Neural-type Cells Research Articles

Monitoring nanosecond electric pulse-evoked catecholamine secretion in adrenal chromaffin cells using a novel switching system for obtaining amperometric measurements

We have been investigating the potential for nanosecond electric pulses (NEPs) to serve as a new electrostimulation approach for neuromodulation, using isolated adrenal chromaffin cells as a model of neural-type cells for these investigations. While fluorescence imaging techniques have established the efficacy of NEPs for evoking catecholamine release, the present goal is to employ carbon fiber amperometry, a quantitative technique that measures the amount of secreted catecholamines in real-time with high sensitivity.

Differentiation of neural-type cells on multi-scale ordered collagen-silica bionanocomposites.

Cells respond to biophysical and biochemical signals. We developed a composite filament from collagen and silica particles modified to interact with collagen and/or present a laminin epitope (IKVAV) crucial for cell-matrix adhesion and signal transduction. This combines scaffolding and signaling and shows that local tuning of collagen organization enhances cell differentiation.

Shaping of Drosophila Neural Cell Lineages Through Coordination of Cell Proliferation and Cell Fate by the BTB-ZF Transcription Factor Tramtrack-69.

Cell diversity in multicellular organisms relies on coordination between cell proliferation and the acquisition of cell identity. The equilibrium between these two processes is essential to assure the correct number of determined cells at a given time at a given place. Using genetic approaches and correlative microscopy, we show that Tramtrack-69 (Ttk69, a Broad-complex, Tramtrack and Bric-à-brac - Zinc Finger (BTB-ZF) transcription factor ortholog of the human promyelocytic leukemia zinc finger factor) plays an essential role in controlling this balance. In the Drosophila bristle cell lineage, which produces the external sensory organs composed by a neuron and accessory cells, we show that ttk69 loss-of-function leads to supplementary neural-type cells at the expense of accessory cells. Our data indicate that Ttk69 (1) promotes cell cycle exit of newborn terminal cells by downregulating CycE, the principal cyclin involved in S-phase entry, and (2) regulates cell-fate acquisition and terminal differentiation, by downregulating the expression of hamlet and upregulating that of Suppressor of Hairless, two transcription factors involved in neural-fate acquisition and accessory cell differentiation, respectively. Thus, Ttk69 plays a central role in shaping neural cell lineages by integrating molecular mechanisms that regulate progenitor cell cycle exit and cell-fate commitment.

Identification of jellyfish neuropeptides that act directly as oocyte maturation-inducing hormones.

Oocyte meiotic maturation is crucial for sexually reproducing animals, and its core cytoplasmic regulators are highly conserved between species. By contrast, the few known maturation-inducing hormones (MIHs) that act on oocytes to initiate this process are highly variable in their molecular nature. Using the hydrozoan jellyfish species Clytia and Cladonema, which undergo oocyte maturation in response to dark-light and light-dark transitions, respectively, we deduced amidated tetrapeptide sequences from gonad transcriptome data and found that synthetic peptides could induce maturation of isolated oocytes at nanomolar concentrations. Antibody preabsorption experiments conclusively demonstrated that these W/RPRPamide-related neuropeptides account for endogenous MIH activity produced by isolated gonads. We show that the MIH peptides are synthesised by neural-type cells in the gonad, are released following dark-light/light-dark transitions, and probably act on the oocyte surface. They are produced by male as well as female jellyfish and can trigger both sperm and egg release, suggesting a role in spawning coordination. We propose an evolutionary link between hydrozoan MIHs and the neuropeptide hormones that regulate reproduction upstream of MIHs in bilaterian species.

