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N L Ostrowski

Publications and source records attributed to N L Ostrowski.

18 recordsLinked to original sources

Distribution of V1a and V2 vasopressin receptor messenger ribonucleic acids in rat liver, kidney, pituitary and brain.

The hepatic, vascular-type (V1aR) and the renal, antidiuretic-type (V2R) vasopressin receptor cDNAs were recently cloned from rat liver and kidney libraries, respectively. DNA fragments containing the region encoding the putative 5/6 transmembrane loops of these receptors were subcloned, separately, into RNA polymerase promoter-containing vectors from which 35S-labeled sense and antisense riboprobes were synthesized. In situ hybridization histochemistry showed high levels of V1aR transcripts in the liver and the renal medulla among the vascular bundles. Sparser labeling was found in the renal cortex, but there were no grains over the glomeruli. V1aR mRNA was detected in many brain areas, including the hippocampal formation, central amygdala, dorsolateral septum, lateral hypothalamus, suprachiasmatic, ventromedial, dorsomedial, and arcuate nuclei of the hypothalamus, nucleus of the solitary tract, cerebellum, spinal nucleus of the trigeminal tract, reticular formation, inferior olivary nucleus, and choroid plexus. Rare labeled cells were seen along the periphery of the posterior pituitary. V2R transcripts were not detected in the liver or brain, but were present in high amounts in the inner and outer renal medulla, primarily associated with collecting ducts. Sparser labeling was found in the renal cortex, and no grains were seen over the glomeruli. These data confirm the expression of the V1a vasopressin receptor in liver and brain and demonstrate that kidney expresses mRNAs encoding V1a and V2 vasopressin receptors.

Animals

Correlation between locomotor stimulation and the electrophysiological effects of low doses of morphine on substantia nigra dopamine neurons. I. Acute drug administration.

Parallel experiments were done to determine whether the behavioral effects of low doses of morphine correlated with changes in the electrophysiological activity of two subpopulations of mesencephalic dopamine neurons in rats. Acute administration of morphine sulfate (0.01 or 0.20 mg/kg i.v.) produced a naloxone antagonizable increase in locomotion and a corresponding increase in the discharge rates of Type A dopamine neurons. Morphine sulfate led to a relatively long-latency (120-200 sec) loss of spontaneous discharge activity in the majority of Type B dopamine neurons and this was blocked by a high dose of naloxone (5.0 mg/kg). Naloxone (0.10 or 5.0 mg/kg i.v.), combined with morphine, led to a paradoxical behavioral suppression. These data suggest that morphine produces opposite effects on two functionally distinct subtypes of neurons in the substantia nigra pars compacta and that behavioral output may reflect an interaction between these changes in discharge activity in mesencephalic dopamine pathways.

Animals

Gastrin-releasing peptide stimulates glycoconjugate release from feline trachea.

The effect of gastrin-releasing peptide (GRP) on respiratory glycoconjugate (RGC) secretion was investigated in a feline tracheal organ culture model. RGC secretion was stimulated by GRP in a dose-dependent fashion at concentrations from 10(-8) to 10(-5) M (range 15-38% increase above control) with a peak effect within 0.5-1 h of incubation. GRP-(14-27), the receptor binding portion of GRP, and the related molecule, bombesin, also stimulated RGC secretion by approximately 20% above control. Acetyl-GRP-(20-27) stimulated RGC release by 10%, whereas GRP-(1-16) was inactive. Autoradiographic studies with 125I-GRP revealed that specific binding was restricted to the submucosal glands and the surface epithelium. A specific radioimmunoassay showed the content of GRP in feline trachea after extraction with ethanol-acetic acid to be 156 +/- 91 fmol/g wet wt. Indirect immunohistochemistry indicated that ganglion cells located just outside the cartilage contained GRP-immunoreactive materials. GRP is a novel mucus secretagogue that may participate in regulating airway mucosal gland secretion.

Animals

The involvement of the benzodiazepine receptor in hepatic encephalopathy: evidence for the presence of a benzodiazepine receptor ligand.

