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Biomedical subjects

N Ben-Jonathan

Publications and source records attributed to N Ben-Jonathan.

At least 91 records · Page 5Linked to original sources

The posterior pituitary: regulation of anterior pituitary prolactin secretion.

Removal of the posterior pituitary from anesthetized male rats results in a prompt and significant increase in circulating prolactin that is reversed by the injection of dopamine. Posterior pituitary extracts, which contain high concentrations of endogenous dopamine, inhibit prolactin secretion from isolated anterior pituitary cells. This inhibition is prevented by incubation of the cells with the dopamine receptor antagonist (+)-butaclamol. The data show that posterior pituitary dopamine reaches the anterior pituitary via the short hypophysial portal vessels and participates in the regulation of prolactin secretion.

Animals↗

Preovulatory depletion of ovarian catecholamines in the rat.

The purpose of the present investigation was to correlate changes in ovarian catecholamines and serum pituitary hormones and sex steroids. As the experimental model we used the prepubertal rat injected with 7.5 IU PMS on day 28 at 0900 h. Within 24 h after the injection, ovarian weight increased, and on day 30 at 2400 h, it was twice that of controls. On day 31, all of the PMS-treated rats but none of the controls had ovulated, and the average number of ova was 9. Serum estradiol in these rats was elevated significantly on the afternoon of day 30 and was followed by a preovulatory surge of LH and FSH. Serum progesterone was also increased on the evening of day 30, and an elevation in serum PRL on the morning of day 30 in the PMS-treated rats was evident, whereas serum GH was unchanged. Ovarian norepinephrine (NE) dropped from 65.5 +/- 6.9 pg/mg ovary (mean +/- SE) before the PMS injection of 14.4 +/- 1.5 pg/mg ovary on the evening of day 30. This depletion proceeded in two apparent phases: an initial drop within 12 h after PMS injection, and a further reduction coincident with the gonadotropin surge. Throughout this period, ovarian NE in the controls increased 50--60%. There was a 2-fold reduction in ovarian dopamine in the PMS-primed rats, but the levels were 10-fold lower than those of NE. The data document that the hormonal profiles and the number of ovulating follicles of the prepubertal rat primed with a low dose of PMS resemble those of the normal cycling rat. A significant depletion of ovarian catecholamines was observed during the preovulatory period, which is temporally related to increases in serum gonadotropins.

Animals↗

Fast axoplasmic transport of noradrenaline and dopamine in mammalian peripheral nerve.

A fast transport of noradrenaline (NA) at a velocity of 392 mm/day was found in cat peroneal nerve using a double-ligation technique and a new sensitive radioenzymatic assay for catecholamines. The velocity of transport of NA is sufficiently close to that of 410 mm/day found for labelled proteins and polypeptides to be considered as moving down within the nerve fibres by the same transport mechanism. In addition, dopamine (DA) was also found to be moved down by anterograde transport at a fast, but not well defined, rate. Disulfiram, a dopamine-beta-hydroxylase inhibiting agent, reduced NA levels and increased DA both in control nerve segments and within the portion of the nerve isolated by ligations where NA-containing densecore vesicles are present. The results are considered to support a dynamic turnover of NA and DA in the dense-core vesicles as they are trasnported in the axons.

Animals↗

Elevation of dopamine in fetal plasma and the amniotic fluid during gestation.

Catecholamines were determined by a radioenzymatic assay in maternal and fetal rat plasma and in the amniotic fluid during the last five days of gestation. Dopamine was significantly higher in fetal than in maternal plasma whereas norepinephrine was the same. Epinephrine was sighificantly lower in fetal than in maternal plasma on days 18 and 19, but was the same during the remainder of gestation. Dopamine was the predominant catecholamine in the amniotic fluid on days 20-22 of gestation showing the largest percent increase prior to parturition. The data suggest that dopamine may have a peripheral hormonal role during fetal development and parturition.

Amniotic Fluid↗

Dopamine in hypophysial portal plasma of the rat during the estrous cycle and throughout pregnancy.

