RESUMEN
The activity of N-acetyltransferase in the rat pineal gland is more than 15 times higher at night than during the day. This circadian rhythm persists in complete darkness, or in blinded animals, and is suppressed in constant lighting. The N-acetyltransferase rhythm is 180 degrees out of phase with the serotonin rhythm and is similar to the norepinephrine and melatonin rhythms. Experiments in vitro indicate that norepinephrine, not serotonin, regulates the activity of N-acetyl-transferase through a highly specific receptor.
Asunto(s)
Ritmo Circadiano , Glándula Pineal/enzimología , Serotonina/metabolismo , Transferasas/metabolismo , Compuestos de Anilina/farmacología , Animales , Isótopos de Carbono , Epinefrina/farmacología , Luz , Masculino , Melatonina/biosíntesis , Norepinefrina/farmacología , Octopamina/farmacología , Glándula Pineal/metabolismo , Efectos de la Radiación , Ratas , Estimulación QuímicaRESUMEN
Light acting by way of the eye causes the dark-induced activity of serotonin N-acetyltransferase in the pineal gland of the rat to decrease with a halving time of about 3 minutes. This effect, which is one of the more rapid physiological changes known to occur in the activity of any enzyme that metabolizes biogenic amines, appears to explain the rapid increase in the concentration of pineal serotonin that is caused by light exposure at night.
Asunto(s)
Aciltransferasas/metabolismo , Luz , Glándula Pineal/enzimología , Adaptación Ocular , Adaptación Fisiológica , Animales , Ceguera , Ritmo Circadiano , Activación Enzimática , Masculino , Estimulación Luminosa , Ratas , Ratas Endogámicas , Serotonina , Factores de TiempoRESUMEN
Pineal glands in organ culture synthesize and release biopterin and are able to maintain concentrations of biopterin occurring in vivo for up to 54 hours in vitro. The intracellular biopterin content is reduced 50 percent by treatment with l-norepinephrine or cyclic adenosine monophosphate derivatives, but not by d-norepinephrine. This is an indication that biopterin levels are regulated by an adrenergic cyclic adenosine monophosphate-dependent mechanism. The decline in tissue biopterin content, produced mainly by inhibited of biosynthesis, is maximal at 6 hours and is not associated with either an increase in biopterin release or a shift in the reduction state of the biopterin.
Asunto(s)
Biopterinas/biosíntesis , Bucladesina/farmacología , Norepinefrina/farmacología , Glándula Pineal/metabolismo , Pteridinas/biosíntesis , 1-Metil-3-Isobutilxantina/farmacología , Animales , Guanosina Trifosfato/metabolismo , Técnicas de Cultivo de Órganos , RatasRESUMEN
Pineal serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase, or AA-NAT) generates the large circadian rhythm in melatonin, the hormone that coordinates daily and seasonal physiology in some mammals. Complementary DNA encoding ovine AA-NAT was cloned. The abundance of AA-NAT messenger RNA (mRNA) during the day was high in the ovine pineal gland and somewhat lower in retina. AA-NAT mRNA was found unexpectedly in the pituitary gland and in some brain regions. The night-to-day ratio of ovine pineal AA-NAT mRNA is less than 2. In contrast, the ratio exceeds 150 in rats. AA-NAT represents a family within a large superfamily of acetyltransferases.
Asunto(s)
Arilamina N-Acetiltransferasa/genética , Clonación Molecular , Glándula Pineal/enzimología , Secuencia de Aminoácidos , Animales , Arilamina N-Acetiltransferasa/metabolismo , Encéfalo/metabolismo , Línea Celular , Ritmo Circadiano , ADN Complementario/genética , Datos de Secuencia Molecular , Glándula Pineal/metabolismo , Hipófisis/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Retina/metabolismo , Alineación de Secuencia , Ovinos , TransfecciónRESUMEN
The nocturnal increase in circulating melatonin in vertebrates is regulated by 10- to 100-fold increases in pineal serotonin N-acetyltransferase (AA-NAT) activity. Changes in the amount of AA-NAT protein were shown to parallel changes in AA-NAT activity. When neural stimulation was switched off by either light exposure or L-propranolol-induced beta-adrenergic blockade, both AA-NAT activity and protein decreased rapidly. Effects of L-propranolol were blocked in vitro by dibutyryl adenosine 3',5'-monophosphate (cAMP) or inhibitors of proteasomal proteolysis. This result indicates that adrenergic-cAMP regulation of AA-NAT is mediated by rapid reversible control of selective proteasomal proteolysis. Similar proteasome-based mechanisms may function widely as selective molecular switches in vertebrate neural systems.
