Catecholaminergic Neurons Research Articles

A Review of Three Commonly Used Herbs Which Enhance Memory and New Evidences Which Show Their Combination Could Improve Memory in Young Animals.

This review looks into the herbs Gingko biloba, Polygala tenuifolia, and Lycii fructus for their widely studied neuroprotective properties. In particular, we investigated memory enhancing effect of these herbs, and their potential synergetic effect on memory with new data. Sixmonth treated mice demonstrated shorter escape latency in water maze and shorter arrival time in a consolidated memory task. Immunochemistry showed evident increase in superoxide dismutase activities in the prefrontal cortex, implying protection against free radicals during aging. Discrete increase of catecholaminergic neurons was found in the prefrontal cortex, hippocampus, corpus striatum, and midbrain, suggesting better memory and better control on mood and behavior. Necrotic cells in the brain decreased as indicated by immunocytochemistry of lactic dehydrogenase. Terminal deoxynucleotidyl transferase dUTP nick end labeling showed no apoptotic cells in most brain areas in high dose group. Biochemistry revealed increase of dopaminergic cells in treatment groups at prefrontal cortex, and in the hippocampus and cerebellum of the high dose group. Most 6-month groups showed increase of serotonin in all three areas. For the high dose group, GABA increased in the hippocampus but not prefrontal cortex, which would help induce sleep at night. Protein kinase C increased in most groups at prefrontal cortex, hippocampus and cerebellum, signifying increase of possible signal transduction pathways for memory or other nervous activations. Our results intimate that the interaction of the three herbs exerts beneficial effects on memory, associated cognitive function, and necrosis. Future investigations based on the present data shall aid development of clinically relevant medication.

Non-motor multiple system atrophy associated with sudden death: pathological observations of autonomic nuclei

Multiple system atrophy (MSA) manifests as a combination of dysautonomia and motor symptoms/signs. However, rare cases presenting with autonomic failures in absence of motor symptoms/signs until their deaths have been reported and are referred to as non-motor MSA. To clarify pathological findings underlying non-motor MSA patients, we analyzed consecutively autopsied 161 patients with MSA. In results, four patients were identified as having non-motor MSA, who showed isolated autonomic disorders throughout their lives and had minimal pathological changes in the motor systems. We also identified two patients with pathologically minimal MSA, who had minimal pathological involvement in the motor systems and presented with definite parkinsonism and dysautonomia. Survival durations of the non-motor MSA patients were much shorter (1.3-2.0years) than those of the classical MSA patients (3.0-7.0years), and the causes of death were all sudden death. The medullary serotonergic neurons were severely involved in the non-motor MSA patients in comparison with the classical MSA patients. Also, one of the pathologically minimal MSA patients had died suddenly and exhibited marked involvement of the medullary serotonergic neurons. The involvement of the medullary catecholaminergic or cholinergic neurons did not differ in severities among the groups. We conclude that non-motor MSA may be a pathological variant of MSA that preferentially involves the medullary serotonergic neurons and autonomic systems in association with poor prognosis.

Nuclear organisation of cholinergic, catecholaminergic, serotonergic and orexinergic neurons in two relatively large-brained rodent species—The springhare (Pedetes capensis) and Beecroft’s scaly-tailed squirrel (Anomalurus beecrofti)

The present study describes the nuclear organization of the cholinergic, catecholaminergic, serotonergic and orexinergic systems in the brains of the springhare and Beecroft’s scaly-tailed squirrel following immunohistochemical labelling. We aimed to investigate any differences in the nuclear organization of these neural systems when compared to previous data on other species of rodents, as these two rodent species have relatively large brains − 1.2 to 1.4 times larger than would be expected for mammals of their body mass and 1.7–1.9 times larger than would be expected for rodents of their body mass. A series of coronal sections were taken through two brains of each species and immunohistochemically labelled with antibodies against choline acetyltransferase, tyrosine hydroxylase, serotonin and orexin-A. Generally, the nuclear complement of these systems revealed extensive similarities between both species and to previously studied rodents. While no differences were observed in the nuclear complement of the serotonergic and orexinergic systems, some differences were observed in the nuclear complement of the cholinergic and catecholaminergic systems. These include the presence of cholinergic neurons in the cerebral cortex and nucleus of the trapezoid body in the springhare; while the Beecroft’s scaly-tailed squirrel exhibited cholinergic neurons in the pretectal area of the midbrain. For the catecholaminergic system it was observed that Beecroft’s scaly-tailed squirrel possessed immunoreactive neurons in the accessory olfactory bulb. Despite these four differences, most not previously observed in rodents, the remaining complement of cholinergic and catecholaminergic nuclei were identical to that observed in other rodents, including the presence of the rodent specific catecholaminergic rostral dorsal midline medullary (C3) nucleus in the medulla oblongata. Thus, even with a significant increase in relative brain size, the overall complement of nuclei forming these systems shows minimal changes in complexity within a specific mammalian order.

