Biomedical subjects

A C Gore

Publications and source records attributed to A C Gore.

At least 19 recordsLinked to original sources

PubMed1998

Effects of propranolol on phosphatidate phosphohydrolase and mitogen-activated protein kinase activities in A7r5 vascular smooth muscle cells.

High doses of propranolol inhibit phosphatidate phosphohydrolase (PAP) activity in intact cells, thus blocking metabolism of phosphatidic acid (PA), product of the phospholipase D (PLD) reaction. Vasopressin and phorbol ester activate PLD and ERK (extracellular signal-regulated protein kinase) mitogen-activated protein kinases in A7r5, a rat vascular smooth muscle cell line. Propranolol increased PA levels in intact A7r5 cells and inhibited cytosolic PAP and membrane calcium-independent phospholipase A2 but did not activate PLD or enhance agonist-induced PA accumulation. Incubation of cells with 200 microM propranolol for 10-45 min markedly elevated PA but caused only partial activation of ERKs. Propranolol and other lipophilic amines caused a time- and dose-dependent detachment of cells from their substrate. These results confirm that elevation of PA is not a strong signal for ERK activation and emphasize that caution should be exercised in using propranolol as a PAP inhibitor in intact cells.

Adrenergic beta-Agonists↗

PubMed1998

Insulin-like growth factor-I effects on gonadotropin-releasing hormone biosynthesis in GT1-7 cells.

The immortalized GT1-7 cell line synthesizes and secretes GnRH, the key hormone of reproduction. However, GT1-7 cells lack the normal inputs from neurotransmitters, growth factors, and steroids, which are involved in the maturation and maintenance of GnRH neurons in the brain. We examined the effects of the neurotrophic factor insulin-like growth factor-I (IGF-I) on GnRH gene expression and the mechanism for these changes. Initially, effects of IGF-I on GnRH gene expression were determined by ribonuclease protection assay. In time-course experiments, IGF-I treatment caused significant increases in nuclear GnRH primary transcript levels, an index of GnRH gene transcription, 4 and 8 h after initiation of IGF-I treatment. GnRH messenger RNA (mRNA) levels in the cytoplasm were stimulated by IGF-I at 24 h of treatment. IGF-I also affected GT1-7 cell morphology, with an increase in process extension and cell-cell contacts. In contrast, GnRH peptide levels in the medium were initially stimulated and then suppressed by IGF-I, indicating an uncoupling of biosynthesis and secretion. The increase in GnRH mRNA levels induced by IGF-I is probably caused by a transcriptional mechanism, as evidenced by the increase in GnRH primary transcript levels before a change in GnRH mRNA levels, as well as our finding of a similar GnRH mRNA half-life for both control and IGF-I-treated cells. Interestingly, GT1-7 cells themselves were observed to express IGF-I immunoreactivity, suggesting the possibility of autoregulation by this neurotrophic factor. It is concluded that IGF-I is an important modulator of GnRH gene expression and release in the GT1-7 cell line. The reported stimulatory effects of IGF-I in vivo, and its hypothesized role in the development of GnRH neurons in the brain, suggest that IGF-I may make the GT1-7 cells line more like a mature GnRH neuron, as a model for future studies.

Animals↗

PubMed1998

The role of calcium in the transcriptional and posttranscriptional regulation of the gonadotropin-releasing hormone gene in GT1-7 cells.

The role of calcium in the regulation of GnRH gene expression and the mechanism for its effects were examined in the present study. Using the immortalized hypothalamic GT1-7 cell line, which synthesizes and secretes GnRH, we demonstrated by ribonuclease protection assay and Northern blot analysis that these cells respond to treatment with the calcium ionophores ionomycin and A23187 with an inhibition of transcription of the GnRH gene and decreases in GnRH messenger RNA (mRNA) levels. Ionomycin treatment caused the GnRH mRNA half-life to decrease from 25 to 9 h, concomitant with a decrease in mRNA poly(A) tail length, suggesting that ionomycin causes a decrease in GnRH mRNA stability. The ionomycin inhibitory effect on GnRH cytoplasmic mRNA levels was significantly inhibited in the presence of cycloheximide or the RNA synthesis inhibitor 5,6-dichloro-1beta-ribofuranosylbenzimidazole, indicating that novel protein/RNA synthesis is obligatory for this effect. We conclude that an increase in calcium levels caused by ionomycin inhibits GnRH gene expression at multiple levels, including GnRH gene transcription and mRNA stability in GT1-7 cells.

