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V D Ramirez

Publications and source records attributed to V D Ramirez.

At least 91 records · Page 5Linked to original sources
PubMed1986

Prolactin induces yawning and the stretch-yawning syndrome in young adult male rats.

Herein we report that subcutaneous injection of low doses of ovine prolactin (oPRL) induce yawning in young adult male rats. The most effective dose of oPRL in evoking yawning was 0.25 microgram/kg body weight (5.2 yawns/60 min at 1000 hr vs 0.3 in control animals). Doses of 0.025, 0.05, 2.5, 25, and 250 micrograms/kg were less effective. Interestingly, yawning in response to oPRL changes over the course of one circadian cycle with highest frequency at 1600 hr (11 yawns/80 min vs 2 yawns/80 min in animals injected with boiled oPRL). The onset of yawning in most oPRL-treated rats began approximately 40 min after oPRL injection, whereas with apomorphine the latency to the response was about 10 min. These results indicate that oPRL in addition to other hypophysial peptides such as ACTH and MSH can stimulate yawning. It is proposed that PRL after initial activation of the nigrostriatal dopamine system secondarily induces yawning by inhibition of this system via an autoreceptor-mediated negative feedback mechanism. This may explain the long latency to the response.

Animals↗
PubMed1986

Push-pull perfusion of the hypothalamus and the caudate nucleus in conscious, unrestrained animals.

The present results further confirm the usefulness of the push-pull cannula to study the in vivo activity of several neural systems in conscious, freely moving animals. However, it is evident that the inflammatory reaction to the local injury induced by the cannula may have different consequences in the function of a particular brain area. In the caudate nucleus, the responsiveness of the dopaminergic terminals of the nigrostriatal DA system to local infusion of AMPH was clearly reduced after the first two to three weeks post cannula implant. In contrast, in the hypothalamus of the rat and the rabbit, the spontaneous fluctuating activity of the LHRH pulse generator remained unaffected during a two to three month period when several perfusions could be performed in the same animal. Further studies will be required to establish the reasons for such marked differences in response to injury of these two brain structures. Various neurochemicals can be measured simultaneously from the hypothalamus of conscious rabbits. The present results indicate that NE hypothalamic impulse flow increases during the afternoon whereas the impulse flow of DA (as estimated by DOPAC output) decreases or does not change. In rabbit No. 2, simultaneous measurement of hypothalamic NE release and LHRH release indicates an absence of correlation. In sharp contrast, the simultaneous measurement of prostaglandin E2 release and LHRH release from the hypothalamus of rabbit 2-ER revealed a closer association between these two neuronal events. Apparently the push-pull technique can be used to examine long-term seasonal as well as circadian (extended 24 hr perfusion) spontaneous changes in the activity of the LHRH pulse generator in individual animals. It seems that seasonal changes characterize the function of the rabbit LHRH pulse generator. Lastly, physiological doses of progesterone infused for short times directly into the hypothalamus of female rabbit stimulate the LHRH pulse generator of this species with marked increases in the amplitude of the LHRH signal. In conclusion, the PPP technique coupled to HPLC-EC and RIA procedures to measure localized release of neurotransmitters, metabolites, and neuropeptides simultaneously in the same perfusate samples from discrete brain areas appears to be a powerful technique in furthering our knowledge of the in vivo local neurochemistry of the brain of conscious, freely moving animals.

Animals↗
PubMed1986

In vitro LHRH release from superfused hypothalamus as a function of the rat estrous cycle: effect of progesterone.

The present study examines the effect of progesterone (P) on in vitro LHRH release from hypothalamic fragments from intact adult rats throughout the estrous cycle. Estrous cyclicity was monitored by daily vaginal smears, and animals which exhibited at least two consecutive 4-day estrous cycles were used. Animals were sacrificed between 10.00 and 13.00 h and the mediobasal hypothalamic-preoptic area-suprachiasmatic nucleus units were removed and transferred into superfusion chambers (one unit/chamber). Following a 2-hour control period, in which the spontaneous LHRH release was established, P (10 ng/ml) was infused in an intermittent mode (10 min-on, 20 min-off). Effluents were collected at 10-min intervals and LHRH concentrations were determined by RIA. The spontaneous LHRH release from control preparations was episodic throughout all stages of the estrous cycle with a significant low release rate and low LHRH amplitude only during estrus. Interestingly, intermittent infusion of P significantly stimulated LHRH release solely in hypothalamic fragments derived from proestrous rats. The P-stimulated LHRH release during the 1st-hour period after P infusion was significantly higher (p less than 0.05) than that observed in the control preparations during the same time period as well as from its own basal pre-P values (1-hour post-P: 4.26 +/- 0.96 vs. 1-hour control and pre-P: 2.34 +/- 0.38 and 2.32 +/- 0.57 pg/10 min, respectively). P administration did not stimulate in vitro LHRH release during the other stages of the estrous cycle.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

In vivo LH-RH output of ovariectomized rats following estrogen treatment.

