Biomedical subjects

V D Ramirez

Publications and source records attributed to V D Ramirez.

At least 55 records · Page 3Linked to original sources

PubMed1990

Effects of progesterone and its metabolites on neuronal membranes.

Evidence supporting a membrane site of action for progesterone includes the rapidity of its effects when directly infused into tissue containing mainly nerve terminals, the absence of functional intracellular progesterone receptors in vitro and the fact that progesterone conjugated to bovine serum albumin (BSA) in the C-3 position (P-3-BSA) activates the release of hypothalamic luteinizing hormone releasing hormone (LHRH) or modulates amphetamine-evoked striatal dopamine release. In addition, P2 membrane fractions from different areas of the CNS but not P1 fractions or P2 membranes from peripheral progesterone targets have specific binding sites for P-11-125I-BSA. Among several BSA-conjugated steroids tested for competition displacement P-3-BSA had the highest affinity with an estimated inhibition constant of 28.5 +/- 2.1 nM. This binding depends on the presence of cations such as Ca2+ and Mg2+ and after chemical depolarization of the P2 membranes the binding curve of P-3-BSA shifts to the right. While progesterone is effective in releasing LHRH from the hypothalamus, 5 beta-pregnan-3 beta-ol-20-one (a 5 beta reduced metabolite) is at least 1000-fold more potent than the parent compound when tested in vitro and in vivo. This action is indirect because tetrodotoxin at 10(-6) M blocks the LHRH releasing action, although 5 beta-prenan-3 beta-ol-20-one is still capable of releasing noradrenaline. Although 5 beta-pregnan-3 beta-ol-20-one can replace progesterone in activating the LHRH neural apparatus this is not true for the nigro-striatal dopamine system where only progesterone or P-3-BSA is effective, an action which is also indirect since tetrodotoxin blocks the effect of either compound. These results indicate that progesterone acts at membrane sites to modulate specific functions of the CNS and that site-specific mechanisms exist within the CNS which may differentially control its conversion to more active compounds.

Animals↗

PubMed1990

Binding of progesterone to nerve cell membranes of rat brain using progesterone conjugated to 125I-bovine serum albumin as a ligand.

Radioiodinated bovine serum albumin conjugated to progesterone was used as a probe to examine binding parameters of steroids to membrane preparations from rat brain tissue. The binding of 11 alpha-hydroxyprogesterone-11-hemisuccinate-125I-bovine serum albumin conjugate reached saturation after 30 min of incubation at 5 degrees C. Several bovine serum albumin-conjugated steroids were then tested for competition displacement studies. Among these steroid conjugates, the bovine serum albumin conjugate at position 3 of progesterone had the highest affinity, with an estimated inhibition constant of 28.5 +/- 2.1 nM (n = 3), whereas bovine serum albumin itself and the 17 beta-estradiol 6-(O-carboxy-methyl)oxime-bovine serum albumin conjugate showed no specific displacement. In addition, the binding sites were localized in an axolemma-enriched fraction of rat brainstem. Specific binding was obtained in tissues from cerebral cortex, brainstem, cerebellum, corpus striatum, and hypothalamus, but little or no binding occurred in uterus, ovary, liver, and spleen. The present data indicate that progesterone-125I-bovine serum albumin conjugate can be used as a ligand to study progesterone-membrane receptor interactions.

Animals↗

PubMed1990

In vitro progesterone modulates amphetamine-stimulated dopamine release from the corpus striatum of castrated male rats treated with estrogen.

Previous work from our laboratory has indicated that a direct infusion of progesterone (P) into superfusion chambers containing corpus striatum (CS) tissue fragments of ovariectomized/estrogen-treated female rats augmented amphetamine-stimulated dopamine release in vitro. In this report, we examine whether this phenomenon is also present in the male rat. Only in castrated male rats treated with estrogen were we able to demonstrate an increase in amphetamine-stimulated dopamine release in response to a direct infusion of P. Intact and castrated male rats treated with testosterone propionate or vehicle failed to show any significant differences in response to P. These results demonstrate that the CS of male rats can respond to P only if males are exposed to an estrogenic hormonal milieu and indicate an absence of a sexually dimorphic response of the CS to P.

Amphetamine↗

PubMed1990

Infusion of progestins into the hypothalamus of female New Zealand white rabbits: effect on in vivo luteinizing hormone-releasing hormone release as determined with push-pull perfusion.