Animals↗

PubMed1997

Protein synthesis-dependent and -independent mechanisms for the regulation of GnRH RNA transcript levels in GT1 cells.

The cellular mechanism for the suppression of GnRH gene expression by the phorbol ester PMA was investigated in GT1 cells. The protein synthesis inhibitor cycloheximide decreased GnRH primary transcript levels, indicating a protein synthesis requirement for basal GnRH transcription. PMA decreased GnRH primary transcript levels even in the presence of cycloheximide, indicating that the PMA suppression of GnRH gene transcription is protein synthesis-independent. In contrast, the PMA-inhibitory effect on GnRH cytoplasmic mRNA levels was significantly reduced or inhibited in the presence of cycloheximide or RNA synthesis inhibitors given within 4 h of PMA, suggesting a protein/RNA synthesis-dependent mechanism for the regulation of GnRH mRNA levels by PMA. Thus, the mechanism for the PMA inhibition of GnRH primary transcript is mediated through a protein and RNA synthesis-independent mechanism, while the decrease in GnRH mRNA levels occurs through a mechanism that involves the induction of new RNA and protein synthesis that happens within 4 h of PMA administration.

Animals↗

PubMed1997

Regulation of gonadotropin-releasing hormone gene expression in vivo and in vitro.

The pulsatile release of gonadotropin-releasing hormone (GnRH) into the portal vasculature is responsible for the maintenance of reproductive function. Levels of GnRH decapeptide available for this process can be regulated at transcriptional, posttranscriptional, and posttranslational levels. In the immortalized neuronal GT1 cell lines which synthesize and secrete GnRH, regulation of GnRH biosynthesis has been studied using activators of the protein kinase A (PKA), protein kinase C (PKC), and calcium second messenger systems. These substances, while stimulating GnRH release, cause a universal inhibition of all biosynthetic indices measured to date, including decreases in transcription of the proGnRH gene, GnRH mRNA levels, mRNA stability, and translational efficiency. In contrast, in the animal, the mechanism for the regulation of GnRH gene expression appears to be primarily posttranscriptional, since changes in GnRH mRNA levels often occur in the absence of changes in GnRH primary transcript levels an index of GnRH gene transcription. For example, GnRH mRNA levels increase in response to stimulation with glutamate analogs, while GnRH primary transcript levels are unchanged. However, parallel changes in GnRH mRNA and primary transcript have been observed on proestrus prior to the LH/GnRH surge, suggesting that the regulation of GnRH mRNA levels in vivo involves a complex interplay of transcriptional and posttranscriptional processes.

Animals↗

PubMed1997

Post-transcriptional regulation of the gonadotropin-releasing hormone gene in GT1-7 cells.

GT1-7 cells respond to treatment with the phorbol ester, phorbol 12-myristate 13-acetate (PMA), with an inhibition of transcription of the proGnRH gene and decreases in GnRH mRNA levels. However, the timing of this decrease in GnRH mRNA levels suggests that a decrease in GnRH mRNA stability may be involved in addition to an inhibition of transcription of the proGnRH gene. To address this possibility, we treated GT1-7 cells with 100 nM PMA for 4 h and then monitored GnRH mRNA levels over time after blockade of GnRH gene transcription with DRB. PMA treatment caused GnRH mRNA half-life to decrease from 30 to 11 h. Then, to verify this observation, we examined changes in GnRH mRNA poly (A) tail length, which may be a reflection of mRNA turnover, following treatment of GT1-7 cells with PMA or vehicle for 0, 4, 8 or 24 h. The poly (A) tail was removed from half of the GT1 cytoplasmic RNA sample by digestion with RNase H and the difference in GnRH mRNA size with and without RNase H treatment was determined by Northern hybridization. PMA treatment (4 and 8 h) resulted in a significant decrease in the length of the GnRH mRNA poly (A) tail, consistent with a decrease in GnRH mRNA stability. This finding suggests that GnRH mRNA turnover is inducible by substances such as PMA. Our study indicates that a change in mRNA stability is one of a multiplicity of levels at which GnRH gene expression is regulated.

