Rat Lower Brainstem Research Articles

Distribution of catecholaminergic presympathetic-premotor neurons in the rat lower brainstem

We previously characterized the organization of presympathetic-premotor neurons (PSPMNs), which send descending poly-synaptic projections with collaterals to skeletal muscle and the adrenal gland. Such neurons may play a role in shaping integrated adaptive responses, and many of them were found within well-characterized regions of noradrenergic cell populations suggesting that some of the PSPMNs are catecholaminergic. To address this issue, we used retrograde trans-synaptic tract-tracing with attenuated pseudorabies virus (PRV) recombinants combined with multi-label immunofluorescence to identify PSPMNs expressing tyrosine hydroxylase (TH). Our findings indicate that TH-immunoreactive (ir) PSPMNs are present throughout the brainstem within multiple cell populations, including the A1, C1, C2, C3, A5 and A7 cell groups along with the locus coeruleus (LC) and the nucleus subcoeruleus (SubC). The largest numbers of TH-ir PSPMNs were located within the LC and SubC. Within SubC and the A7 cell group, about 70% of TH-ir neurons were PSPMNs, which was a significantly greater fraction of neurons than in the other brain regions we examined. These findings indicate that TH-ir neurons near the pontomesencephalic junction that are distributed across the LC, SubC, and the A7 may play a prominent role in somatomotor–sympathetic integration, and that the major functional role of the A7 and SubC noradrenergic cell groups maybe in the coordination of concomitant activation of somatomotor and sympathetic outflows. These neurons may participate in mediating homeostatic adaptations that require simultaneous activation of sympathetic and somatomotor nerves in the periphery.

Localization of Glucokinase-Like Immunoreactivity in the Rat Lower Brain Stem: For Possible Location of Brain Glucose-Sensing Mechanisms

Localization of Glucokinase-Like Immunoreactivity in the Rat Lower Brain Stem: For Possible Location of Brain Glucose-Sensing Mechanisms

Localization of glucokinase-like immunoreactivity in the rat lower brain stem: for possible location of brain glucose-sensing mechanisms.

Pancreatic glucokinase (GK) is considered an important element of the glucose-sensing unit in pancreatic beta-cells. It is possible that the brain uses similar glucose-sensing units, and we employed GK immunohistochemistry and confocal microscopy to examine the anatomical distribution of GK-like immunoreactivities in the rat brain. We found strong GK-like immunoreactivities in the ependymocytes, endothelial cells, and many serotonergic neurons. In the ependymocytes, the GK-like immunoreactivity was located in the cytoplasmic area, but not in the nucleus. The GK-positive ependymocytes were found to have glucose transporter-2 (GLUT2)-like immunoreactivities on the cilia. In addition, the ependymocytes had GLUT1-like immunoreactivity on the cilia and GLUT4-like immunoreactivity densely in the cytoplasmic area and slightly in the plasma membrane. In serotonergic neurons, GK-like immunoreactivity was found in the cytoplasm and their processes. The present results raise the possibility that these GK-like immunopositive cells comprise a part of a vast glucose-sensing mechanism in the brain.

The embryonic and post-natal expression of the nicotinic receptor alpha 3-subunit in rat lower brainstem.

The mRNA expression of the alpha 3 nicotinic acetylcholine receptor subunit in the rat lower brainstem is more widespread in the embryo and at birth than in the mature brain, but the major cell groups expressing alpha 3 are generally the same, with two exceptions. The transcript for the alpha 3-subunit is transiently expressed in the ventral cochlear nucleus (VCN). Expression was very high in the embryonic and early post-natal VCN, but was significantly decreased in the VCN late in the post-natal period (about post-natal day 15). Conversely, alpha 3 mRNA was not expressed in the motor trigeminal nucleus until about PN3.

The endothelin ETA receptor exists in the caudal solitary tract nucleus of the rat brain.

1. The receptor autoradiographic method done on the rat lower brain stem and cerebellum plus 125I-endothelin-1, BQ-123, an antagonist for the endothelin ETA receptor, and sarafotoxin S6c, an agonist for the ETB receptor, revealed minute amounts of the ETA receptor coexisting with the ETB receptor in the caudal solitary tract nucleus of the rat lower brain stem. 2. The ETB receptor is present predominantly in other parts of the lower brain stem. 3. Knowledge of the heterogeneous distribution of the central endothelin receptor subtypes aids in understanding the neurophysiology of endothelins.

