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1.
Y. O. Nikishina A. R. Murtazina A. Ya. Sapronova V. I. Melnikova N. S. Bondarenko M. V. Ugryumov 《Russian Journal of Developmental Biology》2016,47(5):260-268
The goal of the present study was to verify our hypothesis of humoral interaction between the norepinephrine secreting organs in the perinatal period of ontogenesis that is aimed at the sustaining of physiologically active concentration of norepinephrine in blood. The objects of the study were the transitory organs, such as brain, organ of Zuckerkandl, and adrenals, the permanent endocrine organ of rats that releases norepinephrine into the bloodstream. To reach this goal, we assessed the adrenal secretory activity (norepinephrine level) and activity of the Zuckerkandl’s organ under the conditions of destructed noradrenergic neurons of brain caused by (1) their selective death induced by introduction of a hybrid molecular complex, which consisted of antibodies against dopamine-β-hydroxylase (DBH) conjugated with saporin cytotoxin (anti-DBH-saporin) into the lateral brain ventricles of neonatal rats; and (2) microsurgical in utero destruction of embryo’s brain (in utero encephalectomy). It was observed that 72 h after either pharmacological or microsurgical norepinephrine synthesis deprivation in the newborn rat’s brain, the level of norepinephrine was increased in adrenals and, conversely, decreased in the Zuckerkandl’s organ. Therefore, the experiments with models of chronical inhibition of norepinephrine synthesis in prenatal and early postnatal rat’s brain revealed changes in the secretory activity of peripheral norepinephrine sources. This, apparently, favors the sustaining of physiologically active norepinephrine level in the bloodstream. 相似文献
2.
N. S. Bondarenko L. K. Dilmukhametova A. Yu. Kurina A. R. Murtazina A. Ya. Sapronova A. P. Sysoeva M. V. Ugrumov 《Biochemistry. Biokhimii?a》2017,82(3):373-379
The morphogenesis of individual organs and the whole organism occurs under the control of intercellular chemical signals mainly during the perinatal period of ontogenesis in rodents. In this study, we tested our hypothesis that the biologically active concentration of noradrenaline (NA) in blood in perinatal ontogenesis of rats is maintained due to humoral interaction between its central and peripheral sources based on their plasticity. As one of the mechanisms of plasticity, we examined changes in the secretory activity (spontaneous and stimulated release of NA) of NA-producing organs under deficiency of its synthesis in the brain. The destruction of NA-ergic neurons was provoked by administration of a hybrid molecular complex–antibodies against dopamine-β-hydroxylase associated with the cytotoxin saporin–into the lateral cerebral ventricles of neonatal rats. We found that 72 h after the inhibition of NA synthesis in the brain, its spontaneous release from hypothalamus increased, which was most likely due to a compensatory increase of NA secretion from surviving neurons and can be considered as one of the mechanisms of neuroplasticity aimed at the maintenance of its physiological concentration in peripheral blood. Noradrenaline secretion from peripheral sources (adrenal glands and the organ of Zuckerkandl) also showed a compensatory increase in this model. Thus, during the critical period of morphogenesis, the brain is integrated into the system of NA-producing organs and participates in their reciprocal humoral regulation as manifested in compensatory enhancement of NA secretion in each of the studied sources of NA under specific inhibition of NA production in the brain. 相似文献
3.