Blotting, Northern↗

PubMed1996

Gonadotropin-releasing hormone and NMDA receptor gene expression and colocalization change during puberty in female rats.

During development, an increase in gonadotropin-releasing hormone (GnRH) release occurs that is critical for the initiation of puberty. This increase is attributable, at least in part, to activation of the GnRH neurosecretory system by inputs from neurotransmitters, such as glutamate, acting via NMDA receptors. We examined changes in GnRH and NMDA-R1 gene expression by RNase protection assay of preoptic area-anterior hypothalamic (POA-AH) dissections of female rats undergoing normal puberty or in which precocious puberty was induced by treatment with the glutamate agonist NMA. GnRH mRNA levels increased significantly throughout normal development; this was accelerated by treatment with NMA. NMDA-R1 mRNA levels increased only between P10 and P20. The acceleration of the elevation in GnRH mRNA levels by NMDA suggests that a stimulation of GnRH gene expression may be a rate-limiting factor for the onset of puberty. This is attributable to a post-transcriptional mechanism because GnRH primary transcript levels, an index of proGnRH gene transcription, were not observed to change during puberty. Alterations in the colocalization of GnRH neurons with the NMDA-R1 subunit during puberty also were assessed immunocytochemically. The percentage of GnRH neurons that double-labeled with NMDA-R1 was 2% in prepubertal rats and 3% in pubertal rats; this increased to 19% in postpubertal rats. Taken together, these studies suggest that an increase in glutamatergic input to GnRH neurons plays a role in the increase in GnRH release and gene expression that occurs at the initiation of puberty.

Animals↗

PubMed1996

Activation of cJun NH2-terminal kinase/stress-activated protein kinase by insulin.

One of insulin's many biological effects is the increased transcription of AP-1-regulated genes. cJun is the principal component of the AP-1 transcription complex, which is regulated by the newly discovered members of the MAPK superfamily referred to as cJun NH2-terminal kinases (JNKs) or stress-activated protein kinases (SAPKs). We show that insulin stimulates a dose- and time-dependent increase in JNK activity in Rat 1 fibroblasts overexpressing human insulin receptors (Rat 1 HIR cells). Using two different polyclonal anti-JNK antibodies, JNK activity was measured after immunoprecipitation from whole cell extracts by phosphorylation of GSTcJun(1-79). Peak activation occurred 15 min after insulin addition, resulting in a 2.5-fold increase in GSTcJun(1-79) phosphorylation over unstimulated controls. Maximal JNK activation correlated with the onset of AP-1 DNA binding activity. Both insulin-stimulated JNK activity and insulin-induced AP-1 transcriptional activity were found to be Ras-dependent. These data suggest that in Rat 1 cells, JNK activation may play a role in insulin-regulated AP-1 transcriptional activity leading to a mitogenic response.

Amino Acid Sequence↗

PubMed1996

Effects of adrenal medulla transplantation into the third ventricle on the onset of puberty in female rhesus monkeys.

To test the hypothesis that prepubertal exposure of LHRH neurons to a source of catecholamines and neuropeptides accelerates the onset of puberty, we examined the effects of autologous adrenal transplantation into the base of the third ventricle of the brain in juvenile female rhesus monkeys at 11-13 months of age. The adrenal medulla, which contains catecholamines and neuropeptide Y (NPY), was cut into small pieces and mixed with gelfoam in artificial CSF and injected into the third ventricle, adjacent to LHRH neurons and their neuroterminals. Sham control monkeys received artificial CSF with gelfoam alone. Animals were monitored for signs of pubertal development. While menarche was not altered by adrenal transplantation, the timing of first and second ovulations occurred significantly (P < 0.05) earlier in adrenal-transplanted monkeys. Histological examination indicated that the grafts survived in all transplanted monkeys. The presence of catecholamines and NPY in graft tissue was confirmed by tyrosine-hydroxylase-positive, dopamine beta-hydroxylase-positive, and NPY-positive cells. Endogenous LHRH fibers were observed innervating the graft tissue. We conclude that: (1) adrenal medulla transplantation into the third ventricle accelerates the age of first ovulation; (2) this is likely due to neuroactive substances (e.g., catecholamines and NPY) from the graft tissue; and (3) grafted adrenal medulla tissue can survive for at least 30-40 months. However, the age of menarche was not accelerated by this grafting, suggesting that an additional mechanism (e.g., removal of tonic inhibition) may be necessary for the onset of puberty.

