SEARCH · Search PubMed

Results for “TMB”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

SourceResearch connectionFrom yearThrough year

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 919 records · Page 51Linked to original sources

PubMed1996

Simplified PCR-SSO procedure for DQA and DQB typing. Simultaneous hybridization of two probes to one membrane.

To simplify DQA and DQB oligotyping, we applied our improved PCR-SSO procedure for DR typing. We used 12 oligonucleotide probes for DQA typing and 18 for DQB typing. Oligonucleotide probes that require the same hybridization and stringent washing conditions were selected as pairs for simultaneous hybridization to a dot-blot membrane containing various DNA samples. One probe of each pair was labeled with digoxigenin and the other with biotin. After hybridization, the dot-blot membranes were incubated with a mixture of conjugates. Specific binding of the corresponding DNA probes was visualized on an X-ray film using a chemiluminescent substrate (CSPD) and by staining using a chromogenic substrate (TMB). This approach, previously employed for DR typing, is also suitable for DQA and DQB oligotyping and significantly reduces the labor inherent in PCR-SSO typing.

Genotype↗

PubMed1985

Adrenergic stimulation of prostacyclin production in the rat tail artery. II. The role of calcium.

This investigation studied the role of extracellular and intracellular calcium in the stimulation of prostacyclin (PGI2) production by norepinephrine (NE) in the rat tail artery in vitro. Incubation of the artery in zero calcium medium inhibited the response to NE as well as the response to the calcium ionophore A23187. Increasing the extracellular calcium concentration had no effect on basal or NE-stimulated PGI2 (measured by radioimmunoassay of the stable metabolite, 6-keto-PGF1 alpha). Several different calcium entry blocking drugs had a partial inhibitory effect on the NE response, but it was noted that the dihydropyridine derivatives, nifedipine and nimodipine, also decreased the stimulatory effect of added exogenous arachidonic acid (AA). This effect on AA conversion was also seen with W-7, a calcium-calmodulin inhibitor, and with TMB-8, an inhibitor of intracellular calcium mobilization. However, two phenothiazine calmodulin inhibitors, trifluoperazine and pimozide, did not affect AA conversion and were effective in reducing the stimulatory response to NE. Trifluoperazine, particularly at a high dose of 10(-4) M, had an independent stimulatory effect on PGI2 production in this tissue. The data suggests that many of the antagonist probes of extracellular-intracellular calcium metabolism may have variable effects on more than one point in the prostaglandin synthetic pathway. Nevertheless, the results are consistent with the hypothesis that NE stimulates PGI2 in the rat tail artery by activation of a calcium-calmodulin dependent phospholipase and that at least some of the calcium is of extracellular origin.

6-Ketoprostaglandin F1 alpha↗

PubMed1986

Mobilization of intracellular calcium as a mechanism for the positive inotropic effect by prostaglandin F2 alpha in isolated guinea pig atria.

The mechanism of the positive inotropic effect of prostaglandin F2 alpha (PGF2 alpha) was investigated using spontaneously beating isolated guinea pig atria. PGF2 alpha increased the contractile force in a concentration dependent fashion in a range between 10(-7)M to 10(-5)M. The maximal response to PGF2 alpha was obtained within 5 min and continued for at least 20 min. This positive inotropic effect of PGF2 alpha was not affected by verapamil, but was inhibited by procaine or TMB-8, potent inhibitors of Ca2+ release from intracellular Ca2+ compartments. Increase in extracellular Ca2+ concentration failed to provoke a significant positive inotropic effect by PGF2 alpha, suggesting that this response is relatively independent of extracellular Ca2+. These results suggest that the PGF2 alpha-mediated positive inotropic effect is produced by a mobilization of intracellular Ca2+.

Animals↗

PubMed1992

Effect of 2-acetylaminofluorene on intracellular free Ca2+ in isolated rat hepatocytes.