Endothelin (ET) receptor binding sites and expression of ET mRNAs in the rat lower brainstem

Endothelin (ET) receptor binding sites and expression of ET mRNAs in the rat lower brainstem

Regional distribution of messenger RNA encoding the insulin-like growth factor type 2 receptor in the rat lower brainstem.

Distribution of messenger RNA (mRNA) encoding the insulin-like growth factor (IGF) type 2 receptor (IGF2R) is investigated in the rat lower brainstem by in situ hybridization histochemistry. Cells with IGF2R mRNA are distributed widely in a region-specific manner. It is expressed in: (1) motor nuclei such as the oculomotor nucleus, trochlear nucleus, motor trigeminal nucleus, abducens nucleus, facial nucleus, ambiguus nucleus, dorsal motor nucleus of vagus and hypoglossal nucleus; (2) several sensory-related nuclei like the mesencephalic trigeminal nucleus, ventral nucleus of the lateral lemniscus, lateral and spinal vestibular nuclei, ventral and dorsal cochlear nuclei and nucleus of the trapezoid body; and (3) other regions such as the red nucleus, dorsal raphe nucleus, pontine nuclei, three cerebellar nuclei (medial, interposed and lateral), Purkinje cells, cells in the granular layer of the cerebellum, locus coeruleus, several areas of the reticular nucleus and area postrema.

Altered L-DOPA systems for blood pressure regulation in the lower brainstem of spontaneously hypertensive rats.

Recent findings have enhanced our understanding of the roles played by the L-DOPA system in the baroreceptor reflex and in blood pressure regulation in the lower brainstem. L-DOPA is probably a neurotransmitter of primary baroreceptor afferents terminating in depressor sites of the nucleus tractus solitarii (NTS). It also seems to be a neurotransmitter in depressor sites of the caudal ventrolateral medulla (CVLM) and in pressor sites of the rostral ventrolateral medulla (RVLM) of normotensive Wistar rats. We have explored whether or not presynaptic and postsynaptic functions of the L-DOPA system in these areas are altered to maintain hypertension in adult spontaneously hypertensive rats, as compared with age-matched Wistar Kyoto rats. In this review article, we survey the roles of the L-DOPA system in the baroreceptor reflex and in blood pressure regulation in the rat lower brainstem.

Cold exposure elevates thyrotropin-releasing hormone gene expression in medullary raphe nuclei: Relationship with vagally mediated gastric erosions

The stimulation of thyrotropin release by cold is associated with an increase in thyrotropin-releasing hormone gene expression in the paraventricular nucleus of the hypothalamus. Cold exposure also stimulates autonomic outflow to viscera. There is evidence that caudal raphe nuclei are involved in autonomic regulation through thyrotropin-releasing hormone projections to the dorsal vagal complex and spinal cord. To determine whether cold modulates thyrotropin-releasing hormone gene expression in the caudal raphe nuclei, the effect of cold exposure on thyrotropin-releasing hormone messenger RNA levels in the rat lower brainstem was examined by quantitative Northern blot analysis and thyrotropin-releasing hormone messenger RNA was localized by in situ hybridization. The gastric responses to cold exposure were also assessed in sham or vagotomized rats with pylorus ligation. Thyrotropin-releasing hormone messenger RNA signal was detected in the RNA extracted from the medulla and hypothalamus but not from the amygdala, periaqueductal gray or cerebellum. Cold exposure (4°C) for 1 or 3 h increased thyrotropin-releasing hormone messenger RNA levels in the medulla by77 ± 37 and142 ± 39% respectively. In situ hybridization histochemistry showed that the increase in silver grain density occurred exclusively in the raphe pallidus and raphe obscurus. Exposure to cold stress for 2 h stimulated gastric acid secretion and resulted in gastric lesion formation in sham but not vagotomized rats. There are established thyrotropin-releasing hormone projections from the raphe pallidus and obscurus to the dorsal vagal complex. Thyrotropin-releasing hormone is known to exert an excitatory effect on neurons of the dorsal motor nucleus of the vagus, leading to vagally mediated stimulation of gastric function and erosions. The increased thyrotropin-releasing hormone gene expression in the raphe pallidus and obscurus induced by cold supports an important role of endogenous thyrotropin-releasing hormone contained in caudal raphe nuclei-dorsal vagal complex pathways in vagally mediated changes in gastric function induced by cold.