Previously, we have reported that intermittent infusion of progesterone (P4; 10 ng/ml) into the hypothalamus of conscious unrestrained female New Zealand White rabbits stimulates LHRH release in vivo. To further investigate this phenomenon, in the present studies, the effect of pulsatile (six pulses; 10 min on, 30 min off) and continuous infusion of P4 (Exp I) and 4-pregnen-20 alpha-ol-3-one or 20 alpha-hydroxyprogesterone (20 alpha-OH-P; Exp II) on LHRH release were studied in vivo using push-pull cannulae. Furthermore, the effect of pulsatile infusion (six pulses; 10 min on, 30 min off) of low doses of the following three progestins [5 beta-pregnan-3 beta-ol-20-one (pregnanolone), 5 alpha-pregnan-3 alpha-ol-20-one (3 alpha-5 alpha-P), and 20 alpha-OH-P] into the hypothalamus of does using push-pull cannulae were examined in Exp III. In Exp I, continuous infusion of P4 at 10 ng/ml was unable to stimulate mean LHRH release. However, pulses of P4 at 0.01 ng/ml (n = 4) were found to significantly increase the amplitude of the largest LHRH pulse (control period, 1.18 +/- 0.41; versus treatment period, 3.15 +/- 0.75 pg; P less than 0.035) as well as the frequency of LHRH pulses (control period, 0.72 +/- 0.26; treatment period, 1.37 +/- 0.12 pulses/h; P less than 0.035). On the other hand, there was no effect of pulses of P4 at 0.001 ng/ml (n = 4) on the activity of the LHRH neural apparatus. In Exp II, pulses of 20 alpha-OH-P at 10 ng/ml (n = 4) were found to significantly increase the mean LHRH release (control period, 1.24 +/- 0.10; treatment period, 2.07 +/- 0.52 pg/10 min) as well as the amplitude of the largest LHRH pulse (control period, 0.99 +/- 0.36; treatment period, 8.15 +/- 3.75 pg). Interestingly, continuous infusion of 20 alpha-OH-P (10 ng/ml) also significantly increased the mean amplitude (control period, 0.58 +/- 0.34; treatment period, 1.90 +/- 0.29 pg) as well as the amplitude of the largest LHRH pulse (control period, 0.64 +/- 0.37; treatment period, 4.30 +/- 1.97 pg).(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1989

Progesterone enhances L-dopa-stimulated dopamine release from the caudate nucleus of freely behaving ovariectomized-estrogen-primed rats.

In the present experiment we examined the effect of progesterone upon dopamine (DA) release induced by a direct infusion of unlabeled L-dihydroxyphenylalanine (L-DOPA) into the caudate nucleus of freely behaving rats. Ovariectomized rats were implanted with a push-pull cannula directed at the caudate nucleus and subjected to perfusion under 3 different hormonal conditions: (1) following 4 days of treatment with estradiol benzoate (EB), (2) following 4 days of treatment with estradiol benzoate plus progesterone at 4-6 h prior to perfusion (EB + P-4-6 h) and (3) following 4 days of treatment with estradiol benzoate plus progesterone at 28 h prior to perfusion (EB + P-28 h). During each perfusion session and under each of the 3 hormonal treatment conditions, L-DOPA was infused through the push side of the cannula. Three increasing doses of L-DOPA (10(-6), 10(-5) and 10(-4) M) were infused with a 45-75 min interval between infusions. Regardless of hormonal treatment condition, a clear dose-response increase in DA and 3,4-dihydroxyphenylacetic acid (DOPAC), but not 5-HIAA, output was observed in response to the increasing doses of L-DOPA infusion. For each of the 3 doses of L-DOPA, maximal DA output was observed for animals tested under the EB + P-4-6 h hormonal condition, with statistically significant differences in the areas under the L-DOPA-stimulated DA response curves obtained following the 10(-6) and 10(-5) M doses of L-DOPA infusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1989

Effects of prolactin on tyrosine hydroxylase activity of central dopaminergic neurons of male rats.

Previous work from our laboratory has indicated that ovine prolactin can produce a rapid stimulation (within 1 h) of dopamine release from nigrostriatal and mesolimbic dopaminergic neurons of male rats. In the present experiment we attempted to determine whether this effect of prolactin was a function of an increase in the activation of dopamine synthesis. To examine this possibility we used the drug NSD 1015. This drug is a decarboxylase inhibitor that leads to dopa accumulation and is used as an index of in vivo tyrosine hydroxylase activity. Our results demonstrated that ovine prolactin increased turnover of dopamine but had no effect upon tyrosine hydroxylase activation (up to 4 h) in both dorsal and ventral striatal dopaminergic neurons. In contrast, ovine prolacting had a clear effect (within 4 h) on tyrosine hydroxylase activity in mediobasal hypothalamic dopaminergic neurons. These results suggest that ovine prolactin can differentially alter synthesis/release processes of dopamine from these sites.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1989

Studies of the in vivo catabolism of exogenous dopamine as infused through a push-pull cannula implanted in the rat caudate nucleus.