Asunto(s)
Arilamina N-Acetiltransferasa/metabolismo , Cisteína Endopeptidasas/metabolismo , Melatonina/biosíntesis , Complejos Multienzimáticos/metabolismo , Glándula Pineal/metabolismo , Agonistas Adrenérgicos beta/farmacología , Antagonistas Adrenérgicos beta/farmacología , Animales , Bucladesina/farmacología , AMP Cíclico/metabolismo , Inhibidores de Cisteína Proteinasa/farmacología , Isoproterenol/farmacología , Luz , Glándula Pineal/citología , Glándula Pineal/efectos de los fármacos , Glándula Pineal/enzimología , Propranolol/farmacología , Complejo de la Endopetidasa Proteasomal , Ratas , Receptores Adrenérgicos beta/fisiologíaRESUMEN
We isolated a new pea mutant that was selected on the basis of pale color and elongated internodes in a screen under white light. The mutant was designated pcd1 for phytochrome chromophore deficient. Light-grown pcd1 plants have yellow-green foliage with a reduced chlorophyll (Chl) content and an abnormally high Chl a/Chl b ratio. Etiolated pcd1 seedlings are developmentally insensitive to far-red light, show a reduced response to red light, and have no spectrophotometrically detectable phytochrome. The phytochrome A apoprotein is present at the wild-type level in etiolated pcd1 seedlings but is not depleted by red light treatment. Crude phytochrome preparations from etiolated pcd1 tissue also lack spectral activity but can be assembled with phycocyanobilin, an analog of the endogenous phytochrome chromophore phytochromobilin, to yield a difference spectrum characteristic of an apophytochrome-phycocyanobilin adduct. These results indicate that the pcd1-conferred phenotype results from a deficiency in phytochrome chromophore synthesis. Furthermore, etioplast preparations from pcd1 seedlings can metabolize biliverdin (BV) IX[alpha] but not heme to phytochromobilin, indicating that pcd1 plants are severely impaired in their ability to convert heme to BV IX[alpha]. This provides clear evidence that the conversion of heme to BV IX[alpha] is an enzymatic process in higher plants and that it is required for synthesis of the phytochrome chromophore and hence for normal photomorphogenesis.
RESUMEN
In garden pea (Pisum sativum L.), a long-day plant, long photoperiods promote flowering by reducing the synthesis or transport of a graft-transmissible inhibitor of flowering. Previous physiological studies have indicated that this promotive effect is predominantly achieved through a response that requires long exposures to light and for which far-red (FR) light is the most effective. These characteristics implicate the action of phytochrome A (phyA). To investigate this matter further, we screened ethylmethane sulfonate-mutagenized pea seedlings for FR-unresponsive, potentially phyA-deficient mutants. Two allelic, recessive mutants were isolated and were designated fun1 for FR unresponsive. The fun1-1 mutant is specifically deficient in the PHYA apoprotein and has a seedling phenotype indistinguishable from wild type when grown under white light. However, fun1-1 plants grown to maturity under long photoperiods show a highly pleiotropic phenotype, with short internodes, thickened stems, delayed flowering and senescence, longer peduncles, and higher seed yield. This phenotype results in large part from an inability of fun1-1 to detect day extensions. These results establish a crucial role for phyA in the control of flowering in pea, and show that phyA mediates responses to both red and FR light. Furthermore, grafting and epistasis studies with fun1 and dne, a mutant deficient in the floral inhibitor, show that the roles of phyA in seedling deetiolation and in day-length detection are genetically separable and that the phyA-mediated promotion of flowering results from a reduction in the synthesis or transport of the floral inhibitor.
