Biomedical subjects

D K Sarkar

Publications and source records attributed to D K Sarkar.

At least 55 records · Page 3Linked to original sources

PubMed1994

Comparison of the effects of alcohol and acetaldehyde on proopiomelanocortin mRNA levels and beta-endorphin secretion from hypothalamic neurons in primary cultures.

The effects of acute and chronic treatments with ethanol and acute treatments with an ethanol metabolite, acetaldehyde, on proopiomelanocortin (POMC) mRNA expression were compared with those of these agents on the secretion of a POMC gene product, beta-endorphin (beta-EP) peptide. The level of POMC mRNA in cultured cells was determined using an RNase protection assay, and the accumulation of immunoreactive beta-EP (IR-beta-EP) peptide in the culture medium was measured by radioimmunoassay. Treatment of hypothalamic cells with 25-, 50-, and 100-mM doses of ethanol or 12.5 and 25 microM acetaldehyde for 3 h increased POMC mRNA levels. The stimulatory effect of ethanol on POMC mRNA levels lasted for a period of 12 h, although the percentage increase of the ethanol-stimulated mRNA level was gradually reduced over time. Acute treatments with ethanol and acetaldehyde also elevated IR-beta-EP secretion from the cultured neurons for a period of 12 h, and the IR-beta-EP secretory response developed desensitization after 24 h of ethanol incubation. The close association between the ethanol-induced IR-beta-EP secretion and ethanol-regulated POMC mRNA expression suggests that ethanol regulates both secretion and production of beta-EP peptide in the hypothalamic neurons.

Acetaldehyde↗

PubMed1994

Effects of chronic alcohol on immunoreactive beta-endorphin secretion from hypothalamic neurons in primary cultures: evidence for alcohol tolerance, withdrawal, and sensitization responses.

Endogenous opioid peptides are known to be involved in the alcohol tolerance and dependence following alcohol abuse. However, the cellular mechanisms involved in the ethanol tolerance and dependence are not well established. We have previously shown that low concentrations of ethanol stimulate immunoreactive beta-endorphin (IR-beta-EP) release from the cultured hypothalamic neurons and that chronic ethanol exposure desensitizes these neurons to ethanol challenges. In this study, we determined the IR-beta-EP response to increasing doses of ethanol during the desensitizing phase of moderate ethanol doses to test whether the cultured IR-beta-EP-secreting neurons develop tolerance to ethanol following constant exposure. We also determined IR-beta-EP responses following withdrawal from chronic ethanol challenge and compared the IR-beta-EP secretory response to various doses of ethanol in ethanol-naive and ethanol-preexposed cultures. The IR-beta-EP responses to increasing doses of ethanol (50-150 mM) were markedly reduced in the cultures preexposed to a 50 mM dose of ethanol when compared with those that were naive to ethanol. The ethanol-exposed cultures showed hypersecretion of IR-beta-EP after removal from 48 hr of constant ethanol, as compared with ethanol-naive cultures. When ethanol-preexposed cultures were challenged with various doses of ethanol 4 days after ethanol withdrawal, the cultures showed higher IR-beta-EP secretory responses than did the ethanol-naive cultures. These data suggest that IR-beta-EP secretory neurons in primary cultures develop tolerance to chronic ethanol, show withdrawal response after removal of chronic ethanol exposure, and develop sensitization following repeated ethanol challenges.

Alcohol Withdrawal Delirium↗

PubMed1993

Effect of alcohol, acetaldehyde, and salsolinol on beta-endorphin secretion from the hypothalamic neurons in primary cultures.

The effect of ethanol, acetaldehyde, and salsolinol on hypothalamic beta-endorphin secreting neurons is studied by using rat fetal hypothalamic neurons in primary culture. Exposure of these neuronal cells to different concentrations of ethanol (12.5-50 mM) and acetaldehyde (12.5-50 microM) caused a concentration-dependent increase in the secretion of beta-endorphin. Salsolinol (12.5-50 microM) did not cause any significant change in the secretion of beta-endorphin. Ethanol's effect was short-lasting (2 hr). Acetaldehyde's effect on beta-endorphin secretion was greater and longer lasting, as compared with ethanol. Ethanol and salsolinol do not have any effect on cell viability, whereas higher concentrations of acetaldehyde appear to reduce the number of viable cells after 6 hr of treatment. None of the above treatments has any effect on cellular DNA content. These results suggest that ethanol is a potent stimulator of hypothalamic beta-endorphin. These results also show for the first time that ethanol's metabolite acetaldehyde is more potent in stimulating beta-endorphin secretion and may be significant in the ethanol regulated beta-endorphin secretion.