The effect of 2-acetylaminofluorene (2-AAF) on the intracellular free Ca2+ ([Ca2+]i) and viability of isolated rat hepatocytes has been investigated using the fluorescent probes quin 2 and propidium iodide respectively. At the highest concentration tested (224 microM), 2-AAF produces an elevation of [Ca2+]i which shows a biphasic profile. A small initial increase is observed during the first 5 min; this is followed by a considerable rise which reaches up to 2.5 times the control value at 15 min. These changes in intracellular calcium are not accompanied by detectable alterations in cell viability. In order to determine the mechanisms by which this effect of 2-AAF takes place, three calcium antagonists, namely verapamil, TMB-8 (8-(diethylamino)-octyl-3,4,5-trimethoxybenzoate) and ruthenium red (RuR), have been used. The results suggest that the first phase is dependent upon internal Ca2+ store mobilization, while the second phase seems to be related to Ca2+ entry from the extracellular space. The data obtained with RuR further indicate that mitochondria may be involved in the perturbation of calcium homeostasis caused by 2-AAF. In addition, in the experiments involving antagonists, no consistent pattern emerges that suggests a close relationship between intracellular Ca2+ levels and cell viability. The present study provides further information on the mechanisms by which these well-known hepatotoxin 2-AAF may interact with liver cells. It also shows that when these cells are exposed to a toxin, short-term changes in [Ca2+]i may not be accompanied by loss of cell viability, and conversely, that changes in cell viability may occur without alterations in [Ca2+]i.

2-Acetylaminofluorene↗

PubMed1988

Angiotensin II-stimulated changes in calcium metabolism in cultured glomerulosa cells.

Studies were performed to evaluate the relationships between the effects of angiotensin II on calcium metabolism and cytosolic free calcium concentration in primary monolayer cultures of bovine adrenal glomerulosa cells. As noted previously (Kramer (1988) Am. J. Physiol. (in press], angiotensin II produced rapid dose-dependent increases in cytosolic calcium characterized by both an initial transient component and a secondary sustained component. In the absence of extracellular calcium, angiotensin II produced an initial increase in cytosolic calcium comparable to that produced in the presence of calcium, but failed to maintain a sustained calcium signal. The initial, angiotensin-stimulated increase in cytosolic calcium was inhibited by dantrolene and 8-(N,N-diethylamino)-octyl-3,4,5-trimethoxybenzoate (TMB-8) in a concentration-dependent fashion. The onset of the angiotensin-stimulated calcium signal was accompanied by a dose-dependent increase in the rate of calcium efflux that achieved a maximum within 2-3 min and then declined to a level 2.5-3 times that from control cells. The initial rate of calcium influx was also increased about 2.5-fold by angiotensin II, an effect that was only apparent in cells that had been treated with the peptide for at least 5 min. These results indicate that the calcium signal produced by angiotensin II is initiated by the rapid mobilization of calcium from an intracellular site(s) and sustained by the continued uptake of extracellular calcium. Moreover, the kinetics of the calcium signal as well as the final, sustained calcium concentration achieved reflect the balance between intracellular calcium release, calcium influx and calcium efflux.

Angiotensin II↗

PubMed1992

Characterization of the gonadotrophin-releasing hormone calcium response in single alpha T3-1 pituitary gonadotroph cells.

Intracellular calcium ([Ca2+]i) was measured in single immortalized gonadotroph alpha T3-1 cells using dual wavelength fluorescence microscopy combined with dynamic video imaging. Gonadotrophin-releasing hormone (GnRH, 10(-8) M) produced a biphasic rise in [Ca2+]i which could be abolished by a GnRH antagonist. The initial calcium transient was complete within seconds while the smaller secondary plateau phase lasted several minutes. The calcium spike was reduced by nifedipine (10(-6) M), a calcium channel blocker, and thapsigargin (10(-6) M) which inhibits inositol 1,4,5-trisphosphate (IP3) mediated release of [Ca2+]i but abolished by the intracellular calcium antagonist TMB-8 (10(-6) M). The secondary phase was reduced following pretreatment with either nifedipine or the protein kinase C (PKC) antagonist, H-7 (10(-6) M). The PKC agonist PMA (phorbol 12-myristate 13-acetate, 10(-6) M) produced a small rise in basal [Ca2+]i and abolished the GnRH calcium response. The initial calcium response to GnRH therefore involves both an IP3-mediated rise in cytosolic calcium due to the release from intracellular stores and an influx of extracellular calcium through second messenger-operated calcium channels. In contrast the secondary calcium response mainly involves the influx of extracellular calcium through PKC-activated calcium channels.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1981

The median subcallosal fasciculus in the monkey: a unique prefrontal corticostriate and corticocortical pathway revealed by anterogradely transported horseradish peroxidase.