Adrenal Medulla↗

PubMed1996

Characterization of gonadotropin-releasing hormone gene transcripts in a mouse hypothalamic neuronal GT1 cell line.

We have characterized the nuclear and cytoplasmic RNA transcripts derived from the gonadotropin releasing hormone (GnRH) gene in a mouse hypothalamic neuronal GT1 cell line. Analyses of nuclear GnRH RNA precursors present in the GT1 cells by RNase protection assay show that there is no particular order of intron excision, suggesting the existence of multiple processing pathways. A similar pattern is observed in mouse preoptic area-anterior hypothalamus (POA-AH). In GT1 cells, approximately 5% of the total GnRH RNA transcripts are found in the nucleus. In contrast, in the POA-AH of mice, nuclear transcripts comprise 40% of the total GnRH transcripts. Thus the GT1 cells, while similar in overall GnRH RNA processing to mouse hypothalamic GnRH neurons, do not exhibit the high abundance of nuclear GnRH RNA transcripts seen in the rodent GnRH neuron in vivo. Quantitative analysis of the nuclear RNA species shows that the GnRH primary transcript comprises more than 90% of the total nuclear GnRH mRNA precursors in both GT1 cells and mouse POA-AH and thus GnRH processing intermediates account for fewer than 10% of these precursors. Using these probes, we have examined changes in GnRH primary transcript expression in GT1-7 cells. In the presence of RNA synthesis inhibitors, the half-life of the GnRH primary transcript was found to be quite short, approximately 18 min, suggesting that the level of primary transcript would reflect levels of GnRH gene transcription. When GT1-7 cells are treated with the phorbol ester PMA (phorbol, 12-myristate, 13-acetate) for 1 h, GnRH primary transcript levels decrease by approximately 70%. Supporting the hypothesis that GnRH primary transcript is a good indicator of GnRH gene transcription is the finding that 1 h of PMA treatment results in a similar (approximately 50%) decrease in GnRH gene transcription, as assayed by nuclear run-on assay. Our observation that GT1 cells resemble mouse hypothalamic GnRH neurons in their pattern of intron excision and in the ratio of primary transcript to other nuclear transcripts emphasizes the utility of these cells for studying the regulation of GnRH gene expression in this immortalized hypothalamic cell line.

Animals↗

PubMed1996

Adenohypophysial allografts releasing prolactin decrease prolactin mRNA concentration in the host hamster's adenohypophysis in situ.

The inhibitory effects of pituitary allografts on the prolactin (PRL)-secretory system are presumed to be consequences of the unabated release of PRL by the allografts. In the present studies we used pituitary allografts in the Golden Syrian hamster to address the following questions: (a) Do allografts of adult adenohypophysial tissue which elevate serum PRL levels decrease the concentration of PRL mRNA in the host's adenohypophysis? (b) Is this effect shared by allografts of neonatal hypophysial tissue or neonatal muscle tissue which do not elevate serum PRL levels? (c) Do any of these types of allograft alter growth hormone mRNA in the host's adenohypophysis? Prolactin mRNA concentration, but not growth hormone mRNA concentration, was decreased in the adenohypophyses in situ in the hosts bearing adult adenohypophysial allografts in which serum PRL levels were elevated. In contrast, serum PRL in hosts with neonatal hypophysial or muscle allografts were not elevated and PRL mRNA levels in the adenohypophysis in situ were not decreased when compared to the levels measured in hamsters with sham transplants. Prolactin mRNA levels in hosts with neonatal muscle allografts were not different from levels in hosts with neonatal hypophysial allografts but were increased when compared to the levels measured in hamsters with sham transplants. There were no differences in PRL concentration in the adenohypophyses in situ between any of the groups. Also, PRL concentrations in neonatal hypophysial allografts were similar to those in adult adenohypophysial allografts. To our knowledge these observations are the first demonstrating that short-loop feed-back of PRL includes a decrease in PRL mRNA concentration. The observations also support the working hypothesis that PRL and not another pituitary factor exerts the negative feedback.