Structural Organization and Interaction of Intrapancreatic Ganglia with the Myenteric Nerve Plexus of the Duodenum at the Early Stages of Postnatal Ontogeny in Rats

Studies using immunohistochemical markers – synaptophysin (SP), protein gene product 9.5 (PGP 9.5), and tyrosine hydroxylase (TH) – addressed the innervation of the pancreas and the myenteric nerve plexus of the duodenum (duodenum) in neonatal rats (n = 4) and demonstrated a tight link between the nerve plexuses in these organs. Some elements of the myenteric plexus penetrate the pancreas from the walls of the duodenum. Morphological and biochemical similarity was demonstrated between microganglia in the myenteric plexus of the duodenum and pancreas. Most differentiated neurons and bundles of axons in both plexuses were part of the parasympathetic compartment of the autonomic nervous system. There were no catecholaminergic neurons in the pancreas, and bundles of sympathetic fibers entering the organ from outside were few in number and were involved mainly in innervating blood vessels. This is the first report of the use of immunohistochemical reactions for PGP 9.5, SP, and TH to detect two types of nerve fibers in ganglia in the pancreas whose varicose terminals form cholinergic and catecholaminergic synapses around nerve cell bodies.

Conditional Deletion of Prnp Rescues Behavioral and Synaptic Deficits after Disease Onset in Transgenic Alzheimer's Disease.

Biochemical and genetic evidence implicate soluble oligomeric amyloid-β (Aβo) in triggering Alzheimer's disease (AD) pathophysiology. Moreover, constitutive deletion of the Aβo-binding cellular prion protein (PrPC) prevents development of memory deficits in APPswe/PS1ΔE9 mice, a model of familial AD. Here, we define the role of PrPC to rescue or halt established AD endophenotypes in a therapeutic disease-modifying time window after symptom onset. Deletion of Prnp at either 12 or 16 months of age fully reverses hippocampal synapse loss and completely rescues preexisting behavioral deficits by 17 months. In contrast, but consistent with a neuronal function for Aβo/PrPC signaling, plaque density, microgliosis, and astrocytosis are not altered. Degeneration of catecholaminergic neurons remains unchanged by PrPC reduction after disease onset. These results define the potential of targeting PrPC as a disease-modifying therapy for certain AD-related phenotypes after disease onset.SIGNIFICANCE STATEMENT The study presented here further elucidates our understanding of the soluble oligomeric amyloid-β-Aβo-binding cellular prion protein (PrPC) signaling pathway in a familial form of Alzheimer's disease (AD) by implicating PrPC as a potential therapeutic target for AD. In particular, genetic deletion of Prnp rescued several familial AD (FAD)-associated phenotypes after disease onset in a mouse model of FAD. This study underscores the therapeutic potential of PrPC deletion given that patients already present symptoms at the time of diagnosis.