RESUMEN
The lv-1 mutant of pea (Pisum sativum L.) is deficient in responses regulated by phytochrome B (phyB) in other species but has normal levels of spectrally active phyB. We have characterized three further lv mutants (lv-2, lv-3, and lv-4), which are all elongated under red (R) and white light but are indistinguishable from wild type under far-red light. The phyB apoprotein present in the lv-1 mutant was undetectable in all three new lv mutants. The identification of allelic mutants with and without phyB apoprotein suggests that Lv may be a structural gene for a B-type phytochrome. Furthermore, it indicates that the lv-1 mutation results specifically in the loss of normal biological activity of this phytochrome. Red-light-pulse and fluence-rate-response experiments suggest that lv plants are deficient in the low-fluence response (LFR) but retain a normal very-low-fluence-rate-dependent response for leaflet expansion and inhibition of stem elongation. Comparison of lv alleles of differing severity indicates that the LFR for stem elongation can be mediated by a lower level of phyB than the LFR for leaflet expansion. The retention of a strong response to continuous low-fluence-rate R in all four lv mutants suggests that there may be an additional phytochrome controlling responses to R in pea. The kinetics of phytochrome destruction and reaccumulation in the lv mutant indicate that phyB may be involved in the light regulation of phyA levels.
RESUMEN
In fish, individual photoreceptor cells in the pineal organ and retina contain complete melatonin rhythm generating systems. In the pike and seabream, this includes a photodetector, circadian clock, and melatonin synthesis machinery; the trout lacks a functional clock. The melatonin rhythm is due in part to a nocturnal increase in the activity of the arylalkylamine N-acetyltransferase (AANAT) which is inhibited by light. Two AANATs have been identified in fish: AANAT1, more closely related to AANATs found in higher vertebrates, is specifically expressed in the retina; AANAT2 is specifically expressed in the pineal organ. We show that there is a physiological day/night rhythm in pineal AANAT2 protein in the pike, and that light exposure at midnight decreases the abundance of AANAT2 protein and activity. In culture, this decrease is blocked by inhibitors of the proteasomal degradation pathway. If glands are maintained under light at night, treatment with these inhibitors increases AANAT2 activity and protein. Organ culture studies with the trout and seabream also indicate that the light-induced decrease of AANAT2 activity is prevented when proteasomal proteolysis is blocked. A cAMP-dependent pathway protects AANAT2 protein from degradation. These results provide a clue to understanding how light regulates the daily rhythm in melatonin secretion in fish photoreceptor cells and provides evidence that proteasomal proteolysis is a conserved element in the regulation of AANAT in vertebrates.
Asunto(s)
Arilamina N-Acetiltransferasa/genética , Cisteína Endopeptidasas/fisiología , Peces/metabolismo , Regulación Enzimológica de la Expresión Génica , Isoenzimas/genética , Complejos Multienzimáticos/fisiología , Glándula Pineal/enzimología , Animales , Arilamina N-Acetiltransferasa/metabolismo , Ritmo Circadiano , AMP Cíclico/fisiología , Femenino , Luz , Masculino , Técnicas de Cultivo de Órganos , Complejo de la Endopetidasa ProteasomalRESUMEN
In vertebrates, the circadian rhythm in the activity of serotonin N-acetyltransferase [arylalkylamine N-acetyltransferase (AA-NAT); EC 2.3.1.87] drives the daily rhythm in circulating melatonin. We have discovered that expression of the AA-NAT gene in the rat pineal gland is essentially turned off during the day and turned on at night, resulting in a more than 150-fold rhythm. Expression is regulated by a photoneural system that acts through an adrenergic-cAMP mechanism in pinealocytes, probably involving cAMP response element-binding protein phosphorylation. Turning off AA-NAT expression appears to involve de novo synthesis of a protein that attenuates transcription. A approximately 10-fold night/day rhythm in AA-NAT messenger RNA occurs in the retina, and AA-NAT messenger RNA is also detected at low levels in the brain.