Nanosecond pulsed electric fields activate intracellular signaling pathways

In cellular electrochemistry, ions respond to stimuli by constantly shuffling across cellular membranes to perform their physiological roles. This flow of ions, the electromotive force, leaves cells vulnerable to exogenous electromagnetic fields that can stimulate and/or modulate cellular activity. An irreparable link exists between changes in ionic concentration and the electric gradient of the cell (or its potential energy). Consequently, we can manipulate the physiology of the cell by altering its permeability to various ions, thereby modulating its electrical gradient. Only a few millivolts in excess of the resting membrane potential can stimulate a dramatic change in ion distribution within the cellular microenvironment. In excitable neural-type cells, electrical-stimulation-induced changes in membrane potential lead to the generation or inactivation of action potentials (AP). These AP trigger activities, such as nerve impulses in

Differentiation of B16-BL6 Melanoma Cells into Microtubule-Associated Protein 2-Positive Cells after Treatment with the Histone Deacetylase Inhibitors Butyrate and Trichostatin A

Inhibition of histone deacetylase (HDAC) modulates the expression of many genes and induces cell cycle arrest, apoptosis, and differentiation in several cancer cell lines. Melanoma is a malignant phase of cutaneous melanocytes originally derived from the neural crest and is highly metastatic. A therapeutic agent capable of inducing differentiation of metastatic melanoma cells into a nonmalignant stage would be useful to prevent metastasis. We examined whether HDAC inhibitors can induce differentiation of the murine melanoma cell line B16-BL6 into neural-type cells in vitro. A morphologic change accompanied by extended dendrites was induced in melanoma cells after treatment with the HDAC inhibitors butyrate and trichostatin A (TSA). The altered morphology was similar to that of neural cells. Many of the extended dendrites fused with other dendrites of neighboring cells and had synapse-like knobs on their dendrites. These dendrites showed positive labeling with anti-α/β-tubulin and anti-L1 cell adhesion molecule (L1CAM) antibodies but were seldom stained with phalloidin, suggesting that the neural cell-related proteins were major components of the extended dendrites but that actin stress fibers were not. Furthermore, the mature neuron-specific cytoskeletal protein, microtubule-associated protein 2 (MAP2), was detected with the specific antibody in the extended dendrites of cells treated with butyrate and TSA. Butyrate treatment increased the levels of MAP2 and neural cell adhesion molecule (NCAM) mRNA expression in the cells. However, the treatment did not alter the expression of neurofilament light chain (NF-L) mRNA. The observations suggest that the differentiated cells were neural-type cells but not complete neural cells.

Patient-centred asthma education in the emergency department: the case in favour

<h3>Abstract</h3> Oocyte meiotic maturation is a critical process for sexually reproducing animals, and its core cytoplasmic regulators are highly conserved between species. In contrast, the few known Maturation Inducing Hormones (MIHs) that act on oocytes to initiate this process have highly variable molecular natures. Using the hydrozoan jellyfish species <i>Clytia</i> and <i>Cladonema</i>, which undergo oocyte maturation in response to dark-light and light-dark transitions respectively, we deduced from gonad transcriptome data amidated tetrapeptide sequences and found that synthetic peptides could induce maturation of isolated oocytes at nanomolar concentrations. Antibody preabsorption experiments conclusively demonstrated that these W/RPRPamide-related neuropeptides account for endogenous MIH activity produced by isolated gonads. We further showed that the MIH peptides are synthesised by neural-type cells in the gonad, are released following dark-light / light-dark transitions, and probably act on the oocyte surface. They are produced by male as well as female jellyfish and can trigger both sperm and egg release, suggesting a role in spawning coordination. We propose an evolutionary link between hydrozoan MIH and the neuropeptide hormones that regulate reproduction upstream of MIH in bilaterian species.

Design, Characterization, and Optimization of a Broadband Mini Exposure Chamber for Studying Catecholamine Release From Chromaffin Cells Exposed to Microwave Radiation: Finite-Difference Time-Domain Technique

