Biomedical subjects

D K Sundberg

Publications and source records attributed to D K Sundberg.

At least 19 recordsLinked to original sources

PubMed1993

Sinoaortic denervation alters the molecular and endocrine responses to salt loading.

Neuroanatomical and physiological evidence indicates that baroreceptors influence hypothalamic vasopressin (VP) and oxytocin (OT) neurons. We evaluated the effects of sinoaortic denervation (SAD) on the molecular and endocrine response to salt loading. Sham-operated or SAD rats were given a 2% NaCl solution to drink for 72 h. A group with limited salt water intake was included as a second control because the denervated rats consumed less salt than the controls. Plasma VP, OT and osmolality and posterior pituitary peptide content were measured. Brains were processed for evaluation of VP and OT mRNA expression using in situ hybridization with computer quantitation. Salt loading produced equivalent increases in plasma VP and OT in the control and SAD groups, however, there was a greater depletion of posterior pituitary peptides in the denervated animals. Salt loading produced significant decreases in pituitary VP and OT in the SAD animals, 69.8 +/- 8.4% and 68.3 +/- 4.0%, respectively. In the control groups, there was no decrease in VP content and a decrease in OT only in the control ad lib group. The peptide mRNA response to salt loading was also altered in the denervated rats. There was a significant increase in the area and intensity of the labeling for OT mRNA in the PVN in the SAD salt group. The control salt rats showed an increase in the SON and the salt-limited group showed no changes. For VP mRNA, the only change noted was in the SON of the salt-loaded SAD animals. These results show that chronic denervation of arterial baroreceptors augments the hypothalamic VP and OT response to salt loading.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Excitotoxin paraventricular nucleus lesions: stress and endocrine reactivity and oxytocin mRNA levels.

Electrolytic lesion of the paraventricular nucleus (PVN) of the hypothalamus blocks the tachycardia response to stress. The current study examined the effects of chemical lesion of PVN parvocellular neurons on the cardiovascular and endocrine responses to stress and on the content of hypothalamic oxytocin (OT) mRNA levels. Acute footshock stress increased heart rate in both ibotenic acid lesion and control groups of animals; however, the tachycardia was significantly lower in animals with a PVN lesion than the controls. Lesion of the PVN also attenuated the increase in plasma OT induced by stress, 4-fold in the lesion group versus 20-fold for the controls. There was not a generalized decrease in hormonal responsiveness since the OT response to an osmotic challenge was exaggerated in the lesion group. There was no difference between the groups in the arterial pressure and vasopressin responses to acute stress. Neurotoxin lesions of the PVN also resulted in significant depletions of VP and OT in all levels of the spinal cord and decreased OT levels in the dorsal brainstem. Ibotenic acid lesions of the PVN resulted in no significant changes in OT mRNA in the PVN, SON and PP. In addition, the 48-h dehydration resulted in a significant increase in plasma OT and OT mRNA in the PVN. These data indicate that the parvocellular neurons of the PVN play a role in integration of cardiovascular and endocrine responses to both stressful and osmotic stimuli and provide further evidence that parvocellular OT and VP neurons project to the brainstem and spinal cord.

Animals↗

PubMed1992

Central and systemic oxytocin release: a study of the paraventricular nucleus by in vivo microdialysis.

The mechanisms controlling central and systemic oxytocin (OT) release were examined using in vivo microdialysis of the paraventricular (PVN) region. Dialysate and plasma samples were collected from conscious male rats and stimuli were administered via the dialysate fluid. Characterization studies showed that microdialysis was a viable technique for the study of peptide secretion in the conscious animal. OT was consistently detected in the PVN dialysate and a partially purified extract crossreacted in parallel fashion with the synthetic peptide. In vitro studies showed that peptide recovery was positively correlated with the pore size of the dialysis membrane and that there was an inverse relationship between flow rate and recovery. Hypertonic saline administered centrally caused an increase in dialysate and plasma oxytocin while the intravenous injection affected only plasma oxytocin. The excitatory amino acid, glutamate (0.05-0.5 M), caused an increase in plasma, but not dialysate oxytocin, while depolarization with potassium chloride (0.05-0.15 M) had no significant effects. Histological examination showed that the dialysis probe was located in the rostral, lateral PVN. Our results show that in vivo microdialysis provides a method for the delivery of drugs into specific brain regions as well as a useful technique for the evaluation of in vivo neuropeptide release.