In this report, using a push-pull perfusion technique, we examined in vivo the effects of exogenous dopamine (DA) on the output of neurochemical substances from the caudate nucleus (CN) of freely behaving rats. Exogenous DA, at concentrations of 10(-5) M, 5 x 10(-5) M and 10(-4) M, in modified Krebs-Ringer phosphate medium (KRP) was infused directly into the CN for 15 min each. Exogenous DA at the doses tested elicited increases in 3,4-dihydroxyphenylacetic acid (DOPAC) output in a dose-dependent manner. In addition, the higher two doses of exogenous DA also induced increases in homovanillic acid (HVA) output from the rat CN. The increases in DOPAC output by 5 x 10(-5) M DA was partially blocked by the inclusion of 10(-3) M nomifensine in KRP. Interestingly, exogenous DA-induced increases in HVA output were little affected by the nomifensine treatment. However, the catabolism of exogenous DA was almost completely eliminated by a prior 6-hydroxydopamine lesion in the ipsilateral substantia nigra. Furthermore, infusions of exogenous DA did not change 5-hydroxyindoleacetic acid output from the CN. In conclusion, our results confirm in vivo that (a) DA catabolic pathway via DOPAC intermediate predominates over the alternative pathway via 3-methoxytyramine, (b) increases in extracellular DA will lead to increases in DOPAC and HVA levels in extracellular space and (c) the majority of the DA is oxidized by intradopaminergic monoamine oxidase.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1989

Progesterone effects upon dopamine release from the corpus striatum of female rats. I. Evidence for interneuronal control.

In the present experiment, we examined whether progesterone modifies dopamine (DA) release from superfused corpus striatal (CS) tissue fragments of ovariectomized estrogen-primed rats as result of a direct interaction with dopaminergic nerve terminals or through interneurons within the CS. In Expt. I, an in vitro pulsatile infusion of progesterone (2 ng/ml) directly into superfusion chambers containing CS fragments resulted in a statistically significant increase in DA release (P less than 0.008). No such stimulation was obtained following a continuous progesterone infusion or during control superfusions receiving no progesterone. In an effort to examine a direct action of progesterone on DA terminals in Expt. II, tetrodotoxin (TTX, 1 microM) was added to the Superfusion medium to block the activity of interneurons within the CS. TTX completely abolished the effect of pulsatile progesterone on DA release. These results indicate that the stimulatory action of progesterone on in vitro DA release from CS fragments is apparently exerted through activation of interneurons which discriminately respond to a specific (pulsatile) mode of progesterone infusion.

Amphetamine↗

PubMed1989

Progesterone effects upon dopamine release from the corpus striatum of female rats. II. Evidence for a membrane site of action and the role of albumin.

In the present experiment we used immobilized progesterone linked to bovine serum albumin (P4-3-BSA) as a probe to examine whether the effects of a direct in vitro infusion of progesterone upon dopamine (DA) release from corpus striatal (CS) tissue fragments from ovariectomized estrogen-treated rats may be attributable to a surface membrane site of action. In Expt. I, a direct in vitro pulsatile infusion of P4-3-BSA resulted in two discrete episodes of stimulated DA release which were not observed in superfusions receiving a continuous infusion of P4-3-BSA or compared to data of control superfusions. In contrast to that of a pulsatile administration, a continuous infusion of P4-3-BSA completely abolished the amphetamine-stimulated response from these tissue preparations with significantly lower DA release rates compared to the pulsatile P4-3-BSA (P less than 0.02) and control (P less than 0.04) conditions. In Expt. II, the addition of tetrodotoxin (TTX, 1 microM) to the superfusion medium abolished the discriminatory response between pulsatile and continuous administration of immobilized progesterone. These results indicate that the action of progesterone on DA release from the CS is mediated primarily through a surface membrane site of an interneuron(s) which can discriminately respond to a specific infusion mode of this steroid.

Albumins↗

PubMed1989

The effect of L-dopa upon in vitro dopamine release from the corpus striatum of young and old male rats.