The involvement of GABAergic systems in the pathogenesis of HE was supported by electrophysiologic studies of single Purkinje neurons from rabbits with HE which demonstrated the hypersensitivity of these neurons to depression by GABA and BZ receptor agonists. In contrast, these neurons were excited by BZ receptor antagonists. At concentrations which had no effect on neuronal activity, BZ receptor antagonists also reversed the hypersensitivity of HE neurons to depression by muscimol. This combination of neuronal responses is consistent with an increase in the concentration or availability of a ligand for the BZ receptor with agonist properties in the brains of rabbits with HE. Subsequent neurochemical studies support these electrophysiologic observations. Autoradiographic techniques indicated the presence of a reversible inhibitor of [3H]Ro 15-1788 and [3H]flunitrazepam binding to the cerebral and cerebellar cortices of rabbits with HE. The ability of this substance to inhibit [3H]flunitrazepam binding to HE rabbit brain sections was further enhanced in the presence of NaCl and GABA. The autoradiographic studies suggested that the density and affinity of the components of the GABA-BZ receptor complex are unaltered in this animal model of HE. This inference is fully supported by the subsequent studies of radioligand binding to well-washed membrane preparations. Finally, extracts of HE rabbit brains yielded a family of substances with the properties of BZ receptor agonists. These substances may include, but are not limited to, diazepam, oxazepam and desmethyldiazepam, but do not include substances commonly elevated in the plasma and CSF of patients with HE4. The positive identification of these substances awaits confirmation by mass-spectroscopic analysis. However, the precedent for the presence of a family of benzodiazepines in animals that were not administered these drugs has been set. The origin of these substances is a matter of ongoing research. Several studies have shown the presence of benzodiazepines in plant and animal materials. It is possible that these "endogenous" benzodiazepines are the result of contamination of the food chain. A normally functioning liver would capture and metabolize these compounds after their absorption from the gut. This function of the liver would be impaired in liver failure, thus allowing sufficient levels of BZ receptor agonists to accumulate in the CNS, contributing to the pathogenesis of HE. However, studies by DeBlas and coworkers have reported that 1,4 benzodiazepines are present in human brains preserved prior to the commercial use of these compounds. Further, they have found benzodiazepines in cell lines cultured without potential exogenous sources of benzodiazepines.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

The GABAA receptor complex in hepatic encephalopathy. Autoradiographic evidence for the presence of elevated levels of a benzodiazepine receptor ligand.

Autoradiographic analysis was used to examine radioligand binding to benzodiazepine (BZ) and GABAA receptors in the brains of rabbits with hepatic encephalopathy (HE). Thin sections of whole brain from normal rabbits and rabbits with HE were mounted on slides and subdivided into two groups. One group was washed before incubation with radioligand, while the second group was not prewashed. [3H]Flunitrazepam binding to BZ receptors was decreased by 22% to 42% (p less than 0.05) in the cerebral cortex, superior and inferior colliculi, and cerebellum of unwashed sections from rabbits with HE compared to all other groups. The binding of [3H]Ro 15-1788 to unwashed sections from rabbits with HE was reduced by a similar degree (18% to 37%, p less than 0.05) in the cerebral cortex, hippocampus, superior colliculus, and cerebellar cortex. Incubation of sections with the GABA-mimetic muscimol and NaCl produced an additional decrease in [3H]flunitrazepam binding to the cortex and hippocampus (25% to 31%, p less than 0.05) in unwashed HE rabbit brain, but increased radioligand binding (27% to 71%, p less than 0.05) to several regions in control rabbits. No changes in radioligand binding to either GABAA or peripheral benzodiazepine receptors was observed between HE and control rabbit sections. These findings are consistent with previous electrophysiologic and neurochemical observations indicating no significant changes in either the function or density of GABAA or BZ receptors in this model of HE. Further, they indicate that a reversible BZ receptor ligand with agonist properties is present in the brain in HE. This substance may contribute to the enhancement of GABAergic tone observed in this syndrome.

Animals

Sexual behavior suppresses the primary antibody response in the golden hamster.