In the present experiment we examined the effect of estrogen upon in vivo luteinizing hormone releasing hormone (LH-RH) release from the medial basal hypothalamus of freely moving ovariectomized female rats during the period of the LH surge. Ovariectomized females (10-20 day) received two subcutaneous injections of oil or estradiol benzoate (EB; 25 micrograms/rat) at 48 and 24 h prior to push-pull perfusion and were perfused between 10:00 and 18:00 h on the 3rd, day. Unexpectedly, the activity of the LH-RH pulse generator in ovariectomized rats was characterized by a very low mean LH-RH output. Administration of EB increased the activity of the LH-RH pulse generator as indicated by a significant increase in overall mean LH-RH release of EB- versus oil-treated females. In addition, EB-treated females demonstrated a diurnal variation in LH-RH release with marginal, but significant, increases in LH-RH release during the period of the LH surge (14:10-18:00 h) as compared with levels obtained between 10:00 and 14:00 h. This increase in the overall mean release during the period between 14:10 and 18:00 h appears to be attributable to an EB-induced increase in the frequency of LH-RH release, since the amplitude did not change between the two 4-hour sampling periods.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Transient changes in the in vitro activity of the luteinizing hormone-releasing hormone pulse generator after ovariectomy in rats.

We examined in vitro LHRH release as a function of time after ovariectomy (OVX). Adult female rats (diestrous day 1 and 11-16, 61-68, and more than 120 days post-OVX) were decapitated, and the suprachiasmatic-medial preoptic area-medial basal hypothalamus was removed and superfused in vitro. There was a statistically significant (P less than 0.05) 2-fold decrease in mean in vitro LHRH release 11-16 days post-OVX that returned to levels observed in diestrous day 1 females 60 days post-OVX. LHRH cycle amplitudes of 11- to 16-day ovariectomized rats were reduced and significantly lower (P less than 0.05) than those of more than 120-day ovariectomized rats. These results indicate a complex relationship between ovarian steroid hormone removal and LHRH pulse generator function that is only revealed after direct measurement of LHRH release.

Animals↗
PubMed1986

Functional characteristics of the luteinizing hormone-releasing hormone pulse generator in conscious unrestrained female rabbits: activation by norepinephrine.

We probed the activity of the neural LHRH apparatus by means of push-pull cannulae implanted in the hypothalamus of conscious intact anestrous unrestrained New Zealand White female rabbits raised under natural light. The data revealed that the neuropeptide LHRH is released in a pulsatile manner (one pulse per 46.4 +/- 3.9 min; amplitude, 1.63 +/- 0.39 pg; n = 7) from a rather extensive area within the basal hypothalamus of the rabbit covering the rostral, medial, and posterior regions in a rostro-caudal direction and about 1 mm lateral. A remarkable change in mean LHRH output was observed during the year, and two distinct types of LHRH output were detected: low (0.72 +/- 0.07 pg/10 min; n = 7) when these reflex ovulators were perfused during late spring to early summer and high (2.92 +/- 0.29 pg/10 min; n = 5) when perfused during late summer, early fall to spring months. This 4-fold annual change in LHRH output was mainly due to changes in the amplitude of the LHRH signal and was not correlated with the position of the push-pull cannula within a restricted area in the hypothalamus. These changes may be related to the state of the ovaries, since rabbits with high LHRH output showed larger ovaries with well developed follicles than rabbits with low LHRH output. Norepinephrine, a hypothalamic neurotransmitter known to be involved in the mounting-evoked ovulation reflex in this induced ovulator, elicited a 2-fold greater mean LHRH release (P less than 0.05) when infused in doses of 10(-7) - 10(-6) M directly into the hypothalamic perfusion area. The data indicate that the activity of the LHRH pulse generator in this species presents remarkable annual variations in its mean LHRH output mainly due to a greater amplitude of the LHRH signal and that norepinephrine is a potent stimulator of LHRH release in conscious unrestrained female rabbits.

