scholarly journals Stereological Estimation of Dopaminergic Neuron Number in the Mouse Substantia Nigra Using the Optical Fractionator and Standard Microscopy Equipment

10.3791/56103 ◽  
2017 ◽  
Author(s):  
Chi Wang Ip ◽  
David Cheong ◽  
Jens Volkmann
Keyword(s):  
Substantia Nigra ◽  
Dopaminergic Neuron ◽  
Neuron Number ◽  
Optical Fractionator

scholarly journals Regulation of dopaminergic neuron firing by heterogeneous dopamine autoreceptors in the substantia nigra pars compacta

2011 ◽  
Vol 116 (6) ◽  
pp. 966-974 ◽  
Author(s):  
Jin Young Jang ◽  
Miae Jang ◽  
Shin Hye Kim ◽  
Ki Bum Um ◽  
Yun Kyung Kang ◽  
...  
Keyword(s):  
Substantia Nigra ◽  
Dopaminergic Neuron ◽  
Substantia Nigra Pars Compacta ◽  
Dopamine Autoreceptors ◽  
Neuron Firing

scholarly journals Camptothecin regulates microglia polarization and exerts neuroprotective effects via the AKT/Nrf2/HO-1-NF-κB signal axis in vivo and in vitro

2020 ◽  
Author(s):  
dewei he ◽  
dianfeng liu ◽  
ang zhou ◽  
xiyu gao ◽  
yufei zhang ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Substantia Nigra ◽  
Cell Line ◽  
Dopaminergic Neuron ◽  
Inflammatory Responses ◽  
Neuroprotective Effects ◽  
Microglia Polarization

Abstract Background Parkinson's disease (PD), the second largest neurodegenerative disease seriously affects human health. Microglia, the main immune cells in the brain participate in the innate immune response in the central nervous system (CNS). Studies have shown that microglia can be polarized into pro-inflammatory M1 and anti-inflammatory M2 phenotypes. Accumulated evidences suggest that over-activated M1 microglia release pro-inflammatory mediators that damage neurons and lead to Parkinson's disease (PD). In contrast, M2 microglia release neuroprotective factors and exert the effects of neuroprotection. Camptothecin (CPT), an extract of the plant Camptotheca acuminate, has been reported to have anti-inflammation and antitumor effects. However the effect of CPT on microglia polarization and microglia-mediated inflammation responses has not been reported. Therefore, we aim to explore the effect of CPT on microglia polarization and its underlying mechanism on neuroinflammation. Methods C57BL/6 mice (25–30 g) were injected LPS or PBS into the substantia nigra (SN). Open-Field Test and Immunohistochemistry were performed to test the dyskinesia of mice and the loss of neurons in the substantia nigra (SN). Microglia cell line BV-2, the neuroblastoma SH-SY5Y and dopaminergic neuron MN9D cell were cultured. Cytotoxicity assay, reverse transcription quantitative real-time polymerase chain reaction (RT-PCR), Western blot, ELISA and Immunofluorescence staining were performed. All results were presented with mean ± SD. Results In vivo, CPT improved dyskinesia of mice, reduced the loss of neurons in the substantia nigra (SN) and inhibited neuro-inflammatory responses in LPS-injected mice. In vitro, CPT inhibited M1 polarization of microglia and promotes M2 polarization via the AKT/Nrf2/HO-1-NF-κB signal axis. Furthermore, CPT protected the neuroblastoma cell line SH-SY5Y and dopaminergic neuron cell line MN9D from neurotoxicity of mediated by microglia activation. Conclusion CPT regulates the microglia polarization phenotype via the AKT/Nrf2/HO-1-NF-κB signal axis, inhibits neuro-inflammatory responses and exerts neuroprotective effects in vivo and in vitro.