Animals↗

PubMed1995

Glutamate regulation of GDNF gene expression in the striatum and primary striatal astrocytes.

The aim of this study was to investigate the regulation of glial cell line-derived neurotrophic factor (GDNF) mRNA by activation of glutamate receptors in the rat striatum. We observed an increase in GDNF mRNA levels in the adult rat striatum after administration of subseizure doses of N-methyl-D,L-aspartic acid (NMA) and kainic acid. Since it is unclear whether the upregulation of GDNF occurred in neurons or astrocytes within the striatum, we further investigated whether GDNF gene expression in primary striatal astrocytes in culture could be regulated by glutamate receptor activation. We found that treatment of the cultures with NMA and kainic acid similarly upregulated GDNF gene expression as observed in vivo, suggesting that striatal astrocytes express functional glutamate receptors. Immunocytochemical and nuclease protection analysis revealed that striatal astrocytes expressed the NMDA-R1 subunit. These findings suggest the regulation of GDNF mRNA in the striatum may be mediated by excitation of glutamate receptors via glutamatergic cortical afferents.

Animals↗

PubMed1995

Regulation of gonadotropin-releasing hormone gene expression in the rat during the luteinizing hormone surge.

In the present study we characterized GnRH gene expression in intact female rats across the estrous cycle and during a steroid-induced LH surge in ovariectomized (OVX) rats, using the quantitative ribonuclease protection assay. We measured cytoplasmic messenger RNA (mRNA) levels and nuclear primary transcript levels as an index of transcription. In Exp I, cycling rats were killed at 1100 or 1800 h on estrus, diestrus day 1, or diestrus day 2 (D2) or at 1100, 1500, 1800, or 2100 h on proestrus (P). In Exp II, proestrous rats were killed at the same time points or injected with pentobarbital (Pb) at the onset of the LH surge and killed on that day or the following day. In Exp III, a LH surge was induced in OVX rats treated with estradiol benzoate plus progesterone. Rats were killed at 1200, 1500, 1600, 1700, or 2100 h on the day of the surge. For all experiments, blood samples were collected and frozen for quantitation of LH by RIA. The preoptic area-anterior hypothalamus was dissected, and cytoplasmic and nuclear RNA were extracted and assayed separately by ribonuclease protection assay. In Exp I, cytoplasmic mRNA levels exhibited two significant peaks, one on D2 and another at 1500h on P. Primary transcript levels were significantly elevated only at 1500 h on P. In Exp II, proestrous rats and rats given Pb and killed the next day had a peak in cytoplasmic mRNA levels at 1500 h, which was blocked in rats given Pb and killed the same day. In Exp III with OVX rats, no significant changes in mRNA or primary transcript levels were observed between steroid or control groups. We hypothesize that the increase in cytoplasmic mRNA levels in cycling rats on D2 is probably due to a posttranscriptional mechanism, because it was not paralleled by changes in primary transcript levels, which would be expected if a transcriptional mechanism were involved. On P, both cytoplasmic mRNA and primary transcript levels changed, suggesting a transcriptional mechanism at this time.

Animals↗

PubMed1995

Translational efficiency of gonadotropin-releasing hormone messenger ribonucleic acid is negatively regulated by phorbol ester in GT1-7 cells.