Catecholamine levels in hypophysial portal plasma were determined in pregnant and non-pregnant female rats as well as in intact and castrated male rats, using a radioenzymatic assay for the simultaneous determination of dopamine, norepinephrine, and epinephrine in 50 mul of plasma. Portal and arterial blood were collected from anesthetized rats at 7 mul/min for 60 min. During the collection, blood was kept at 0 C, a temperature at which endogenous catecholamines were relatively stable. Dopamine was present in high concentrations in hypophysial portal plasma thorughout pregnancy, attaining a level near 20 ng/ml on the 20th day of gestation. Dopamine levels in arterial plasma from the same rats were low or undetectable (0.4--0.8 ng/ml1. Norepinephrine and epinephrine was undetectable (less than 0.6 ng/ml) in portal as well as arterial plasma from these rats. The major catecholamine in extracts of the hypothalamus from pregnant rats was norepinephrine, whereas that in the posterior pituitary was dopamine. Dopamine levels in portal plasma collected during proestrus, estrus, diestrus 1, and diestrus 2, were 1.32 +/- 0.21 (mean +/- SE), 3.87 +/- 0.96, 3.11 +/- 0.73, and 2.3 +/- 0.45, respectively. Dopamine in portal plasma from intact and from castrated male rats was approximately 0.6 ng/ml. Norepinephrine and epinephrine were not detectable in either portal or arterial plasma from these animals. It is concluded 1) that dopamine is secreted into hypophysial portal blood in significant quantities during pregnancy, 2) that hypothalamic secretion of dopamine in cyclic rats is greatest during the day of estrus and early diestrus and at least on the day of proestrus, and 3) that these findings support the view that dopamine of hypothalamic origin may have an important role in the regulation of anterior pituitary function.

Animals↗

A sensitive radioenzymatic assay for dopamine, norepinephrine, and epinephrine in plasma and tissue.

A double-isotope, radioenzymatic assay for measuring dopamine, norepinephrine, and epinephrine in one sample is described. The assay procedure includes incubation, solvent extraction, and thin-layer chromatography. Dopamine, norepinephrine, and epinephrine were incubated with catechol-O-methyl transferase (COMT) and [3H]S-acenosyl methionine ([3H]SAM) and were converted to the O-methylated tritiated derivatives: [3H]methoxytyramine, [3H]normetanephrine, and [3H]metanephrine, respectively. After several extraction steps the O-methylated products were purified by means of two-dimensional, thin-layer chromatography using silica gel. The thin-layer chromatographic system resulted complete separation of the three O-methylated compounds with an overlap of only 1-2%. The assay was linear from 0 to 5 ng for each catecholamine and had a sensitivity of 10-30 pg. The addition of large amounts of plasma reduced the activity of COMT, but increasing the magnesium concentration in the incubation mixture and the addition of EGTA to plasma samples improved the recoveries. Each sample was corrected for losses incurred during extraction and chromatography by using [14C]methoxytyramine, [14C]normetanephrine, and [14Ci1metanephrine that were added at the end of incubation. Several catechol compounds known to be O-methylated by COMT were examined for crossreactivity. Of the substances tested, only dihydroxyphenylalanine (DOPA) exhibited cross-reactivity. However, the apparent 30% cross-reactivity of DOPA with dopamine was due to the presence of decarboxylase activity in the COMT preparation. As little as 50 mul of trunk plasma from decapitated rats was sufficient for the determination of the three catecholamines.

Adrenal Glands↗

Differential sub-cellular compartmentalization of thyrotropin releasing hormone (TRH) and gonadotropin releasing hormone (LRH) in hypothalamic tissue.

Homogenates of male rat hypothalami were fractionated by differential centrifugation. Of the total quantity of TRH and LRH in the homogenate, about 50% was in particles sedimenting at 11,500 X g. and 15-20% was in particles sedimenting between 20,000 and 105,000 X g. No LRH or TRH was recovered in the 105,000 X g supernatant fluid. When the 900 X g supernatant fluid was subjected to continuous sucrose density gradient centrifugation, TRH-containing particles distributed as two peaks located at 0.9 and 1.1 M sucrose. On the other hand, LRH-containing particles distributed as only one peak located at 1.2 M sucrose. The 11,500 X g pellet contained those particles comprising the 1.2 M peak of LRH and the 1.1 M peak of TRH. Acid phosphatase activity was found in the gradient fractions containing TRH and LRH, whereas NADPH-cytochrome c reductase and cytochrome c oxidase activities were separated from the peaks of TRH and LRH. Norepinephrine, dopamine, and TRH distributed identically on the gradient. Hypo-osmotic treatment changed the gradient profile of TRH but not that of LRH. Most of the TRH was found near the top of the gradient, but a small amount of TRH was associated with particles which were lighter than those previously noted. The peak of LRH was reduced but its location on the gradient was unchanged. It is concluded that in hypothalamic homogenates TRH and LRH are localized in synaptosome-like particles which have different physical properties.