Analysis of Variance↗

PubMed1990

Extracellular oxytocin in the paraventricular nucleus: hyperosmotic stimulation by in vivo microdialysis.

The effect of central osmotic stimulation on oxytocin (OT) secretion from the paraventricular nucleus (PVN) was examined using a newly developed in vivo microdialysis technique. A dialysis probe was inserted into the PVN region and microdialysis was performed in conscious animals. Hyperosmotic solutions were delivered via the dialysis probe, and perfusate and blood samples were collected. OT was consistently detected in the PVN dialysate. Hyperosmotic sodium chloride (1 M) produced a significant increase in dialysate and plasma OT, whereas D-mannitol (2 M) had no effect. These results suggest that (1) in vivo microdialysis may provide a useful technique for the evaluation of neuropeptide secretion from specific brain regions and (2) there are sodium-sensitive cells in the PVN region which respond to increases in extracellular sodium, resulting in an increase in central and peripheral oxytocin secretion.

Animals↗

PubMed1989

Cultured hypothalamic explants from spontaneously hypertensive rats have decreased vasopressin and oxytocin content and release.

To investigate the central nervous system (CNS) changes in the spontaneously hypertensive rat (SHR), a tissue culture model was used to examine the content and release (24 hour) of the peptide hormones, vasopressin (VP) and oxytocin (OT), from brain explants. Nuclear regions consisting of the paraventricular (PVN) or supraoptic (SON) nuclei were microdissected from prehypertensive SHR and Wistar-Kyoto (WKY) rats. Media levels of VP and OT were measured at 1, 3, 4, and 7 days of culture. After three days of culture, the PVN explants from SHR secreted significantly less VP and OT (both reduced 80%) than did those from WKY. Release of both VP and OT in the SON explants was significantly lower (approximately 50% lower) in the SHR only at seven days of culture. Additionally, tissue content of the peptides was measured after 0, 1, 4, and 7 days of culture. Tissue content of VP and OT was decreased (40% or more) in the SHR in both nuclear regions after four and seven days of culture. In addition, nicotine was found to stimulate the release of VP from SON, but not PVN, cultures in both SHR and WKY explants. Immunohistochemical data showed that there was not a preferential loss of VP or OT neurons in explants from the SHR. Therefore, this in vitro model would indicate that there is a difference in the ability of cultured explants of PVN and SON from SHR and WKY (four-week-old) to synthesize and/or release the peptide hormones VP and OT.

Animals↗

PubMed1988

Effect of central hypertonic stimulation on plasma atrial natriuretic peptide and oxytocin in conscious rats.

The effect of the intracerebroventricular (i.c.v.) injection of hypertonic sodium chloride on plasma atrial natriuretic peptide (ANP) and oxytocin (OT) was evaluated in conscious freely moving rats. A hypertonic or isotonic NaCl solution was injected into the third ventricle. Blood pressure and heart rate were monitored and blood samples were collected. I.c.v. injection of the hypertonic solution resulted in a significant increase in mean arterial pressure (105.3 +/- 2.9 mmHg at time 0 to 124.2 +/- 4.4 mmHg at 5 min, P less than 0.01) and heart rate (350.0 +/- 25.0 bpm at time 0 to 420.8 +/- 13.6 bpm at 20 min, P less than 0.01). Plasma OT increased 4-fold over the basal values 5 min after the injection (4.5 +/- 1.1 to 20.1 +/- 3.2 pg/ml, P less than 0.01), while there was no significant change in plasma ANP (37.3 +/- 9.1 to 46.6 +/- 12.6 pg/ml, n.s.). The control injection produced no significant changes in any parameters. These results show that hemodynamic changes are not necessarily associated with alterations in plasma ANP. Furthermore they suggest that central osmoreceptors are not involved in the control of ANP secretion.

Animals↗

PubMed1988

Plasma catecholamine levels in patients transported by helicopter for acute myocardial infarction and unstable angina pectoris.