The possibility that sexual behavior triggers neural or endocrine events that influence immune function was explored in male Golden Hamsters (Mesocricetus auratus). Sexually experienced males showed a 62.8 and 64.9% decrease in antibody response to TNP-ovalbumin compared to virgin and sham-mated males, respectively, as measured by ELISA. Antibody deficiency was associated with copulation on the day of immunization and was highly correlated with ejaculatory behavior and the incidence of prolonged intromissions, an indicator of sexual satiety. Although a single sexual encounter was not sufficient to inhibit antibody levels, prolonged intromissions were inversely correlated with titers. Changes in antibody production were not related to plasma levels of cortisol, corticosteroid-binding globulin activity, or testosterone. These results suggest that sexual behavior is immunosuppressive and may play a role in the communicability of sexually transmitted diseases.

Animals

Mating suppresses splenic natural killer cell activity in male golden hamsters.

The possibility that sexual behavior is associated with changes in natural killer cell activity was explored using a 4-h chromium-51 release assay. Mated male Golden Hamsters (Mesocricetus auratus) showed a significant suppression of natural killer cell activity 2 h after sexual activity relative to matched Virgin controls. Natural killer cell activity returned to control levels by 16 h postmating. The suppression of natural killer cell activity did not correlate with changes in plasma cortisol, transcortin, or testosterone levels, or with ejaculatory or intromissive behaviors. Administration of 0.40 mg/100 g of testosterone also suppressed natural killer activity at 2 h.

Animals

Galanin antagonizes acetylcholine on a memory task in basal forebrain-lesioned rats.

Galanin coexists with acetylcholine in medial septal neurons projecting to the ventral hippocampus, a projection thought to modulate memory functions. Neurochemical lesions of the nucleus basalis-medial septal area in rats impaired choice accuracy on a delayed alternation t-maze task. Acetylcholine (7.5 or 10 micrograms intraventricularly or 1 micrograms micro-injected into the ventral hippocampus) significantly improved performance in the lesioned rats. Atropine (5 mg/kg intraperitoneally or 10 micrograms intraventricularly), but not mecamylamine (3 mg/kg intraperitoneally or 20 micrograms intraventricularly), blocked this action of acetylcholine, suggesting involvement of a muscarinic receptor. Galanin (100-500 ng intraventricularly or 200 ng into the ventral hippocampus) attenuated the ability of acetylcholine to reverse the deficit in working memory in the lesioned rats. The antagonistic interaction between galanin and acetylcholine suggests that endogenous galanin may inhibit cholinergic function in memory processes, particularly in pathologies such as Alzheimer disease that involve degeneration of basal forebrain neurons.

Acetylcholine

Autoradiographic visualization of sex differences in the pattern and density of opiate receptors in hamster hypothalamus.

Slide-mounted brain sections were used to visualize the distribution of opiate receptors in the hypothalamus of male and female hamsters using the vitro film autoradiography. Sex differences were found in the binding density and patterns of [3H]naloxone-labeled receptors. The distribution and density of [3H][D-Ala2,D-Leu5]enkephalin-labeled delta-receptors in adjacent brain sections were similar in males and females. Male hamsters showed a U-shaped pattern of [3H]naloxone binding in the sexually dimorphic nuclear complex with 28% and 34% greater labeling of the sexually dimorphic nucleus (SDN) and bed nucleus/stria terminalis (BNST), respectively, than periovulatory estrous females. Estrous and diestrous females showed a V-shaped pattern of [3H]naloxone binding in the same region, but binding density was higher at diestrus. Greater specific [3H]naloxone binding in diestrous females was also evident following extensive prewashing of slide-mounted tissue sections indicating that residual endogenous opioids were not occupying receptors, and thus, reducing the labeling of receptors in tissue from estrous females. An estrous-linked change in the affinity of hypothalamic opiate receptors was suggested by findings that [3H]naloxone binding density was greater in tissue from diestrous females when incubations were conducted in the presence of a 1-nM, but not a 10-nM, concentration of the labeled antagonist. Finally, a dense are of [3H]naloxone binding was discovered in the dorso-suprachiasmatic region of the hypothalamus. These data provide evidence for a sexual dimorphism in the distribution and density of opiate receptors in hamsters. The data suggest that mu- or kappa-receptors are more likely than delta-receptors to be involved in mediating hypothalamic effects of endogenous and exogenous opioids on reproductive functions in this species.

Animals

The pattern of [3H]cyclofoxy retention in rat brain after in vivo injection corresponds to the in vitro opiate receptor distribution.