Animals↗
PubMed1985

Dibutyryl cyclic adenosine monophosphate stimulates in vitro luteinizing hormone-releasing hormone release only from median eminence derived from ovariectomized, estradiol-primed rats.

The present study examined the effect of intermittent infusion of dibutyryl cyclic AMP (dbcAMP; 10(-7) M; 10 min on, 20 min off) on in vitro luteinizing hormone-releasing hormone (LH-RH) release from the rat median eminence (ME) derived from immature rats: intact females, intact males, ovariectomized (OVX) females, castrated (CAST) males, ovariectomized, estradiol primed (OVX + E2) females and castrated, estradiol primed (CAST + E2) males. In intact, OVX and CAST conditions, spontaneous LH-RH release from MEs was not modified by dbcAMP infusion. However, E2 implants in OVX and CAST rats selectively affected the responsiveness of MEs to dbcAMP: ME from OVX + E2 were highly responsive to dbcAMP; contrarily, MEs from CAST + E2 were unresponsive to this nucleotide. Therefore, these differences in MEs responsiveness to dbcAMP-induced LH-RH release appear to be dependent upon a critical effect of E2 priming on this tissue in female but not in male rats.

Animals↗
PubMed1985

Responsiveness of immature versus adult male rat hypothalami to dibutyryl cyclic AMP- and forskolin-induced LHRH release in vitro.

In the present study, we have investigated the effects of intermittent dibutyryl cyclic AMP (dbcAMP, 5 X 10(-8) M), butyrate (5 X 10(-8) M) and forskolin (10(-4) M) on immunoreactive luteinizing hormone-releasing hormone (LHRH) release from superfused hypothalamic fragments from intact male rats of age 25, 30, 45, or 60-75 day (adult). The results indicate that at 25 days of age, male rat hypothalami were most responsive to cyclic AMP (162% of preinfusion basal LHRH release); by 30 days of age, dbcAMP also elicited increased LHRH release (120% of basal). By 45 days of age, the dbcAMP effect on in vitro LHRH release was slightly inhibitory (78% of basal); however, by adulthood, the effect of this cyclic nucleotide on LHRH release was minimal (92% of basal). Butyrate also induced age-dependent modifications in in vitro LHRH release from male rat hypothalami, with slight increases following butyrate delivery at 25 days of age (115%), slight decreases at 30 (82%) and 45 days of age (68%), and little change in adulthood (94%). This latter finding emphasizes the importance of using butyrate as a control for butyryl derivatives of cyclic AMP, which are known to liberate butyric acid as a product of hydrolysis of parent compounds. To address the possibility that the lack of effectiveness of dbcAMP in the older animal preparations was solely due to an increasing sensitivity of male rat medio-basal hypothalamus fragments to butyrate, we examined the effect of forskolin (10(-4) M), an adenylate cyclase stimulator, on LHRH release.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1985

In vitro dopamine release from the rat striatum: diurnal rhythm and its modification by the estrous cycle.

In the present experiment we examined spontaneous endogenous in vitro dopamine release from the corpus striatum of female rats during the morning (09.00-09.30 h) and afternoon (15.00-15.30 h) photoperiod on each day of the estrous cycle. In the morning, the spontaneous dopamine release rates of D-1, D-2 and proestrous female rats were characterized by initial low values which gradually increased (approximately 3-fold) over the 2.5-hour in vitro perifusion. In the afternoon, spontaneous release rates of D-1, D-2 and estrous females gradually declined (approximately 2.5-fold) over the perifusion period. This rhythmic diurnal fluctuation was disrupted in the afternoon of proestrus and morning of estrus when release rate profiles remained stable over the entire perifusion period. These results suggest that changes in spontaneous in vitro dopamine release of corpus striata derived from rats in different phases of the estrous cycle may reflect novel in vivo interactions of both photoperiodic and hormonal cues.

Animals↗
PubMed1985

In vitro luteinizing hormone-releasing hormone release from superfused rat hypothalami: site of action of progesterone and effect of estrogen priming.

