scholarly journals CYTOLOGICAL STUDIES OF ORGANOTYPIC CULTURES OF RAT DORSAL ROOT GANGLIA FOLLOWING X-IRRADIATION IN VITRO

1967 ◽  
Vol 32 (2) ◽  
pp. 467-496 ◽  
Author(s):  
Edmund B. Masurovsky ◽  
Mary Bartlett Bunge ◽  
Richard P. Bunge
Keyword(s):  
Endoplasmic Reticulum ◽  
Dorsal Root Ganglia ◽  
Satellite Cells ◽  
Dorsal Root ◽  
Nervous Tissue ◽  
X Rays ◽  
Organotypic Cultures ◽  
X Irradiation

Long-term organotypic cultures of rat dorsal root ganglia were exposed to a single 40 kR dose of 184 kvp X-rays and studied in the living and fixed states by light or electron microscopy at 1–14 day intervals thereafter. Within the first 4 days following irradiation, over 30% of the neurons display chromatolytic reactions (eccentric nuclei, peripheral dispersal of Nissl substance, central granular zone) as well as abnormal nucleolar changes and dissociation of ribosomes from endoplasmic reticulum cisternae. Some satellite cells undergo retraction or acute degeneration, leaving only basement membrane to cover the neuron in these areas. 8 days after irradiation, neurons also exhibit (a) areas in which ribosomes are substantially reduced, (b) regions of cytoplasmic sequestration, (c) extensive vacuolization of granular endoplasmic reticulum and Golgi complex, and (d) diversely altered mitochondria (including the presence of ribosome-like particles or association with abnormal glycogen and lipid deposits). Nucleolar components become altered or reoriented and may form abnormal projections and ringlike configurations. Sizeable areas of the neuronal soma are now denuded of satellite cells; underlying these areas, nerve processes are found abnormally invaginated into the neuronal cytoplasm. By the 14th day following irradiation, most neurons display marked degenerative changes including extensive regions of ribosome depletion, sequestration, vacuolization, autolysis, and, in some areas, swirls of filaments, myelin figures, and heterogeneous dense bodies. These observations demonstrate that X-irradiation produces profound cytopathological changes in nervous tissue isolated from the host and that many of these changes resemble the effects of radiation on nervous tissue in vivo.

Neurotoxicity of Seven MEIC Chemicals Evaluated in Organotypic Cultures of Chick Embryonic Dorsal Root Ganglia

1997 ◽  
Vol 25 (3) ◽  
pp. 303-309
Author(s):  
Václav Mandys ◽  
Katerina Jirsová ◽  
Jirí Vrana
Keyword(s):  
Toxic Effect ◽  
Neurite Outgrowth ◽  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Growth Parameters ◽  
In Vitro Cytotoxicity ◽  
Organotypic Cultures ◽  
Response Curves ◽  
Agar Culture

The neurotoxic effects of seven selected Multicenter Evaluation of In Vitro Cytotoxicity programme chemicals (methanol, ethanol, isopropanol, sodium chloride, potassium chloride, iron [II] sulphate and chloroform) were evaluated in organotypic cultures of chick embryonic dorsal root ganglia (DRG), maintained in a soft agar culture medium. Two growth parameters of neurite outgrowth from the ganglia — the mean radial length of neurites and the area of neurite outgrowth — were used to evaluate the toxicities of the chemicals. Dose-dependent decreases of both parameters were observed in all experiments. IC50 values (the concentration causing 50% inhibition of growth) were calculated from the dose-response curves established at three time-points during culture, i.e. 24, 48 and 72 hours. The lowest toxic effect was observed in cultures exposed to methanol (the IC50 ranging from 580mM to 1020mM). The highest toxic effect was observed in cultures exposed to iron (II) sulphate (the IC50 ranging from 1.2mM to 1.7mM). The results of other recent experiments suggest that organotypic cultures of DRG can be used during in vitro studies on target organ toxicity within the peripheral nervous system. Moreover, these cultures preserve the internal organisation of the tissue, maintain intercellular contacts, and thus reflect the in vitro situation, more precisely than other cell cultures.