Cyclofoxy, a fluorinated analog of naltrexone, has been designed specifically to permit in vivo labeling of opiate receptors in experimental animals and ultimately, humans. Recently, using positron emission tomography (PET), 3-[18F]acetylcyclofoxy was shown to accumulate in opiate receptor-rich brain regions of a live baboon and to be stereospecifically displaced by injections of (-)-naloxone but not (+)-naloxone. Autoradiographic evidence is presented here that the unacetylated compound, [3H]cyclofoxy, labels a population of opiate receptors in brain after in vivo injections that is virtually identical to that labeled by [3H]naloxone. The in vivo binding patterns of [3H]cyclofoxy in brain were similar to those obtained following incubation of slide-mounted brain sections in vitro. Intravenous injections of [3H]cyclofoxy to rats yielded high (greater than 4:1) striatal and thalamic to cerebellar binding ratios in brain homogenates, supernatants and in 24 micron-thick brain sections 60 min after 30 mu Ci per animal (spec. act. = 16.4 Ci/mmol). Autoradiographs revealed the typical opiate antagonist binding profile with marked retention of [3H]cyclofoxy in the striatal patches, subcallosal streak, medial habenula and central thalamus with little retention of label in cerebellum. [3H]Cyclofoxy binding was reversible since the radiolabeled drug disappeared from brain tissue within 2 h after injections to rats, or could be removed from brain slices in vitro by washing slide-mounted tissue sections for 45 min. In addition, after in vitro washing the same brain sections again bound [3H]cyclofoxy or [3H]naloxone in the same pattern. When pre-washed brain sections were incubated with [3H]cyclofoxy in the presence of unlabeled naloxone, [3H]cyclofoxy binding was reduced to background levels. These data show that [3H]cyclofoxy labels-sensitive opiate receptors in vivo and in vitro. The present results combined with evidence that cyclofoxy demonstrates a low level of toxicity in animals suggest that cyclofoxy is an excellent tool with which to study the physiological role of opiate receptors in living animals using in vivo autoradiography, and in humans using PET.

Animals

Aldosterone-reversible decrease in the density of renal peripheral benzodiazepine receptors in the rat after adrenalectomy.

A statistically significant decrease in the density of peripheral benzodiazepine receptors was observed in renal membranes of rats beginning 2 weeks after adrenalectomy when compared with sham-operated controls. This decrease in peripheral benzodiazepine receptor density was manifest as a decrease in the maximum binding of two ligands, [3H]Ro 5-4864 and [3H]PK 11195, without accompanying changes in their Kd for this site. Similar changes were not seen in another aldosterone-sensitive organ, the submandibular salivary gland. The decrease in peripheral benzodiazepine receptor density observed in adrenalectomized rat renal membranes was restored to control levels after 1 week of aldosterone administration using a dose (12.5 micrograms/kg/day) that had no effect on peripheral benzodiazepine receptor density in sham-operated animals. In contrast, dexamethasone administration (50 micrograms/kg/day, 1 week) had no effect on renal peripheral benzodiazepine receptor density when administered to either adrenalectomized or sham-operated rats. Further, adrenal demedullation had no effect on renal peripheral benzodiazepine receptor density or affinity. The decrease in peripheral benzodiazepine receptor density was localized to the renal cortex and the outer stripe of the medulla by gross dissection of renal slices and renal tissue section autoradiography. The specific effect of adrenalectomy on renal peripheral benzodiazepine receptor density, the lack of direct effect of aldosterone on [3H] Ro 5-4864 binding and the localization of the change in peripheral benzodiazepine receptor density to the renal cortex and outer stripe suggest that these changes may reflect an adaptation of the renal nephron (possibly the distal convoluted tubule, intermediate tubule and/or the collecting duct) to the loss of mineralocorticoid hormones.

Adrenal Cortex

Parasympathetic ganglia: naloxone antagonizes inhibition by leucine-enkephalin and GABA.

Synaptic transmission in parasympathetic ganglia of the cat urinary bladder was depressed by low doses (0.1-10 micrograms i.a.) of Leu- or Met-enkephalin but only by larger doses (10 micrograms-1 mg i.a.) of morphine. Naloxone blocked the depressant effects of the opiates as well as the depression produced by GABA, but did not block the depressant effects of norepinephrine. Intracellular recording revealed that Leu-enkephalin reduced EPSP-amplitude and lowered the probability of synaptically evoked firing without altering postsynaptic membrane potential or resistance. These findings suggest that enkephalinergic inhibition in bladder ganglia is mediated at least in part by a presynaptic site of action on delta opiate receptors.