Fourteen patients with either acute myocardial infarction or unstable angina pectoris were transported by helicopter air ambulance to North Carolina Baptist Hospital during a 1-month period. Six patients had preflight and inflight plasma epinephrine and norepinephrine levels determined. All 14 patients were monitored for ventricular arrhythmias. The mean inflight plasma epinephrine level was significantly higher than the mean preflight level (3455 v 841 pg/mL, P less than .005). There was also a trend toward higher inflight norepinephrine levels; however, this increase was not statistically significant. No patient had a monitored ventricular arrhythmia. These findings suggest that helicopter transport of cardiac patients may be associated with significant patient stress, as reflected by high inflight catecholamine levels. Further study with a larger population of patients is needed to determine whether or not an increased incidence of inflight ventricular arrhythmias is associated with these catecholamine changes.

Aged↗

PubMed1988

Comparison of hormonal responses to hypotension in mature and immature fetal lambs.

We studied norepinephrine (NE) and plasma renin activity (PRA) responses to sodium nitroprusside (NP)-induced hypotension in seven chronically catheterized fetal lambs 0.79-0.94 gestation (mature) and in seven fetuses 0.64-0.72 gestation (immature) 4 or 5 days after surgery. We infused intravenously 5% dextrose in water (DW) or NP in DW to reduce arterial pressure 30% in fetuses for 10 min. Initial infusion choice was random, and the two infusions were separated by 24-48 h. In both groups, basal NE levels were similar and doubled in response to hypotension. In mature fetuses, PRA basal levels were 6.89 +/- 1.80 ng.ml-1.h-1 and increased two- to threefold with hypotension. In immature fetuses, PRA basal levels were 2.42 +/- 0.86 ng.ml-1.h-1 and did not change with hypotension. No changes were observed with DW infusion in either group. Arterial blood gases were normal and remained unchanged. We conclude that in the lamb fetus, NE responses to hypotension are present before and are independent of the development of PRA responses and that before 0.72 gestation there is a functional deficit in the renin-angiotensin system.

Animals↗

PubMed1987

Responses to converting-enzyme inhibition and hemorrhage in newborn lambs and adult sheep.

We compared the cardiovascular and hormonal responses to angiotensin converting enzyme inhibition and hemorrhage of 20% of blood volume in chronically instrumented unanesthetized newborn lambs and adult sheep. Administration of the nonsulfhydryl-containing converting-enzyme inhibitor enalapril reduced mean arterial pressure in the newborn but not in the adult animals. Blood pressure fell in both age groups after hemorrhage, and the hemorrhage-induced fall in blood pressure, integrated over the period of hypovolemia, was more pronounced when converting-enzyme inhibition was present in the lambs. This was not observed in the adults. Cardiac output fell following hemorrhage in both age groups, and the fall was greater when enalapril was present in the lambs, but this was not the case in the adults. Hemorrhage increased plasma renin activity in both groups, and enalapril augmented this increase. Plasma concentrations of vasopressin and catecholamines increased following hemorrhage within and between groups. Taken together these data suggest that the renin-angiotensin system plays a more important role in the maintenance of cardiovascular homeostasis in newborn lambs than it does in adult sheep, and catecholamine and vasopressin responses to volume loss can occur in the presence of blockade of the renin-angiotensin system.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed1987

Effect of chronic alcohol consumption on the pregnant mare serum gonadotropin-induced luteinizing hormone surge.

The studies reported in this paper were undertaken to investigate the effect of chronic (10 day) alcohol consumption on female pituitary-gonadal function. The immature female rat model treated with pregnant mare serum gonadotropin (PMSG) was used since it results in a highly reproducible luteinizing hormone (LH) surge and ovulation. Twenty-day-old female rats were placed on diets that were either (1) unrestricted (ad libitum); (2) contained 5% ethanol in a liquid diet, or (3) isocalorically pair-fed with the liquid diet to the ethanol group. After 10 days on their respective diets, the groups were subdivided and given either 8 IU of PMSG in 0.1 ml saline or 0.1 ml saline s.c. between 10.00 and 11.00 h. The animals were sacrificed by decapitation at 24, 48, 52, 54, 56, 58, 60 and 62 h after injection. Trunk blood was obtained for serum measurements of LH. The uteri were weighed and prepared for the histological study. In all dietary groups, serum LH levels were significantly higher in the PMSG-treated animals when compared to the saline controls at all time intervals with the exception of the alcohol 58-hour group. In the ad libitum animals, plasma LH concentrations were highest at 52 h following hormone administration. The serum LH concentrations were highest at 56 h after hormone administration in the pair-fed group and were significantly less than the ad libitum group at 52, 54, 56, and 58 h after PMSG stimulation. No significant plasma LH surge was observed in the alcohol group and the LH concentrations were significantly less than the pair-fed rats at 56 and 58 h after PMSG treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Alcoholism↗