In the present experiments, we examined the effects of superfusion of L-DOPA upon in vitro dopamine (DA) release from the corpus striatum (CS) in young (2-4 months) and old (24 months) male rats. In Experiment I, responses to two successive increasing doses of L-DOPA (1.0 and 10 microM) indicated a clear age difference with DA release from CS of young rats significantly greater than that of old rats in response to the 10 microM dose of L-DOPA. Interestingly, CS fragments of young and old male rats responded marginally to the 1.0 microM L-DOPA infusion, and no significant differences between these two age groups were obtained. Since we have previously observed that the addition of naloxone to the superfusion medium restored the potassium stimulated DA release of old rats to that of young rats, in Experiment II, responses of CS tissue fragments of young and old rats superfused in medium with or without naloxone (100 microM) were tested with a single 10 microM L-DOPA infusion. The results of Experiment II indicated that the addition of naloxone to the superfusion medium did not alter the responses of CS tissue to L-DOPA in either young or old rats; however, the overall response of the young rats remained significantly greater than that of the old rats. Taken together, these results demonstrate a significant age-related decrement in the capacity of 10 microM L-DOPA to stimulate DA release from CS fragments.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

PubMed1989

Receptive female rats stimulate norepinephrine release from olfactory bulbs of freely behaving male rats.

In vivo perfusion of the olfactory bulbs (OB) of freely behaving male rats by means of miniaturized push-pull cannulae (PPC) revealed consistently detectable and relatively stable levels of norepinephrine (NE) output. Introduction of a receptive female rat into the male's cage on three separate occasions resulted in a threefold increase in NE output following the second and third, but not the first exposure. No increase in NE was obtained when perfused males were similarly exposed to male rats or in the absence of social stimulation. These results demonstrate a complex, discriminatory in vivo response of the OB noradrenergic system in male rats which may be critically involved in the receipt and processing of socially stimulated memory/recognition cues related to reproduction.

Animals↗

PubMed1989

Spontaneous changes in LHRH release during the rat estrous cycle, as measured with repetitive push-pull perfusions of the pituitary gland in the same female rats.

In the present study, we examined the in vivo release profiles of luteinizing hormone-releasing hormone (LHRH) during the rat estrous cycle by utilizing repetitive push-pull perfusions of the anterior pituitary in the same freely behaving animal. Throughout the estrous cycle, LHRH levels were higher than those reported from previous work using a push-pull cannula placed in the mediobasal hypothalamus. A significant increase in LHRH input to the anterior pituitary was obtained in proestrus (Pro), though the magnitude of increment was different among animals. This LHRH increment in Pro was attributable to a significant increase in the amplitude of LHRH pulses. Interestingly, no changes were observed in the frequency of LHRH pulses (about 1 pulse/50 min) throughout the estrous cycle. The data demonstrate that the anterior pituitary of female rats receives an increased amount of LHRH during Pro due to changes in the amplitude of the LHRH signal without changes in the frequency.

Animals↗

PubMed1988

A potent dopamine-releasing factor is present in high concentrations in the rat adrenal gland.

The present study shows that a novel, protease-sensitive factor present in a partially purified preparation from the rat adrenal gland selectively stimulates the release of dopamine from the rat striatal tissue superfused in vitro in a dose-dependent manner. Biological activity is also found to be present in much lower concentrations in the neocortex, striatum and cerebellum but absent in liver or spleen. This putative dopamine releasing factor is probably a glycoprotein, resistent to boiling, partially inactivated by trypsin and completely inactivated by the non-specific protease pronase E.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1988

Comparison of the effect of prolactin on dopamine release from the rat dorsal and ventral striatum and from the mediobasal hypothalamus superfused in vitro.

In this paper, by means of a superfusion technique, we have examined the effect of ovine prolactin on dopaminergic neurons innervating the dorsal striatum, ventral striatum and the mediobasal hypothalamus of male rats. Fragments from dorsal striatum were superfused with ovine prolactin dissolved in normal medium (Krebs-Ringer phosphate) or medium containing tetrodotoxin (TTX, 10(-6) M). Ovine prolactin stimulated the in vitro release of dopamine from dorsal and ventral striatal fragments. In dorsal striatal fragments a linear dose-dependent dopamine release was observed only when fragments were superfused with Krebs-Ringer phosphate-TTX medium. In addition, [Leu5]enkephalin (10(-6) and 10(-5) M) decreased the prolactin-induced in vitro dopamine release from dorsal striatal fragments superfused with Krebs-Ringer phosphate-TTX medium. Ovine prolactin (10(-9)-10(-5) M) did not elicit changes in dopamine, 3,4-dihydroxyphenylacetic acid or 5-hydroxyindoleacetic acid outputs from mediobasal hypothalamic fragments superfused with Krebs-Ringer phosphate medium containing TTX. The possible regulatory mechanisms of ovine prolactin on dopaminergic neurons are discussed.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1988

The importance of striatal interneurons in age-related effects upon potassium- and amphetamine-stimulated dopamine release.

