Cultured Rat Pineal Glands Research Articles

Retinoic Acid Signalling in the Pineal Gland Is Conserved across Mammalian Species and Its Transcriptional Activity Is Inhibited by Melatonin.

The pineal gland is integral to the circadian timing system due to its role in nightly melatonin production. Retinoic acid (RA) is a potent regulator of gene transcription and has previously been found to exhibit diurnal changes in synthesis and signalling in the rat pineal gland. This study investigated the potential for the interaction of these two systems. PCR was used to study gene expression in mouse and human pineal glands, ex-vivo organotypic cultured rat pineal gland and cell lines. The mouse and human pineal glands were both found to express the necessary components required for RA signalling. RA influences the circadian clock in the brain, therefore the short-term effect of RA on clock gene expression was determined in ex vivo rat pineal glands but was not found to rapidly regulate Per1, Per2, Bmal1, or Cry1. The interaction between RA and melatonin was also investigated and, unexpectedly, melatonin was found to suppress the induction of gene transcription by RA. This study demonstrates that pineal expression of the RA signalling system is conserved across mammalian species. There is no short-term regulation of the circadian clock but an inhibitory effect of melatonin on RA transcriptional activity was demonstrated, suggesting that there may be functional cross-talk between these systems.

Norepinephrine activates NF-κB transcription factor in cultured rat pineal gland

AimsThe circadian rhythm in mammalian pineal melatonin secretion is modulated by norepinephrine (NE) released at night. NE interaction with β1-adrenoceptors activates PKA that phosphorylates the transcription factor CREB, leading to the transcription and translation of the arylalkylamine-N-acetyltransferase (AANAT) enzyme. Several studies have reported the interplay between CREB and the nuclear factor-κB (NF-κB) and a circadian rhythm for this transcription factor was recently described in the rat pineal gland. In this work we studied a direct effect of NE on NF-κB activation and the role played by this factor on melatonin synthesis and Aanat transcription and activity. Main methodsCultured rat pineal glands were incubated in the presence of two different NF-κB inhibitors, pyrrolidine-dithiocarbamate or sodium salicylate, and stimulated with NE. Melatonin content was quantified by HPLC with electrochemical detection. AANAT activity was measured by a radiometric assay and the expression of Aanat mRNA was analyzed by real-time PCR. Gel shift assay was performed to study the NF-κB activation in cultured rat pineal glands stimulated by NE. Key findingsOur results showed that the p50/p50 homodimer of NF-κB is activated by NE and that it has a role in melatonin synthesis, acting on Aanat transcription and activity. SignificanceHere we present evidence that NF-κB is an important transcription factor that acts, directly or indirectly, on Aanat transcription and activity leading to a modulation of melatonin synthesis. NE plays a role in the translocation of NF-κB p50/p50 homodimer to the nucleus of pinealocytes, thus probably influencing the nocturnal pineal melatonin synthesis.

Leptin Modulates Norepinephrine-Mediated Melatonin Synthesis in Cultured Rat Pineal Gland

Pineal melatonin synthesis can be modulated by many peptides, including insulin. Because melatonin appears to alter leptin synthesis, in this work we aimed to investigate whether leptin would have a role on norepinephrine- (NE-)mediated melatonin synthesis in cultured rat pineal glands. According to our data, cultured rat pineal glands express leptin receptor isoform b (Ob-Rb). Pineal expression of Ob-Rb mRNA was also observed in vivo. Administration of leptin (1 nM) associated with NE (1 µM) reduced melatonin content as well as arylalkylamine-N-acetyl transferase (AANAT) activity and expression in cultured pineal glands. Leptin treatment per se induced the expression of STAT3 in cultured pineal glands, but STAT3 does not participate in the leptin modulation of NE-mediated pineal melatonin synthesis. In addition, the expression of inducible cAMP early repressor (ICER) was further induced by leptin challenge when associated with NE. In conclusion, leptin inhibition of pineal melatonin synthesis appears to be mediated by a reduction in AANAT activity and expression as well as by increased expression of Icer mRNA. Peptidergic signaling within the pineal gland appears to be one of the most important signals which modulates melatonin synthesis; leptin, as a member of this system, is not an exception.