Using the remarkable capability of tetramethylbenzidine (TMB) neurohistochemistry to delinate anterogradely transported horseradish peroxidase (HRP) following HRP gel implants in the prearcuate prefrontal cortex of macaque and cebus monkeys, this study revealed bilateral projections to the caudate nucleus and inferior parietal lobule that followed the median subcallosal fasciculus, a rostrocaudally running, midline trajectory located between the body of the corpus callosum and the fornix. This seemingly aberrant fascicle runs through the septum pellucidum from the genu to the splenium of the corpus callosum, and gives off fibers bilaterally to the body of the caudate nucleus and inferior parietal lobule. The description of this unique pathway re-emphasizes the close relationship that exists between the granular frontal and inferior parietal cortices, and the striatum, and explains earlier observations based upon silver degeneration and autoradiographic studies, of prefrontal corticofugal projections to the septum and fornix.

Animals↗

PubMed1987

Central brainstem projections of the superior vagal ganglion of the cat.

The central projections of the cat superior vagal (jugular) ganglion (SVG) cells were determined using anterograde transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). WGA-HRP was injected into the SVG of 5 cats, 3 of which had the vagal nerve sectioned distal to the jugular ganglion several days previously. Following 72 h, the animals were sacrificed and the brainstem and rostral cervical spinal cord sectioned and processed with tetramethylbenzidine (TMB) and examined for anterograde transport. The vagal root afferent fibers entered the brainstem at the level of the caudal facial motor nucleus and bifurcated into descending tracts. Terminal label was identified in: the dorsal lateral subnucleus of the nucleus tractus solitarius (NTS); nucleus interpolaris and nucleus caudalis of spinal V; the ventrolateral aspect of the cuneate nucleus and; the superficial laminae (I, II) of the rostral C1-dorsal horn. To our knowledge this is the first report describing central vagal afferent terminations following injection of current axonal tract tracing substances into the isolated superior vagal ganglion. The projection of jugular ganglion cells to a region of the NTS associated with cardiovascular function is of particular interest, and may be related to ganglion cells known to innervate the cerebral vasculature and meninges.

Animals↗

PubMed1988

Muscarinic stimulation of guinea pig adrenal chromaffin cells stimulates catecholamine secretion without significant increase in Ca2+ uptake.

In isolated guinea pig adrenal chromaffin cells, not only nicotine, but also muscarine stimulated catecholamine (CA) secretion, the stimulation by muscarine being the greater. The secretions of CA by muscarine and nicotine were both dependent on the presence of Ca2+ in the medium, but only the latter was associated with a rapid increase in 45Ca2+ uptake. Experiments with the fluorescent Ca2+ indicator quin 2, showed that muscarine caused an increase in cytoplasmic free Ca2+ concentration [( Ca2+]i). Moreover, the intracellular Ca2+ antagonist 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) inhibited both CA secretion and increase in [Ca2+]i induced by muscarine. These results indicate that in isolated guinea pig adrenal chromaffin cells, nicotine stimulated CA secretion by increasing Ca2+ uptake by the cells, whereas muscarine stimulated CA secretion by mobilizing Ca2+ from the intracellular pool.

Adrenal Medulla↗

PubMed1989

Triwaglerin: a potent platelet aggregation inducer purified from Trimeresurus wagleri snake venom.

Trimeresurus wagleri venom is the most potent inducer of platelet aggregation among the seven Trimeresurus snake venoms tested. By means of CM-Sephadex C-50 column chromatography, T. wagleri venom was separated into 19 fractions. Fraction XVI possessed the strongest aggregating activity and was further purified by Sephadex G-75 and on heparin-agarose columns, and finally Triwaglerin, with a molecular weight of 68000, was obtained. Its aggregating and ATP-releasing activity was dose-dependent and 10-times more potent than the crude venom. Triwaglerin was devoid of any of the enzymatic activities possessed by the crude venom. Triwaglerin-induced aggregation was not affected by indomethacin, creatine phosphate/creatine phosphokinase (CP/CPK), platelet-activating factor (PAF) antagonists, verapamil or heparin, but was inhibited completely by mepacrine, imipramine and forskolin and markedly by tetracaine and sodium nitroprusside. Thromboxane B2 formation caused by Triwaglerin was suppressed by mepacrine, imipramine and indomethacin. R59022 and TMB-8 caused a synergistic inhibitory effect against Triwaglerin-induced aggregation. These data suggest that Triwaglerin activates platelets in a unique action which is independent of formation of thromboxane A2 and PAF, or release of ADP.