Asunto(s)
Arilamina N-Acetiltransferasa/biosíntesis , Encéfalo/enzimología , Ritmo Circadiano , Melatonina/biosíntesis , Glándula Pineal/enzimología , Transcripción Genética , Agonistas Adrenérgicos beta/farmacología , Antagonistas Adrenérgicos beta/farmacología , Secuencia de Aminoácidos , Animales , Arilamina N-Acetiltransferasa/química , Secuencia de Consenso , ADN Complementario , Oscuridad , Luz , Masculino , Datos de Secuencia Molecular , Especificidad de Órganos , Fenilefrina/farmacología , Glándula Pineal/fisiología , Reacción en Cadena de la Polimerasa , Propranolol/farmacología , ARN Mensajero/análisis , ARN Mensajero/biosíntesis , Ratas , Ratas Sprague-Dawley , Receptores Adrenérgicos beta/fisiología , Homología de Secuencia de Aminoácido , OvinosRESUMEN
Recent evidence indicates that melatonin production is controlled by norepinephrine acting via alpha 1-and beta 1-adrenoceptors on pinealocytes; activation of alpha 1-adrenoceptors appears to potentiate the effects of beta 1-adrenoceptor activation. However, alpha-adrenergic potentiation of beta 1-adrenergic activation has been demonstrated with only one alpha-adrenergic agonist. For this reason, this issue was reinvestigated using two other alpha-adrenergic agonists, 6-fluoronorepinephrine and cirazoline. Both compounds, which were found to have a high affinity for pineal alpha 1-adrenoceptors, potentiated the stimulatory effects of isoproterenol on pineal N-acetyltransferase. 6-Fluoronorepinephrine also potentiated the stimulation of N-acetyltransferase activity produced by another beta-adrenergic agonist, 2-fluoronorepinephrine. These findings support the hypothesis that pineal N-acetyltransferase activity is regulated by norepinephrine acting through both alpha 1- and beta 1-adrenoceptors.
Asunto(s)
Acetiltransferasas/análisis , Agonistas alfa-Adrenérgicos/farmacología , Agonistas Adrenérgicos beta/farmacología , Imidazoles/farmacología , Norepinefrina/análogos & derivados , Glándula Pineal/enzimología , Animales , Sinergismo Farmacológico , Técnicas In Vitro , Masculino , Norepinefrina/farmacología , Norepinefrina/fisiología , Ratas , Ratas Endogámicas , Receptores Adrenérgicos alfa/metabolismoRESUMEN
Recent studies suggest that a common theme links the diverse elements of pineal photoneuroendocrine transduction--regulation via binding to 14-3-3 proteins. The elements include photoreception, neurotransmission, signal transduction and the synthesis of melatonin from tryptophan. We review general aspects of 14-3-3 proteins and their biological function as binding partners, and also focus on their roles in pineal photoneuroendocrine transduction.