Tryptophan hydroxylase is modulated by L-type calcium channels in the rat pineal gland

Calcium is an important second messenger in the rat pineal gland, as well as cAMP. They both contribute to melatonin synthesis mediated by the three main enzymes of the melatonin synthesis pathway: tryptophan hydroxylase, arylalkylamine N-acetyltransferase and hydroxyindole- O-methyltransferase. The cytosolic calcium is elevated in pinealocytes following α 1-adrenergic stimulation, through IP 3-and membrane calcium channels activation. Nifedipine, an L-type calcium channel blocker, reduces melatonin synthesis in rat pineal glands in vitro. With the purpose of investigating the mechanisms involved in melatonin synthesis regulation by the L-type calcium channel, we studied the effects of nifedipine on noradrenergic stimulated cultured rat pineal glands. Tryptophan hydroxylase, arylalkylamine N-acetyltransferase and hydroxyindole- O-methyltransferase activities were quantified by radiometric assays and 5-hydroxytryptophan, serotonin, N-acetylserotonin and melatonin contents were quantified by HPLC with electrochemical detection. The data showed that calcium influx blockaded by nifedipine caused a decrease in tryptophan hydroxylase activity, but did not change either arylalkylamine N-acetyltransferase or hydroxyindole- O-methyltransferase activities. Moreover, there was a reduction of 5-hydroxytryptophan, serotonin, N-acetylserotonin and melatonin intracellular content, as well as a reduction of serotonin and melatonin secretion. Thus, it seems that the calcium influx through L-type high voltage-activated calcium channels is essential for the full activation of tryptophan hydroxylase leading to melatonin synthesis in the pineal gland.

Insulin modulates norepinephrine-mediated melatonin synthesis in cultured rat pineal gland

The mammalian pineal gland synthesizes melatonin in a circadian manner, peaking during the dark phase. This synthesis is primarily regulated by sympathetic innervations via noradrenergic fibers, but is also modulated by many peptidergic and hormonal systems. A growing number of studies reveal a complex role for melatonin in influencing various physiological processes, including modulation of insulin secretion and action. In contrast, a role for insulin as a modulator of melatonin synthesis has not been investigated previously. The aim of the current study was to determine whether insulin modulates norepinephrine (NE)-mediated melatonin synthesis. The results demonstrate that insulin (10 − 8M) potentiated norepinephrine-mediated melatonin synthesis and tryptophan hydroxylase (TPOH) activity in ex vivo incubated pineal glands. When ex vivo incubated pineal glands were synchronized (12h NE-stimulation, followed by 12h incubation in the absence of NE), insulin potentiated NE-mediated melatonin synthesis and arylalkylamine- N-acetyltransferase (AANAT) activity. Insulin did not affect the activity of hydroxyindole- O-methyltranferase (HIOMT), nor the gene expression of tpoh, aanat, or hiomt, under any of the conditions investigated. We conclude that insulin potentiates NE-mediated melatonin synthesis in cultured rat pineal gland, potentially through post-transcriptional events.

Analysis of gene expression following norepinephrine stimulation in the rat pineal gland using DNA microarray technique.

Several studies have demonstrated that norepinephrine (NE) is the critical neurotransmitter for the regulation of gene expression in the pineal gland. We studied the acute effect of NE stimulation in cultured rat pineal glands using Affymetrix rat genome microarray GeneChip probe arrays. Our data demonstrate that NE stimulation affects regulation of several genes; 44 and 29 genes were up- or down-regulated more than 2.5-fold, respectively. As shown in previous studies, arylalkylamine N-acetyltransferase, cyclic AMP responsive element modulator, jun-B and c-fos mRNA levels were increased by NE stimulation. Genes that were not previously reported and increased by NE stimulation in the pineal gland were protein tyrosine phosphatase, nuclear receptors, and activity and neurotransmitter-induced early genes. Unlike up-regulated genes, most of the down-regulated genes were not reported previously. Genes encoding enzymes involved in metabolism and structural proteins were decreased following NE stimulation. Identification of genes affected by NE stimulation would provide valuable information to understanding pineal biology fully.