A free-space in vitro exposure system for identifying specific microwave (MW) parameters in the frequency range of 1-6 GHz that can induce nonthermal effects on exocytosis, which is the process by which neurotransmitter release occurs, has been designed, constructed, characterized, and optimized. The exposure system is placed within an anechoic chamber and incorporates continuous online monitoring of basal and stimulated catecholamine release from cultured bovine adrenal-medullary chromaffin cells, which are well-established models of neural-type cells. The cells are immobilized inside a cell-perfusion apparatus (CPA) and are continuously superfused with temperature-controlled balanced salt solution, with the entire CPA placed within a mini exposure chamber (MEC) constructed out of a MW-absorbing material. All relevant equipment for carrying out the experiments is shielded from the MW field by being housed in an aluminum conductor box located behind the MEC. Detailed distributions of the electric field and the specific absorption rate (SAR) at the location of the cells within the MEC were computed using the finite-difference time-domain (FDTD) method. FDTD computations were also used for optimizing the exposure system, so that the highest intensity of electric field could be delivered under dynamic temperature control and with an acceptable degree of field homogeneity (to within 30%) over the entire frequency range of 1-6 GHz. A major finding is that maintaining an acceptable level of homogeneity of the electric field and SAR for exposing cells to 1-6 GHz MW fields requires a different distribution of the cells within the CPA for exposures carried out at the lower versus the higher end of the frequency range of interest

A Novel Waveguide-Based Radio Frequency/Microwave Exposure System for Studying Nonthermal Effects on Neurotransmitter Release—Finite-Difference Time-Domain Modeling

A research effort is underway to identify specific radio frequency/microwave parameters in the frequency range 0.75-1.12 GHz that can produce nonthermal effects on the release of catecholamines from cultured bovine adrenal medullary chromaffin cells, an established in vitro model of neural-type cells. A well-characterized exposure system is crucial for interpreting the biological outcomes of experiments. This paper describes a novel waveguide-based exposure system that permits perfusion of the cells with a temperature-controlled balanced salt solution for online monitoring of catecholamine release from the cells during radio frequency/microwave exposure. The finite-difference time-domain method was used to optimize the exposure conditions, the goal being to achieve the maximum possible homogeneity in the distribution of the specific absorption rate at the location of the cells in the waveguide. At a frequency of 1 GHz, optimal coupling of the radio frequency/microwave field into the region containing the cells was obtained only when the cells were placed at the location of the electric field maximum of the standing wave pattern, with the electric field parallel to the plane containing the cells. In this case, the specific absorption rate distribution was found to be the highest as well as homogeneous to within 30%.

Compact low-power calibration mini-DACs for neural arrays with programmable weights

This paper considers the viability of compact low-resolution low-power mini digital-to-analog converters (mini-DACs) for use in large arrays of neural type cells, where programmable weights are required. Transistors are biased in weak inversion in order to yield small currents and low power consumptions, a necessity when building large size arrays. One important drawback of weak inversion operation is poor matching between transistors. The resulting effective precision of a fabricated array of 50 DACs turned out to be 47% (1.1 bits), due to transistor mismatch. However, it is possible to combine them two by two in order to build calibrated DACs, thus compensating for inter-DAC mismatch. It is shown experimentally that the precision can be improved easily by a factor of 10 (4.8% or 4.4 bits), which makes these DACs viable for low-resolution applications such as massive arrays of neural processing circuits. A design methodology is provided, and illustrated through examples, to obtain calibrated mini-DACs of a given target precision. As an example application, we show simulation results of using this technique to calibrate an array of digitally controlled integrate-and-fire neurons.

Catecholamine release from cultured bovine adrenal medullary chromaffin cells in the presence of 60-Hz magnetic fields

Effects of powerline frequency (50/60 Hz) electric and magnetic fields on the central nervous system may involve altered neurotransmitter release. This possibility was addressed by determining whether 60-Hz linearly polarized sinusoidal magnetic fields (MFs) alter the release of catecholamines from cultured bovine adrenal chromaffin cells, a well-characterized model of neural-type cells. Dishes of cells were placed in the center of each of two four-coil Merritt exposure systems that were enclosed within mu-metal chambers in matched incubators for simultaneous sham and MF exposure. Following 15-min MF exposure of the cells to flux densities of 0.01, 0.1, 1.0 or 2 mT, norepinephrine and epinephrine release were quantified by high-performance liquid chromatography (HPLC) coupled with electrochemical detection. No significant differences in the release of either norepinephrine or epinephrine were detected between sham-exposed cells and cells exposed to MFs in either the absence or presence of Bay K-8644 (2 μM) or dimethylphenylpiperazinium (DMPP, 10 μM). Consistent with these null findings is the lack of effect of MF exposure on calcium influx. We conclude that catecholamine release from chromaffin cells is not sensitive to 60-Hz MFs at magnetic flux densities in the 0.01–2 mT range.