Neuroprotective Effects of Intranasal IGF-1 against Neonatal Lipopolysaccharide-Induced Neurobehavioral Deficits and Neuronal Inflammation in the Substantia Nigra and Locus Coeruleus of Juvenile Rats

Neonatal lipopolysaccharide (LPS) exposure-induced brain inflammation resulted in motor dysfunction and brain dopaminergic neuronal injury, and increased the risks of neurodegenerative disorders in adult rats. Our previous studies showed that intranasal administration of insulin-like growth factor-1 (IGF-1) protects against LPS-induced white matter injury in the developing rat brain. To further examine whether IGF-1 protects against LPS-induced brain neuronal injury and neurobehavioral dysfunction, recombinant human IGF-1 (rhIGF-1) at a dose of 50 µg/pup was administered intranasally 1 h following intracerebral injection of LPS (1 mg/kg) in postnatal day 5 (P5) Sprague-Dawley rat pups. Neurobehavioral tests were carried out from P7 to P21, and brain neuronal injury was examined at P21. Our results showed that LPS exposure resulted in disturbances of motor behaviors in juvenile rats. Moreover, LPS exposure caused injury to central catecholaminergic neurons, as indicated by a reduction of tyrosine hydroxylase (TH) immunoreactivity in the substantia nigra (SN), ventral tegmental area (VTA) and olfactory bulb (OB), and brain noradrenergic neurons, as indicated by a reduction of TH immunoreactivity in the locus coeruleus (LC) of the P21 rat brain. The LPS-induced reduction of TH+ cells was observed at a greater degree in the SN and LC of the P21 rat brain. Intranasal rhIGF-1 treatment attenuated LPS-induced central catecholaminergic neuronal injury and motor behavioral disturbances, including locomotion, beam walking test and gait analysis. Intranasal rhIGF-1 administration also attenuated LPS-induced elevation of IL-1β levels and numbers of activated microglia, and cyclooxygenase-2+ cells, which were double labeled with TH+ cells in the SN, VTA, OB and LC of the P21 rat brain. These results suggest that IGF-1 may provide protection against neonatal LPS exposure-induced central catecholaminergic neuronal injury and motor behavioral disturbances, and that the protective effects are associated with the inhibition of microglia activation and the reduction of neuronal oxidative stress by the suppression of the neuronal cyclooxygenase-2 expression.

Self-administration of methamphetamine alters gut biomarkers of toxicity.

Methamphetamine (METH) is a highly abused psychostimulant that is associated with an increased risk for developing Parkinson's disease (PD). This enhanced vulnerability likely relates to the toxic effects of METH that overlap with PD pathology, for example, aberrant functioning of α-synuclein and parkin. In PD, peripheral factors are thought to contribute to central nervous system (CNS) degeneration. For example, α-synuclein levels in the enteric nervous system (ENS) are elevated, and this precedes the onset of motor symptoms. It remains unclear whether neurons of the ENS, particularly catecholaminergic neurons, exhibit signs of METH-induced toxicity as seen in the CNS. The aim of this study was to determine whether self-administered METH altered the levels of α-synuclein, parkin, tyrosine hydroxylase (TH), and dopamine-β-hydroxylase (DβH) in the myenteric plexus of the distal colon ENS. Young adult male Sprague-Dawley rats self-administered METH for 3h per day for 14days and controls were saline-yoked. Distal colon tissue was collected at 1, 14, or 56days after the last operant session. Levels of α-synuclein were increased, while levels of parkin, TH, and DβH were decreased in the myenteric plexus in the METH-exposed rats at 1day following the last operant session and returned to the control levels after 14 or 56days of forced abstinence. The changes were not confined to neurofilament-positive neurons. These results suggest that colon biomarkers may provide early indications of METH-induced neurotoxicity, particularly in young chronic METH users who may be more susceptible to progression to PD later in life.

Acute action of rotenone on excitability of catecholaminergic neurons in rostral ventrolateral medulla.

The degeneration of the rostral ventrolateral medulla (RVLM) catecholaminergic neurons was responsible for some cardiovascular symptoms in Parkinson's disease (PD). Our previous study had observed the impairment of these neurons in the early stage of PD in the rotenone-induced PD rat model, but the related mechanisms remain unclear. Rotenone is a mitochondrial inhibitor, influencing the neuronal electrophysiological activity through activation of K-ATP channels that potentially participate in cell death processes. In the present study, effects of rotenone on electrophysiological properties of RVLM catecholaminergic neurons and its underlying mechanisms were investigated. In coronal slices of brain containing the RVLM through patch clamp technique, rotenone (0.5μM) induced gradual postsynaptic inhibition on the spontaneous firing and cell membrane hyperpolarization with outward currents of catecholaminergic neurons. The electrophysiological changes were blocked by glibenclamide (30μM), a blocker of K-ATP channels, and were nearly unchanged by diazoxide (100μM), an opener of K-ATP channels. Our results also showed that effects of rotenone on catecholaminergic neurons including reactive oxygen species (ROS) generation were prevented by pretreatment of coenzyme Q10 (CoQ10, 100μM), a scavenger of ROS. These suggest that rotenone-induced electrophysiological changes of RVLM catecholaminergic neurons are caused by the opening of K-ATP channels, which are partly related to ROS generation. The changes of K-ATP channels might account for the vulnerability of RVLM catecholaminergic neurons.

