scholarly journals Bidirectional Cycling Dynamics of Living Neuronal Networks in Vitro

2017 ◽  
Author(s):  
Arseniy Gladkov ◽  
Oleg Grinchuk ◽  
Pigareva Yana ◽  
Irina Mukhina ◽  
Victor Kazantsev ◽  
...  
Keyword(s):  
Binary Sequence ◽  
Rhythmic Activity ◽  
Time Base ◽  
Cellular Density ◽  
Self Organized ◽  
High Frequency Oscillations ◽  
Random Switching ◽  
Cultured Networks

AbstractPhenomena of synchronization, rhythmogenesis and coherence found in brain networks are believed to be a dynamical substrate for cognitive functions such as learning and memory. However, it is still debated whether the rhythmic activity emerges from network morphology developed in neurogenesis or as a result of neuronal dynamics realized under certain conditions. In this research we found, that in neural networks formed in mature hippocampal cultures with high cellular density the spiking activity self-organized and converged to long, complex and rhythmically repeated superbursts. The superburst lasted tens of seconds and consisted of hundreds of short (50-100 ms) small bursts with a high spiking rate of 139.0 ± 78.6 Hz that can be associated with high-frequency oscillations in the hippocampus. In turn, the interval between peak burst activities in the range of 100-150 ms can be treated as a theta rhythm (11.2 ± 1.5 Hz). Distribution of spikes within the bursts was non-random, representing a set of well-defined space-time base patterns or motifs. We found that the long superburst can be classified into two types. Each type was associated with a unique direction of spike propagation and, hence, can be encoded by a binary sequence with random switching between the two “functional” states. Such precisely structured bidirectional rhythmic activity developed in self-organizing cultured networks were quite similar to what observed in the in vivo experiments.

Nasal Trigeminal Inputs Release the A5 Inhibition Received by the Respiratory Rhythm Generator of the Mouse Neonate

2004 ◽  
Vol 91 (2) ◽  
pp. 746-758 ◽  
Author(s):  
Jean-Charles Viemari ◽  
Michelle Bévengut ◽  
Patrice Coulon ◽  
Gérard Hilaire
Keyword(s):  
Trigeminal Nerve ◽  
Respiratory Rhythm ◽  
Rhythmic Activity ◽  
Electrolytic Lesion ◽  
Burst Frequency ◽  
Neuronal Tracing ◽  
Rhythm Generator ◽  
Trigeminal Nerve Stimulation

Experiments were performed on neonatal mice to analyze why, in vitro, the respiratory rhythm generator (RRG) was silent and how it could be activated. We demonstrated that in vitro the RRG in intact brain stems is silenced by a powerful inhibition arising from the pontine A5 neurons through medullary α2 adrenoceptors and that in vivo nasal trigeminal inputs facilitate the RRG as nasal continuous positive airway pressure increases the breathing frequency, whereas nasal occlusion and nasal afferent anesthesia depress it. Because nasal trigeminal afferents project to the A5 nuclei, we applied single trains of negative electric shocks to the trigeminal nerve in inactive ponto-medullary preparations. They induced rhythmic phrenic bursts during the stimulation and for 2–3 min afterward, whereas repetitive trains produced on-going rhythmic activity up to the end of the experiments. Electrolytic lesion or pharmacological inactivation of the ipsilateral A5 neurons altered both the phrenic burst frequency and occurrence after the stimulation. Extracellular unitary recordings and trans-neuronal tracing experiments with the rabies virus show that the medullary lateral reticular area contains respiratory-modulated neurons, not necessary for respiratory rhythmogenesis, but that may provide an excitatory pathway from the trigeminal inputs to the RRG as their electrolytic lesion suppresses any phrenic activity induced by the trigeminal nerve stimulation. The results lead to the hypothesis that the trigeminal afferents in the mouse neonate involve at least two pathways to activate the RRG, one that may act through the medullary lateral reticular area and one that releases the A5 inhibition received by the RRG.

scholarly journals Decreased Fast Ripples in the Hippocampus of Rats with Spontaneous Recurrent Seizures Treated with Carbenoxolone and Quinine

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Consuelo Ventura-Mejía ◽  
Laura Medina-Ceja
Keyword(s):  
Gap Junctions ◽  
Gap Junction ◽  
High Frequency Oscillations ◽  
The Mean ◽  
Eeg Recordings ◽  
The Right ◽  
Spontaneous Recurrent Seizures