Alpha 2A-adrenergic receptors are present in lower brainstem catecholaminergic and serotonergic neurons innervating spinal cord

A subtype-specific polyclonal antibody was used for the immunohistochemical detection of alpha 2A-adrenergic receptors (α 2A-ARs) in the rat lower brainstem (medulla and pons). Using dual-label fluorescence histochemistry, punctate α 2A-AR-like immunoreactivity (α 2A-AR-LIR) was identified in noradrenergic, adrenergic, and serotonergic neurons of the pontomedullary region. Confocal microscopic examination of material simulataneously labeled for TH-LIR and α 2A-LIR revealed that the clusters of α 2A-LIR were located intracellularly. Lower medullary neurons with spinal protections to segment T 3 were retrogradely labeled using FITC-conjugated microbeads and the material was processed for simulatneous detection of α 2A-LIR and either TH-LIR or 5-HT-LIR. Using this triple-label approach, we found that virtually all medullary serotonergic cells (raphe pallidus, raphe obscurus and parapyramidal area) including those with identified spinal projections contain α 2A-AR-LIR. In contrast, fewer than 10% of dorsal raphe serotonergic cells examined for comparison were immunoreactive. The triple labeling approach also indicated that more than 95% of the TH-immunoreactive cells of the dorsal and ventrolateral medulla, including those with demonstrable spinal projections (A5 noradrenergic and C1/C3 adrenergic) had detectable amounts of α 2A-AR-LIR. The presence of α 2A-ARs in a large fraction of bulbospinal pre-sympathetic neurons (noradrenergic A5, adrenergic C1 and C3 and serotonergic raphe cells) could explain the powerful and relatively selective effect of clonidine and other centrally acting α 2A-AR agonists on sympathetic efferent activity and hypertension.

Isolation and characterization of periaxolemmal and axolemmal enriched membrane fractions from the rat central nervous system.

In this report, we describe the fractionation of crude axolemmal fractions from rat lower brainstem into subfractions enriched in markers for either periaxolemmal myelin or axolemma. These subfractions were isolated on density gradients as bands layering on 0.8M and 1.0M sucrose. Both subfractions consisted of unilamellar vesicles. Relative to myelin purified from the same starting material, the 0.8M subfraction was enriched in MAG, CNPase, carbonic anhydrase and Na+, K+ ATPase but was extremely low in PLP and MBP. In addition, this fraction exhibited a protein profile distinct from myelin. The 1.0M fraction was also highly enriched in Na+, K+ ATPase and had an overall composition similar to the 0.8M subfraction. However, it differed from the 0.8M subfraction by being low in MAG, CNPase, and carbonic anhydrase, but enriched in voltage-dependent Na+ channel, axon-specific fodrin, and MAP-1B. Based on these characteristics we concluded that the 0.8M and 1.0M subfractions were highly enriched in periaxolemmal myelin and axolemmal membrane, respectively. Plasmolipin10 was unique with equally high levels in myelin and in the 0.8M and 1.0M subfractions. Both subfractions were enriched, relative to myelin, in the alpha subunit of the GTP binding protein, Go, and the alpha subunit common to all G proteins, GA/1. Electrophysiology with membrane subfractions fused to lipid bilayers showed that both membranes contained sets of K+ and Cl- channels, which based on channel sizes and open times, are largely distinct from one another.

Postnatal development of preproenkephalin mRNA containing neurons in the rat lower brainstem

Postnatal developmental changes of preproenkephalin (PPE) gene expression in rat brainstem neurons were studied by in situ hybridization histochemistry. On the basis of PPE mRNA expression, brainstem neurons were categorized into three types: 1) type I neurons were characterized by constant or increasing expression of PPE mRNA during postnatal development; 2) type II neurons started to express PPE mRNA several days after birth and continued to do so thereafter; and 3) type III neurons showed transient expression of PPE mRNA or stopped expressing the mRNA during early postnatal development. Type I PPE neurons were observed in diverse brainstem structures including the mesencephalic and pontine central gray matter, various reticular and raphe nuclei, the ventral tegmental area of Tsai, the interpeduncular nucleus, the nucleus of the brachium of the inferior colliculus, the ventral and dorsal tegmental nuclei of Gudden, the sphenoid nucleus, the laterodorsal tegmental nucleus, Barrington's nucleus, the parabrachial region, the lateral lemniscus and its related nuclei, the trapezoid nucleus, the rostral and ventromedial periolivary nuclei, the mesencephalic trigeminal and principal sensory trigeminal nuclei, the locus coeruleus, the subcoeruleus nucleus, the medial and spinal vestibular nuclei, the dorsal and ventral cochlear nuclei, the medial and lateral cerebellar nuclei, the Roller nucleus, and the intermedius nucleus of the medulla. Type II PPE neurons were found in the superior colliculus, the inferior colliculus, the central part of the dorsal tegmental nucleus, and as Golgi neurons in the granular layer of the cerebellum. Type III PPE neurons were located in the substantia nigra, the red nucleus, the superior olive, the motor trigeminal nucleus, the facial nucleus, the inferior olive, the dorsal motor nucleus of the vagus, and the hypoglossal nucleus. Such region-specific expression of the PPE gene during postnatal ontogeny suggests that rat brainstem PPE neurons may be involved in a variety of developmental events, such as cell proliferation, differentiation, and migration.