Distribution of tyrosine hydroxylase-immunoreactive neurons in the brain of the viviparous fish Gambusia affinis

Tyrosine hydroxylase (TH) is the common precursor enzyme involved in the biosynthetic pathway of the catecholaminergic neurotransmitters, dopamine and norepinephrine. In this investigation, the neuroanatomical distribution of TH-immunoreactivity was studied in the brain of the female mosquitofish Gambusia affinis. Numerous intensely stained TH-immunoreactive (ir) neurons were scattered in the olfactory bulb with their fibres extending towards the medial olfactory tract, whereas few telencephalic TH-ir cells with distinct fibres were observed in the dorsal nucleus of area ventralis telencephali and the posterior nucleus of area ventralis telencephali regions. Large TH-ir cell populations were seen in the suprachiasmatic nucleus and the nucleus dorsomedialis thalami regions of the diencephalon. Distinct TH-ir cells with long fibres were found at the preoptic area and the nucleus preopticus pars magnocellularis as well as the nucleus preopticus pars parvocellularis regions. Numerous intensely stained TH-ir cells were observed in the paraventricular organ and the nucleus posterior tuberis regions, whereas moderately stained cells were present in the nucleus of recessus lateralis medialis. Several TH-ir neurons were detected in medial and lateral subdivisions of the nucleus lateralis tuberis. Furthermore, the projections of the TH-ir fibres were seen in the proximal pars distalis region of the pituitary gland, where gonadotropin-secreting cells are located, suggesting the communication between TH cells and gonadotrope cells. In the rostral spinal cord, dense aggregations of the TH-ir fibres were noticed. Overall, the widespread distribution of the TH-ir neurons throughout the brain and their fibres in the spinal cord and the pituitary gland suggests diverse roles for the catecholaminergic neurons in various physiological functions including reproduction in the mosquitofish.

Ciliomotor circuitry underlying whole-body coordination of ciliary activity in the Platynereis larva.

Ciliated surfaces harbouring synchronously beating cilia can generate fluid flow or drive locomotion. In ciliary swimmers, ciliary beating, arrests, and changes in beat frequency are often coordinated across extended or discontinuous surfaces. To understand how such coordination is achieved, we studied the ciliated larvae of Platynereis dumerilii, a marine annelid. Platynereis larvae have segmental multiciliated cells that regularly display spontaneous coordinated ciliary arrests. We used whole-body connectomics, activity imaging, transgenesis, and neuron ablation to characterize the ciliomotor circuitry. We identified cholinergic, serotonergic, and catecholaminergic ciliomotor neurons. The synchronous rhythmic activation of cholinergic cells drives the coordinated arrests of all cilia. The serotonergic cells are active when cilia are beating. Serotonin inhibits the cholinergic rhythm, and increases ciliary beat frequency. Based on their connectivity and alternating activity, the catecholaminergic cells may generate the rhythm. The ciliomotor circuitry thus constitutes a stop-and-go pacemaker system for the whole-body coordination of ciliary locomotion.