All-trans-retinoic acid-mediated modulation of p53 during neural differentiation in murine embryonic stem cells.

All-trans-retinoic acid (RA) plays an important physiological role in embryonic development and is teratogenic in large doses in almost all species. p53, a tumor suppressor gene encodes phosphoproteins, which regulate cellular proliferation, differentiation, and apoptosis. Temporal modulation of p53 by retinoic acid was investigated in murine embryonic stem cells during differentiation and apoptosis. Undifferentiated embryonic stem cells express a high level of p53 mRNA and protein followed by a decrease in p53 levels as differentiation proceeds. The addition of retinoic acid during 8-10 days of differentiation increased the levels of p53 mRNA and protein, accompanied by accelerated neural differentiation and apoptosis. Marked increase in apoptosis was observed at 10-20 h after retinoic acid treatment when compared with untreated controls. Retinoic acid-induced morphological differentiation resulted in predominantly neural-type cells. Maximum increase in p53 mRNA in retinoic acid-treated cells occurred on day 17, whereas maximum protein synthesis occurred on days 14-17, which coincided with increased neural differentiation and proliferation. Increased p53 levels did not induce p21 transactivation, interestingly a decrease in p21 was observed on day 17 on exposure to retinoic acid. The level of p53 declined by day 21 of differentiation. The results demonstrated that retinoic acid-mediated apoptosis preceded the changes in p53 expression, suggesting that p53 induction does not initiate retinoic acid-induced apoptosis during development. However, retinoic acid accelerated neural differentiation and increased the expression of p53 in proliferating neural cells, corroborated by decreased p21 levels, indicating the importance of cell type and stage specificity of p53 function.

VLSI implementation of synaptic weighting and summing in pulse coded neural-type cells

Presents the hardware realization for synaptic weighting and summing using pulse-coded neural-type cells (NTCs). The basic information processing element (NTC) encodes the information into the form of pulse duty cycles using voltage-controlled resistors, for which a pulse duty cycle modulation technique is proposed. Summation is executed by a simple capacitor circuit as a current integrator. Layouts and measurements on a fabricated integrated design are included.

Differentiation of membrane excitability in isolated cleavage‐arrested blastomeres from early ascidian embryos.

1. Differentiation of excitable cells was studied electrophysiologically and histochemically in cleavage-arrested blastomeres isolated from early ascidian embryos. Blastomeres were isolated at the 4- or 8-cell stage, and cultured in sea water containing cytochalasin B until the time of hatching of control larvae. Electrical responses, immunoreactivity to epidermis-specific monoclonal antibody (2C5) and activity of muscle-specific acetylcholinesterase were examined. 2. All cleavage-arrested blastomeres isolated from an 8-cell embryo differentiated to elicit either muscular- or epidermal-type action potentials, but no neural-type action potentials were observed in these blastomeres. The anterior-animal and the posterior-animal blastomeres developed only epidermal-type action potentials, which involved expression of Ca2+ channels and immunoreactivity to 2C5. One-third of anterior-vegetal blastomeres developed epidermal-type action potentials which are mediated by Ca2+ channels though the immunoreactivity to 2C5 was absent. A majority of remaining blastomeres showed action potentials composed of Ca2+ currents and TEA-sensitive delayed K+ currents (type I response), and a few of them had fast transient K+ currents (A-currents) in addition (type II response). One-third of posterior-vegetal blastomeres developed epidermal-type action potentials without expression of the immunoreactivity to 2C5. The remainder differentiated into muscular-type cells, which expressed Ca2+ currents, TEA-sensitive and TEA-insensitive delayed K+ currents, and showed acetylcholinesterase activity. 3. Cleavage-arrested blastomeres isolated from a 4-cell embryo also differentiated into epidermal- or muscular-type cells, but not neural-type cells. The anterior blastomere, which is the parent cell of anterior-animal and anterior-vegetal blastomeres of an 8-cell embryo, developed epidermal-type, type I or type II responses, as was the case in the anterior-vegetal blastomere isolated from an 8-cell embryo. The posterior blastomere, which was the parent cell of posterior-animal and posterior-vegetal blastomeres of an 8-cell embryo, differentiated into either epidermal-type or muscular-type cells in terms of both membrane excitability and immunochemical reactivity. 4. Cleavage-arrested 1-cell embryos differentiated exclusively into epidermal-type cells in terms of membrane excitability and 2C5 immunoreactivity, even when the cytochalasin B concentration was decreased below 0.1 microgram/ml.(ABSTRACT TRUNCATED AT 400 WORDS)