Background. In models of temporal lobe epilepsy and in patients with this pathology, high frequency oscillations called fast ripples (FRs, 250–600 Hz) can be observed. FRs are considered potential biomarkers for epilepsy and, in the light of manyin vitroandin silicostudies, we thought that electrical synapses mediated by gap junctions might possibly modulate FRsin vivo.Methods. Animals with spontaneous recurrent seizures induced by pilocarpine administration were implanted with movable microelectrodes in the right anterior and posterior hippocampus to evaluate the effects of gap junction blockers administered in the entorhinal cortex. The effects of carbenoxolone (50 nmoles) and quinine (35 pmoles) on the mean number of spontaneous FR events (occurrence of FRs), as well as on the mean number of oscillation cycles per FR event and their frequency, were assessed using a specific algorithm to analyze FRs in intracranial EEG recordings.Results. We found that these gap junction blockers decreased the mean number of FRs and the mean number of oscillation cycles per FR event in the hippocampus, both during and at different times after carbenoxolone and quinine administration.Conclusion. These data suggest that FRs may be modulated by gap junctions, although additional experimentsin vivowill be necessary to determine the precise role of gap junctions in this pathological activity associated with epileptogenesis.

scholarly journals Placenta Defects and Embryonic Lethality Resulting from Disruption of Mouse Hydroxysteroid (17-β) Dehydrogenase 2 Gene

2008 ◽  
Vol 22 (3) ◽  
pp. 665-675 ◽  
Author(s):  
Pia Rantakari ◽  
Leena Strauss ◽  
Riku Kiviranta ◽  
Heidi Lagerbohm ◽  
Jenni Paviala ◽  
...  
Keyword(s):  
Embryonic Stem ◽  
Basal Layer ◽  
Embryonic Lethality ◽  
Cellular Density ◽  
Targeted Gene Disruption ◽  
Structural Abnormalities ◽  
The Brain

Abstract Hydroxysteroid (17-β) dehydrogenase 2 (HSD17B2) is a member of aldo-keto reductase superfamily, known to catalyze the inactivation of 17β-hydroxysteroids to less active 17-keto forms and catalyze the conversion of 20α-hydroxyprogesterone to progesterone in vitro. To examine the role of HSD17B2 in vivo, we generated mice deficient in Hsd17b2 [HSD17B2 knockout (KO)] by a targeted gene disruption in embryonic stem cells. From the homozygous mice carrying the disrupted Hsd17b2, 70% showed embryonic lethality appearing at the age of embryonic d 11.5 onward. The embryonic lethality was associated with reduced placental size measured at embryonic d 17.5. The HSD17B2KO mice placentas presented with structural abnormalities in all three major layers: the decidua, spongiotrophoblast, and labyrinth. Most notable was the disruption of the spongiotrophoblast and labyrinthine layers, together with liquid-filled cysts in the junctional region and the basal layer. Treatments with an antiestrogen or progesterone did not rescue the embryonic lethality or the placenta defect in the homozygous mice. In hybrid background used, 24% of HSD17B2KO mice survived through the fetal period but were born growth retarded and displayed a phenotype in the brain with enlargement of ventricles, abnormal laminar organization, and increased cellular density in the cortex. Furthermore, the HSD17B2KO mice had unilateral renal degeneration, the affected kidney frequently appearing as a fluid-filled sac. Our results provide evidence for a role for HSD17B2 enzyme in the cellular organization of the mouse placenta.

scholarly journals Quantifying the correlation between spatially defined oxygen gradients and cell fate in an engineered three-dimensional culture model

2014 ◽  
Vol 11 (98) ◽  
pp. 20140501 ◽  
Author(s):  
Amir G. Ardakani ◽  
Umber Cheema ◽  
Robert A. Brown ◽  
Rebecca J. Shipley
Keyword(s):  
Cell Fate ◽  
Cellular Proliferation ◽  
Three Dimensional ◽  
Quantitative Information ◽  
Cell Number ◽  
Cellular Density ◽  
Spatially Resolved ◽  
Migratory Speed

A challenge in three-dimensional tissue culture remains the lack of quantitative information linking nutrient delivery and cellular distribution. Both in vivo and in vitro , oxygen is delivered by diffusion from its source (blood vessel or the construct margins). The oxygen level at a defined distance from its source depends critically on the balance of diffusion and cellular metabolism. Cells may respond to this oxygen environment through proliferation, death and chemotaxis, resulting in spatially resolved gradients in cellular density. This study extracts novel spatially resolved and simultaneous data on tissue oxygenation, cellular proliferation, viability and chemotaxis in three-dimensional spiralled, cellular collagen constructs. Oxygen concentration gradients drove preferential cellular proliferation rates and viability in the higher oxygen zones and induced chemotaxis along the spiral of the collagen construct; an oxygen gradient of 1.03 mmHg mm −1 in the spiral direction induced a mean migratory speed of 1015 μm day −1 . Although this movement was modest, it was effective in balancing the system to a stable cell density distribution, and provided insights into the natural cell mechanism for adapting cell number and activity to a prevailing oxygen regime.

