boguszewski ◽  
Leszek Kaczmarek ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Hippo Pathway ◽  
Dendritic Tree ◽  
Cell Contact ◽  
Cellular Polarity ◽  
Dendritic Trees ◽  
Conditional Deletion ◽  
The Central Nervous System ◽  
Cerebellar Size

ABSTRACTThe Amot-Yap1 complex plays a major role in the regulation of cell contact inhibition, cellular polarity and growth. However, the function of Angiomotin (Amot) and Hippo pathway transcription co-activator Yap1 in the central nervous system remains unclear. In this study, we demonstrate that Amot is a critical mediator of dendritic morphogenesis in cultured hippocampal cells and Purkinje cells in the brain. Amot function in developing hippocampal neurons depends on interactions with Yap1, which is also indispensable for dendrite growth and arborization in vitro. Conditional deletion of Amot or Yap1 in neurons leads to impaired morphogenesis of Purkinje cell dendritic trees, decreased cerebellar size, and causes defects in locomotor coordination of mutant animals. Thus, our studies identified Amot and Yap1 as novel regulators of dendritic tree morphogenesis.

scholarly journals Erythropoietin affects GABAergic transmission in hippocampal neurons in vitro

2008 ◽  
Vol 13 (4) ◽  
Author(s):  
Tomasz Wójtowicz ◽  
Jerzy Mozrzymas
Keyword(s):  
Hippocampal Neurons ◽  
Term Treatment ◽  
Current Frequency ◽  
Gabaergic Transmission ◽  
Endogenous Factors ◽  
Long Term Treatment ◽  
The Central Nervous System ◽  
Neuronal Functions ◽  
The Impact

AbstractErythropoietin is a potent regulator of erythropoiesis. It acts via the specific membrane receptor (EpoR). Erythropoietin is also known to be present in the central nervous system, and its concentration and the expression of EpoR change during development, which raises the possibility that this modulator might be involved in the regulation of neuronal functions in the developing brain. The GABAergic system undergoes profound changes during development and is particularly susceptible to modulation by endogenous factors. Therefore, we decided to investigate the impact of Epo on GABAergic transmission in hippocampal neurons developing in vitro. An analysis of miniature IPSCs (mIPSCs) revealed that a long-term treatment with Epo (48 or 72 h) resulted in a major acceleration of the decaying phase of these currents while the amplitude and current frequency remained unchanged. Interestingly, this effect was restricted to the youngest considered age group (6-8 DIV), indicating that Epomediated modulation of mIPSCs depends on the developmental stage of the neurons. We conclude that Epo may exert a modulatory action on GABAergic transmission in developing neural networks.

scholarly journals What Can We Learn from FGF-2 Isoform-Specific Mouse Mutants? Differential Insights into FGF-2 Physiology In Vivo

2020 ◽  
Vol 22 (1) ◽  
pp. 390
Author(s):  
Friederike Freiin von Hövel ◽  
Ekaterini Kefalakes ◽  
Claudia Grothe
Keyword(s):  
Molecular Weight ◽  
Intracellular Localization ◽  
Comprehensive Overview ◽  
Knock Out ◽  
Mouse Mutants ◽  
Maturation In Vitro ◽  
Organ Systems ◽  
The Central Nervous System

Fibroblast growth factor 2 (FGF-2), ubiquitously expressed in humans and mice, is functionally involved in cell growth, migration and maturation in vitro and in vivo. Based on the same mRNA, an 18-kilo Dalton (kDa) FGF-2 isoform named FGF-2 low molecular weight (FGF-2LMW) isoform is translated in humans and rodents. Additionally, two larger isoforms weighing 21 and 22 kDa also exist, summarized as the FGF-2 high molecular weight (FGF-2HMW) isoform. Meanwhile, the human FGF-2HMW comprises a 22, 23, 24 and 34 kDa protein. Independent studies verified a specific intracellular localization, mode of action and tissue-specific spatiotemporal expression of the FGF-2 isoforms, increasing the complexity of their physiological and pathophysiological roles. In order to analyze their spectrum of effects, FGF-2LMW knock out (ko) and FGF-2HMWko mice have been generated, as well as mice specifically overexpressing either FGF-2LMW or FGF-2HMW. So far, the development and functionality of the cardiovascular system, bone formation and regeneration as well as their impact on the central nervous system including disease models of neurodegeneration, have been examined. This review provides a summary of the studies characterizing the in vivo effects modulated by the FGF-2 isoforms and, thus, offers a comprehensive overview of its actions in the aforementioned organ systems.

Effect of [Asu,]eel calcitonin on prolactin release in normal subjects and patients with prolactinoma

1985 ◽  
Vol 108 (3) ◽  
pp. 297-304 ◽  
Author(s):  
Hidesuke Kaji ◽  
Kazuo Chihara ◽  
Naoto Minamitani ◽  
Hitoshi Kodama ◽  
Tetsuya Kita ◽  
...  
Keyword(s):  
Body Weight ◽  
Bolus Injection ◽  
Normal Subjects ◽  
Regular Insulin ◽  
Prolactin Release ◽  
Saline Administration ◽  
The Central Nervous System ◽  
Plasma Prl ◽  
Male Subjects

Abstract. The effect of [Asu]eel calcitonin (ECT), an equipotent analogue of eel CT, on prolactin (Prl) secretion was examined in 12 healthy male subjects and in 6 patients with prolactinoma. In healthy subjects, ECT (0.5 μg/kg body weight · h) or saline was infused for 2 h and TRH was injected iv as a bolus of 500 μg at 1 h of ECT or saline administration. ECT did not affect basal Prl levels during 1 h of infusion. TRH caused a significant increase of plasma Prl with peak values of 75.2 ± 11.6 ng/ml in ECT-infused subjects, which did not differ from those infused with saline (68.5 ± 8.3 ng/ml). Next, an iv bolus injection of regular insulin (0.1 U/kg body weight) was followed by an infusion of ECT or saline alone. Plasma Prl peaks after hypoglycaemic stress were significantly lower in ECT-infused subjects than those in saline-injected controls (ECT, 16.5 ± 3.1 vs 33.5 ± 9.6 ng/ml, P < 0.05). In patients with prolactinoma, basal levels of plasma Prl ranging from 42.0–4130 ng/ml failed to change during iv infusion of ECT. Moreover, ECT (10−9–10−6m) did not affect Prl release from prolactinoma tissues perifused in vitro. These findings suggest that ECT may not act directly on the pituitary to modify Prl release. Rather, peripherally administered ECT appears to suppress Prl release via the central nervous system.

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