Characterisation of P2Y 1-like receptor in cultured rat pineal glands

The rat pineal gland possesses P2 receptors which potentiate the effect of noradrenaline-induced N′-acetyl-5-hydroxytryptamine ( N′-acetyl-5-HT) production. In the current study, this receptor was characterised according to agonist selectivity and signal transduction mechanisms. 2-MethylthioATP (2MeSATP), 2-chloroATP (2-ClATP), adenosine 5′- O-2-thiodiphosphate, (ADPβS), ATP and ADP, but not UTP, potentiated noradrenaline-induced N′-acetyl-5-HT production in a concentration-dependent manner. 2MeSATP neither induced the production of adenosine 3′:5′-cyclic monophosphate (cyclic AMP), nor inhibited its formation when the glands were stimulated by forskolin. The phospholipase C inhibitor 1-[6-[[(17β)-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-1 H-pyrrole-2,5-dione (U73122), but not the inactive analogue, 1-[6-[[(17β)-3-Methoxyestra-1,3,5(10)-trien-17-yl]amino]hexyl]-2,5-pyrrolidinedione (U73343), blocked the 2MeSATP effect. The P2 receptor antagonist pyridoxalphosphate-6-azophenyl-2′,4′-dissulphonic acid (PPADS), which inhibits phospholipase C-coupled P2Y 1 receptors, blocked the 2MeSATP effect. In conclusion, our data strongly suggest that the P2-like receptor that is present in rat pinealocytes and which is responsible for the potentiation of noradrenaline-induced N′-acetyl-5-HT production is a P2Y 1-like receptor, coupled to a G protein which stimulates phospholipase C.

Pretranslational regulation of rhythmic type II iodothyronine deiodinase expression by beta-adrenergic mechanism in the rat pineal gland.

It has been demonstrated that type II iodothyronine deiodinase is present in rat pineal gland, and the deiodinase activity markedly increases during the hours of darkness, primarily through beta-adrenergic mechanism. We have studied the relationship between pineal type II iodothyronine deiodinase messenger RNA (mRNA) and the deiodinase activity to elucidate the mechanisms involved in the nocturnal rise in pineal deiodinase activity. Northern analysis has demonstrated that type II iodothyronine deiodinase mRNA is expressed in rat pineal gland, and the mRNA markedly increases during the hours of darkness. The nocturnal increase in pineal type II iodothyronine deiodinase activity is preceded by the increase in its mRNA. Daytime isoproterenol administration resulted in a rapid increase in pineal type II iodothyronine deiodinase mRNA followed by the increase in deiodinase activity. Propranolol treatment, bilateral superior cervical ganglionectomy, or constant light exposure significantly suppressed the nocturnal rise in type II iodothyronine deiodinase mRNA as well as the deiodinase activity. Moreover, isoproterenol or (Bu)2AMP stimulated type II iodothyronine deiodinase mRNA and the deiodinase activity in cultured rat pineal glands. These results suggest that the rhythmic change in pineal type II iodothyronine deiodinase activity is regulated at least in part at the pretranslational level by a beta-adrenergic mechanism transmitted through superior cervical ganglia.

Morphometry of pineal synaptic ribbon profile numbers after cytochalasin D treatment.

Synaptic ribbons (SRs) are electron-dense, plate-shaped synaptic organelles, to which electron-lucent synaptic vesicles (SVs) are attached by tiny stalks. In the mammalian pineal gland SRs are dynamic organelles, waxing and warning in number under different physiological and experimental conditions. The way in which SRs are formed, or catabolized, is not known. Since it has been suggested that actin may be part of SRs, in the present study the effect of the actin-disrupting drug cytochalasin D (CD, 1 microgram/ml, for 4 h) was examined in cultured guinea pig and rat pineal glands. The glands were preincubated for 48 h so that intra-pineal sympathetic nerve fibres degenerate and no longer release noradrenaline which may distort the results. CD had no effect on SR profile numbers in guinea pigs, but decreased them in rats (p > 0.05). The nonsignificant depressive effect of CD in rats was verified in a second experiment. To clarify the issue, acutely cultured rat pineal glands were treated with CD for 4 h, without effect. The results taken together suggest to us that CD has no major effect on pineal SR profile numbers, but that in rats preincubation for 48 h makes them vulnerable to catabolic processes. In all the experiments, the electron-dense plate of the SRs was qualitatively unaffected. However, the SVs were often larger and more irregular in shape and the stalks linking the SVs with the SRs were less frequently seen in CD-treated glands. In guinea pig pineals, in which SRs frequently lie in groups and parallel to each other, neither the distance between neighboring SRs nor the thickness of individual SRs were affected by CD. It is concluded that actin is not a major component of the SRs and the connecting stalks.