Spinal 5-HT7 Receptors Are Critical for Alternating Activity During Locomotion: In Vitro Neonatal and In Vivo Adult Studies Using 5-HT7 Receptor Knockout Mice

2009 ◽  
Vol 102 (1) ◽  
pp. 337-348 ◽  
Author(s):  
Jun Liu ◽  
Turgay Akay ◽  
Peter B. Hedlund ◽  
Keir G. Pearson ◽  
Larry M. Jordan
Keyword(s):  
Spinal Cord ◽  
Knockout Mice ◽  
Rhythmic Activity ◽  
Neonatal Mouse ◽  
Emg Activity ◽  
In Vivo Experiments ◽  
Adult Mice ◽  
Ankle Joints

5-HT7 receptors have been implicated in the control of locomotion. Here we use 5-HT7 receptor knockout mice to rigorously test whether 5-HT acts at the 5-HT7 receptor to control locomotor-like activity in the neonatal mouse spinal cord in vitro and voluntary locomotion in adult mice. We found that 5-HT applied onto in vitro spinal cords of 5-HT7+/+ mice produced locomotor-like activity that was disrupted and subsequently blocked by the 5-HT7 receptor antagonist SB-269970. In spinal cords isolated from 5-HT7−/− mice, 5-HT produced either uncoordinated rhythmic activity or resulted in synchronous discharges of the ventral roots. SB-269970 had no effect on 5-HT-induced rhythmic activity in the 5-HT7−/− mice. In adult in vivo experiments, SB-269970 applied directly to the spinal cord consistently disrupted locomotion and produced prolonged-extension of the hindlimbs in 5-HT7+/+ but not 5-HT7−/− mice. Disrupted EMG activity produced by SB-269970 in vivo was similar to the uncoordinated rhythmic activity produced by the drug in vitro. Moreover, 5-HT7−/− mice displayed greater maximal extension at the hip and ankle joints than 5-HT7+/+ animals during voluntary locomotion. These results suggest that spinal 5-HT7 receptors are required for the production and coordination of 5-HT-induced locomotor-like activity in the neonatal mouse and are important for the coordination of voluntary locomotion in adult mice. We conclude that spinal 5-HT7 receptors are critical for alternating activity during locomotion.

scholarly journals Neurosteroids and Focal Epileptic Disorders

2020 ◽  
Vol 21 (24) ◽  
pp. 9391
Author(s):  
Maxime Lévesque ◽  
Giuseppe Biagini ◽  
Massimo Avoli
Keyword(s):  
Brain Slices ◽  
Mesial Temporal Lobe Epilepsy ◽  
Epileptic Activity ◽  
Network Activity ◽  
Interictal Spikes ◽  
In Vivo Models ◽  
High Frequency Oscillations ◽  
Spontaneous Seizures

Neurosteroids are a family of compounds that are synthesized in principal excitatory neurons and glial cells, and derive from the transformation of cholesterol into pregnenolone. The most studied neurosteroids—allopregnanolone and allotetrahydrodeoxycorticosterone (THDOC)—are known to modulate GABAA receptor-mediated transmission, thus playing a role in controlling neuronal network excitability. Given the role of GABAA signaling in epileptic disorders, neurosteroids have profound effects on seizure generation and play a role in the development of chronic epileptic conditions (i.e., epileptogenesis). We review here studies showing the effects induced by neurosteroids on epileptiform synchronization in in vitro brain slices, on epileptic activity in in vivo models, i.e., in animals that were made epileptic with chemoconvulsant treatment, and in epileptic patients. These studies reveal that neurosteroids can modulate ictogenesis and the occurrence of pathological network activity such as interictal spikes and high-frequency oscillations (80–500 Hz). Moreover, they can delay the onset of spontaneous seizures in animal models of mesial temporal lobe epilepsy. Overall, this evidence suggests that neurosteroids represent a new target for the treatment of focal epileptic disorders.

Medullary regions for neurogenesis of gasping: noeud vital or noeuds vitals?