PubMed1986

Distribution and development of proenkephalin-like immunoreactivity in the lumbar spinal cord of the chicken.

Met5-enkephalin- (Met-ENK), Leu5-enkephalin-, Met5-enkephalin-Arg6-Phe7-, metorphamide- and BAM 22P-like peptides could be detected in the lumbar spinal cord of the chicken by immunocytochemistry and/or high performance liquid chromatography. However, a peptide identical to Met5-enkephalin-Arg6-Gly7-Leu8 could not be detected in the lumbar spinal cord of the chicken using an antiserum that was capable of detecting the octapeptide in mammalian tissues. Nerve fiber- and terminal-like processes containing proenkephalin-derived peptides were concentrated in the superficial laminae of the dorsal horn and along the midline rostral to the central canal. A lesser concentration of processes containing proenkephalin-derived peptides occurred in the medial and lateral motor columns of the ventral horn. The level of total radioimmunoassayable Met-ENK in the lumbar spinal cord of the chicken embryo increased more than 1000-fold between day 4.5 and day 18. A schedule of curare administration that had previously been shown to prevent naturally occurring somatic motoneuron death in the chicken lumbar spinal cord resulted in a two-fold increase in total radioimmunoassayable Met-ENK in the lumbar spinal cord.

Animals↗

PubMed1986

A tissue culture model for the study of peptide synthesis and secretion from microdissected hypothalamic explants.

Using established tissue culture methodology, a protocol has been developed for the culture of specific hypothalamic regions. A microdissection technique is used to remove the paraventricular and supraoptic nuclear regions from fresh tissue. These explants are maintained in culture and appear to be viable on the basis of several criteria: the continuing presence of peptides in both tissue and media, the ability to incorporate a labeled amino acid precursor into proteins and peptides, the histological appearance using phase contrast microscopy, and the immunocytochemical identification of peptide neurons. This tissue culture model may prove useful in the study of neuroendocrine mechanisms in specific brain regions.

Catecholamines↗

PubMed1986

Methods for estimating the contribution of proenkephalin A and proenkephalin B input to neuronal areas.

Final expression of the proenkephalin A and proenkephalin B (prodynorphin) gene should yield a fixed ratio of enkephalin-like peptides depending on their precursor origin. If a tissue contained peptides only from proenkephalin A this ratio would be 4/1/1/1 (Met-Enk/Leu-Enk/Met-Enk-Arg6-Phe7/Met-Enk-Arg6-Gly7-Leu8). On the other hand, if the tissue-derived opiates were from prodynorphin, only Leu-enkephalin should be found. The techniques described here combining oxidation of the Met-Enk species, reverse-phase HPLC, and measurement with a radioimmunoassay which recognized many of the opiate-like peptides have been found useful for determining the precursor origin of enkephalins within different biological sources.

Animals↗

PubMed1986

Biochemical and immunochemical studies of supraoptic and paraventricular cultures.

A tissue culture model was established for the study of hypothalamic peptide synthesis and secretion. Microdissected explants of the paraventricular and supraoptic regions from Sprague-Dawley rats (neonates or young rats) were maintained in culture for up to 3 weeks. Studies were performed to evaluate vasopressin and oxytocin content (medium and tissue levels), immunocytochemical localization, and biosynthetic activity. Immunocytochemical staining for oxytocin, neurophysin, and neuron-specific enolase showed positive neurons in both the paraventricular and supraoptic cultures. In many cases, the neurons were large (30-40 microns) and bipolar, resembling the classic magnocellular neuron. Measurement of tissue and medium content showed the continued presence of vasopressin and oxytocin in the cultured explants. Even after 3 weeks, there were significant amounts of vasopressin present. Biosynthesis was evaluated by determining the incorporation of 35S-labeled cystine or cysteine into proteins and peptides. The medium and tissue extracts were separated by reverse-phase high-performance liquid chromatography. Results showed that most of the labeled peptides were released into the medium rather than stored. There were two labeled peaks in the medium, which chromatographically resembled native vasopressin and oxytocin. Treatment with a protein synthesis inhibitor, either puromycin or cycloheximide, resulted in a decrease in labeled peptides. A comparison of 35S-labeled cystine and cysteine showed that the latter was the label of choice, with significantly greater incorporation.

