Cellular localization of tyrosine hydroxylase by                immunohistochemistry.

The enzyme tyrosine hydroxylase (TH) was immunohistochemically localized by the peroxidase-antiperoxidase method in rat to chromaffin cells of the adrenal medulla, large neurons and small darkly staining cells of the superior cervical ganglia and noradrenergic and dopaminergic neurons in brain. As compared with the conjugated peroxidase or immunofluorescence techniques, the peroxidase-antiperoxidase method gave the most selective and specific cytoplasmic localization of TH antisera in every tissue examined. The peroxidase staining with the TH antisera was more intense in dopaminergic than in noradrenergic neurons of the central nervous system. While TH was visualized in cell bodies of both dopaminergic and noradrenergic neurons, it could only be detected in axons and terminals in the dopaminergic system. The perikarya of noradrenergic neurons could be distinguished from dopaminergic neurons by the immunohistochemical demonstration of the enzyme dopamine-beta-hydroxylase only in the former.

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Pituitary Adenylate Cyclase-Activating Polypeptide Induces Gene Expression of the Catecholamine Synthesizing Enzymes, Tyrosine Hydroxylase, and Dopamine beta-Hydroxylase in Cultured Porcine Adrenal Medullary Chromaffin Cells

Annals of the New York Academy of Sciences ◽

10.1111/j.1749-6632.1996.tb17504.x ◽

2006 ◽

Vol 805 (1) ◽

pp. 464-469 ◽

Cited By ~ 4

Author(s):

KAZUMASA ISOBE ◽

NOBUYASU YUKIMASA ◽

TOSHIAKI NAKAI ◽

YOH TAKUWA

Keyword(s):

Gene Expression ◽

Tyrosine Hydroxylase ◽

Adenylate Cyclase ◽

Chromaffin Cells ◽

Dopamine Beta Hydroxylase ◽

Pituitary Adenylate Cyclase ◽

Catecholamine Synthesizing Enzymes

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Cardiac Tyrosine Hydroxylase Activation and Mb-Comt in Dyskinetic Monkeys

10.21203/rs.3.rs-126537/v1 ◽

2021 ◽

Author(s):

Lorena Cuenca-Bermejo ◽

Pilar Almela ◽

Pablo Gallo-Soljancic ◽

José Yuste ◽

Vicente de Pablos ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Tyrosine Hydroxylase ◽

Dopaminergic System ◽

Nigrostriatal Pathway ◽

Noradrenergic Neurons ◽

Methyl Transferase ◽

Mptp Treatment ◽

The Impact

Abstract The impact of age-associated disorders is increasing as the life expectancy of the population increments. Cardiovascular diseases and neurodegenerative disorders, such as Parkinson’s disease, have the highest social and economic burden and increasing evidence show interrelations between them. Particularly, dysfunction of the cardiovascular nervous system is part of the dysautonomic symptoms of Parkinson’s disease, although more studies are needed to elucidate the role of cardiac function on it. We analyzed the dopaminergic system in the nigrostriatal pathway of Parkinsonian and dyskinetic monkeys and the expression of some key proteins in the metabolism and synthesis of catecholamines in the heart: total and phosphorylated (phospho) tyrosine hydroxylase (TH), and membrane (MB) and soluble (S) isoforms of catechol-O-methyl transferase (COMT). The dopaminergic system was significantly depleted in all MPTP-intoxicated monkeys. MPTP- and MPTP+L-DOPA-treated animals also showed a decrease in total TH expression in both right (RV) and left ventricle (LV). We found a significant increase of phospho-TH in both groups (MPTP and MPTP+L-DOPA) in the LV, while this increase was only observed in MPTP-treated monkeys in the RV. MB-COMT analysis showed a very significant increase of this isoform in the LV of MPTP- and MPTP+L-DOPA-treated animals, with no significant differences in S-COMT levels. These data suggest that MB-COMT is the main isoform implicated in the cardiac noradrenergic changes observed after MPTP treatment, suggesting an increase in NA metabolism. Moreover, the increase of TH activity indicates that cardiac noradrenergic neurons still respond despite MPTP treatment.

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The Effect of Perinatal Hypoxic/Ischemic Injury on Tyrosine Hydroxylase Expression in the Locus Coeruleus of the Human Neonate

Developmental Neuroscience ◽

10.1159/000439270 ◽

2015 ◽

Vol 38 (1) ◽

pp. 41-53 ◽

Cited By ~ 3

Author(s):

Marianna A. Pagida ◽

Anastasia E. Konstantinidou ◽

Anna Korelidou ◽

Dimitra Katsika ◽

Effrosini Tsekoura ◽

...

Keyword(s):

Tyrosine Hydroxylase ◽

Locus Coeruleus ◽

Dopaminergic Neurons ◽

Ischemic Injury ◽

Selective Vulnerability ◽

Noradrenergic Neurons ◽

Critical Periods ◽

Human Neonate ◽

Almost All ◽

Hypoxic Ischemic Injury

We have previously shown that perinatal hypoxic/ischemic injury (HII) may cause selective vulnerability of the mesencephalic dopaminergic neurons of human neonate. In the present study, we investigated the effect of perinatal HII on the noradrenergic neurons of the locus coeruleus (LC) of the same sample. We studied immunohistochemically the expression of tyrosine hydroxylase (TH, first limiting enzyme for catecholamine synthesis) in LC neurons of 15 autopsied infants (brains collected from the Greek Brain Bank) in relation to the neuropathological changes of acute or chronic HII of the neonatal brain. Our results showed that perinatal HII appears to affect the expression of TH and the size of LC neurons of the human neonate. In subjects with neuropathological lesions consistent with abrupt/severe HII, intense TH immunoreactivity was found in almost all neurons of the LC. In most of the neonates with neuropathological changes of prolonged or older injury, however, reduction in cell size and a decrease or absence of TH staining were observed in the LC. Intense TH immunoreactivity was found in the LC of 3 infants of the latter group, who interestingly had a longer survival time and had been treated with anticonvulsant drugs. Based on our observations and in view of experimental evidence indicating that the reduction of TH-immunoreactive neurons occurring in the LC after perinatal hypoxic insults persists into adulthood, we suggest that a dysregulation of monoaminergic neurotransmission in critical periods of brain development in humans is likely to predispose the survivors of perinatal HII, in combination with genetic susceptibility, to psychiatric and/or neurological disorders later in life.