Our laboratory and others have reported that treatment of GT1-7 cells with the phorbol ester, phorbol 12-myristate 13-acetate (PMA), inhibits transcription of the pro-GnRH gene and decreases messenger RNA (mRNA) levels. We were interested in whether translation of the existing GnRH mRNA decreases in parallel with these other indexes of biosynthesis after PMA treatment. GT1-7 cells were treated with PMA (100 nM) or vehicle for 0, 1, or 4 h. The cytosolic ribosome-associated RNA was isolated and layered on a continuous (10-40%) sucrose gradient, and fractions were analyzed for the distribution of ribosome-associated GnRH mRNA through the gradient by ribonuclease protection assay. The mRNA found in the lighter fractions is associated with fewer ribosomes per RNA, suggesting that these fractions are translated less efficiently, and RNA recovered from heavier fractions has a higher number of ribosomes per mRNA, representing mRNA that is more actively translated. We found that the distribution of the ribosome-associated GnRH mRNA was shifted into lighter fractions (i.e. fewer ribosomes per mRNA) after PMA treatment, indicating that a decrease in the translational efficiency of GnRH mRNA occurs after PMA treatment. Thus, PMA exerts inhibitory effects on translation of GnRH mRNA as well as on gene transcription, mRNA stability, and mRNA levels.

Animals↗

PubMed1994

Regulation of gonadotropin-releasing hormone gene expression by the excitatory amino acids kainic acid and N-methyl-D,L-aspartate in the male rat.

The glutamate analogs N-methyl-D,L-aspartate (NMA) and kainic acid are involved in the regulation of GnRH and LH release in mammals. It has recently been reported that the increase in GnRH release induced by NMA is accompanied by an increase in GnRH mRNA levels, as measured by in situ hybridization. In the present study we assessed the effects of NMA and kainic acid on cytoplasmic mRNA levels using the more quantitative solution hybridization/RNase protection assay. To address the mechanism responsible for these mRNA changes, we also examined changes in heteronuclear RNA transcripts as a reflection of gene transcription. Adult male rats were implanted with a jugular catheter, and 1-2 days later, NMA (14 mg/kg BW), kainic acid (2 mg/kg BW), both NMA and kainic acid, or saline vehicle were injected through the cannula. Rats were killed 15 min or 1 h later by decapitation, blood samples were collected for RIA of LH, brains were removed, and the preoptic area was dissected and frozen. Cytoplasmic and nuclear RNA were extracted and assayed separately by RNase protection assay. Treatment with NMA or NMA plus kainic acid resulted in significantly elevated cytoplasmic mRNA levels 15 min and 1 h later compared to saline control values, with no differences between the two drug treatments observed. Kainic acid stimulated mRNA levels 1 h, but not 15 min after injection. Nuclear RNA transcripts were unaffected by all drug or vehicle treatments. As nuclear primary transcript levels presumably reflect GnRH gene transcription, and these levels are unaltered, the present study indicates that the regulation of GnRH gene expression by excitatory amino acids occurs at a posttranscriptional level. The increase in cytoplasmic GnRH mRNA levels also does not result from an increased translocation of the relatively large nuclear GnRH mRNA pool into the cytoplasm, because nuclear GnRH mRNA levels are also unchanged. Therefore, the elevation of cytoplasmic mRNA levels after excitatory amino acid treatment is probably due to an increase in mRNA stability.

Animals↗

PubMed1993

A possible role of neuropeptide Y in the control of the onset of puberty in female rhesus monkeys.

