scholarly journals Comparison of Rat Primary Midbrain Neurons Cultured in DMEM/F12 and Neurobasal Mediums

2021 ◽  
Vol 12 (2) ◽  
pp. 205-212
Author(s):  
Neda Valian ◽  
◽  
Mansooreh Heravi ◽  
Abolhassan Ahmadiani ◽  
Leila Dargahi ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Dopaminergic Neurons ◽  
Motor Behavior ◽  
Cell Number ◽  
High Price ◽  
Rat Embryo ◽  
Brain Functions ◽  
Midbrain Neurons ◽  
Neurobasal Medium ◽  
Learning And Cognition

Introduction: Midbrain dopaminergic neurons are involved in various brain functions, including motor behavior, reinforcement, motivation, learning, and cognition. Primary dopaminergic neurons and also several lines of these cells are extensively used in cell culture studies. Primary dopaminergic neurons prepared from rodents have been cultured in both DMEM/F12 and neurobasal mediums in several studies. However, there is no document reporting the comparison of these two mediums. So in this study, we evaluated the neurons and astroglial cells in primary midbrain neurons from rat embryos cultured in DMEM/F12 and neurobasal mediums. Methods: Primary mesencephalon cells were prepared from the E14.5 rat embryo. Then they were seeded in two different mediums ( Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 [DMEM/F12] and neurobasal). On day 3 and day 5, half of the medium was replaced with a fresh medium. On day 7, β3-tubulin-, GFAP (Glial fibrillary acidic protein)- and Tyrosine Hydroxylase TH-positive cells were characterized as neurons, astrocytes, and dopaminergic neurons, respectively, using immunohistochemistry. Furthermore, the morphology of the cells in both mediums was observed under light microscopy on days 1, 3, and 5. Results: The cells cultured in both mediums were similar under light microscopy regarding the cell number, but in a neurobasal medium, the cells have aggregated and formed clustering structures. Although GFAP-immunoreactive cells were lower in neurobasal compared to DMEM/F12, the number of β3-tubulin- and TH-positive cells in both cultures was the same. Conclusion: This study’s findings demonstrated that primary midbrain cells from the E14.5 rat embryo could grow in both DMEM/F12 and neurobasal mediums. Therefore, considering the high price of a neurobasal medium, it can be replaced with DMEM/F12 for culturing primary dopaminergic neurons.

The endogenous cannabinoid receptor ligand, anandamide, inhibits the motor behavior: role of nigrostriatal dopaminergic neurons

Life Sciences ◽  
1995 ◽  
Vol 56 (23-24) ◽  
pp. 2033-2040 ◽  
Author(s):  
J. Romero ◽  
L. Garcia ◽  
M. Cebeira ◽  
D. Zadrozny ◽  
J.J. Fernández-Ruiz ◽  
...  
Keyword(s):  
Dopaminergic Neurons ◽  
Cannabinoid Receptor ◽  
Motor Behavior ◽  
Receptor Ligand ◽  
Endogenous Cannabinoid

Combining Neuroprotective Treatment of Embryonic Nigral Donor Tissue with Mild Hypothermia of the Graft Recipient

2005 ◽  
Vol 14 (5) ◽  
pp. 301-309 ◽  
Author(s):  
Jenny Karlsson ◽  
ÅSa Petersén ◽  
Gunilla Gidö ◽  
Tadeusz Wieloch ◽  
Patrik Brundin
Keyword(s):  
Graft Survival ◽  
Dopaminergic Neurons ◽  
Mild Hypothermia ◽  
Combined Treatment ◽  
Cell Number ◽  
Behavioral Recovery ◽  
Donor Tissue ◽  
Before And After ◽  
Neuroprotective Treatment ◽  
Graft Implantation

Around 80–95% of the immature dopaminergic neurons die when embryonic ventral mesencephalic tissue is transplanted. Cell death occurs both during the preparation of donor tissue and after graft implantation, but the effect of combining successful neuroprotective treatments before and after transplantation has not been extensively investigated. We therefore treated embryonic rat mesencephalic tissue with a combination of the lipid peroxidation inhibitor tirilazad mesylate (3 μM) and the caspase inhibitor Ac.YVAD.cmk (500 μM) and transplanted the tissue into hemiparkinsonian rats kept hypothermic (32–33°C) or normothermic (37°C) during, and 90 min following, graft surgery. Suspension cell number did not differ between untreated or tirilazad/YVAD-treated preparations prior to transplantation. When graft survival was evaluated 6 weeks after implantation, both tirilazad/YVAD pretreatment and mild hypothermia increased the survival of transplanted dopaminergic neurons. Approximately 50–57% of the embryonic dopaminergic neurons survived the dissociation and grafting procedure in rats rendered hypothermic, but there was no significant additive effect on graft survival with a combined treatment. All groups of rats exhibited behavioral recovery in the amphetamine-induced rotation test. There was a significantly enhanced functional capacity of grafts placed in hypothermic as compared to normothermic rats. However, tirilazad/YVAD pretreated implants did not afford greater behavioral improvement than control-treated grafts. Our results suggest that neuroprotective treatments administered prior to and immediately after neural graft implantation may under certain conditions rescue, at least in part, the same subset of dopaminergic neurons. The study also emphasizes the importance of the immediate time after grafting for transplant survival, with relevance both for primary mesencephalic implants and stem cell grafts.