Yukio Oomori Sachiko Okumo Hitoshi Fujisawa Hiromichi Iuchi Katsushi Ishikawa Yohichi Satoh Kazuyuki Ono 《Cell and tissue research》1994,275(2):201-213
Immunohistochemistry has been used to demonstrate tyrosine hydroxylase (TH), dopamine--hydroxylase (DBH), phenylethanolamine N-methyltransferase (PNMT), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) immunoreactivities, and acetylcholinesterase (AChE) activity was demonstrated in rat adrenal glands. The TH, DBH, NPY and VIP immunoreactivities and AChE activity were observed in both the large ganglion cells and the small chromaffin cells whereas PNMT immunoreactivity was found only in chromaffin cells, and not in ganglion cells. Most intraadrenal ganglion cells showed NPY immunoreactivity and a few were VIP immunoreactive. Numerous NPY-immunoreactive ganglion cells were also immunoreactive for TH and DBH; these cells were localized as single cells or groups of several cells in the adrenal cortex and medulla. Use of serial sections, or double and triple staining techniques, showed that all TH- and DBH-immunoreactive ganglion cells also showed NPY immunoreactivity, whereas some NPY-immunoreactive ganglion cells were TH and DBH immunonegative. NPY-immunoreactive ganglion cells showed no VIP immunoreactivity. AChE activity was seen in VIP-immunopositive and VIP-immunonegative ganglion cells. These results suggest that ganglion cells containing noradrenaline and NPY, or NPY only, or VIP and acetylcholine occur in the rat adrenal gland; they may project within the adrenal gland or to other target organs. TH, DBH, NPY, and VIP were colocalized in numerous immunoreactive nerve fibres, which were distributed in the superficial adrenal cortex, while TH-, DBH- and NPY-immunoreactive ganglion cells and nerve fibres were different from VIP-immunoreactive ganglion cells and nerve fibres in the medulla. This suggests that the immunoreactive nerve fibres in the superficial cortex may be mainly extrinsic in origin and may be different from those in the medulla. 相似文献
4.
—Exposure of rats to 3°C for up to 30 min leads to a decrease of 30 per cent in the dopamine-β-hydroxylase activity of the vesicular pellet of the heart; this is greater than can be accounted for by loss of soluble DBH from the two populations of noradrenaline storage vesicles known to be present in the heart. Cold exposure in the presence of α-methyltyrosine causes a much smaller reduction in dopamine-β-hydroxylase activity; this suggests that there is a decrease in transmitter release when synthesis is inhibited. The noradrenaline concentration of the vesicular pellet rises briefly during cold exposure and is then maintained at control levels; the early rise is absent in the presence of α-methyltyrosine. The use of the noradrenaline : dopamine-β-hydroxylase ratio as an index of saturation of vesicular storage capacity suggests that during cold exposure an increased synthesis rate leads to increased filling of vesicles. 相似文献
5.
The adrenal medulla of the hedgehog (E. europaeus L.) was studied by electron microscopy and biochemical methods for a characterization of the seasonal changes in the cells and subcellular organelles responsible for catecholamine (CA) storage.In the prehibernating state noradrenaline accounted for about 25% of the total CA; these glands contained a number of cells with small, electron-dense cored vesicles not seen in hibernation when adrenalin was the only detectable amine.Upon entrance into hibernation the hedgehog adrenal medulla increased in total CA, ATP, and dopamine-β-hydroxylase (DBH) concomitant with a decline in weight of the medulla.In hibernation the adrenomedullary cells contained granules with cores of moderate electron density. However, most of the cytoplasmic space was occupied by lipid droplets and large, membrane-enclosed follicles. These were disrupted by homogenization concomitant with a relative increase in particle-free, soluble DBH activity.The present findings indicate that the hedgehog stores a reserve of soluble DBH within the adrenomedullary cells during hibernation, thereby appearing well prepared for a rapid switching-on of adrenomedullary CA synthesis upon arousal from hypothermia. 相似文献
6.
N. S. Bondarenko A. R. Murtazina L. K. Dil’mukhametova M. A. Ikonopistseva E. V. Volina M. V. Ugrumov 《Doklady. Biochemistry and biophysics》2016,467(1):153-156
Spontaneous and K+-stimulated release of noradrenaline from the hypothalamus, adrenal gland, and organ of Zuckerkandl under their flowing incubation was investigated in the perinatal period of ontogenesis of rats. The results suggest that, during the investigated period of ontogenesis, adrenal glands are the main source of noradrenaline in the blood, whereas the contributions of the organ of Zuckerkandl and the brain are not as significant and change during this period. 相似文献
7.