Animals↗

Transformation of 3H-dopamine during transport from CSF to hypophysial portal blood.

The capacity of the hypothalamus to transport dopamine from brain tissue into hypophysial portal blood was investigated. One hour after injection 10 muCi of 3H-dopamine into a lateral ventricle of male rats, radioactivity was present in the brain, in the anterior pituitary gland, and in systemic plasma. The highest concentration of radioactivity was found in the hypothalamus, whereas the lowest was found in systemic plasma. The cerebrum and systemic plasma contained over 50% of the dose injected, whereas the anterior pituitary gland contained no more than 0.2%. Three doses of 3H-dopamine (0.1, 1, 1nd 10 muCi) were injected into a lateral ventricle, and hypophysial portal and arterial blood were collected at 7 mu1/min for two hours. Ten min after injection, radioactivity was detected in hypophysial portal blood, reached a peak within 15-20 min, and then declined gradually to attain a concentration similar to that in arterial blood. Portal and arterial plasma and tissue extracts from rats injected intraventricularly with 3H-dopamine were subjected to gel filtration on a Sephadex G-10 comumn and to paper electrophoresis. The hypothalamus as well as the cerebrum contained several radiolabeled substances one of which appeared to behave like 3H-dopamine. However, no free 3H-dopamine was evident at this time either in blood or in the anterior pituitary gland. The results of chromatography and electrophoresis of dialyzed portal plasma indicate that much of the radioactivity was bound to macromolecules (probably plasma proteins) larger than 10,000-12,000 molecular weight. When the radioactive compound was dissociated from the macromolecule with perchloric acid, the dissociated compound did not behave as free 3H-dopamine as judged by gel filtration and electrophoresis. It is concluded that a transformation of 3H-dopamine occurred during passage from the CSF to blood and that no 3H-dopamine was present in portal blood. A large portion of the radioactivity was bound to plasma macromolecules.

Animals↗

Transport of thyrotropin-releasing hormone from cerebrospinal fluid to hypophysial portal blood and the release of thyrotropin.

The capacity of the medium eminence to transport thyrotropin-releasing hormone (TRH) from cerebrospinal fluid (CSF) to hypophysial portal blood, and the ability of TRH when introduced into a lateral ventricle to stimulate TSH release from the pituitary gland were investigated. Male rats were injected either intraventricularly or intravenously with 0, 1, 10, or 100 ng of TRH, and plasma TSH concentrations were determined at various times thereafter. TRH administration via both routes resulted in substantial release of TSH. Following intraventricular injection of TRH, there was a delay in reached maximal TSH concentration when compared with the faster elevation and faster decline in TSH concentrations which followed intravenous injection of the same dose of TRH. In a second experiment, 7 muCi of [3H]TRH were introduced intraventricularly or intravenously, and hypophysial portal and arterial blood were simultaneously collected and examined for the presence of radioactivity. The intraventricular injection of [3H]TRH resulted in a peak of radioactivity in portal blood within minutes, which was maintained for 20--30 min and then declined. The concentration of radioactivity in arterial blood from the same animals was considerably lower than that in portal blood. The intravenous administration of [3H]TRH resulted in radioactive peaks in both portal and arterial blood with a higher concentration of radioactive substances in arterial blood. However, the level of radioactivity in portal blood following intravenous injection of [3H]TRH comprised no more than 5--10% of that found following intraventricular administration of the saem dose. The data support the view that TRH is able to cross the medium eminence from CSF into hypophysial portal blood and that it is capable of stimulating the pituitary gland to release TSH.

Animals↗