Animals↗

PubMed1983

Hypothalamic catecholamine biosynthesis and pituitary gonadotropin secretion in vitro: effect of hyperprolactinemia.

The effect of hyperprolactinemia on central catecholamine biosynthesis and anterior pituitary hormone release was studied using an in vitro methodology. The incorporation of [3H]tyrosine into hypothalamic and neurohypophyseal catecholamines was determined using a new method which combines high performance liquid chromatography (HPLC) with amperometric detection (LCEC). Elevated plasma prolactin levels, induced by pituitary transplants, resulted in increased in vitro biosynthesis of medial basal hypothalamic (MBH) dopamine (DA), but not norepinephrine (NE). Neurohypophyseal DA biosynthesis (including the intermediate lobe) was not affected. Plasma LH levels were depressed by hyperprolactinemia although the content of hypothalamic luteinizing hormone-releasing hormone (LHRH) was not changed. In parallel studies, the anterior pituitaries from these animals were incubated in vitro using a paired-half technique and LH and PRL release measured. While the basal release of prolactin was not altered by hyperprolactinemia, LH release was significantly decreased. Gonadotroph responsiveness to LHRH was significantly increased, while the inhibition of prolactin by dopamine was not altered. There was a decrease in pituitary prolactin content with normal LH levels. These experiments confirm several in vivo reports which show that hypothalamic dopaminergic but not noradrenergic activity is increased by prolactin. This action is specifically localized in the tuberoinfundibular dopaminergic neurons. Furthermore, these experiments suggest that these central changes result in alterations in both gonadotroph and mammotroph function.

Animals↗

PubMed1983

Hypothalamic catecholamine biosynthesis and neuropeptides.

In conclusion, both steady state and non-steady state methods for investigating the turnover of central catecholamines can provide valuable information regarding the central control of neuroendocrine function. The system described here, utilizing HPLC with electrochemical detection, offers a relatively easy and reliable method for determining steady state catecholamine biosynthesis in the hypothalamus and other brain areas.

Animals↗

PubMed1983

Methods for investigating peptide precursors in the hypothalamus.

In conclusion, as in the aminergic nervous system, the hypothalamic neuropeptides "turn over" at a rate sufficient to maintain a "steady state" concentration within the hypothalamus. However, unlike catecholamine-secreting neurons, the brain peptides are synthesized and secreted by a much more circuitous mechanism, involving ribosomal biosynthesis as larger prohormones, axonal transport to the site of secretion, and enzymic processing to the biologically active hormones. The methods described here, including in vitro incubation and high-performance size-exclusion and reverse-phase chromatography, have been found to be useful for examining and comparing the rate and mechanism of biosynthesis of several hypothalamic neuropeptides.

Amino Acids↗

PubMed1982

Patterns of radioactivity in the eyes of rats after injection of iodinated prolactin.

The present study involves the intracardial injection of iodinated ovine prolactin into albino rats and the autoradiographic demonstration of patterns of isotopic incorporation into ocular tissues, including the retina, choroid coat and ciliary body. Control procedures include the utilization of competitive hormonal binding, a comparison of the radioactive pattern in eyes after the injection of iodinated beef serum albumen and an autoradiographic evaluation of the thyroid glands of all groups. The competitive uptake of iodinated prolactin into ocular tissues also was examined by liquid scintillation spectrometry. In normal rats, radioactivity was localized over the cells of the choroid coat and ciliary body of all groups receiving iodinated prolactin. The inner segments of the photoreceptors were radioactive in rats at 15 min after injection, but were unlabeled at only 5 min after injection.

Animals↗