A. R. Murtazina Y. O. Nikishina N. S. Bondarenko A. Ja. Sapronova M. V. Ugrumov 《Doklady. Biochemistry and biophysics》2016,466(1):74-76
Using the method of high performance liquid chromatography with electrochemical detection, the age dynamics of the content of noradrenaline (NA) in the brain, adrenal gland, and the organ of Zuckerkandl in prenatal (18th and 21st days of embryogenesis) and early postnatal (3, 7, 15, and 30th days) periods of development was studied. The potential contribution of these organs to the formation of physiologically active concentration of noradrenalin in the blood was also assessed. The results suggest that, during the development of the organism, the activity of the sources of noradrenaline in the general circulation changes, which gives a reason to assume the existence of humoral interaction between NA-producing organs in the perinatal period of ontogenesis. 相似文献
8.
A. R. Murtazina Yu. O. Nikishina N. S. Bondarenko A. Ya. Sapronova E. V. Volina M. V. Ugrumov 《Doklady. Biochemistry and biophysics》2017,474(1):200-203
Gene expression and content of the key enzymes involved in the synthesis of noradrenaline—tyrosine hydroxylase and dopamine beta-hydroxylase—was evaluated in the organ of Zuckerkandl of rats in the critical period of morphogenesis. High levels of mRNA and protein of both enzymes in the perinatal period of development and their sharp decline on day 30 of postnatal development were detected. These data indicate that the synthesis of noradrenaline in the organ of Zuckerkandl is maximum during the critical period of morphogenesis and decreases during the involution of this paraganglion. 相似文献
9.
Proliferation and distribution of cells that transiently express a catecholaminergic phenotype during development in mice and rats 总被引:2,自引:0,他引:2
Cells that transiently express a catecholaminergic phenotype have previously been shown to appear in the rat gut during development. In the present study the immunocytochemical demonstration of the enzymes, tyrosine hydroxylase (TH) and dopamine-β-hydroxylase (DBH), were used as markers to examine tissues of rats and mice for catecholaminergic cells. The simultaneous radioautographic demonstration of labeling of identified catecholaminergic cells by tritiated thymidine was used to assess their ability to proliferate. Transient catecholaminergic cells were not limited to rat gut. They were also found in the gut of the mouse where they were present by 10 days' gestation and disappeared before Day 13. Similar cells were found in the mouse kidney, the mantle layer of the sacral spinal cord, and the dorsal mesentery. In mice, transient catecholaminergic cells contained TH but did not react with antiserum to DBH. Transient catecholaminergic cells in the rat gut and other locations synthesized DNA. We conclude that transient catecholaminergic cells (1) occur in both rat and mouse embryos, although the cells of mice may not contain DBH; (2) appear in other organs as well as the gut; (3) are able to proliferate. The ultimate fate of these cells remains to be demonstrated. 相似文献
10.
EFFECT OF DIBUTYRYL-CYCLIC AMP and DEXAMETHASONE ON NORADRENALINE SYNTHESIS IN ISOLATED SUPERIOR CERVICAL GANGLIA 总被引:4,自引:1,他引:3
Abstract— Incubation with dibutyryl-cyclic AMP increased levels of both noradrenaline and dopamine- β -hydroxylase in isolated rat superior cervical ganglia. Dexamethasone also increased the dopamine- β -hydroxylase content but did not affect noradrenaline levels. Cycloheximide blocked the effect of dibutyryl-cyclic AMP on ganglion dopamine- β -hydroxylase but did not affect the rise in noradrenaline content.
Dibutyryl-cyclic AMP increased the synthesis of noradrenaline from [14 C]tyrosine but not from [3 H]DOPA.
The results are discussed in terms of a possible role for cyclic AMP in the control of noradrenaline synthesis in sympathetic ganglia. 相似文献
Dibutyryl-cyclic AMP increased the synthesis of noradrenaline from [
The results are discussed in terms of a possible role for cyclic AMP in the control of noradrenaline synthesis in sympathetic ganglia. 相似文献
11.