Neuronal intermediate filament expression in rat dorsal root ganglia sensory neurons: An in vivo and in vitro study

Neuroscience ◽  
2008 ◽  
Vol 153 (4) ◽  
pp. 1153-1163 ◽  
Author(s):  
M. Fornaro ◽  
J.M. Lee ◽  
S. Raimondo ◽  
S. Nicolino ◽  
S. Geuna ◽  
...  
Keyword(s):  
Sensory Neurons ◽  
Dorsal Root Ganglia ◽  
Intermediate Filament ◽  
Dorsal Root ◽  
In Vitro Study ◽  
Vitro Study

Systematic investigation and comparison of US FDA-approved immunosuppressive drugs FK506, cyclosporine and rapamycin for neuromuscular regeneration following chronic nerve compression injury

2021 ◽  
Vol 16 (11) ◽  
pp. 989-1003
Author(s):  
Lucas Degrugillier ◽  
Katharina M Prautsch ◽  
Dirk J Schaefer ◽  
Raphael Guzman ◽  
Daniel F Kalbermatten ◽  
...  
Keyword(s):  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Nerve Compression ◽  
In Vivo Model ◽  
Compression Injury ◽  
Potent Effect ◽  
Therapeutic Benefits ◽  
Chronic Nerve Compression

Aim: To compare therapeutic benefits of different immunophilin ligands for treating nerve injuries. Materials & methods: Cyclosporine, FK506 and rapamycin, were evaluated first in vitro on a serum-free culture of embryonic dorsal root ganglia followed by a new in vivo model of chronic nerve compression. Results: Outcomes of the in vitro study have shown a potent effect of cyclosporine and FK506, on dorsal root ganglia axonal outgrowth, comparable to the effect of nerve growth factor. Rapamycin exhibited only a moderate effect. The in vivo study revealed the beneficial effects of cyclosporine, FK506 and rapamycin for neuromuscular regeneration. Cyclosporine showed the better maintenance of the tissues and function. Conclusion: Cyclosporine, FK506 and rapamycin drugs showed potential for treating peripheral nerve chronic compression injuries.

scholarly journals Hydrogen Peroxide Induces Muscle Nociception via Transient Receptor Potential Ankyrin 1 Receptors

2017 ◽  
Vol 127 (4) ◽  
pp. 695-708 ◽  
Author(s):  
Daisuke Sugiyama ◽  
Sinyoung Kang ◽  
Nicholas Arpey ◽  
Preeyaphan Arunakul ◽  
Yuriy M. Usachev ◽  
...  
Keyword(s):  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Transient Receptor Potential ◽  
Receptor Potential ◽  
Deep Muscle ◽  
Plantar Incision ◽  
Dorsal Root Ganglia Neurons ◽  
Transient Receptor

Abstract Background H2O2 has a variety of actions in skin wounds but has been rarely studied in deep muscle tissue. Based on response to the transient receptor potential ankyrin 1 antagonists after plantar incision, we hypothesized that H2O2 exerts nociceptive effects via the transient receptor potential ankyrin 1 in muscle. Methods Nociceptive behaviors in rats (n = 269) and mice (n = 16) were evaluated after various concentrations and volumes of H2O2 were injected into the gastrocnemius muscle or subcutaneous tissue. The effects of H2O2 on in vivo spinal dorsal horn neuronal activity and lumbar dorsal root ganglia neurons in vitro were evaluated from 26 rats and 6 mice. Results Intramuscular (mean ± SD: 1,436 ± 513 s) but not subcutaneous (40 ± 58 s) injection of H2O2 (100 mM, 0.6 ml) increased nociceptive time. Conditioned place aversion was evident after intramuscular (–143 ± 81 s) but not subcutaneous (–2 ± 111 s) injection of H2O2. These H2O2-induced behaviors were blocked by transient receptor potential ankyrin 1 antagonists. Intramuscular injection of H2O2 caused sustained in vivo activity of dorsal horn neurons, and H2O2 activated a subset of dorsal root ganglia neurons in vitro. Capsaicin nerve block decreased guarding after plantar incision and reduced nociceptive time after intramuscular H2O2. Nociceptive time after intramuscular H2O2 in transient receptor potential ankyrin 1 knockout mice was shorter (173 ± 156 s) compared with wild-type mice (931 ± 629 s). Conclusions The greater response of muscle tissue to H2O2 may help explain why incision that includes deep muscle but not skin incision alone produces spontaneous activity in nociceptive pathways.