Effect of an n‐3 Fatty Acid‐Deficient Diet on the Adenosine‐Dependent Melatonin Release in Cultured Rat Pineal

We studied the effect of a diet deficient in n-3 fatty acids on the adenosine-dependent melatonin release from cultured rat pineal gland after stimulation by 5'-N-ethylcarboxamidoadenosine (NECA), an A2 adenosine agonist. Experiments were conducted with 2-month-old rats raised on semipurified diets containing either peanut oil (n-3 deficients) or peanut plus rapeseed oil (controls). The proportion of docosahexaenoic acid (22:6 n-3) in the pineal total lipid fraction and in phosphatidylcholine and phosphatidylethanolamine was significantly decreased in n-3-deficient rats. This was compensated for partially by an increase in 22:4 n-6 and 22:5 n-6 levels. The activity of the cultured rat pineal, in terms of cyclic AMP content and N-acetylserotonin and melatonin release in the medium, was lower after stimulation by 10(-5) mol/L NECA in the group fed peanut oil than in the group fed peanut plus rapeseed oil. The increased ratio of n-6/n-3 fatty acids in pineal total lipids and the major glycerophospholipids (phosphatidylcholine and phosphatidylethanolamine) may have an important influence on the rat pineal responses. The results are discussed in the context of changes in membrane-bound proteins, including enzymes and/or receptors involved in the rat pineal gland function.

Beta-adrenergic and peptide N-terminal histidine and C-terminal isoleucine stimulation of N-acetyl-transferase activity and melatonin production in the cultured rat pineal gland.

The purpose of this investigation was to compare the effect of peptide N-terminal histidine and C-terminal isoleucine (PHI) with that of the beta-adrenergic agonist isoproterenol (ISO) on N-acetyltransferase (NAT) activity and melatonin production in the cultured rat pineal gland. Pineal glands were removed and placed in organ culture containing PHI, ISO, or PHI plus ISO at different dosages, and subsequent changes in NAT activity and melatonin levels were measured. PHI stimulated these parameters in both a time- and dose-dependent manner. Combined treatment with PHI (10(-6) M) and high doses of ISO (either 10(-7) or 10(-8) M) did not potentiate the effect of the peptide in terms of either NAT activity or melatonin levels in the pineal gland. However, at a lower concentration, ISO (10(-9) M) had additive effects to those of PHI in both cultured pineals and medium. The results suggest that PHI modulates melatonin synthesis in the rat pineal gland. Furthermore, stimulation of the pineal with both PHI and ISO demonstrates an additive effect rather than a synergistic action of these compounds. It is presumed that ISO and PHI stimulate pineal melatonin production via separate receptors, but they probably use the same intracellular second messenger, cAMP, to do so. This is the first study showing an effect of the peptide PHI on pineal melatonin production in any vertebrate.

Beta-adrenergic and peptide N-terminal histidine and C-terminal isoleucine stimulation of N-acetyl-transferase activity and melatonin production in the cultured rat pineal gland

Beta-adrenergic and peptide N-terminal histidine and C-terminal isoleucine stimulation of N-acetyl-transferase activity and melatonin production in the cultured rat pineal gland

Evidence that interferon-γ alters pineal metabolism both indirectly via sympathetic nerves and directly on the pinealocytes

1. 1. Interferon-γ (IFN-γ) has been shown to suppress N-acetyltransferase (NAT) activity in cultured rat pineal glands when stimulated with isoproterenol (ISO). 2. 2. Conversely, IFN-γ has also been shown to increase the melatonin content of the rat pineal gland in organ culture. 3. 3. Circumstantial evidence leads to a hypothesis that the NAT suppressive effect may be due to the action of IFN-γ on the sympathetic nerve terminals. 4. 4. To test this hypothesis, pineal glands from intact (INT) and superior cervical ganglionectomized (SCGX) rats, which had been operated 5 days earlier, were cultured with either ISO or ISO + IFN-γ. 5. 5. The concentration of ISO was 10 −8 M and that of IFN-γ was 300 antiviral units/ml. 6. 6. The pineals were incubated for a total period of 5.5 hr, after which the activities of NAT and hydroxyindole- O-methyltransferase (HIOMT) and the levels of melatonin and cAMP were estimated. 7. 7. Suppression of NAT by IFN-γ was observed in the pineals from INT rats, but not in those from SCGX animals. 8. 8. IFN-γ significantly enhanced melatonin levels over those in ISO-stimulated pineals and culture media from the SCGX animals, but not from the INT animals. 9. 9. IFN-γ treatment had no effect on either the HIOMT activity or cAMP levels. 10. 10. The results indicate that the IFN-γ-induced NAT suppression requires the integrity of the sympathetic nerve terminals and the IFN-γ-induced enhancement of melatonin production is accomplished through its direct action on pinealocytes.