Animals↗

PubMed1990

Evidence for the mechanism of the inhibitory action of jatrophone in the isolated rat uterine muscle.

1. Jatrophone (JAT), a diterpene isolated from the plant Jatropha elliptica (1-300 microM), caused a concentration-dependent relaxation effect against acetylcholine (Ach)-oxytocin (Ot)- and KCl-induced uterine sustained contraction. The relative potency order was: Ach greater than Ot greater than KCl. 2. The relaxant effect of JAT was not modified by phorbol ester, forskolin, MIX, TMB-8 and W-7. The increase concentration of calcium (0.2-2 mM) in the medium did not reverse the inhibitory effect caused by JAT. 3. Pre-incubation of the preparations with JAT (16-32 microM) for 20 min, caused a concentration-dependent inhibition of KCl-induced contractile response. At 30 microM, JAT inhibited in an apparently non-competitive manner CaCl2-induced contraction in K+-depolarized preparations. High concentrations of JAT (100 microM) also caused a time-dependent relaxation in CaCl2-induced sustained uterine contraction (T1/2 = approx. 15 min). 4. JAT (30 microM) inhibited the dihydropyridine calcium channel agonist Bay K 8644-induced uterine contraction in an apparently non-competitive fashion, while verapamil (0.1 microM) caused an rightward displacement of Bay K 8644 contraction and marked inhibition of the maximal response.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1991

Organic Ca(2+)-antagonist-resistant response to FMRF-NH2 on the molluscan smooth muscle.

1. FMRF-amide (10(-7)-10(-5) M) contracted molluscan anterior byssus retractor muscle in a concentration-dependent fashion. 2. The concentration-response curve of FMRF-amide was shifted rightward by an analogue of FMRF-amide, FMRf-amide ([D-Phe4]FMRF-amide, putative FMRF-amide receptor antagonist) in a parallel manner (pA2 = 4.87 +/- 0.04). 3. Although a contractile response to KCl was reduced by the organic Ca2+ antagonists (verapamil, diltiazem and high concentration of nifedipine and nicardipine). FMRF-amide-induced contraction was not markedly reduced by them. 4. In the Ca(2+)-free medium, FMRF-amide-induced contraction was diminished. The response was also reduced by TMB-8 (10(-4) M), suggesting that FMRF-amide-induced contraction might be partly dependent on intracellular Ca2+. 5. An inorganic Ca(2+)-antagonist, MnCl2, markedly reduced the FMRF-amide- and KCl-induced contraction. The results show that FMRF-amide-induced contraction might be dependent on extracellular Ca2+. 6. These findings suggest that FMRF-amide-induced contraction might be mediated through an action on FMRF-amide receptors and not through the activation of organic Ca2+ antagonist-sensitive Ca2+ channels.

Animals↗

PubMed1992

Mediators involved in the rat uterus contraction in calcium-free solution.

1. The effect of (Na+ + K+)-ATPase inhibitor ouabain (10(-5)-3 x 10(-4) M), and the (Ca2+ + Mg2+)-ATPase inhibitors vanadate (6 x 10(-6)-6 x 10(-4) M), oxytocin (2 x 10(-9)-4 x 10(-8) M, and prostaglandin F2 alpha (PGF2 alpha, 10(-7)-6 x 10(-6) M) were assayed on rat uterus incubated in Ca-free medium. 2. Vanadate, oxytocin and PGF2 alpha, but not ouabain, induced contractions in a dose-dependent way (ED50: 7.5 +/- 0.03 x 10(-5) M; 6.5 +/- 0.064 x 10(-9) M and 3.8 +/- 0.085 x 10(-7) M). 3. Vanadate (3 x 10(-4) M) and oxytocin (OT, 10 mU/ml = 2 x 10(-8) M)-induced tonic contraction were not modified by nifedipine (10(-10)-10(-6) M), monensin (10(-5)-3 x 10(-4) M) or amiloride (10(-5)-10(-3) M). 4. The intracellular calcium release inhibitors TMB-8 (10(-6)-10(-4) M) and dantrolene (3 x 10(-6)-10(-4) M), and the prostaglandin release inhibitor indomethacin (3 x 10(-8)-6 x 10(-5) M) relaxed the vanadate and OT-induced tonic contractions. 5. The calmodulin inhibitors trifluoperazine (3 x 10(-5)-3 x 10(-4) M), bepridil (10(-8)-3 x 10(-4) M), calmidazolium (10(-7)-10(-4) M) and W-7 (10(-7)-10(-5) M) also relaxed the vanadate and OT-induced tonic contractions. 6. Our results suggest that oxytocin and vanadate-induced contractions on rat uterus in Ca-free medium could be produced by release of prostaglandins and intracellular calcium, and mediated by calmodulin.