Animals

Morphine antagonists and consummatory behaviors.

Opiate antagonists were tested for their effects upon either drinking or eating in eight experiments. Naloxone, nalorphine, and the active isomer of WIN 44,441 all reduce drinking. Neither an analog of nalorphine that does not cross the blood-brain barrier, nor the inactive isomer of WIN 44,441 is effective in reducing water intake. These data provide support for the conclusion that these antagonists ahve stereospecific effects within the central nervous system. Naloxone suppresses drinking following procedures inducing osmotic, volemic, or hormonal thirst. Naloxone suppresses eating following procedures inducing glucoprivation but does not alter eating elicited by tail-pressure. Collectively, these data lead to the conclusion that endorphins play a role in the organization of ingestive behavior following challenges to homeostasis.

Angiotensin II

Opiate antagonists and sexual behavior in female hamsters.

Mating or administration of morphine to female hamsters reliably decreases lateral displacement, a sensitive index of female sexual responsitivity. Morphine's effects are antagonized by naloxone. We asked whether endogenous opiates are significantly involved in the mating-induced inhibition of sexual responsitivity by testing whether naloxone or naltrexone attenuated the mating-induced decreases in lateral displacement. Naloxone (4 mg/kg) increased lateral displacement in only one of three tests in females before mating. Naloxone did not attenuate the mating-induced decreases in lateral displacement or lordosis behavior in either ovariectomized, hormonally supplemented or intact females. Large doses of naltrexone produced no reliable effects on sexual behavior during estrus in unmated females, nor did it attenuate the mating-induced decreases in sexual responding, regardless of the time of day of mating. Naloxone often increases the variability of sexual responding. We conclude that naloxone-sensitive mechanisms do not play a critical role in the expression of sexual behavior in female hamsters.

Animals

Naloxone reduces fluid intake: effects of water and food deprivation.

Food and fluid deprived and nondeprived male rats showed 36% and 46% decreases, respectively, in sucrose consumption 15-min after injection with 2 mg/kg of naloxone in one hr tests. The magnitude of this decrease was not correlated with an index of naloxone's ability to produce a sickness, as measured by the conditioned taste aversion test. Tests with animals scheduled to drink water in a 15-min daily session showed naloxone had similar effects in reducing water intake in 23-hr and 47-hr water deprived rats. Morphine, when self-administered, produced an increase in water intake during 6-hr sessions. The data support the idea that naloxone disrupts a component of normal regulation of ingestion.

Animals

Morphine and naloxone's effects on sexual behavior of the female golden hamster.

The effects of morphine and naloxone were observed after administration to female golden hamsters (Mesocricetus auratus). Large doses of morphine, 80 mg/kg, consistently produced sedation and behavioral depression of responses to nociceptive stimuli. Smaller doses of morphine (e.g., 10 mg/kg), that produced few other behavioral changes, suppressed a measure of female sexual responding. The suppressive effects on sexual behavior were reversed by 4 mg/kg of naloxone. Morphine administered intracerebroventricularly had little effect on sexual responding, even at doses which produced other side effects. Doses of 4 and 8 mg/kg of naloxone in opioid-naive subjects did not reliably alter sexual responding up to 2 hr after administration. These observations lead to the suggestion that morphine produces effects which are incompatible with full sexual functioning in female hamsters.

Animals

The effects of low doses of morphine on the activity of dopamine-containing cells and on behavior.

Intravenous (IV) administration of 10 ug/kg of morphine sulfate increased the firing rate of Type A dopamine neurons but decreased the extracellular activity of Type B dopamine cells in the substantia nigra pars compacta and ventral tegmental area of anesthetized male rats. The morphine-produced increase in Type A cells was antagonized by 0.10 mg/kg of naloxone, whereas the suppression of B cells was blocked only by pretreatment with 5.0 mg/kg of naloxone. This same low dose of morphine (IV) also produced increased locomotor activity in awake male rats within 10 min of administration.

Animals