Distribution and number of gonadotrophic cells in the pituitary gland of prepubertal female rats

1982 ◽  
Vol 94 (3) ◽  
pp. 381-NP ◽  
Author(s):  
H. G. Jansen
Keyword(s):  
Light Microscopy ◽  
Pituitary Gland ◽  
Unit Volume ◽  
Cell Number ◽  
Female Rats ◽  
Pars Distalis ◽  
Pituitary Tissue ◽  
Lh Cells ◽  
Gonadotrophic Cells

The localization and number of LH- and FSH-containing cells (LH cells and FSH cells) in the pituitary gland of prepubertal female rats aged 5, 10, 15, 20, 25 and 30 days were determined by means of immunocytochemical light microscopy of 5 μm horizontal sections. Anti-rat LHβ and anti-rat FSHβ were used in combination with the unlabelled peroxidase–antiperoxidase method. The gonadotrophic cells were regularly distributed throughout the pars distalis but at 5 and 10 days of age fewer LH and FSH cells were found in lateral regions. The numbers of LH and FSH cells per pituitary gland increased with age. The number of LH cells per unit volume of pituitary tissue reached a maximum at 20 days; the number of FSH cells reached a maximum at 15 days and then decreased with increasing age. At all ages except 5 days more LH than FSH cells were counted per unit volume. Some cells reacting with both anti-rat LHβ and anti-rat FSHβ were detected. Comparison of the data obtained on the dynamics of cell number with data previously obtained on pituitary gonadotrophin content suggests that at successive prepubertal ages LH and FSH cells contain increasing amounts of their respective hormones.

scholarly journals Information topology of gene expression profile in dopaminergic neurons

2017 ◽  
Author(s):  
Mónica Tapia Pacheco ◽  
Pierre Baudot ◽  
Martial A. Dufour ◽  
Christine Formisano-Tréziny ◽  
Simone Temporal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dopaminergic Neurons ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Combinatorial Structure ◽  
Maximum Information ◽  
Midbrain Neurons ◽  
High Degree ◽  
Multivariate Mutual Information ◽  
Gene Expression Levels

SUMMARY PARAGRAPHExtracting high-degree interactions and dependences between variables (pairs, triplets, … k-tuples) is a challenge posed by all omics approaches1, 2. Here we used multivariate mutual information (Ik) analysis3 on single-cell retro-transcription quantitative PCR (sc-RTqPCR) data obtained from midbrain neurons to estimate the k-dimensional topology of their gene expression profiles. 41 mRNAs were quantified and statistical dependences in gene expression levels could be fully described for 21 genes: Ik analysis revealed a complex combinatorial structure including modules of pairs, triplets (up to 6-tuples) sharing strong positive, negative or zero Ik, corresponding to co-varying, clustering and independent sets of genes, respectively. Therefore, Ik analysis simultaneously identified heterogeneity (negative Ik) of the cell population under study and regulatory principles conserved across the population (homogeneity, positive Ik). Moreover, maximum information paths enabled to determine the size and stability of such transcriptional modules. Ik analysis represents a new topological and statistical method of data analysis.

scholarly journals Dopaminergic axon tracts within a hyaluronic acid hydrogel encasement for implantation to restore the nigrostriatal pathway

2021 ◽  
Author(s):  
Wisberty J. Gordián-Vélez ◽  
Kevin D. Browne ◽  
Jonathan H. Galarraga ◽  
John E. Duda ◽  
Rodrigo A. España ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Dopaminergic Neurons ◽  
Dopamine Release ◽  
Neurite Growth ◽  
Nigrostriatal Pathway ◽  
Cell Therapies ◽  
The Core ◽  
Midbrain Neurons ◽  
Hyaluronic Acid Hydrogel ◽  
Deep Brain