Acetaldehyde↗

PubMed1993

Beta-endorphin regulation of LHRH release at the median eminence level: immunocytochemical and physiological evidence in hens.

We studied the effect that beta-endorphin (beta END) might have at the median eminence (ME) on luteinizing hormone (LH)-releasing hormone (LHRH) during the ovulatory cycle of domestic hens. Thus, we assessed (a) the immunocytochemical distribution of beta END and LHRH in the hen ME, (b) the temporal changes in ME and preoptic area (POA) LHRH and beta END content, in both a spontaneous and a premature C2 ovulatory model. The premature C2 ovulation occurs 6-7 h after the administration of progesterone (P4) injected 14 h before the spontaneous second (C2) ovulation of a sequence and therefore 7-8 h earlier than expected, (c) the ME in vitro release of beta END in both models, and (d) the effect of beta END and naloxone on in vitro ME-LHRH release in the two models. In the hen, beta END cell bodies are located in the periarcuate area with axons projecting to both the ME and the POA. LHRH perikarya are located in the medial POA and anterior hypothalamus and project to the ME and infundibulum through the ventrolateral hypothalamus. In the spontaneous C2 ovulatory model, both beta END and LHRH content in the ME remained unchanged during the 14 h preceding the C2 ovulation. However, POA-LHRH content was increasing at the time of the LH surge (4 h before the expected C2 ovulation) and remained elevated until the C2 ovulation occurred. In contrast, POA-beta END content was lowest at the time of the LH surge and remained low until the C2 ovulation occurred.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Feed restriction in prepubertal lambs: effect on puberty onset and on in vivo release of luteinizing-hormone-releasing hormone, neuropeptide Y and beta-endorphin from the posterior-lateral median eminence.

The exact nature of the interaction between energy balance and reproduction is still elusive. Theoretically, nutrition-related variables must reach the hypothalamic luteinizing-hormone-releasing hormone (LHRH) network and/or its neuronal inputs, to alter plasma luteinizing hormone (LH) and therefore reproductive activity. In an attempt to assess the potential mechanism of such interaction at the median eminence (ME) level, the area of hypophysiotropic LHRH neuronal terminals and release, we used a decreased caloric intake lamb model which delays the onset of puberty. Thus, we determined the in vivo release of neuropeptides, by push-pull cannula (PPC) sampling from the posterior-lateral ME, in feed-restricted (FR) ewe lambs and in full-fed (FF), age-matched, contemporary control animals. Specifically, we assessed: (1) serum LH and ME in vivo release of LHRH, beta-endorphin (beta-END) and neuropeptide Y (NPY); beta-END and NPY are two putative neuronal inputs to LHRH neuronal terminals at the ME, reported to be involved in the control of both reproduction and feed intake; (2) the effect that exogenous infusion of beta-END through the PPC might have on the release of ME LHRH and NPY, and on plasma LH. In contrast to other works, the present results were obtained in lambs with intact ovaries. Furthermore, FR lambs were always compared statistically with FF contemporary paired controls that had attained puberty. Feed restriction decreased ME LHRH release, lowered plasma LH and prevented the onset of puberty. The changes induced by feed restriction in both LHRH and LH release were associated predominantly with decreases in pulse amplitude, rather than alterations in pulse frequency. The decreased LHRH and LH release occurred in the presence of a decreased beta-END but unchanged NPY release from the ME. Exogenous infusion of beta-END into the posterior-lateral ME decreased both LHRH and NPY release from this site and decreased plasma LH. In conclusion, decreased caloric intake lowers LH release and prevents puberty onset by decreasing the amplitude of the LHRH output from the hypothalamic hypophysiotropic network. A compensatory but unsuccessful mechanism for the FR status might be a lower beta-END-inhibitory tone on ME LHRH neuronal terminals. The unchanged release of NPY at this site supports the specificity of the changes induced by feed restriction on LHRH and beta-END in vivo release.