Distribution and postnatal development of somatostatinergic neuron system in rat lower brainstem—In situ hybridization histochemistry

Distribution and postnatal development of somatostatinergic neuron system in rat lower brainstem—In situ hybridization histochemistry

Immunocytochemical distribution of met-enkephalin-Arg6-Gly7-Leu8 in the rat lower brainstem.

The distribution of methionine-enkephalin-Arg6-Gly7-Leu8, a unique peptide derived from proenkephalin A in the rat brainstem, was studied immunocytochemically by using a highly specific antiserum to this octapeptide sequence. Immunoreactive perikarya with various shapes and sizes were detected in many regions of the rat brainstem. Dense accumulation of immunoreactive perikarya and fibers was seen in the nuclei associated with special sensory and visceral functions, such as the interpeduncular nucleus, the parabrachial nucleus, the nucleus of the solitary tract, and the nucleus of the spinal tract of the trigeminal nerve. Clusters of methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactive perikarya and fibers were observed in certain areas considered to play a role in nociception and analgesia, such as the central gray of the midbrain central gray and the raphe magnus nucleus. Some methionine-enkephalin-Arg6-Gly7-Leu8-like immunoreactive perikarya were distributed in the lateral reticular nucleus, the nucleus of the solitary tract, and the raphe magnus nucleus, where monoaminergic neurons were also detected. In addition to the previously reported enkephalinergic cells, we found many methionine-enkephalin-Arg6-Gly7-Leu8 containing neurons; the rostral and caudal linear nucleus of raphe, the median raphe nucleus, entire length of the raphe magnus nucleus, the medial longitudinal fasciculus, the cuneate nucleus, the external cuneate nucleus, the gracile nucleus, and the area postrema. The wide distribution of this octapeptide-like immunoreactivity reflected neurons expressing the preproenkephalin A gene distributed more widely than previously reported and that innervated many regions.

Quantitative autoradiographic localization of neuropeptide Y receptors in the rat lower brainstem

Using quantitative autoradiography, we have characterized the binding of 125I-Bolton-Hunter coupled neuropeptide Y ([ 125I]NPY) and observed the localization of 125I-NPY receptors in the rat lower brainstem. [ 125I]NPY bound to the receptors in a specific, saturable and reversible manner with high affinity. The binding was blocked only by unlabeled NPY but not by NPY-related peptides i.e. peptide YY, pancreatic polypeptide (avian and human), nor by neurotensin. [ 125I]NPY receptors were revealed to be coupled to the guanosine triphosphate (GTP)-binding protein. Regional distribution study showed that [ 125I]NPY has a distinctive pattern of distribution in the rat lower brainstem, being particularly concentrated in the area postrema and the medial subnucleus of the nucleus tractus solitarii. These results suggest that such NPY receptors have an important role in cardiovascular regulation.

O-219 - Autoradiographic localization of 125I-NPY receptors in the rat lower brainstem

O-219 - Autoradiographic localization of 125I-NPY receptors in the rat lower brainstem

Distribution of dopamine, noradrenaline and adrenaline in coronal sections of the rat lower brainstem