Role of the locus coeruleus catecholaminergic neurons in the chemosensory control of breathing in a Parkinson's disease model

A previous study has demonstrated that in the 6-hydroxydopamine (6-OHDA)-model of Parkinson's disease (PD) there is a reduction in the number of Phox2b neurons in the retrotrapezoid nucleus (RTN) and a decrease in the respiratory response to hypercapnia 40days after PD-induction. The functional deficiency is restored 60days after 6-OHDA injection and here we tested the hypothesis that the locus coeruleus (LC) could be a candidate to restore the breathing deficiency. Minute Ventilation (VE) in response to hypercapnia (7% CO2) was assessed one day before, and then 40 and 60days after bilateral 6-OHDA (24μg/μL) or vehicle injections into the LC in control or PD-induced male Wistar rats. Bilateral injections of 6-OHDA decreased catecholaminergic neurons by 86% and 83% in the substantia nigra pars compacta (SNpc) and LC, respectively. As already described, in animals with lesions to the SNpc (N=6/group), the reduction in the ventilatory response to hypercapnia was restored 60days after PD (1257±81 vs. vehicle: 1185±49mL/kg/min). However, in animals with PD and lesion in the LC, the ventilation was blunted (674±39mL/kg/min). In another group of PD rats, we observed a reduction in the number of hypercapnia-induced-fos+ cells in the RTN region (40days: 38±3 and 60days: 8.5±0.9 vs. vehicle 78±3 cells) and an increase in the LC (40days: 46±4 and 60days: 94±22 vs. vehicle 1±1 cells). Our data suggest that LC catecholaminergic neurons can be a candidate structure mediating chemoreceptor function in a model of PD.

Regional distribution of cholinergic, catecholaminergic, serotonergic and orexinergic neurons in the brain of two carnivore species: The feliform banded mongoose (Mungos mungo) and the caniform domestic ferret (Mustela putorius furo)

The nuclear organization of the cholinergic, catecholaminergic, serotonergic and orexinergic neurons in the brains of two species of carnivore, the banded mongoose (Mungos mungo) and domestic ferret (Mustela putorius furo), is presented. The banded mongoose belongs to the feliform suborder and the domestic ferret to the caniform suborder, having last shared a common ancestor approximately 53 million years ago; however, they have a very similar overall morphology and life history, presenting an interesting opportunity to examine the extent of evolutionary plasticity in these systems. The brains of the two carnivore species were coronally sectioned and immunohistochemically stained with antibodies against choline acetyltransferase, tyrosine hydroxylase, serotonin and orexin-A. The overall organization and complement of the nuclei of these systems was identical between the two species, although minor differences were noted. Moreover, this overall organization is identical to other studies undertaken in the domestic cat and dog. While for the most part the nuclei forming these systems are similar to those observed in other mammals, two species differences, which appear to be carnivore-specific, were noted. First, cholinergic neurons were observed in the lateral septal nucleus of both species, an apparently carnivore specific feature not recorded previously in other mammals. Second, the serotonergic neurons of the peripheral division of the dorsal raphe complex exhibited a significant caudad expansion, intermingling with the cholinergic and catecholaminergic nuclei of the pons, a carnivore specific feature. These carnivore specific features likely have functional consequences related to coping with stress and the expression of sleep.

Thalamic dopaminergic neurons projects to the paraventricular nucleus-rostral ventrolateral medulla/C1 neural circuit.

Paraventricular nuclei (PVN) projections to the rostral ventrolateral medulla (RVLM)/C1 catecholaminergic neuron group constitute the pre-autonomic sympathetic center involved in the neural control of systemic cardiovascular function. However, the role of extra-hypothalamic and thalamic dopaminergic (DA) inputs in this circuit remains underexplored. Using retrograde neuroanatomical tracing and high contrast confocal imaging methods, we investigated the projections and morphology of the discrete thalamic DA neuron groups in the dorsal hypothalamic area and their contribution to the PVN-RVLM neural circuit. We found that DA neuron subgroups in the Zona Incerta (Zi; 60%) and Reuniens thalamic nuclei (Re; 40%) were labeled comparably to the PVN (85%) after a retrograde tracer was injected into the RVLM/C1 (P < 0.01 mean ± SEM). The Re/Zi DA neuron subgroups were characterized by angulated cell bodies, superiomedial and inferiomedial projections reaching the contralateral Re/Zi and ipsilateral PVN DA neurons respectively. Ultimately, we deduced that the DA projections of the Re/Zi to the PVN contribute to the PVN-RVLM/C1 neural circuit. As a result of these connections, the Re/Zi DA neuron groups may regulate preautonomic sympathetic events associated with the PVN-RVLM pathway. Anat Rec, 300:1307-1314, 2017. © 2016 Wiley Periodicals, Inc.