Animals↗

PubMed1993

Transforming growth factor beta 1-like immunoreactivity in the pituitary gland of the rat: effect of estrogen.

We have recently shown that transforming growth factor-beta 1 (TGF beta 1) is produced in the rat pituitary gland and inhibits the secretion of PRL and the estrogen-induced growth of lactotropes. In this study, we sought to ascertain whether TGF beta 1 inhibits lactotropic functions by an autocrine or paracrine mechanism. Our techniques consisted of localizing the pituitary distribution of the growth factor immunoreactivity and measuring changes in pituitary TGF beta 1 and PRL levels in the presence and absence of estrogen in ovariectomized animals. With the use of standard immunohistochemical techniques, we observed that in cyclic females and ovariectomized rats, 60 +/- 7% of the cells exhibiting TGF beta 1-like immunoreactivity in the anterior pituitary were lactotropes. In addition, the melanotropes in the intermediate lobe appeared to be TGF beta 1 immunopositive. Treatment with estrogen for 7 days reduced the number of TGF beta 1-immunoreactive lactotropes. Immunoreactive TGF beta 1 was also detectable in anterior pituitary extracts using a specific RIA. Estrogen treatment decreased the level of TGF beta 1 in the anterior pituitary extracts from ovariectomized rats. TGF beta 1 immunoreactivity was inversely proportional to the overall size of the anterior pituitary and the concentrations of PRL, as measured in both the anterior lobe extracts and plasma. These results suggest that lactotropes may serve as a site of TGF beta 1 synthesis and that the production of TGF beta 1 in these cells can be negatively influenced by PRL stimulation through the action of the lactotrope-proliferating hormone, estrogen. Furthermore, these data support the notion that TGF beta 1 controls lactotropic function by an autocrine mechanism.

Animals↗

PubMed1992

Pituitary portal plasma levels of oxytocin during the estrous cycle, lactation, and hyperprolactinemia.

The median eminence receives fibers from both parvocellular and magnocellular OT neurons in the hypothalamus. The OT neuronal terminal in the median eminence is secretory and is the major source of the neuropeptide in the blood of pituitary portal vessels. The OT secretion into pituitary portal plasma increases by ovarian steroids, PRL, and the suckling stimulus. The OT secretion into the blood of pituitary portal vessels changes in parallel with the altered secretion of PRL from the pituitary. Because of correlative association between pituitary portal plasma OT and systemic plasma PRL and much abundant evidence for a direct stimulatory action of OT on PRL release, we propose that the pituitary portal vascular system serves as the window for the central OT neurotransmission to the pituitary lactotropes.

Anesthesia↗

PubMed1992

Central administration of neuropeptide Y induces precocious puberty in female rats.

To examine the role of neuropeptide Y (NPY) in the regulation of puberty, the effects of various doses of NPY or saline injected into the 3rd ventricle of peripubertal female rats were determined. A single injection of NPY at a dose range of 10-20 micrograms into the 3rd ventricle of 30-day-old female rats advanced the time of vaginal opening (VO) and first ovulation by 4 days, as compared to saline-treated controls. Ova number and ovarian, adrenal and uterine weights at first estrous were similar in all animals regardless of treatment. The NPY-treated precocious animals showed lower body weight than did the VO-matched controls, but showed similar body weight as that of age-matched controls. Determination of plasma luteinizing hormone (LH) levels in the peripubertal female rats revealed that plasma LH was increased transiently immediately after NPY administration. Also, NPY-treated rats which had precocious puberty showed elevated pituitary and plasma LH levels on the day of VO. These results suggest that NPY can activate the hypothalamic-pituitary-gonadal axis to precociously initiate puberty.

Age Factors↗

PubMed1992

Transforming growth factor-beta 1 messenger RNA and protein expression in the pituitary gland: its action on prolactin secretion and lactotropic growth.