The study examined the effect of estrogen priming on progesterone (P4)-induced LHRH release, the tissue site of action of P4, and the effect of 5 alpha-dihydroxyprogesterone (5 alpha-DHP) on LHRH release from hypothalamic fragments superfused in vitro. Immature female rats were ovariectomized (OVX) and, at 28 days of age, Silastic capsules containing estradiol (E2) were implanted. Two days later, animals were killed and hypothalamic fragments were removed and transferred to superfusion chambers. The hypothalamic units received P4 or 5 alpha-DHP delivered in an intermittent mode (10-min on, 20-min off). LHRH was determined in perfusates by RIA. After the input characteristics of different infusion modes (single pulses, intermittent, and continuous) of P4 infused into superfusion chambers were assessed, an intermittent infusion mode (10-min on, 20-min off) was selected for further examinations. In the mediobasal hypothalamic-anterior hypothalamic-preoptic area (MBH-AHA-POA) tissue preparations, we observed: 1) an infusion of 5 alpha-DHP was ineffective in stimulating LHRH release; 2) the release pattern of LHRH in response to three different P4 doses (10, 20, and 50 ng/ml) was similar in terms of percent changes (202% to 219% over control values); and 3) E2 priming was absolutely required for P4-stimulated LHRH release, and this requirement appeared to be dose dependent. Upon an examination of three hypothalamic tissue boundaries [the MBH, the POA-suprachiasmatic nuclei (POA-SCN), and the median eminence (ME)] to better delineate the in vitro site of action of P4 on LHRH release, it was demonstrated that the MBH responded upon P4 infusion, whereas the POA-SCN was unable to do so. The ME also responded upon P4 infusion, and LHRH release followed closely the pulsatile administration of P4 since upon each challenge of the steroid at the concentration of 10 or 20 ng/ml, a significant rise in LHRH release occurred. However, the temporal patterns of LHRH release from the ME appears to be different from those obtained from the MBH as well as the MBH-AHA-POA. These observations demonstrate that an intermittent infusion of P4, but not 5 alpha-DHP, is effective in activating the neural LHRH apparatus. Estrogen is an obligatory requirement for this P4-stimulated LHRH release, and the neural site of action of P4 resides within the MBH. However, this steroid also can act directly upon the ME nerve terminals to release LHRH.

Algestone↗
PubMed1985

In-vivo activity of the LH-releasing hormone pulse generator in castrated and intact male rats.

The in-vivo LH-releasing hormone (LHRH) output from the mediobasal hypothalamus of conscious freely moving sham-castrated, acutely castrated (immediately after castration) and long-term castrated (greater than 20 days after castration) adult male rats has been studied. Five rats in each of the three conditions were perfused for an 8-h period (11.00-19.00 h). One animal in each condition was perfused for a 24-h period. In the sham-castrated and acutely castrated, but not in the long-term castrated rats, apparently random surges of LHRH output (greater than 5.6 pmol/l), separated by periods of non-detectable activity, were observed throughout the perfusion period. The LHRH output of the long-term castrated rats was characterized by an extremely low overall mean release and markedly attenuated pulse amplitudes, with both parameters significantly lower than those of the sham-and acutely castrated rats. These results indicate that the raised blood levels of LH in long-term castrated rats do not appear to be the consequence of high amplitude and high overall release of LHRH.

Animals↗
PubMed1984

Poststimulatory endocytosis, microvesicle repopulation and changes in smooth endoplasmic reticulum in nerve endings of the median eminence superfused in vitro.

Mediobasal hypothalami of adult rats were superfused in vitro. A single 5 min pulse of 60 mM KCl-containing medium was infused, followed by either 15, 30, 45, 60 or 75 min superfusions with standard medium. In some experiments, 5 or 10% dextran was added followed by a 15 min recovery. Morphologically, two recovery phases were recognized. The early phase (15-30 min) was characterized by two features: (1) A clear-cut increase in the quantity of large, pleomorphic vacuoles occupying the axoplasm of nerve endings; these vacuoles were observed to be connected to caveolae of the same diameter in the axolemma and they were either coated or uncoated. (2) Progressive increase in the quantity of microvesicles (synaptic vesicles) from an initial depleted state. The vacuoles were found to contain dextran aggregates. Microvesicle-like protrusions bulged from the membrane of vacuoles. The late phase, from 45 min poststimulation onward, was typically identified after the appearance of tubules of smooth endoplasmic reticulum at the most distal segments of the nerve terminals. During this period, large vacuoles tended to decrease in quantity. Granular vesicles remained scant during the entire observation period. Images suggesting formation of microvesicles from tubules of smooth endoplasmic reticulum were observed. These results open the possibility that endocytosis of patches of membranes forming large vacuoles be an important mechanism for retrieving the membranes belonging to microvesicles and granular vesicles. Some of these large vacuoles may contribute to the early regeneration of microvesicles. More microvesicles could later be produced from the smooth endoplasmic reticulum.