Animals↗

PubMed1993

Contractile responses and calcium movements induced by alpha 1-adrenoceptor stimulant, norepinephrine, in rabbit iris dilator muscle.

1. The mechanisms involved in contraction of rabbit iris dilator muscle induced by norepinephrine (NE) were studied. 2. The concentration-response curve of NE was not influenced by Ca2+ blockers in the normal physiological saline solution (PSS) and removal of Ca2+ from PSS. 3. In 0.01 mM EGTA containing Ca(2+)-free PSS, the NE-induced contraction was phasic, which was suppressed by TMB-8, cyclopiazonic acid, ionomycin and A23187 but still partly remained. 4. In 2 mM EGTA containing Ca(2+)-free PSS, NE increased the intracellular Ca2+ ([Ca2+]i) and muscle tension. Ryanodine abolished the increase in [Ca2+]i induced by NE but slightly inhibited the tension. 5. These results suggest that the NE-induced contraction of rabbit iris dilator in normal PSS is mainly due to the increase in the release of intracellularly sequestered Ca2+ and partly due to the Ca(2+)-independent processes.

Adrenergic alpha-Agonists↗

PubMed1991

Characterization of platelet-activating factor-induced elevation of cytosolic free-calcium level in neurohybrid NCB-20 cells.

The effect of platelet-activating factor on the intracellular cytosolic level of free calcium ([Ca2+]i) was studied in neurohybrid NCB-20 cells. In fura-2-loaded NCB-20 cells, platelet-activating factor induced an immediate and concentration-dependent increase in [Ca2+]i with a maximum increase of 334 +/- 27 nM above a basal value of 147 +/- 6 nM (n = 40). Platelet-activating factor-induced [Ca2+]i mobilization was inhibited by the platelet-activating factor antagonists BN 50739, WEB 2086, SRI 63-441 and BN 52021 in a dose-dependent manner with IC50 values of 12, 38, 897 and 45000 nM, respectively. The calcium-channel blockers nifedipine (10 microM) and diltiazem (10 microM) had no effect on the platelet-activating factor-induced increase in [Ca2+]i; however, extracellular Ca(2+)-depletion caused a 63.6 +/- 4.7% reduction of platelet-activating factor-induced increase in [Ca2+]i (n = 5, P less than 0.001). The remaining 36% contributed from intracellular sources was completely inhibited by 10 microM of 8-(N,N-diethylamine)octyl 3,4,5-trimethoxytenzoate hydrochloride (TMB-8). NCB-20 cells exhibited homologous desensitization to sequential addition of platelet-activating factor, but no heterologous desensitization between platelet-activating factor and bradykinin or ATP was observed. These data suggest that activation of the neuronal platelet-activating factor receptor results in an increase in [Ca2+]i primarily via a receptor-operated rather than a voltage-dependent calcium-channel and to a lesser extent from intracellular Ca2+ release. Our findings may contribute to an understanding of the mechanism of platelet-activating factor actions on neuronal cells.

Animals↗

PubMed1983

Subcortical auditory and somatosensory afferents to hamster superior colliculus.