The onset of puberty is heralded by an increase in pulsatile LHRH release. Since neuropeptide Y (NPY) has been implicated as a major regulator in the control of pulsatile LHRH release in mature monkeys, we have hypothesized that maturational changes in the NPY neuronal system play an important role in puberty. To test this hypothesis, three experiments were conducted in female rhesus monkeys using a push-pull perfusion method. In the first experiment, changes in NPY release in the stalk-median eminence (S-ME) during puberty were determined in 9 prepubertal, 7 early pubertal and 8 midpubertal monkeys. NPY and LHRH levels were measured in aliquots of the same perfusate samples obtained from the S-ME. NPY release was pulsatile in all three groups. Mean NPY release and pulse frequency increased significantly from the prepubertal through the midpubertal stage. These developmental changes in NPY release were parallel to those observed for LHRH release in the same monkeys. In order to examine whether NPY infusion into the S-ME influences LHRH release during puberty, in the second experiment, NPY (10(-6) or 10(-8) M) or vehicle was infused into the S-ME for 10 min at 90-min intervals in 5 prepubertal and 9 midpubertal monkeys. In the midpubertal stage, infusion of NPY at doses of 10(-8) and 10(-6) M resulted in significant (p < 0.01) increases in LHRH release, while vehicle administration had no effect. In contrast, in prepubertal monkeys, neither NPY nor vehicle infusion altered LHRH release. In order to test whether endogenous NPY plays a role in the maintenance of pulsatile LHRH release, in the third experiment, a specific antiserum to NPY (aNPY) was infused into the S-ME of 6 prepubertal and 8 midpubertal monkeys. Infusion of aNPY (1:100, 1:1,000 dilution) significantly suppressed LHRH release in midpubertal but not prepubertal monkeys. The results are summarized as follows. (1) In prepubertal monkeys, NPY release is low, and the presence of NPY in the S-ME does not influence LHRH release. (2) At the onset of puberty, NPY release begins to increase, and NPY probably starts to stimulate LHRH release. (3) In the midpubertal period, NPY release increases further, and NPY in the S-ME is highly stimulatory to LHRH release.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1991

A study of the hypothalamic pulse-generating mechanism responsible for LH release: electrical stimulation of the medial basal hypothalamus in the ovariectomized guinea pig.

In order to examine the neurophysiological properties of the hypothalamus responsible for the pulsatile release of luteinizing hormone (LH) the medial basal hypothalamus (MBH) of the ovariectomized adult female guinea pig was stimulated electrically through an implanted electrode and LH release was monitored as an indication of luteinizing hormone-releasing hormone (LHRH) release. Electrical stimulation (ES) consisting of monophasic square wave pulses with 0.5 ms pulse duration was applied for 2 min at 60, 40, 20, or 10 min intervals. Current intensity (80, 200, 500 microA) and frequency (8, 50, 100 Hz) were varied to determine the parameters of ES which resulted in an LH pulse with amplitude similar to that of a pulse induced by the endogenous pulse-generating mechanism. Blood samples were collected at 5 or 10 min intervals through an indwelling catheter, and LH was measured by RIA.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

A role for norepinephrine in the control of puberty in the female rhesus monkey, Macaca mulatta.

The onset of puberty in female rhesus monkeys is characterized by increases in pulsatile LHRH release. In this study we have tested the hypothesis that changes in input to the LHRH neurosecretory system from noradrenergic neurons contribute to this pubertal increase in LHRH release. In the first experiment, the ability of the LHRH neurosecretory system of prepubertal (12-20 months of age, no signs of puberty evident), early pubertal (24-30 months, premenarchial), and midpubertal (30-45 months, postmenarchial but prior to first ovulation) monkeys to respond to alpha 1-adrenergic stimulation was tested. LHRH release in the stalk-median eminence of conscious monkeys was measured using an in vivo push-pull perfusion method. During push-pull perfusion, perfusates were collected continuously in 10-min fractions, and the alpha 1-adrenergic stimulant methoxamine (MTX, 10(-8), 10(-5) M) or vehicle was infused through the push cannula for 10 min at 90 min intervals. LHRH levels in perfusates were estimated by RIA. Monkeys in all three age groups responded to MTX with significant increases in LHRH release, with the response of the prepubertal group being significantly greater than that of the older age groups. The results indicate that alpha 1-adrenergic receptors are present and functional prior to puberty. In the second experiment, norepinephrine (NE) release in perfusates collected from monkeys in the three age groups was measured by HPLC with electrochemical detection. NE release increased significantly from the pre- and early pubertal to the midpubertal stage. The enhanced sensitivity of prepubertal monkeys to MTX may be due to the absence of high levels of endogenous NE, which results in a situation similar to denervation hypersensitivity. During the early pubertal stage, increases in input from noradrenergic neurons to the LHRH neurosecretory system may occur, thereby resulting in increases in LHRH release, since early pubertal monkeys are highly sensitive to alpha-adrenergic input. Therefore, we propose that the increase in NE release during puberty contributes to the developmental increase in LHRH release.

Aging↗