Norepinephrine suppresses both photoreceptor and neuron-like properties expressed by cultured rat pineal glands

Our previous study showed that pineal glands of the newborn rat express properties of both photoreceptors and endocrine cells, when transferred to cell culture conditions, whereas opsin was not detected in cells of the intact pineal glands of the postnatally developing or mature rats (Araki et al., 1988). In the present study we asked a question why photoreceptor-like cells do not differentiate in vivo, although the pineal cells have the potency as revealed in cell culture conditions. Our hypothesis was that norepinephrine (NE) may suppress the differentiation of cells with photoreceptor properties. This was proved by adding NE into the culture medium, and only few opsin-like immunoreactive cells, if any, were differentiated in the presence of 10 −6 M NE. NE also suppresses the extension of neurite-like processes by serotonin-like immunoreactive (endocrine) cells, another property expressed in cultures. Other aminergic neurotransmitters, such as dopamine and serotonin, did not have such effects. These results imply that the massive innervations of the pineal gland by the sympathetic ganglion may suppress both photoreceptor and neuron-like properties of the mammalian pineal gland in vivo.

Changes in Indole Metabolism in Organ Cultured Rat Pineal Glands Induced by Interferon‐γ

The purpose of this study was to test the effects of recombinant rat interferon-gamma (IFN-gamma) on rat pineal function in organ culture. The glands were pre-incubated with IFN-gamma (50, 100, and 1,000 antiviral units/ml) for two hours, followed by a stimulation with isoproterenol (ISO, 10(-6) M) (in the presence of IFN-gamma) for an additional three hours. The effect of IFN-gamma alone on the pineal function was also tested by adding another group incubated with IFN-gamma (1,000 antiviral units/ml) without ISO stimulation. The following parameters were estimated at the end of the incubation: the activities of N-acetyltransferase (NAT) and hydroxyindole-O-methyltransferase (HIOMT), the levels of melatonin, 5-hydroxytryptophan (5HTP), 5-hydroxytryptamine (5HT), and 5-hydroxyindole acetic acid (5HIAA), and the 5HT/5HIAA ratio. IFN-gamma had no effect on either NAT or HIOMT activities in the pineal glands. At 50 units/ml, IFN-gamma increased the melatonin content over the ISO-stimulated level in the pineal but not in the media. IFN-gamma alone, at 1,000 units/ml, also increased the melatonin content significantly over the control level. The levels of 5HTP and 5HIAA were significantly suppressed by IFN-gamma, either alone or in combination with ISO. The level of 5HT was not altered by any of the treatments. The 5HT/5HIAA ratio was significantly increased by IFN-gamma, either when given alone or when given in combination with ISO. The results suggest that IFN-gamma enhances melatonin production in the pineal gland by suppressing the oxidative deamination of 5HT to 5HIAA and shunting the biosynthetic pathway toward melatonin production.

Effect of 6‐Methoxy‐2‐Benzoxazolinone on the Activities of Rat Pineal N‐Acetyltransferase and Hydroxyindole‐O‐Methyltransferase and on Melatonin Production

A naturally occurring compound, 6-methoxybenzoxazolinone (6-MBOA), present in grasses, has been shown to induce sexual maturation in a number of rodent species. The structural similarity of 6-MBOA and melatonin has led researchers to suspect that 6-MBOA might induce its progonadal effects by directly altering pineal function. Previous studies have shown that 6-MBOA has the properties of a weak beta-adrenergic agonist capable of stimulating rat pineal N-acetyltransferase (NAT) activity at pharmacological concentrations of 10(-3) M. In the present study we have examined the effect of 6-MBOA on both the pineal melatonin synthesizing enzymes, namely, NAT and hydroxyindole-O-methyltransferase (HIOMT) as well as on melatonin production, in organ cultured rat pineal glands. In addition, we have also examined the ability of 6-MBOA to displace a ligand from rat brain alpha- and beta-adrenoceptors. Our results confirm that 6-MBOA stimulates NAT activity and melatonin production at the high concentration of 10(-3) M. It appears to have no effect on HIOMT activity. A competition study shows that 6-MBOA is able of displacing ligands at the alpha- and beta-adrenoceptors but only at concentrations greater than 10(-4) M. Whether such high concentrations of 6-MBOA reach the pineal of rodent in their natural habitat is unknown. However, if 6-MBOA does mediate progonadal effects by altering pineal function it would be expected that 6-MBOA would ultimately inhibit the effects of melatonin. The possibilities are that the high melatonin levels induced by 6-MBOA cause desensitization of melatonin receptors or that 6-MBOA is an antagonist at the level of the melatonin receptor, thus restricting the inhibitory effects of melatonin on the reproductive system.