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Comparison of LR White and Unicryl as embedding media for light and electron immunomicroscopy of chromaffin cells.

Journal of Histochemistry & Cytochemistry ◽

10.1177/44.3.8648090 ◽

1996 ◽

Vol 44 (3) ◽

pp. 289-295 ◽

Cited By ~ 12

Author(s):

G Goping ◽

G A Kuijpers ◽

R Vinet ◽

H B Pollard

Keyword(s):

Electron Microscopy ◽

Tyrosine Hydroxylase ◽

Membrane Fusion ◽

Chromaffin Cells ◽

Chromaffin Cell ◽

Osmium Tetroxide ◽

Immunogold Labeling ◽

Dopamine Beta Hydroxylase ◽

Lr White ◽

Embedding Medium

LR White and Unicryl are members of the same family of acrylic embedding resins and are very suitable for "on grid" postembedding immunogold labeling. We studied the ultrastructure of LR White- and Unicryl-embedded cultured chromaffin cells and the immunolocalization of three chromaffin cell proteins, the enzymes dopamine-beta-hydroxylase (DbetaH) and tyrosine hydroxylase (TH), and the membrane fusion and Ca2+ channel protein synexin (annexin VII). We report here that Unicryl is preferable to LR White as an embedding medium for electron microscopy when osmium tetroxide fixation is omitted. The basis for this distinction is better ultrastructural preservation and improved immunodetection efficiency.

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Cardiac tyrosine hydroxylase activation and MB-COMT in dyskinetic monkeys

Scientific Reports ◽

10.1038/s41598-021-99237-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Lorena Cuenca-Bermejo ◽

Pilar Almela ◽

Pablo Gallo-Soljancic ◽

José E. Yuste ◽

Vicente de Pablos ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Tyrosine Hydroxylase ◽

Dopaminergic System ◽

Nigrostriatal Pathway ◽

Noradrenergic Neurons ◽

Methyl Transferase ◽

Mptp Treatment ◽

The Impact

AbstractThe impact of age-associated disorders is increasing as the life expectancy of the population increments. Cardiovascular diseases and neurodegenerative disorders, such as Parkinson’s disease, have the highest social and economic burden and increasing evidence show interrelations between them. Particularly, dysfunction of the cardiovascular nervous system is part of the dysautonomic symptoms of Parkinson’s disease, although more studies are needed to elucidate the role of cardiac function on it. We analyzed the dopaminergic system in the nigrostriatal pathway of Parkinsonian and dyskinetic monkeys and the expression of some key proteins in the metabolism and synthesis of catecholamines in the heart: total and phosphorylated (phospho) tyrosine hydroxylase (TH), and membrane (MB) and soluble (S) isoforms of catechol-O-methyl transferase (COMT). The dopaminergic system was significantly depleted in all MPTP-intoxicated monkeys. MPTP- and MPTP + L-DOPA-treated animals also showed a decrease in total TH expression in both right (RV) and left ventricle (LV). We found a significant increase of phospho-TH in both groups (MPTP and MPTP + L-DOPA) in the LV, while this increase was only observed in MPTP-treated monkeys in the RV. MB-COMT analysis showed a very significant increase of this isoform in the LV of MPTP- and MPTP + L-DOPA-treated animals, with no significant differences in S-COMT levels. These data suggest that MB-COMT is the main isoform implicated in the cardiac noradrenergic changes observed after MPTP treatment, suggesting an increase in noradrenaline (NA) metabolism. Moreover, the increase of TH activity indicates that cardiac noradrenergic neurons still respond despite MPTP treatment.

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Evidence for a Blood Ganglion Barrier in the Superior Cervical Ganglion of the Rat

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100053681 ◽

1982 ◽

Vol 40 ◽

pp. 204-204

Author(s):

D. M. DePace

Keyword(s):

Blood Vessels ◽

Superior Cervical Ganglion ◽

Peripheral Nerves ◽

Time Schedule ◽

Transmission Electron ◽

Cervical Ganglion ◽

The Central Nervous System ◽

Cervical Ganglia ◽

Capillary Circulation ◽

And Control

The majority of blood vessels in the superior cervical ganglion possess a continuous endothelium with tight junctions. These same features have been associated with the blood brain barrier of the central nervous system and peripheral nerves. These vessels may perform a barrier function between the capillary circulation and the superior cervical ganglion. The permeability of the blood vessels in the superior cervical ganglion of the rat was tested by intravenous injection of horseradish peroxidase (HRP). Three experimental groups of four animals each were given intravenous HRP (Sigma Type II) in a dosage of.08 to.15 mg/gm body weight in.5 ml of.85% saline. The animals were sacrificed at five, ten or 15 minutes following administration of the tracer. Superior cervical ganglia were quickly removed and fixed by immersion in 2.5% glutaraldehyde in Sorenson's.1M phosphate buffer, pH 7.4. Three control animals received,5ml of saline without HRP. These were sacrificed on the same time schedule. Tissues from experimental and control animals were reacted for peroxidase activity and then processed for routine transmission electron microscopy.

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