Transforming growth factor-beta 1 (TGF-beta 1) is known to inhibit cell growth and proliferation of many estrogen-responsive normal and transformed cells. The effect of this polypeptide growth factor on the estrogen-responsive pituitary lactotropes has not been determined. To evaluate the role of TGF-beta 1 in the control of lactotropic growth, the action and production of TGF-beta 1 in the anterior pituitary was studied in rats. The growth factor suppressed basal PRL release from the primary culture of enriched rat lactotropes in a concentration-dependent manner in the range of 2 pg/ml-20 ng/ml. The growth factor did not affect the secretion of other pituitary hormones in the cultures. The inhibitory action of TGF-beta 1 on PRL release was time dependent. The minimum time required to produce a significant effect was 4 h. The growth factor also suppressed estradiol-induced PRL release as well as it inhibited estradiol-induced proliferation of lactotropes. TGF-beta 1 immunoreactivity was detected in the cellular extracts of cultured anterior pituitary cells and in the extracts of anterior pituitary tissue. In addition, the primary culture of enriched rat lactotropes secreted TGF-beta 1. Using Northern blot techniques, a 2.4-kilobase transcript of pro-TGF-beta 1 mRNA was detected both in the anterior pituitary tissue and in the primary culture of anterior pituitary cells. These data suggest that TGF-beta 1 is produced in the pituitary gland and inhibits the secretion of PRL and growth of lactotropes in an autocrine and/or paracrine fashion.

Animals↗

PubMed1990

Effect of acute ethanol on beta-endorphin secretion from rat fetal hypothalamic neurons in primary cultures.

To characterize the effect of ethanol on the hypothalamic beta-endorphin-containing neurons, rat fetal hypothalamic neurons were maintained in primary culture, and the secretion of beta-endorphin (beta-EP) was determined after ethanol challenges. Constant exposure to ethanol at doses of 6-50 mM produced a dose-dependent increase in basal secretion of beta-EP from these cultured cells. These doses of ethanol did not produce any significant effect on cell viability, DNA or protein content. The stimulated secretion of beta-EP following constant ethanol exposure is short-lasting. However, intermittent ethanol exposures maintained the ethanol stimulatory action on beta-EP secretion for a longer time. The magnitude of the beta-EP response to 50 mM ethanol is similar to that of the beta-EP response to 56 mM of potassium. Ethanol-stimulated beta-EP secretion required extracellular calcium and was blocked by a calcium channel blocker; a sodium channel blocker did not affect ethanol-stimulated secretion. These results suggest that the neuron culture system is a useful model for studying the cellular mechanisms involved in the ethanol-regulated hypothalamic opioid secretion.

Animals↗

PubMed1990

Correlative changes of the gonadotropin-releasing hormone and gonadotropin-releasing-hormone-associated peptide immunoreactivities in the pituitary portal plasma in female rats.

The precursor protein of gonadotropin-releasing hormone (GnRH) contains a 56-amino acid peptide, known as GnRH-associated peptide (GAP), and GnRH. Both of these peptides are localized in the same neurons and coprocessed under varieties of physiological conditions. In the present study, we evaluated whether these two peptides are cosecreted into the pituitary portal blood in female rats under the conditions in which the secretion of hypothalamic GnRH and pituitary luteinizing hormone (LH) are known to be altered. The immunoreactivities of GAP-like peptide (IR-GAP-LI) and GnRH (IR-GnRH) in the portal plasma were 2- to 15-fold higher than those observed in peripheral plasma of female rats. In the pubertal females, the preovulatory LH surge which occurred in the afternoon of the day before vaginal opening (puberty) was found to coincide with surges of IR-GAP-LI and IR-GnRH in the pituitary portal plasma. The surges of IR-GAP-LI and IR-GnRH in portal plasma corresponded with a fall in the preoptic and hypothalamic contents of these peptides. In the adult rats, the levels of IR-GAP-LI and IR-GnRH in portal plasma and LH in peripheral plasma were significantly higher during the afternoon of proestrus than those in the afternoon of diestrus. Ovariectomy increased the portal plasma levels of IR-GAP-LI and GnRH and peripheral plasma levels of LH as compared to the level of these hormones in diestrous females. These results indicate that both GnRH and GAP-LI are cosecreted into pituitary portal blood and that changes in the endocrine environment similarly affect both GnRH and GAP secretion.