The ability of superfused corpus striatal tissue fragments to release endogenous dopamine (DA) in response to potassium (K+ 30 mM) stimulation was significantly attenuated in 18- to 24-month- compared to 2- to 4-month-old male rats. These age-related effects on K+ stimulation were completely abolished with the addition of tetrodotoxin (1 microM) to the superfusion medium. Moreover, no difference in stimulated DA release was obtained between these two age groups following amphetamine stimulation (10 microM).

Aging↗

PubMed1988

Effect of chronic administration of progesterone on the naloxone-induced LHRH release from hypothalami of ovariectomized, estradiol-primed prepubertal rats.

To test the hypothesis that progesterone (P) administered in a high dose and for a long duration would influence in vitro LHRH release by naloxone, silastic capsules containing P (50 mg/ml) or vehicle were implanted sc in ovariectomized, estradiol-primed (OVX+E) prepubertal rats. Following a 48 hr exposure to P, rats were sacrificed and mediobasal hypothalamic (MBH) fragments were obtained. In addition, in other OVX+E primed rats, a single injection of P (1 mg) was given sc 6 hr prior to decapitation. Naloxone or control medium were infused into superfusion chambers containing MBH fragments derived from these animals for 2 hr following a 1 hr control period. Infusion of naloxone (1 x 10(-4) M) markedly stimulated in vitro LHRH release from MBH from rats pre-exposed to P for 48 hr, whereas it was unable to do so in other groups examined. These data clearly indicate that the ability of naloxone (1 x 10(-4) M) to stimulate LHRH release is dependent on the duration of P administration.

Animals↗

PubMed1988

Prolactin-induced yawning behavior requires an intact nigro-striatal dopamine system.

Herein, we evaluate the importance of the nigro-striatal dopamine system in prolactin-, apomorphine-, and physostigmine-induced yawning behavior. Bilateral 6-OH-dopamine lesions of the substantia nigra were performed on male rats (2-4 months old). The lesioned as well as control rats were injected with either physiological saline, physostigmine (200 micrograms/kg), apomorphine (50 micrograms/kg), or ovine prolactin (0.25 micrograms/kg) 72 hours after the surgical procedure. The results show that bilateral lesions of the substantia nigra did not affect physostigmine-induced yawning whereas both apomorphine- and prolactin-induced yawning were reduced by the lesion. Following the observation period the caudate nuclei were removed and analyzed for dopamine (DA) and dihydroxyphenylacetic acid (DOPAC) content. The lesions reduced DA and DOPAC content in all treatment groups compared to the respective intact groups. Also, both DA and DOPAC concentrations were lower in the intact apomorphine and prolactin treated groups compared to intact saline controls, at times that were temporally related to the display of yawning behavior suggesting a decrease in dopamine activity following apomorphine and prolactin treatment. Interestingly, DA and DOPAC concentrations were higher in the lesioned apomorphine group compared to lesioned saline controls; however, in the lesioned prolactin group only the DA concentrations were higher when compared to lesioned saline controls. These results indicate that prolactin- and apomorphine-induced yawning require an intact nigro-striatal dopamine system and that these substances induce yawning by different mechanisms.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1988

In vivo dopaminergic activity from nucleus accumbens, substantia nigra and ventral tegmental area in the freely moving rat: basal neurochemical output and prolactin effect.

In this paper, by means of a push-pull perfusion technique, we have examined the local effect of ovine prolactin (oPRL) on the spontaneous in vivo dopaminergic (DAergic) activity of the nucleus accumbens (NAcc), substantia nigra (SN) and ventral tegmental area (VTA) of living male rats. First, we found a 2-fold spontaneous rise of 3,4-dihydroxyphenylacetic acid (DOPAC) output during the afternoon hours of the photoperiod only in the NAcc. In addition, native oPRL (10 ng/microliters for 20 min), but not boiled oPRL, perfused into the NAcc caused a significant elevation in DOPAC output without changes in homovanillic acid or 5-hydroxyindoleacetic acid. In a similar preparation, male rats bearing push-pull cannulae in the SN or VTA were infused locally with oPRL (10 or 50 ng/microliters) for 20 min. The hormone did not alter the efflux of DOPAC from these DA cell body/dendrite areas. These results indicate that oPRL can activate the mesolimbic DA system through local actions on DA presynaptic terminals of the NAcc without affecting the SN or VTA.

3,4-Dihydroxyphenylacetic Acid↗