N-oleoyldopamine modulates activity of midbrain dopaminergic neurons through multiple mechanisms

N-oleoyl-dopamine (OLDA) is an amide of dopamine and oleic acid, synthesized in catecholaminergic neurons. The present study investigates OLDA targets in midbrain dopaminergic (DA) neurons. Substantia Nigra compacta (SNc) DA neurons recorded in brain slices were excited by OLDA in wild type mice. In transient receptor potential vanilloid 1 (TRPV1) knockout (KO) mice, however, SNc DA neurons displayed sustained inhibition of firing. In the presence of the dopamine type 2 receptor (D2R) antagonist sulpiride or the dopamine transporter blocker nomifensine no such inhibition was observed. Under sulpiride OLDA slightly excited SNc DA neurons, an action abolished upon combined application of the cannabinoid1 and 2 receptor antagonists AM251 and AM630. In ventral tegmental area (VTA) DA neurons from TRPV1 KO mice a transient inhibition of firing by OLDA was observed. Thus OLDA modulates the firing of nigrostriatal DA neurons through interactions with TRPV1, cannabinoid receptors and dopamine uptake. These findings suggest further development of OLDA-like tandem molecules for the treatment of movement disorders including Parkinson's disease.

A brain-sparing diphtheria toxin for chemical genetic ablation of peripheral cell lineages

Conditional expression of diphtheria toxin receptor (DTR) is widely used for tissue-specific ablation of cells. However, diphtheria toxin (DT) crosses the blood–brain barrier, which limits its utility for ablating peripheral cells using Cre drivers that are also expressed in the central nervous system (CNS). Here we report the development of a brain-sparing DT, termed BRAINSPAReDT, for tissue-specific genetic ablation of cells outside the CNS. We prevent blood–brain barrier passage of DT through PEGylation, which polarizes the molecule and increases its size. We validate BRAINSPAReDT with regional genetic sympathectomy: BRAINSPAReDT ablates peripheral but not central catecholaminergic neurons, thus avoiding the Parkinson-like phenotype associated with full dopaminergic depletion. Regional sympathectomy compromises adipose tissue thermogenesis, and renders mice susceptible to obesity. We provide a proof of principle that BRAINSPAReDT can be used for Cre/DTR tissue-specific ablation outside the brain using CNS drivers, while consolidating the link between adiposity and the sympathetic nervous system.

Increased BDNF Expression in the Paraventricular Nucleus of the Hypothalamus (PVN) Abolishes Hypotensive Actions of Nucleus of the Solitary Tract (NTS) Catecholaminergic Neurons

Brain derived neurotrophic factor (BDNF) expression in the PVN increases in response to hypertensive stimuli including stress, and we have recently shown that BDNF is a significant pro‐hypertensive mediator in the PVN. However, the downstream mechanisms mediating this effect of BDNF remain to be investigated. BDNF is an important modulator of catecholaminergic neuronal function throughout the central nervous system; therefore it could potentially influence catecholaminergic input into the PVN as well. The majority of catecholaminergic projections to the PVN originate from the NTS, and these neurons have been shown to exert hypotensive actions. Here, we tested the hypothesis that increased BDNF levels in the PVN elevate blood pressure in part by diminishing the hypotensive influence of NTS catecholaminergic neurons projecting to the PVN. Sprague‐Dawley rats received bilateral PVN injections of AAV2 viral vectors expressing either green fluorescent protein (GFP) or BDNF and bilateral NTS injections of phosphate‐buffered saline (PBS) or anti‐dopamine‐β‐hydroxylase‐conjugated saporin (DSAP, 22ng in 100nL), a neurotoxin selective to noradrenaline‐ and adrenaline‐synthesizing neurons. Blood pressure was monitored with telemetry during baseline conditions and during acute restraint stress. Our results showed that BDNF treatment significantly increased resting daytime mean arterial pressure (MAP). MAP was 115±3 mmHg in the BDNF+PBS group compared with 96±2 mmHg in the GFP+PBS group (p&lt;0.01) 3 wks after injections. Lesioning NTS catecholaminergic neurons with DSAP also increased daytime MAP in GFP‐treated rats (GFP+DSAP: 109±3 mmHg, p&lt;0.05 vs GFP+PBS), but failed to induce further MAP increases in the BDNF group (BDNF+DSAP: 115±4 mmHg, p&gt;0.99 vs BDNF+PBS). Furthermore, BDNF overexpression reduced spontaneous baroreflex sensitivity in rats receiving PBS injections into the NTS, and DSAP reduced baroreflex sensitivity in GFP rats. However, DSAP failed to have an effect in the BDNF group. Heart rate was increased by BDNF overexpression, and in contrast to MAP, heart rate was elevated by DSAP both in GFP‐ and BDNF‐treated rats, although the tachycardic effect was reduced in the BDNF group. Restraint stress‐induced pressor responses were significantly elevated by DSAP in GFP rats, whereas the increase in heart rate was reduced. In contrast, DSAP had no effect on either of these parameters in the BDNF group. Finally, immunohistochemistry indicated that BDNF overexpression in the PVN upregulated tyrosine hydroxylase and dopamine‐β‐hydroxylase (DBH) expressions in the NTS, and elevated DBH immunoreactivity in the PVN. In summary, our findings indicate that increased BDNF expression in the PVN abolishes the hypotensive actions of NTS catecholaminergic neurons both under baseline conditions and during stress despite a significant upregulation of catecholamine biosynthesizing enzymes in the NTS. Heart rate is differentially regulated by NTS catecholaminergic neurons during baseline conditions and stress, but these actions are also reduced by elevated BDNF levels in the PVN.Support or Funding InformationSupported by AHA 11SDG7560022 and UVM start‐up funds.