scholarly journals Mineralocorticoid Receptor Blocker Eplerenone Reduces Pain Behaviors In Vivo  and Decreases Excitability in Small-diameter Sensory Neurons from Local Inflamed Dorsal Root Ganglia In Vitro 

2012 ◽  
Vol 117 (5) ◽  
pp. 1102-1112 ◽  
Author(s):  
Fei Dong ◽  
Wenrui Xie ◽  
Judith A. Strong ◽  
Jun-Ming Zhang
Keyword(s):  
Sensory Neurons ◽  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Mineralocorticoid Receptor ◽  
Small Diameter ◽  
Mechanical Hypersensitivity ◽  
Proinflammatory Role ◽  
Immunohistochemical Studies

Background Inflammation of the dorsal root ganglia (DRG) may contribute to low back pain, postherpetic neuralgia, and neuropathic pain. The mineralocorticoid receptor (MR) plays a proinflammatory role in many nonrenal tissues, but its role in peripheral pain at the DRG level is not well studied. Methods Local inflammation of the L5 DRG with the immune activator zymosan rapidly leads to mechanical hypersensitivity and increased excitability of sensory neurons. Using this pain model, the authors applied the MR antagonist eplerenone locally to the inflamed DRG. Excitability of small-diameter sensory neurons was examined in acute primary culture by using patch clamp techniques. Results Local eplerenone significantly reduced the mechanical hypersensitivity and shortened its duration. The same dose was ineffective systemically. Immunohistochemical studies showed the MR was present in most neurons and rapidly translocated to the nucleus 1 day after local DRG inflammation. Activation of satellite glia (defined by expression of glial fibrillary acidic protein) in the inflamed DRG was also reduced by local eplerenone. Increased excitability of small-diameter sensory neurons 1 day after inflammation could be observed in vitro. Eplerenone applied in vitro (8-12 h) could reverse this increased excitability. Eplerenone had no effect in neurons isolated from normal, uninflamed DRG. The MR agonist aldosterone (10 nM) applied in vitro increased excitability of neurons isolated from normal DRG. Conclusions The MR may have a pronociceptive role in the DRG. Some of its effects may be mediated by neuronal MR. The MR may represent a novel therapeutic target in some pain syndromes.

scholarly journals CYTOLOGICAL STUDIES OF ORGANOTYPIC CULTURES OF RAT DORSAL ROOT GANGLIA FOLLOWING X-IRRADIATION IN VITRO

1967 ◽  
Vol 32 (2) ◽  
pp. 497-518 ◽  
Author(s):  
Edmund B. Masurovsky ◽  
Mary Bartlett Bunge ◽  
Richard P. Bunge
Keyword(s):  
Schwann Cells ◽  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Light And Electron Microscopy ◽  
Ultrastructural Changes ◽  
Type I ◽  
X Rays ◽  
Metabolic Disturbances ◽  
Line Type

Under suitable conditions rat dorsal root ganglia differentiate and myelinate in culture, providing an organotypic model of the ganglion (8). Mature cultures of this type were irradiated with a 40 kR dose of 184 kvp X-rays and, after daily observation in the living state, were fixed for light and electron microscopy. Within 24 hr after irradiation, numerous Schwann cells investing unmyelinated axons acutely degenerate. The axons thus denuded display little change. Conversely, few ultrastructural changes develop in Schwann cells investing myelinated axons until after the 4th day. During the 4–14 day period, these Schwann cells and their related myelin sheaths undergo progressive deterioration. Associated axons decrease in diameter but are usually maintained. Myelin deterioration begins as a nodal lengthening and then progresses along two different routes. In intact Schwann cells, fragmentation of myelin begins in a pattern reminiscent of Wallerian degeneration, but its slow breakdown thereafter suggests metabolic disturbances in these Schwann cells. The second pattern of myelin deterioration, occurring after complete degeneration of the related Schwann cell, involves unusual configurational changes in the myelin lamellae. Atypical repeating periods are formed by systematic splitting of lamellae at each major dense line with further splitting at the intraperiod line (Type I) or by splitting in the region of every other intraperiod line (Type II); some sheaths display a compact, wavy, inner zone and an abnormally widened lamellar spacing peripherally (Type III). Extensive blebbing of myelin remnants characterizes the final stages of this extracellular myelin degradation. These observations provide the first description of ultrastructural changes produced by ionizing radiation in nerve fascicles in vitro.