Scanning electron microscopy and X-ray microprobe analysis in detection of acetylcholinesterase in cultured embryonal carcinoma cells.

Mouse F9 cells, induced by retinoic acid and dibutyryl cyclic adenosine monophosphate (cAMP) to differentiate into neural-type cells, were incubated for localization of specific acetylcholinesterase (AChE) activity according to the Karnovsky-Roots method where the final enzymatic reaction product is crystalline cupric ferrocyanide and cuprous thiocholine iodide. By scanning electron microscopy (SEM) neural-type cells with long processes were seen. Most of these cells exhibited crystalline precipitates on their surface that in microprobe analysis contained copper, iron, and sulfur. These elements were also detected in some of the neural-type cells that had no visible surface precipitates. Thus, the X-ray analysis also revealed intracellular enzymatic activity. Undifferentiated rounded cells, devoid of AChE activity at the light microscope level, did not show any surface precipitates by SEM and lacked copper, iron, and sulfur emission peaks in the elementary analysis. These results demonstrate that elementary analysis of cytochemical enzymatic reaction products by SEM can be used in identifying cells.

Myasthénie, thymome et tumeur hypophysaire associant une prolifération adénomateuse hétérogène et un ganglio-neurome de la selle turcique

The authors report a case of myasthenia gravis in which a tumour of the sella turcica, without clinical symptoms, was discovered on X-ray examination of the skull and was removed with success. Histological study showed endocrine and neural type cells with cells of intermediate type. Subsequent thoracic surgery was performed with removal of a thymoma which was not visible on X-ray examination of the chest. The first operation on the sella turcica had no effect on the myasthenia but after the second on the thymoma a definite improvement was noted. The association of myasthenia, thymoma and ganglioneuroma of the sella turcica is unknown. In order to explain this, the authors hypothesize a common origin in the neural crests of both tumours.

High Atropine Dosage in Chronic Epidemic Encephalitis

Cell diversity in multicellular organisms relies on coordination between cell proliferation and the acquisition of cell identity. The equilibrium between these two processes is essential to assure the correct number of determined cells at a given time at a given place. Using genetic approaches and correlative microscopy, we show that Tramtrack-69 (Ttk69, a Broad-complex, Tramtrack and Bric-a-brac - Zinc Finger (BTB-ZF) transcription factor ortholog of the human promyelocytic leukemia zinc finger factor) plays an essential role in controlling this balance. In the Drosophila bristle cell lineage, which produces the external sensory organs composed by a neuron and accessory cells, we show that ttk69 loss-of-function leads to supplementary neural-type cells at the expense of accessory cells. Our data indicate that Ttk69 (1) promotes cell cycle exit of newborn terminal cells by downregulating CycE, the principal cyclin involved in S-phase entry, and (2) regulates cell-fate acquisition and terminal differentiation, by downregulating the expression of hamlet and upregulating that of Suppressor of Hairless, two transcription factors involved in neural-fate acquisition and accessory cell differentiation, respectively. Thus, Ttk69 plays a central role in shaping neural cell lineages by integrating molecular mechanisms that regulate progenitor cell cycle exit and cell-fate commitment.