Asunto(s)
Fototransducción/fisiología , Sistemas Neurosecretores/metabolismo , Glándula Pineal/metabolismo , Tirosina 3-Monooxigenasa/fisiología , Proteínas 14-3-3 , Animales , Arilamina N-Acetiltransferasa/química , Arilamina N-Acetiltransferasa/metabolismo , Luz , Melatonina/metabolismo , Modelos Moleculares , Norepinefrina/fisiología , Glándula Pineal/química , Relación Estructura-Actividad , Tirosina 3-Monooxigenasa/química , Tirosina 3-Monooxigenasa/clasificación , Tirosina 3-Monooxigenasa/metabolismo , Tirosina 3-Monooxigenasa/efectos de la radiaciónRESUMEN
Hydroxyindole-O-methyltransferase (HIOMT) catalyzes the last step in the synthesis of the pineal hormone melatonin. In this study, an HIOMT clone was isolated from a human pineal cDNA library using synthetic oligonucleotide probes based on the bovine HIOMT sequence. The human sequence is unusual because it contains a 3' fragment (84 bp) of the LINE-1 sequence, a highly repetitive sequence in the human genome and the genome of some primates and rodents. Exclusive of this LINE-1 fragment, the human HIOMT clone is 75% and 63% homologous to bovine and avian HIOMT sequences, respectively. The deduced amino acid sequence of the human cDNA clone encodes a 41.6-kD protein. In addition, the sequence is 70% and 57% identical and 81% and 73% similar to bovine and avian HIOMT, respectively. In agreement with the results of earlier studies, it was found that vertebrate HIOMT amino acid sequences are not homologous to any other vertebrate proteins, including several methyltransferases. However, HIOMT exhibits homology with a plant O-methyltransferase and an internal 120-amino-acid region is approximately 35% identical to a region of four bacterial O-methyltransferases. The results of PCR and Southern blot analysis indicate that three species of HIOMT mRNA are typically present in the human pineal gland, only one of which contains the LINE-1 fragment. An antiserum was raised against a mixture of three synthetic peptides, corresponding to three regions of the deduced amino acid sequence of human HIOMT. This antiserum detected a single immunoreactive protein in Western blot analysis of human pineal glands. The size of the protein (approximately 42 kD) is identical to that predicted from the HIOMT clone, including the LINE-1 fragment. The human HIOMT sequence should be useful in further studies of this enzyme and will also be of special importance in evaluating the functional significance of the inclusion of a fragment of the LINE-1 in an mRNA.
Asunto(s)
Acetilserotonina O-Metiltransferasa/genética , Glándula Pineal/fisiología , Secuencias Repetitivas de Ácidos Nucleicos , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bovinos , Pollos , Clonación Molecular , ADN Complementario/genética , Regulación Enzimológica de la Expresión Génica , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Mapeo Restrictivo , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido NucleicoRESUMEN
Two prominent proteins (30 and 33 kD) in a purified preparation of the sheep pineal gland were studied. Amino acid analysis of tryptic peptides indicated that the 33-kD protein was the epsilon isoform of the 14-3-3 family of proteins, and that the 30-kD protein was the zeta isoform. The sheep pineal gland was found to have six other 14-3-3 isoforms in addition to the epsilon and zeta, suggesting that copurification of the epsilon and zeta forms may reflect the existence of homo- or heterodimers comprised of these isoforms. To characterize 14-3-3 proteins further in the pineal gland, the full sequence of the epsilon isoform and a partial sequence of the zeta isoform were cloned from a rat pineal cDNA library and are reported here. Tissue distribution studies using Western blot analysis revealed that rat pineal and retina have levels of 14-3-3 protein similar to those found in brain, and that relatively low levels occur in other tissues. This investigation also revealed the epsilon isoform was present at high levels in the rat pineal gland early in development and decreased steadily thereafter and that 30-kD isoforms exhibited the inverse developmental pattern.
Asunto(s)
Proteínas del Tejido Nervioso/genética , Glándula Pineal/metabolismo , Tirosina 3-Monooxigenasa , Proteínas 14-3-3 , Secuencia de Aminoácidos , Animales , Arilamina N-Acetiltransferasa/metabolismo , Secuencia de Bases , Western Blotting , Tampones (Química) , Clonación Molecular , Citosol/química , ADN , Electroforesis en Gel de Poliacrilamida , Femenino , Humanos , Masculino , Datos de Secuencia Molecular , Proteínas del Tejido Nervioso/aislamiento & purificación , Proteínas del Tejido Nervioso/metabolismo , Glándula Pineal/crecimiento & desarrollo , Homología de Secuencia de Aminoácido , Ovinos , Distribución TisularRESUMEN
The developmental patterns of appearance of 5,6,7,8-tetrahydrobiopterin (BH4) and the first enzyme in BH4 biosynthesis, guanosine-5'-triphosphate cyclohydrolase (GTPcyc), were examined in rat brain and pineal gland. A parallel relationship between BH4 content and GTPcyc activity was evident in both tissues during development. In brain, the maximal content of BH4 and activity of GTPcyc was observed 2 days prior to, and 10 days after, birth. In contrast, both pineal BH4 content and GTPcyc activity became maximal postnatally. The influence of neural input on the developmental appearance of pineal BH4 was examined in rats that had been superior cervical ganglionectomized shortly after birth. It was found that this procedure did not alter the developmental appearance of BH4.