Comparison of isoproterenol and dibutyryl adenosine cyclic 3',5'-monophosphate stimulation of thyroxine 5'-deiodinase activity in cultured pineal glands from euthyroid and hypothyroid rats

Thyroxine 5'-deiodinase was increased by isoproterenol and dibutyryl adenosine cyclic 3',5'-monophosphate in a dose- and time-related manner in cultured rat pineal gland. Basal and stimulated activity was higher in glands from hypothyroid than from euthyroid animals. Our data suggest direct β-adrenergic stimulation of intracellular cyclic AMP may be involved in the regulation of pineal thyroxine 5'-deiodinase activity.

Effect of psychotropic drugs on [ 14c]tryptophan metabolism in cultured rat pineal gland

A potentiated production of [ 14C]melatonin ([ 14C]Mt) with a simultaneous decline in [ 14C]serotonin ([ 14C]5-HT) conversion to its final metabolite [ 14C]5-hydroxyindole acetic acid ([ 14C]5-HIAA) was recorded in a rat pineal medium cultured with [ 14C]tryptophan and either a neuroleptic [haloperidol (HAL) or chloropromazine (CPZ)] or an antidepressant [desipramine (DIP)] drug. A difference between the pharmacological groups of drugs was evident only in [ 14C]5-HT levels, which were found significantly elevated in the presence of the neuroleptic HAL and reduced in the presence of the tricyclic antidepressant DIP. A parallel trend was observed in norepinephrine (NE)-induced [ 14C]tryptophan metabolism in the pineal culture medium. All three drugs tested brought about further enhancement of NE-induced synthesis of MT; HAL produced an elevated, but DIP a reduced, level of [ 14C]5-HT, compared to the NE-induced value.

Norepinephrine-induced elevation of acetyl coenzyme A in rat pineal glands in culture

A micromethod for the determination of acetyl coenzyme A was developed and the relationship between the concentration of acetyl coenzyme A and the activity of rat pineal serotonin N-acetyltransferase was studied. Acetyl coenzyme A was determined by converting it into N-acetylserotonin using rat liver serotonin N-acetyltransferase. Subsequently, the hydroxy group of N-acetylserotonin was O-methylated by hydroxyindole- O-methyltransferase and S-[methyl- 3H]adenosyl- l-methionine to form [ 3H]melatonin, which was then conveniently separated from S-[methyl- 3H]adenosyl- l-methionine by thin-layer chromatography. The amount of radioactivity in melatonin is a measure of acetyl coenzyme A concentration. This method is sensitive and specific, since it can detect as low as 5 pmol of acetyl coenzyme A but not structurally related substances such as coenzyme A, adenosine diphosphate, cysteamine, d-panthothenic acid, or sodium acetate. After treating cultured rat pineal glands with l-norepinephrine (10 μM) for 6 hr, the concentration of acetyl coenzyme A was increased significantly from 3.26 ± 0.37 to 10.24 ± 0.93 pmol/gland, while the activity of serotonin N-acetyltransferase increased 68-fold. This result suggests that acetyl coenzyme A may play an important role in the norepinephrine-induced induction of serotonin N-acetyltransferase. Sensitivity and adaptability of this method can be utilized to measure acetyl coenzyme A in discrete regions of rat brain and in experimental conditions in which micromeasuremenf of acetyl coenzyme A may be required.

Ornithine dec arboxylase—apparent lack of involvement in the induction of n-acetyltransferase in rat pineal gland

The potential role ornithine decarboxylase in the induction of serotonin- N-acetyltransferase in the rat pineal gland has been studied. The injection of isoproterenol (5 mg/kg, s.c.) resulted in an apparent induction of serotonin- N-acetyltransferase within 1 hr with a maximal response at 4 hr. No change in ornithine decarboxylase was observed in the first hour following isoproterenol administration. Rat pineal glands in organ culture responded to addition of 2 μM isoproterenol with a similar induction of N-acetyltransferase. There was a small but significant decrease in ornithine decarboxylase in the cultured rat pineal glands in response to isoproterenol. Additional experiments indicated that isoproterenol also did not elevate ornithine decarboxylase in the superior cervical ganglia under conditions known to induce tyrosine hydroxylase in this organ. The results of this study are consistent with our previous finding that ribosomal RNA synthesis is unaffected during the induction process in the pineal gland.