Optimization of dopaminergic neuron counting in substantia nigra of parkinsonian mice

2010 ◽  
Vol 4 (4) ◽  
pp. 391-398 ◽  
Author(s):  
V. G. Khaindrava ◽  
P. V. Ershov ◽  
V. E. Antsiperov ◽  
Yu. V. Obukhov ◽  
A. K. Nanaev ◽  
...  
Keyword(s):  
Substantia Nigra ◽  
Dopaminergic Neuron

Effects of age-related dopaminergic neuron loss in the substantia nigra on the circadian rhythms of locomotor activity in mice

2012 ◽  
Vol 74 (3-4) ◽  
pp. 210-215 ◽  
Author(s):  
Makoto Tanaka ◽  
Eriko Yamaguchi ◽  
Mami Takahashi ◽  
Kana Hashimura ◽  
Takao Shibata ◽  
...  
Keyword(s):  
Locomotor Activity ◽  
Circadian Rhythms ◽  
Substantia Nigra ◽  
Dopaminergic Neuron ◽  
Neuron Loss ◽  
Age Related ◽  
Dopaminergic Neuron Loss

scholarly journals ESTIMATION OF THE NUMBER OF NEURONS IN THE HIPPOCAMPUS OF RATS WITH PENICILLIN INDUCED EPILEPSY

2011 ◽  
Vol 21 (2) ◽  
pp. 117 ◽  
Author(s):  
Ilgaz Akdogan ◽  
Nedim Unal ◽  
Esat Adiguzel
Keyword(s):  
Pyramidal Neuron ◽  
Epileptic Seizures ◽  
Penicillin G ◽  
Control Group ◽  
Neuron Number ◽  
Optical Disector ◽  
Cell Layers ◽  
The Central Nervous System ◽  
Optical Fractionator ◽  
Experimental Group

Epilepsy is a neurological disease arising from strong and uncontrollable electrical firings of a group of neurons in the central nervous system. Experimental epileptic models have been developed to assess the physiopathology of epileptic seizures. This study was undertaken to estimate the number of neurons in the rat hippocampus with penicillin induced epilepsy, using a stereological method, "the optical fractionator". In the experimental group, 500 IU penicillin-G was injected intra-cortically, and in the control group, the same volume of saline was administered. A week later, the animals were decapitated and their brains were removed by craniatomy. Frozen brains were cut with a thickness of 150 ěm in a cryostat. Sections were collected by systematic random sampling and stained with hematoxylen-eosin. Microscopic images of pyramidal cell layers from hippocampus CA1, CA2 and CA3 subfields were then transferred to a monitor, using a 100x objective (N.A. = 1.25). Using the optical disector method, the neurons were counted in the frames and determined with a fractionator sampling scheme. The total pyramidal neuron number was then estimated using the optical fractionator method. The total pyramidal neuron number was found to be statistically lower in the experimental group (mean = 142,888 ± 11,745) than in the control group (mean = 177,953 ± 10,907) (p < 0.05). The results suggest that a decrease in the hippocampal neuronal number in a penicillin model of epilepsy can be determined objectively and efficiently using the optical fractionator method.

scholarly journals MODERN STEREOLOGICAL EVALUATION IN THE AGING HUMAN SUBSTANTIA NIGRA

2011 ◽  
Vol 22 (2) ◽  
pp. 73 ◽  
Author(s):  
Shuang Y Ma ◽  
Frank M Longo ◽  
Matias Röyttä ◽  
Yrjö Collan
Keyword(s):  
Substantia Nigra ◽  
Quantitative Estimation ◽  
Degenerative Change ◽  
Reduction Rate ◽  
Cell Number ◽  
Aging Brain ◽  
Tissue Sections ◽  
Neuronal Number ◽  
Optical Fractionator ◽  
Human Substantia Nigra

Quantitative estimation of neuronal numbers in the human substantia nigra (SN) can be achieved by a conventional single section (SS) count or by the more modern stereological disector (DS) count. However, counting results from SS counts are potentially biased and might not accurately reflect the total neuronal number in the SN or the changes in the total number of neurons occurring during aging or with neurodegenerative disease. Potential sources of bias include the lack of linearity between cell number per area of section and cell number per volume; the variation in the counting level and orientation of tissue sections; and shrinkage of tissue. Modern stereological DS counting overcomes these problems and has played a crucial role in many recent studies in neuropathology, neuroanatomy, neuropharmacology and neurogenetics. Over the past decades, four stereology based counting methods including physical DS, physical fractionator, optical DS and optical fractionator, have been established for quantitative measurement. Recently, stereological estimates have revealed a linear reduction rate of total nigral neuronal numbers with age of about 10% per decade. These findings suggest that the surviving nigral neurons undergo a degenerative change leading to neuronal dysfunction with aging. Furthermore, as an advanced quantitative tool, modern stereological evaluation may provide new insights into the aging of the human SN thereby enabling us to better understand the pathophysiological processes in aging brain.

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