To determine whether the pattern of superior colliculus (SC) afferents seen in cat is generalizable to other mammalian species, HRP was injected into the SC of twenty hamsters (Mesocricetus auratus). After reaction with TMB, subcortical structures were examined for labeled perikarya. Although most subcortical afferents to SC were similar in cat and hamster, there were differences in auditory and somatosensory SC innervation. In hamster only two putatively auditory structures showed labeled cells, the external nucleus of inferior colliculus and nucleus of the brachium of the inferior colliculus, whereas in cat additional cells are reported in the dorsal cochlear nucleus, trapezoid and superior olivary nuclei, and nucleus of the lateral lemniscus. Again, in hamster the major trigeminal somatosensory input to SC is from spinal trigeminal nucleus caudalis whereas in cat trigeminal input is reported to be from the principalis and oralis portions of the trigeminal nucleus. Thus the hamster possesses a very restricted auditory input to and a different pattern of somatosensory innervation of the SC relative to the cat.

Afferent Pathways↗

PubMed1983

Localization of motoneurons innervating the levator veli palatini muscle in the cat.

Recent investigations of the nucleus ambiguus (NA) have attempted to identify motoneurons associated with the branchiomeric muscles of the larynx and pharynx. However, relatively little attention has been directed to the levator veli palatini muscle (LVP) which is critical in respiration, deglutition and eustachian tube function. Although the consensus is that cranial nerve X (vagus) innervates this muscle, some investigators have suggested that the LVP is innervated by either cranial nerve VII (facial) or IX (glossopharyngeal). The present study was designed to identify the specific location of LVP motoneurons within the brainstem. Horseradish peroxidase (HRP) was injected into the LVP of 18 cats. Following a survival period of 24-48 hours, animals were sacrificed and tissue processed according to Mesulam's TMB procedure. HRP labeled cells were located in the rostral NA both ipsilateral and contralateral to the side of injection and in the ipsilateral retrofacial nucleus (RFN). There were no labeled cells in the facial nucleus. Innervation of the LVP by cranial nerve VII would thus be excluded. This is the first report to definitively localize LVP motoneurons. Although the innervation of LVP by cranial nerve X is generally agreed upon in basic anatomy textbooks, identification of LVP motoneurons within the NA does not exclude innervation by cranial nerve IX.

Animals↗

PubMed1984

Vagal and gastric connections to the central nervous system determined by the transport of horseradish peroxidase.

Horseradish peroxidase (HRP, Sigma Type VI) crystals were encased in a parafilm envelope and applied to the transected central ends of the left and right cervical vagus nerves and the anterior and posterior esophageal vagus nerves of adult male hooded rats. Injections of 30% HRP were made into the muscle wall of the fundus and antrum regions of the stomach. After 48 hr survival time, animals were perfused intracardially with a phosphate buffer plus sucrose wash followed by glutaraldehyde and paraformaldehyde fixative. The brain stem, spinal cord and corresponding dorsal root ganglia, superior cervical sympathetic ganglion, and the nodose ganglion were removed and cut into 50 micron sections. All tissue was processed with tetramethylbenzidine (TMB) for the blue reaction according to Mesulum and counterstained with neutral red. Sequential sections were examined under a microscope. Labeled neurons and nerve terminals were identified using bright and dark field condensers and polarized light. In tissue from animals that had HRP applied to the cervical vagus nerves, retrogradely labeled neurons were identified ipsilaterally in the medulla located in the dorsal motor nucleus of the vagus (DMN) and the nucleus ambiguus (NA). Labeled cells extended from the DMN into the spinal cord in ventral-medial and laminae X regions C1 and C2 of cervical segments. Many neurons were labeled in the nodose ganglion. Anterogradely labeled terminals were observed throughout and adjacent to the solitary nucleus (NTS) dorsal to the DMN and intermixed among labeled neurons located in the DMN. In tissue from animals that had HRP applied to the esophageal vagus nerves, similar labeling was observed. However, fewer neurons were identified in the NA, the nodose ganglion, and only in laminae X of the cervical spinal cord segments C1 and C2. Also, very little terminal labeling was observed in and adjacent to the NTS. Labeled neurons in tissue from animals that had HRP injected into the stomach wall were observed bilaterally in the DMN, nodose ganglion, and only in laminae X at the C1 and C2 levels of the spinal cord. Labeled neurons also were observed in the dorsal root ganglia of the thoracic cord. These data indicate that cervical cord and NA neurons are important in the supradiaphragmatic motor innervation by the vagus. Also, many afferents to the NTS originate above the diaphragm. In addition, some afferents from the stomach enter the central nervous system via the thoracic spinal cord.

Animals↗