Asunto(s)
Aminohidrolasas/metabolismo , Biopterinas/metabolismo , Encéfalo/crecimiento & desarrollo , GTP Ciclohidrolasa/metabolismo , Glándula Pineal/crecimiento & desarrollo , Pteridinas/metabolismo , Envejecimiento , Animales , Biopterinas/análogos & derivados , Encéfalo/metabolismo , Femenino , Masculino , Especificidad de Órganos , Glándula Pineal/metabolismo , Ratas , Ratas Endogámicas , Factores SexualesRESUMEN
Pineal glands convert [3H]tryptophan to [3H]N-acetylserotonin and [3H]melatonin in organ culture. Taurine treatment increases the rate of production of these compounds 40- and 25-fold respectively by stimulating the activity of N-acetyltransferase. This stimulation is blocked stereospecifically by L-propranolol, indicating that taurine is probably acting via beta-adrenergic receptors. Taurine is active in stimulating N-acetyltransferase activity in denervated glands, suggesting that it might interact directly with the beta-adrenergic receptor, and not by causing the release of norepinephrine from nerve terminals.
Asunto(s)
Acetiltransferasas/metabolismo , Melatonina/biosíntesis , Glándula Pineal/enzimología , Receptores Adrenérgicos beta/efectos de los fármacos , Receptores Adrenérgicos/efectos de los fármacos , Taurina/farmacología , Acetilserotonina O-Metiltransferasa/metabolismo , Animales , Técnicas de Cultivo , Desnervación , Masculino , Norepinefrina/farmacología , Propranolol/farmacología , Ratas , Taurina/fisiología , Triptófano/metabolismoRESUMEN
Pineal gamma-aminobutyric acid (GABA) content and glutamic acid decarboxylase (GAD) activity were found not to be influenced by environmental light, catecholamines, sympathetic innervation, or input via the pineal stalk. The observation that GAD activity did not disappear after pineal stalk section, ganglionectomy, or 48 h of organ culture leads us to suggest that GAD activity is not located in nerve processes entering the pineal gland. Treatment in organ culture with an inhibitor of protein synthesis did not greatly influence the slow rate of decrease of GAD activity. This finding is consistent with the conclusion that GAD turnover is slow. Treatment of denervated glands or glands containing functional sympathetic nerve structures with GABA, amino-oxyacetic acid (AOAA) or bicuculline in organ culture did not alter unstimulated levels, or significantly block the adrenergic stimulation of the activity of pineal serotonin N-acetyl transferase (NAT). It is clear from our studies that GABA does not influence or modulate the adrenergic regulation of.pineal NAT activity, and that GABA content and synthesis are not regulated by an adrenergic mechanism. The role of GABA in the pineal gland remains to be discovered.