Parkinson′s disease (PD) affects 10 million patients worldwide, making it the second most prevalent neurodegenerative disease. Motor symptoms emerge from the loss of dopamine in the striatum after the death of dopaminergic neurons and the long-projecting axons of the nigrostriatal pathway. Current treatments, such as dopamine replacement, deep brain stimulation or cell therapies, disregard the loss of this pathway at the core of symptoms. We sought to address this by improving our tissue-engineered nigrostriatal pathway (TE-NSP) technology, which consists of a tubular hydrogel with a collagen/laminin core that encases an aggregate of dopaminergic neurons and their axons in a way that resembles the nigrostriatal pathway. These constructs can be implanted to replace the lost neurons and axons with fidelity to the pathway, and thus provide dopamine according to feedback from the host circuitry. While TE-NSPs have been traditionally fabricated with agarose, here we utilized a hyaluronic acid (HA) hydrogel to expand the functionality of the encasement and our control over its properties. Using rat ventral midbrain neurons, we found that TE-NSPs exhibited longer and faster neurite growth with HA relative to agarose, with no differences observed in electrically-evoked dopamine release. When transplanted, HA hydrogels reduced host neuron loss and inflammation around the implant compared to agarose, and the cells and axons within TE-NSPs survived and maintained their cytoarchitecture for at least 2 weeks.

scholarly journals A Modeling Study Suggesting a Possible Pharmacological Target to Mitigate the Effects of Ethanol on Reward-Related Dopaminergic Signaling

2008 ◽  
Vol 99 (5) ◽  
pp. 2703-2707 ◽  
Author(s):  
Michele Migliore ◽  
Claudio Cannia ◽  
Carmen C. Canavier
Keyword(s):  
Dopaminergic Neurons ◽  
Temporal Structure ◽  
Ionic Current ◽  
Brain Regions ◽  
Point Of View ◽  
Dopamine Level ◽  
Addictive Behaviors ◽  
Brain Functions

Midbrain dopaminergic neurons are involved in several critical brain functions controlling goal-directed behaviors, reinforcing/reward processes, and motivation. Their dysfunctions alter dopamine release and contribute to a vast range of neural disorders, from Parkinson's disease to schizophrenia and addictive behaviors. These neurons have thus been a natural target of pharmacological treatments trying to ameliorate the consequences of several neuropathologies. From this point of view, a clear experimental link has been recently established between the increase in the pacemaker frequency of dopaminergic neurons in vitro after acute ethanol application and a particular ionic current ( Ih). The functional consequences in vivo, however, are not clear and they are very difficult to explore experimentally. Here we use a realistic computational model of dopaminergic neurons in vivo to suggest that ethanol, through its effects on Ih, modifies the temporal structure of the spiking activity. The model predicts that the dopamine level may increase much more during bursting than during pacemaking activity, especially in those brain regions with a slow dopamine clearance rate. The results suggest that a selective pharmacological remedy could thus be devised against the rewarding effects of ethanol that are postulated to mediate alcohol abuse and addiction, targeting the specific HCN genes expressed in dopaminergic neurons.

scholarly journals Neuroprotective Effect of Curcumin on the Nigrostriatal Pathway in a 6-Hydroxydopmine-Induced Rat Model of Parkinson’s Disease is Mediated by α7-Nicotinic Receptors

2020 ◽  
Vol 21 (19) ◽  
pp. 7329
Author(s):  
Eslam El Nebrisi ◽  
Hayate Javed ◽  
Shreesh K Ojha ◽  
Murat Oz ◽  
Safa Shehab
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Rat Model ◽  
Dopaminergic Neurons ◽  
Motor Behavior ◽  
Neurodegenerative Disorder ◽  
Neuroprotective Effect ◽  
Intraperitoneal Administration ◽  
Neuroprotective Effects ◽  
Α7 Nachr