1996 ◽  
Vol 81 (5) ◽  
pp. 1865-1877 ◽  
Author(s):  
Walter M. St. John
Keyword(s):  
Mammalian Species ◽  
Respiratory Rhythm ◽  
Rhythmic Activity ◽  
Neonatal Rat ◽  
Rhythm Generation ◽  
Bötzinger Complex ◽  
Per Se ◽  
Respiratory Rhythm Generation

St. John, Walter M. Medullary regions for neurogenesis of gasping: noeud vital or noeuds vitals? J. Appl. Physiol. 81(5): 1865–1877, 1996.—Gasping is a critical mechanism for survival in that it serves as a mechanism for autoresuscitation when eupnea fails. Eupnea and gasping are separable patterns of automatic ventilatory activity in all mammalian species from the day of birth. The neurogenesis of the gasp is dependent on the discharge of neurons in the rostroventral medulla. This gasping center overlaps a region termed “the pre-Bötzinger complex.” Neuronal activities of this complex, characterized in an in vitro brain stem spinal cord preparation of the neonatal rat, have been hypothesized to underlie respiratory rhythm generation. Yet, the rhythmic activity of this in vitro preparation is markedly different from eupnea but identical with gasping in vivo. In eupnea, medullary neuronal activities generating the gasp and the identical rhythm of the in vitro preparation are incorporated into a portion of the pontomedullary circuit defining eupneic ventilatory activity. However, these medullary neuronal activities do not appear critical for the neurogenesis of eupnea, per se.

scholarly journals Microfluidic Organoids-on-a-Chip: Quantum Leap in Cancer Research

Cancers ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 737 ◽  
Author(s):  
Fahriye Duzagac ◽  
Gloria Saorin ◽  
Lorenzo Memeo ◽  
Vincenzo Canzonieri ◽  
Flavio Rizzolio
Keyword(s):  
Ex Vivo ◽  
Fluid Flows ◽  
Cost Effective ◽  
In Vitro Models ◽  
Organoid Culture ◽  
Self Organized ◽  
Quantum Leap ◽  
High Degree

Organ-like cell clusters, so-called organoids, which exhibit self-organized and similar organ functionality as the tissue of origin, have provided a whole new level of bioinspiration for ex vivo systems. Microfluidic organoid or organs-on-a-chip platforms are a new group of micro-engineered promising models that recapitulate 3D tissue structure and physiology and combines several advantages of current in vivo and in vitro models. Microfluidics technology is used in numerous applications since it allows us to control and manipulate fluid flows with a high degree of accuracy. This system is an emerging tool for understanding disease development and progression, especially for personalized therapeutic strategies for cancer treatment, which provide well-grounded, cost-effective, powerful, fast, and reproducible results. In this review, we highlight how the organoid-on-a-chip models have improved the potential of efficiency and reproducibility of organoid cultures. More widely, we discuss current challenges and development on organoid culture systems together with microfluidic approaches and their limitations. Finally, we describe the recent progress and potential utilization in the organs-on-a-chip practice.

scholarly journals Rapid fabrication of hydrogel micropatterns by projection stereolithography for studying self-organized developmental patterning

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0245634
Author(s):  
Ye Zhu ◽  
Daniel Sazer ◽  
Jordan S. Miller ◽  
Aryeh Warmflash
Keyword(s):  
Embryonic Stem ◽  
Culture Vessel ◽  
Developmental Studies ◽  
Contact Printing ◽  
Self Organized ◽  
Rapid Fabrication ◽  
Micro Features ◽  
Projection Stereolithography

Self-organized patterning of mammalian embryonic stem cells on micropatterned surfaces has previously been established as an in vitro platform for early mammalian developmental studies, complimentary to in vivo studies. Traditional micropatterning methods, such as micro-contact printing (μCP), involve relatively complicated fabrication procedures, which restricts widespread adoption by biologists. Here, we demonstrate a rapid method of micropatterning by printing hydrogel micro-features onto a glass-bottomed culture vessel. The micro-features are printed using a projection stereolithography bioprinter yielding hydrogel structures that geometrically restrict the attachment of cells or proteins. Compared to traditional and physical photomasks, a digitally tunable virtual photomask is used in the projector to generate blue light patterns that enable rapid iteration with minimal cost and effort. We show that a protocol that makes use of this method together with LN521 coating, an extracellular matrix coating, creates a surface suitable for human embryonic stem cell (hESC) attachment and growth with minimal non-specific adhesion. We further demonstrate that self-patterning of hESCs following previously published gastrulation and ectodermal induction protocols achieves results comparable with those obtained with commercially available plates.

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