Proteasome Inhibitor Associated Neuropathy Is Mechanism Based

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2646-2646 ◽  
Author(s):  
Lee Silverman ◽  
Vilmos Csizmadia ◽  
Katherine Brewer ◽  
Chris Simpson ◽  
Carl Alden
Keyword(s):  
Nervous System ◽  
Peripheral Neuropathy ◽  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Proteasome Inhibition ◽  
Ubiquitinated Proteins ◽  
Vitro Data ◽  
In Vitro Data

Abstract Background: Peripheral neuropathy is a common side effect of chemotherapeutics, particularly taxanes, platinums, vinca alkaloids, and proteasome inhibitors (PIs). There are currently three chemical scaffolds of PIs in clinical development: leucine boronates, lactacystin analogs, and epoximicin derivatives. Among these, only a leucine boronate, VELCADE, is currently approved for use in humans. Approximately 39% of patients treated with VELCADE develop peripheral neuropathy. Preclinical data suggest that peripheral neuropathy may develop as a consequence of functional effects in the dorsal root ganglia (Silverman et al. 2006; Toxicologic Pathology34:989). Moreover, in vitro data suggests that these effects may be the result of a reorganization of cytoskeletal elements and an accumulation of ubiquitinated proteins in the neuronal cytoplasm (Csizmadia et al. 2008; Neurotoxicology29:232). In vivo and in vitro data suggest that PI-associated neuropathy develops following treatment with PIs of differing chemical scaffolds, consistent with a mechanism based effect. A better understanding of the mechanism underlying PI-associated peripheral neuropathy could potentially lead to improved management strategies for this adverse event while maintaining therapeutic benefit. Aims: To investigate if PI-associated neuropathy is mechanism based (a consequence of proteasome inhibition), and If mechanism based, to further understand the mechanism by which proteasome inhibition results in peripheral neuropathy. Methods: The effects of treating in vitro neuronal cell cultures (PC-12 rat pheochromocytoma cell line treated with nerve growth factor [NGF] in order to induce neuronal differentiation) with leucine boronate, lactacystin analog, or epoximicin PIs, as well as with taxol and cisplatinum, were examined by Western immunoblot and immunohistochemistry specific for ubiquitinated proteins. Additionally, the morphologic effects on the peripheral nervous system of mice treated with leucine boronate or lactacystin analog PIs were examined. Results: Treatment of PC-12 cells with leucine boronate, lactacystin analog or epoximicin PIs but not with taxol or cisplatinum resulted in perinuclear accumulations of ubiquitinated proteins. In in vivo models, morphologic lesions were observed in the peripheral nervous system of mice treated with both leucine boronate and lactacystin analog PIs. In a time course study with a prototypical PI, dorsal root ganglia was identified as the primary target leading to secondary peripheral nerve degeneration. Routine hematoxylin and eosin histologic analysis showed an accumulation of eosinophilic material within the dorsal root ganglia neurons. Immunohistochemistry showed these accumulations to contain ubiquitinated proteins, consistent with the in vitro observations. The in vivo effects were similar following treatment with either a leucine boronate or a lactacystin analog PI. Summary: Our in vitro and in vivo studies identified a morphologic effect which among the chemotherapeutics which cause peripheral neuropathy is uniquely associated with PIs. Moreover, these findings were seen regardless of the chemical scaffold.

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