Role of Alpha‐adrenergic Inputs to PVN in the Cardiovascular Responses to Acute Hypoxia

Previous studies indicate that catecholaminergic neurons in nucleus of solitary tract (NTS) contribute to intermittent hypoxia hypertension and that the hypothalamic paraventricular nucleus (PVN) plays a role in the cardiovascular response to hypoxia. The present study was designed to examine the role of alpha‐adrenergic receptors in PVN in the cardiovascular responses to acute hypoxia. Rats were anesthetized with chloralose and urethane and mean arterial pressure (MAP), heart rate (HR), renal sympathetic nerve activity (RSNA) and end tidal CO2 measured. The rat was placed in a stereotaxic frame for microinjections in PVN. After paralysis with gallamine, the rat was artificially ventilated with room air. Hypoxia was induced by ventilating the animal with 10% O2 for 30 seconds. When the parameters returned to the baseline, bilateral injections of vehicle or either α1 antagonist prazosin (500nmol) or α2 antagonist idazoxan (5nmol) were made in PVN and responses to hypoxia observed after 20 minutes. At the end of experiment brain was removed for localization of microinjection site. Hypoxia increased MAP with no change in HR or RSNA. The increase in MAP induced by systemic hypoxia was significantly blunted by bilateral administration of either prazosin (36 ± 3mmHg to 19 ± 4mmHg, n=11) or idazoxan (36 ± 2mmHg to 25 ± 5mmHg, n=10) in PVN. Changes in baseline blood pressure were not significant following prazosin (−2 ± 4mmHg) or idazoxan (0 ± 3mmHg). The results suggest that catecholaminergic inputs to PVN play a role in acute hypoxia induced increases in MAP.Support or Funding InformationSupported by PO1 HL88052

Optogenetic stimulation of PVN‐RVLM neurons increases sympathetic nerve activity in anesthetized rats