Animals↗

PubMed1990

Facilitatory role of neuropeptide Y on the onset of puberty: effect of immunoneutralization of neuropeptide Y on the release of luteinizing hormone and luteinizing-hormone-releasing hormone.

To examine the role of neuropeptide Y (NPY) in the first luteinizing hormone (LH) surge of puberty, the effect of passive immunoneutralization of NPY with antiserum against NPY (anti-NPY) injected centrally (third ventricle) or peripherally (jugular vein) was studied in pubertal female rats on the day of first proestrus. Both peripheral and central anti-NPY administration reduced the magnitude of the LH surge during the afternoon of first proestrus; however, the central route of administration appeared to be most effective. Centrally administered anti-NPY also reduced the magnitude of proestrous LH-releasing hormone (LHRH) release into pituitary portal blood in these rats. These results suggest that endogenous NPY plays a facilitatory role in the generation of the LHRH surge necessary for preovulatory gonadotropin release and puberty.

Animals↗

PubMed1990

Characterization of the neurosecretory activity of hypothalamic beta-endorphin-containing neurons in primary culture.

To determine the neurosecretory activity of hypothalamic beta-endorphin (beta EP)-containing neurons, rat fetal hypothalamic cells were mechanically dispersed and maintained in primary cultures for periods up to 24 days; their electrophysiological properties and regulation by depolarization, calcium and sodium channel-active agents were studied. Under culture conditions, the majority of the cells were immunopositive to neurofilament antibody, and a significant number (7-10%) were reactive to beta EP antibody. Cultured cells were often electrically excitable and possessed voltage-activated ionic conductances. In culture, there was a progressive increase in immunoreactive beta EP (IR-beta EP) in both cells and media, reaching maximum values at 12-16 days. The majority of IR-beta EP in both cells and media corresponded to [125I]beta EP on gel chromatography and was similar to the form previously found in the hypothalamus. These findings suggest viability of the beta EP neurons and continuing synthesis of IR-beta EP during the culture period. To evaluate the influence of membrane depolarization on IR-beta EP release, the cells were challenged with 56 mM potassium. This treatment induced a significant increase in medium IR-beta EP. The depolarization-induced IR-beta EP release was dependent upon calcium, since a calcium channel blocker, verapamil (0.1 microM), prevented the release; also a calcium ionophore, A23187 (1 microM), stimulated IR-beta EP release in the cultures. Activation of the sodium channel by veratridine (100 microM) also increased the medium content of IR-beta EP, and this effect was blocked by tetrodotoxin (1 microM). These results suggest that the beta EP neurons in primary culture respond to the well defined physiological challenges and that the culture system can be used in determining the regulation of hypothalamic beta EP activity.

Animals↗

PubMed1989

Evidence for prolactin feedback actions on hypothalamic oxytocin, vasoactive intestinal peptide and dopamine secretion.

Ovariectomized rats, when transplanted with 4 anterior pituitaries (APs) to the kidney capsule for 2-3 weeks, had elevated plasma prolactin (PRL) levels (3.8-fold) and showed decreased in situ AP weights (0.62-fold) and PRL concentrations (0.63-fold). The concentrations of dopamine (DA) and oxytocin (OT) in pituitary portal plasma of hyperprolactinemic rats were increased 1.7- and 1.9-fold, respectively. However, the levels of vasoactive intestinal peptide (VIP) in pituitary portal plasma of these rats were decreased 0.31-fold. The secretion of DA, dihydroxyphenylalanine (DOPA) and OT from fetal hypothalamic cells in primary culture was increased, whereas VIP secretion from these cells was reduced in a dose-dependent fashion following PRL treatment. These data are the first in vivo and in vitro demonstration of a stimulatory action of PRL on OT release and an inhibitory action of PRL on VIP release. Furthermore, these data suggest that a subtle imbalance between the secretion of the PRL-inhibiting factor (DA) and the PRL-releasing factors (VIP and OT) during elevated systemic levels of PRL is responsible for decreased lactotrophic function.

Animals↗

PubMed1989

Chronic inhibition of hypothalamic-pituitary-ovarian axis and body weight gain by brain-directed delivery of estradiol-17 beta in female rats.