Parkinson’s disease (PD) is a common neurodegenerative disorder, characterized by selective degeneration of dopaminergic nigrostriatal neurons. Most of the existing pharmacological approaches in PD consider replenishing striatal dopamine. It has been reported that activation of the cholinergic system has neuroprotective effects on dopaminergic neurons, and human α7-nicotinic acetylcholine receptor (α7-nAChR) stimulation may offer a potential therapeutic approach in PD. Our recent in-vitro studies demonstrated that curcumin causes significant potentiation of the function of α7-nAChRs expressed in Xenopus oocytes. In this study, we conducted in vivo experiments to assess the role of the α7-nAChR on the protective effects of curcumin in an animal model of PD. Intra-striatal injection of 6-hydroxydopmine (6-OHDA) was used to induce Parkinsonism in rats. Our results demonstrated that intragastric curcumin treatment (200 mg/kg) significantly improved the abnormal motor behavior and offered neuroprotection against the reduction of dopaminergic neurons, as determined by tyrosine hydroxylase (TH) immunoreactivity in the substantia nigra and caudoputamen. The intraperitoneal administration of the α7-nAChR-selective antagonist methyllycaconitine (1 µg/kg) reversed the neuroprotective effects of curcumin in terms of both animal behavior and TH immunoreactivity. In conclusion, this study demonstrates that curcumin has a neuroprotective effect in a 6-hydroxydopmine (6-OHDA) rat model of PD via an α7-nAChR-mediated mechanism.

scholarly journals Recent Advances in Imaging of Dopaminergic Neurons for Evaluation of Neuropsychiatric Disorders

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
Lie-Hang Shen ◽  
Mei-Hsiu Liao ◽  
Yu-Chin Tseng
Keyword(s):  
Dopaminergic Neurons ◽  
Motor Behavior ◽  
Single Photon ◽  
Photon Emission ◽  
Neuropsychiatric Disorders ◽  
Clinical Aspects ◽  
Emission Computed Tomography ◽  
Dopamine System ◽  
Dopaminergic Neurotransmission ◽  
Positron Emission

Dopamine is the most intensely studied monoaminergic neurotransmitter. Dopaminergic neurotransmission plays an important role in regulating several aspects of basic brain function, including motor, behavior, motivation, and working memory. To date, there are numerous positron emission tomography (PET) and single photon emission computed tomography (SPECT) radiotracers available for targeting different steps in the process of dopaminergic neurotransmission, which permits us to quantify dopaminergic activity in the living human brain. Degeneration of the nigrostriatal dopamine system causes Parkinson’s disease (PD) and related Parkinsonism. Dopamine is the neurotransmitter that has been classically associated with the reinforcing effects of drug abuse. Abnormalities within the dopamine system in the brain are involved in the pathophysiology of attention deficit hyperactivity disorder (ADHD). Dopamine receptors play an important role in schizophrenia and the effect of neuroleptics is through blockage of dopamine D2receptors. This review will concentrate on the radiotracers that have been developed for imaging dopaminergic neurons, describe the clinical aspects in the assessment of neuropsychiatric disorders, and suggest future directions in the diagnosis and management of such disorders.

scholarly journals Absence of Glia Maturation Factor Protects Dopaminergic Neurons and Improves Motor Behavior in Mouse Model of Parkinsonism

2015 ◽  
Vol 40 (5) ◽  
pp. 980-990 ◽  
Author(s):  
Mohammad Moshahid Khan ◽  
Smita Zaheer ◽  
Ramasamy Thangavel ◽  
Margi Patel ◽  
Duraisamy Kempuraj ◽  
...  
Keyword(s):  
Mouse Model ◽  
Dopaminergic Neurons ◽  
Motor Behavior ◽  
Glia Maturation Factor ◽  
Maturation Factor

Effects of Carbon Tetrachloride on Neuronal Differentiation in Rat Embryo Mid-Brain Micromass Cultures

1989 ◽  
Vol 16 (3) ◽  
pp. 287-292
Author(s):  
Cecilia Clemedson ◽  
Erik Walum ◽  
Oliver Flint
Keyword(s):  
Carbon Tetrachloride ◽  
Cell Survival ◽  
Neuronal Differentiation ◽  
Neuronal Development ◽  
Neutral Red ◽  
Cell Number ◽  
Rat Embryo ◽  
Focus Formation ◽  
Drastic Increase ◽  
Development And Differentiation

The effects of carbon tetrachloride on neuronal development and differentiation have been studied. Rat embryo mid-brain micromass cultures were exposed for five days to 50, 100, 250, 500 or 1000ppm CCl4 in closed chambers. Differentiation was indicated by the formation of neuronal foci in the cultures. Effects on cell survival were estimated using a neutral red staining method. Carbon tetrachloride at 100ppm caused a drastic increase in neuronal differentiation, accompanied by a slight increase in total cell number. At 250ppm the number of differentiated foci was still elevated, but the outgrowth of neurites was markedly reduced. Neuronal differentiation and cell survival were reduced by 50% at a concentration of 430ppm. Using criteria previously defined for the identification of potentially teratogenic substances in the micromass system, CCl4 was classified as a non-teratogen. Nevertheless, the results show that a substance can cause inhibition of differentiation (reduction in neunte formation) without altering focus formation.

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