The hypothalamic paraventricular nucleus (PVN) sends direct projections to the rostral ventrolateral medulla (RVLM). The present study was conducted to examine the effect of optogenetic stimulation of PVN‐RVLM neurons on sympathetic nerve activity in anesthetized rats. Male rats anesthetized with pentobarbital received unilateral microinjection into the PVN with an adeno‐associated virus (AAV) vector that encodes the reporter protein palGFP (control rats, N=4) or the channel rhodopsin variant, ChIEF‐tdTomato (ChIEF rats, N=8). Two‐to‐five weeks later, in the rats anesthetized with a mixture of urethane and α‐chloralose, photostimulation (473 nm wavelength, 8–10 mW) was provided to the RVLM ipsilateral to the PVN into which AAV had been injected. In ChIEF rats, photostimulation of the RVLM at 40 Hz (5‐ms pulse duration) for 2 min significantly (P &lt; 0.05) evoked renal sympathoexcitatory (+126 ± 20% from baseline, means ± SE) and pressor (+13 ± 1 mmHg) responses while this stimulation had no effect in control rats. Moreover, immunofluorescence staining revealed abundant distribution of PVN‐derived, tdTomato‐labeled axons in the ipsilateral RVLM as compared with that in the contralateral RVLM. Confocal imaging further showed close association of tdTomato‐labeled axons with tyrosine hydroxylase‐positive neurons in the RVLM. These observations suggest that excitation of the PVN‐RVLM monosynaptic pathway elicits sympathoexcitation via, at least in part, stimulation of RVLM catecholaminergic neurons, potentially playing a role in sympathetic adjustments to stress such as exercise.Support or Funding InformationJSPS KAKENHI 15H05367 &amp; 16K15190 (SK); 16H05128 &amp; 15H05932 (KN).

Neurochemistry of neurons in the ventrolateral medulla activated by hypotension: Are the same neurons activated by glucoprivation?

Previous studies have demonstrated that a range of stimuli activate neurons, including catecholaminergic neurons, in the ventrolateral medulla. Not all catecholaminergic neurons are activated and other neurochemical content is largely unknown hence whether stimulus specific populations exist is unclear. Here we determine the neurochemistry (using in situ hybridization) of catecholaminergic and noncatecholaminergic neurons which express c-Fos immunoreactivity throughout the rostrocaudal extent of the ventrolateral medulla, in Sprague Dawley rats treated with hydralazine or saline. Distinct neuronal populations containing PPCART, PPPACAP, and PPNPY mRNAs, which were largely catecholaminergic, were activated by hydralazine but not saline. Both catecholaminergic and noncatecholaminergic neurons containing preprotachykinin and prepro-enkephalin (PPE) mRNAs were also activated, with the noncatecholaminergic population located in the rostral C1 region. Few GlyT2 neurons were activated. A subset of these data was then used to compare the neuronal populations activated by 2-deoxyglucose evoked glucoprivation (Brain Structure and Function (2015) 220:117). Hydralazine activated more neurons than 2-deoxyglucose but similar numbers of catecholaminergic neurons. Commonly activated populations expressing PPNPY and PPE mRNAs were defined. These likely include PPNPY expressing catecholaminergic neurons projecting to vasopressinergic and corticotrophin releasing factor neurons in the paraventricular nucleus, which when activated result in elevated plasma vasopressin and corticosterone. Stimulus specific neurons included noncatecholaminergic neurons and a few PPE positive catecholaminergic neuron but neurochemical codes were largely unidentified. Reasons for the lack of identification of stimulus specific neurons, readily detectable using electrophysiology in anaesthetized preparations and for which neural circuits can be defined, are discussed.

Depletion of rostral ventrolateral medullary catecholaminergic neurons impairs the hypoxic ventilatory response in conscious rats

The stimuli that commonly activate the catecholaminergic C1 neurons (nociception, hypotension, and hypoxia) also increase breathing. Pharmacogenetic evidence suggests that catecholaminergic neurons regulate breathing. Therefore, we evaluated whether the loss of C1 cells affects cardiorespiratory control during resting, hypoxic (8% O2) and hypercapnic (7% CO2) conditions. A bilateral injection of the immunotoxin anti-dopamine β-hydroxylase-saporin (anti-DβH-SAP; 2.4ng/100nl) or saline was performed in adult male Wistar rats (270–300g, N=5–8/group). Histology revealed a 60–75% loss of C1 neurons in anti-DβH-SAP-treated rats, but no significant changes or C1 cell loss was observed in sham-treated rats or those with off-target injection sites. Bilateral depletion of C1 neurons did not alter cardiorespiratory variables during rest and hypercapnia (7% CO2), but it did affect the response to hypoxia. Specifically, the increase in ventilation, the number of sighs, and the tachycardia were reduced, but unexpectedly, the mean arterial pressure increased during hypoxia (8% O2). The present study indicates that C1 neurons contribute to cardiorespiratory control during hypoxia rather than at rest or during hypercapnia.