Asunto(s)
Aminobutiratos/fisiología , Glándula Pineal/fisiología , Ácido gamma-Aminobutírico/fisiología , Acetilcolinesterasa/metabolismo , Acetiltransferasas/metabolismo , Animales , Mapeo Encefálico , Acueducto del Mesencéfalo/fisiología , Ventrículos Cerebrales/fisiología , Diencéfalo/fisiología , Glutamato Descarboxilasa/metabolismo , Luz , Masculino , Vías Nerviosas , Norepinefrina/farmacología , Norepinefrina/fisiología , Técnicas de Cultivo de Órganos , Glándula Pineal/enzimología , Ratas , Núcleos Talámicos/fisiologíaRESUMEN
Pineal physiology is controlled by norepinephrine released from sympathetic nerves terminating in the gland. In the present study, the effect of norepinephrine on the labelling of specific proteins was investigated by incubating glands with [35S]methionine and then resolving the proteins by two-dimensional polyacrylamide gel electrophoresis; the patterns were analyzed by computer-assisted image analysis. The most prominent effects of norepinephrine were distinct and consistent increases in the labelling of two proteins (37 kDa, pI = 6.0, 50 kDa, pI = 6.0), designated adrenergically induced protein (AIP 37/6 and AIP 50/6). In both cases, norepinephrine was effective at low concentrations (EC50 = 10 nM). Pharmacological studies indicated that the effects of norepinephrine on both proteins involved a beta-adrenergic receptor, and that cyclic AMP was the second messenger. Pulse-chase labelling experiments revealed that these effects of norepinephrine did not involve post-translational modification of previously labelled precursor proteins, but depended upon de novo synthesis of protein. An inhibitor of mRNA synthesis, actinomycin-D, was found to block the effect of norepinephrine on AIP 50/6 but not on AIP 37/6, suggesting that norepinephrine acted on AIP 50/6 via a transcriptional mechanism and on AIP 37/6 via a translational mechanism. These in vitro studies were extended into in vivo investigations by measuring silver-stained AIP 37/6 in the two-dimensional gels. Changes in the amount of AIP 37/6 in pineal glands were studied in response to treatments which block the adrenergic stimulation of the gland, including exposure to constant lighting or removal of the superior cervical ganglia. Both treatments reduced AIP 37/6 by 50-75% in 8 weeks. These observations, together with those from in vitro studies, suggest that the amount of AIP 37/6 in the pineal gland is regulated by norepinephrine; and further, that norepinephrine acts through a beta-adrenergic-cyclic AMP mechanism to control AIP 37/6 synthesis at a translational level.
Asunto(s)
Fibras Adrenérgicas/fisiología , Proteínas del Tejido Nervioso/biosíntesis , Norepinefrina/fisiología , Glándula Pineal/metabolismo , Fibras Adrenérgicas/metabolismo , Animales , AMP Cíclico/farmacología , Dactinomicina/farmacología , Peso Molecular , Ratas , Ratas Endogámicas , Simpatectomía , Transcripción GenéticaRESUMEN
The adrenergic transmitter norepinephrine (NE) dramatically increases the prominence of only two out of the hundreds of [35S]methionine-labeled pineal proteins resolved by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). One of these regulated proteins is AIP 37/6 (37 kDa, pI approximately 6). The labeling of this protein is increased approximately 100-fold by NE. In the study presented here the identity of AIP 37/6 was investigated. The results of microsequencing, immunochemical analysis of 2D-PAGE blots and size exclusion chromatography indicate that AIP 37/6 is an isoform of cytosolic malate dehydrogenase (cMDH; approximately 36.3 kDa; pI approximately 6.5). Associated studies indicate that this isoform is phosphorylated whereas the bulk of cMDH is not. Cotranslational phosphorylation of cMDH is discussed.
Asunto(s)
Malato Deshidrogenasa/metabolismo , Glándula Pineal/metabolismo , Glándula Pineal/fisiología , Proteínas/metabolismo , Animales , Autorradiografía , Cicloheximida/farmacología , Femenino , Inmunohistoquímica , Masculino , Metionina/farmacología , Fosforilación , Ratas , Ratas Sprague-DawleyRESUMEN
Pineal metabolism is regulated primarily by noradrenergic innervation. Stimulation of the adult gland with norepinephrine elevates both cyclic AMP and cyclic GMP production, through remarkably similar mechanisms requiring activation of both beta- and alpha 1-adrenergic receptors. As described here, however, the adrenergic stimulation of cyclic GMP is first detectable about 2 weeks after the cyclic AMP response can be detected. This indicates there is a profound difference in when cyclic AMP- and cyclic GMP-regulated processes can be adrenergically regulated.