The concentration of the three major catecholamines (CAs) were determined in 500 μm thick coronal sections of the rat medulla oblongata disserted into microcubes. Noradrenaline (NA) concentrations were always found much higher than the levels of the two other CAs in the same microcube. The highest concentrations of the three CAs were found in the dorso-medial region of the lower brainstem, more exactly in the more caudally located parts of the nucleus tractus solitarii (NTS). In the ventro-lateral region, the CA concentrations were lower and, except for adrenaline (A), did not exhibit any substantial change in their rostro-caudal distribution. Conversely, in the dorso-medial region, there was a clear rostro-caudal pattern of distribution of the three CAs. This distribution was similar for the three amines, since only a small difference (about 500 μm) was found between the maximal NA and A concentrations. Since the three CAs are present in highest concentrations within the same dorso-medial or ventro-lateral groups of microcubes, a microdissection of these two areas seems suitable to study simultaneously the metabolism of the three CAs in the rat lower brainstem. These data also suggest a microdissection procedure to study A metabolism within the C2–C3 A cell bodies and within a region more caudally located, rich in A terminals.

Distribution of neuropeptide Y in the lower brainstem: an immunohistochemical analysis

The distribution of neuropeptide Y (NPY)-like immunoreactive (NPYI) structures in the rat lower brainstem was examined by means of indirect immunofluorescence or peroxidase-antiperoxidase methods. In addition to the well known immunoreactive NPYI containing cell groups, the present study demonstrated a much wider distribution of immunoreactive NPYI cells in the lower brainstem, i.e. substantia nigra pars lateralis, interpeduncular nucleus, inferior colliculus, nucleus cuneiformis, dorsal tegmental nucleus of Gudden, nucleus laterodorsalis tegmenti, nucleus vestibularis medialis, nucleus vestibularis inferioris, in the fasciculus longitudinalis medialis and nucleus parvocellularis compacta. We also demonstrated an extensive network of NPYI fibers in various areas of the lower brainstem including the auditory system, viscerosensory system, visceromotor system, raphe nuclei, reticular formation, parabrachial area, locus coeruleus and interpeduncular nucleus, etc. The possible importance of NPY is briefly discussed.

Localization of chick retinal 24,000 dalton protein (visinin)-like immunoreactivity in the rat lower brain stem

The distribution of visinin, a 24,000 dalton peptide, in the lower brain stem of the rat was examined by means of an indirect immunofluorescent method. Visinin-immunoreactive structures were found to be unevenly distributed only in the neuronal elements. The following neuronal systems were strongly labeled by the antiserum; the Purkinje cell system, mammillotegmental system, habenulointerpeduncular system, the second layer of the superior colliculus, ventral tegmental area, substantia nigra pars lateralis, area medial to the medial geniculate body, parabrachial area, dorsal and ventral nuclei of the lateral lemniscus, pontine reticular formation just medial to the trigeminal principal nucleus, superior olivary nucleus, solitarii nucleus, external layer of the inferior colliculus and spinal trigeminal nucleus. The densities of the labeled fibers in these areas paralleled those of the labeled cells. In addition, highly dense visinin-immunoreactive fiber plexuses were seen in the zona compacta of the substantia nigra, lateral portion of the interpeduncular nucleus, ventral tegmental nucleus of Gudden and vestibular nucleus.

Increased feeding in response to decreased glucose utilization in the rat and monkey.

SMITH, GERARD P., AND ALAN N. EPSTEIN. Increased feeding in response to decreased glucose utili<ation ilz the rat and monkey. Am. J Physiol. 217(4) : 1083-1087. 1969.-The glucostatic hypothesis for the control of food intake predicts that decreased glucose utilization leads to increased food intake. Since the glucose analogue, 2-deoxy-D-glucose (Z-DG), produces decreased intracellular glucose utilization of most tissues and particularly of brain, the injection of 2-DG into 5 monkeys and 28 rats provided an explicit test of the predicted relationship. Both monkeys and rats ate more after 2-DG. The most effective systemic doses of Z-DG were 300 mg/kg (iv) in monkeys and 750 mg/kg (ip) in rats. Larger doses ( < 1,000 mg/kg) produced drowsiness, stupor, ataxia, or retching, but not convulsions. When separate groups of rats were tested with either insulin (6 U Iletin) or 2-DG (750 mg/kg), they increased their mean food intake to approximately the same amount, but the rats treated with Z-DG ate sooner. Shorter latency of feeding with 2-DG is interpreted as resulting from the fact that 2-DG produces decreased glucose utilization of brain directly, whereas insulin reduces cerebral glucose utilization indirectly as a consequence of hypoglycemia. Since Z-DG produced marked hyperglycemia during the period of increased feeding, these results show that the abrupt onset of decreased glucose utilization, not hypoglycemia, is a sufficient condition for feeding in mammals.