Animals↗
PubMed1984

In vivo output of dopamine and metabolites from the rat caudate nucleus as estimated with push-pull perfusion on-line with HPLC-EC in unrestrained, conscious rats. I. Chromatographic and biological validation.

In the present experiment we used push-pull perfusion (PPP) on-line with high performance liquid chromatography with electrochemical detection (HPLC-EC) to measure the concentration of neuroactive substances collected in perfusates from the caudate nucleus (CN) of conscious, freely moving rats. To validate the suitability of such an approach, both chromatographic and biological procedures were used. The chromatographic performances of four pure standards, dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxyindole acetic acid (5-HIAA) were examined under the conditions of the experiment and the release of these four chemicals by amphetamine (AMPH) locally infused into the CN or systemically administered to conscious rats used as an index of biological validation. Distinct dose-response curves were obtained for each standard injected into the HPLC-EC singly or mixed together in perfusion medium (modified Krebs-Ringer's Phosphate, KRP , pH 7.4). Moreover, each standard in the chromatogram appeared as a well-defined elution band with a different retention time. The brain perfusate samples did not contain factors interfering with the normal operation of the HPLC-EC or measurement of the concentration of added standards. The recovery of pure DA, DOPAC, 5-HIAA and HVA added to the perfusate samples was 97, 87, 98 and 114%, respectively. No decrements in peak heights were observed in the chromatograms when a 1-ng dose mixture of the four standards dissolved in medium and maintained at 4 degrees C was injected into the HPLC-EC at regular intervals for a 60-min period after initial preparation.(ABSTRACT TRUNCATED AT 250 WORDS)

3,4-Dihydroxyphenylacetic Acid↗
PubMed1984

Bimodal effect of progesterone on in vitro dopamine function of the rat corpus striatum.

In the present experiment we examined the temporal effects of progesterone (P) upon in vitro dopamine (DA) release from superfused corpus striatum (CS) and medial basal hypothalami (MBH) tissue fragments. In ovariectomized-estrogen-primed adult female rats, P was systemically administered at 0.5, 2, 4, 12 or 24 h prior to sacrifice for CS and at 4 or 24 h for MBH superfusions. Control groups receiving either steroid alone and/or the oil vehicle were included. Progesterone at the 2- to 12-hour periods stimulated spontaneous and amphetamine-evoked DA release and increased post-superfusion tissue concentrations of DA from the CS when compared to controls. At 24 h P apparently produced an active inhibition, with all parameters of DA activity (spontaneous and AMPH-induced DA release), significantly decreased compared to controls. In contrast, P failed to alter the spontaneous or marginal amphetamine response of the MBH but did significantly reduce post-superfusion DA concentration at 24 h. We hypothesize that the dual effect of P upon brain dopaminergic systems (facilitation followed by inhibition), in the CS but not in the MBH represents changes in DA synthesis and release which are coupled in the CS but not in the MBH.

Amphetamine↗
PubMed1983

Localized and discrete changes in neuropeptide (LHRH and TRH) and neurotransmitter (NE and DA) concentrations within the olfactory bulbs of male mice as a function of social interaction.

Individually housed male mice were exposed to either an intact male or an ovariectomized female mouse for 1 min and decapitated at 5, 15, or 60 min to examine the hypothesis whether discrete changes in olfactory bulb neuropeptide (LHRH and TRH) and neurotransmitter (NE and DA) concentrations would occur following onset of exposure. A nonexposed control group (decapitated at time 0) was also included. Bilateral olfactory bulbs were dissected into anterior dorsal (ADOB) and posterior dorsal (PDOB) olfactory bulb fragments and prepared for radioimmunoassays (LHRH and TRH) or radioenzymatic assays (NE and DA). Concentrations of LHRH and NE, but not of TRH and DA, from the PDOB were significantly greater than those of ADOB fragments. Exposure to a male resulted in a significant increase of PDOB LHRH at 5 min following exposure and a significant increase in LHRH at 15 min following female exposure. Norepinephrine within the ADOB and PDOB and DA within the PDOB demonstrated a statistically significant increase at 60 min following exposure to an ovariectomized female. In marked contrast, no statistically significant changes were obtained following male exposure. These results not only demonstrate a preferential localization of neuroregulators within the olfactory bulb of male mice but discrete changes in the concentration of these neuroregulators in response to male or female exposure, suggesting the possibility that some processing and coding of chemical cue information during social encounters already occurs at the level of the olfactory bulb.

Animals↗