The effect of preferential delivery of estradiol (E2) into the brain on both the hypothalamic-pituitary-ovarian axis and weight gain was studied in female rats. When E2 was coupled to a lipoidal dihydropyridine-pyridinium carrier, the resulting carrier E2 complex (CE), upon a single intravenous administration to cycling female rats, caused a dose-dependent inhibition of ovulation which lasted 3 times longer than with uncoupled E2. The dose of CE that delayed ovulation for 4 days was one twentieth the amount of E2 needed to produce the same effect. Studies in ovariectomized (OVEX) rats indicated that the prolonged ovulation-blocking action of CE appeared to be related to a sustained storage and release of E2 in the brain, which in turn suppressed the release of hypothalamic luteinizing hormone-releasing hormone (LHRH) and pituitary luteinizing hormone (LH). Upon single intravenous administration in pubertal female rats, CE caused a dose-dependent reduction of body weight gain for a minimum period of 28 days. The inhibitory action of CE on body weight gain was more potent and longer lasting than that of E2 in pubertal rats. When administered in OVEX rats, CE produced a loss of body weight that lasted significantly longer than that produced by uncoupled E2 in these rats. These results suggest that the biological action of E2 can be potentiated by this novel chemical delivery system.

Animals↗

PubMed1988

Immunoneutralization of oxytocin attenuates preovulatory prolactin secretion during proestrus in the rat.

Recent studies have demonstrated that oxytocin (OT) stimulates prolactin (PRL) release from the anterior pituitary gland and that the secretion of OT into pituitary portal blood changes during the rat estrous cycle. To better define the role of OT on PRL release during the reproductive cycle, the effect of administration of antiserum specific for OT on preovulatory PRL secretion in female rats was studied. Intravenous injection of OT antiserum into cyclic female rats between 13.30 and 14.00 h of proestrus neutralized the elevated levels of OT in pituitary portal blood and significantly reduced the subsequent PRL surge. The characteristic proestrous surge of luteinizing hormone (LH) was not affected by the OT antiserum treatment. These data support a physiological role for OT in the regulation of PRL release during the reproductive cycle.

Animals↗

PubMed1988

Neuropeptide Y (NPY): a possible role in the initiation of puberty.

NPY modulates the rat hypothalamic-pituitary-gonadal axis at both the adenohypophysial and central levels. Previously published studies have consistently shown elevations of GnRH content in the preoptic area (POA) and hypothalamus starting with the appearance of GnRH immunoreactivity around fetal day 12-14 until stabilization around the time of puberty. In the present studies, irNPY content of male and female rat hypothalami and POA was examined during days 0 to 36 of postnatal development. Hypothalamic irNPY content rose steadily from 4.54 +/- 0.19 ng/fragment (males) and 2.72 +/- 0.55 ng/fragment (females) at birth to 34.14 +/- 3.94 ng/fragment (males) and 46.79 +/- 6.16 ng/fragment (females) at day 36, corresponding approximately to the time of vaginal opening. A similar elevation of irNPY content was observed in the POA. At day 0, POA content was 1.91 +/- 0.18 ng/fragment (males) and 2.02 +/- 0.25 ng/fragment (females) and progressively increased to 42.26 +/- 3.94 ng/fragment and 41.33 +/- 3.72 ng/fragment by postnatal day 36. In subsequent investigations, hypophysial-portal and peripheral plasma irNPY was determined around the time of vaginal opening, revealing a surge in portal levels of irNPY which preceded the prepubertal LH surge. The progressive postnatal increase in hypothalamic and POA irNPY content culminating in a prepubertal surge of irNPY secretion into the hypophysial-portal circulation suggests involvement of this neuropeptide in reproductive development and the onset of puberty.

Aging↗

PubMed1987

In vivo secretion of LHRH in ovariectomized rats is regulated by a possible autofeedback mechanism.

An episodic secretion of LHRH in pituitary portal blood was observed in ovariectomized and hypophysectomized rats under Saffan anesthesia. This anesthetic did not effect the pulsatile release of LH in ovariectomized rats. Third-ventricular administration of an LHRH agonist, at a concentration which did not cross-react in the LHRH RIA, suppressed both the pulse amplitude and frequency of LHRH release. This inhibitory action of the LHRH agonist on LHRH release was blocked by an LHRH antagonist.

Animals↗