Olavi Eränkö Virginia M. Pickel Matti Härkönen Liisa Eränkö Tong H. Joh Donald J. Reis 《The Histochemical journal》1982,14(3):461-478
Summary Newborn rats were daily injected with 0.2 mg hydrocortisone acetate for seven days. They were killed 1, 7 or 21 days after the last injection, together with untreated controls. Hydrocortisone caused a great increase in the number of the small, intensely fluorescent (SIF) cells and the appearance of similar small cells with intense immunohistochemical reactions for tyrosine hydroxylase (TH), dopamine--hydroxylase (DBH) and phenylethanolamine (noradrenaline)N-methyltransferase (PNMT) in the superior cervical ganglion. At the same time, the adrenaline content and the PNMT activity of the ganglion greatly increased, while no significant changes were observed in the dopamine or noradrenaline content or TH or DBH activity. All these changes essentially disappeared after a recovery period of seven or 21 days.It is concluded that hydrocortisone causes a temporary increase in the number of SIF cells by causing a synthesis of TH, DBH and PNMT in previously existing small, non-fluorescent cells, which start to synthesize and store adrenaline, thus becoming intensely fluorescent SIF cells. These SIF cells are different from the normal SIF cells of the same ganglion, most of which appear at a later stage of postnatal development when response to hydrocortisone is lost, which contain TH but neither DBH nor PNMT, and which permanently remain in the ganglion. 相似文献
12.
RICHARD M. WEINSHILBOUM RICHARD KVETNANSKY JULIUS AXELROD IRWIN J. KOPIN 《Nature: New biology》1971,230(17):287-288
THE formation of the neurotransmitter noradrenaline from 3,4-dihydroxyphenylethylamine (dopamine) is catalysed by the enzyme dopamine-β-hydroxylase (DBH)1. This enzyme is associated both with the catecholamine-containing chromaffin granules in the adrenal medulla2,3 and with the vesicular structures in sympathetic nerve terminals which contain catecholamines4. Furthermore, DBH activity is released with catecholamines into the perfusate after stimulation of either the isolated perfused adrenal gland5 or the isolated perfused spleen6–8. DBH activity has been reported in the serum of both man and the rat9,10. This activity is similar to adrenal and sympathetic nerve DBH activity with regard to cofactor requirements, oxygen requirement and kinetic characteristics9,10. It has been suggested that serum DBH activity might be present as a result of release of enzyme with catecholamines from the adrenal glands and sympathetic nerves. If this is the case, serum DBH activity might be a useful and convenient index of sympathetic-adrenal activity. The work described here was undertaken to investigate both the source of the serum DBH and the effect on this activity of forced immobilization, a procedure which has been used as a model of stress and which has been shown to release catecholamines from the adrenal gland and increase catecholamine excretion11. 相似文献
13.
Expression of the adrenergic phenotype in cultured fetal adrenal medullary cells: role of intrinsic and extrinsic factors 总被引:4,自引:0,他引:4
During embryogenesis of the rat the enzymes tryosine hydroxylase (TH) and dopamine-β-hydroxylase (DBH) are first detected by immunocytochemistry or biochemical assay on the 16th day of gestation (E 16). It is not until E 18 that the enzyme phenylethanolamine-N-methyltransferase (PNMT), which is required for biosynthesis of adrenaline, can be detected cytochemically or biochemically. In this study we sought to determine whether the delayed appearance of PNMT is consequent to invasion of the adrenal medulla by E 18 of cells destined to express PNMT, cues provided by the ingrowing splachnic nerves or the action of corticosterone (CS) secreted by the adrenal cortical anlage, a hormone which regulates PNMT in adult rats. When adrenal glands are removed on E 16 and placed in culture, PNMT cannot be detected cyto- or biochemically until 2 days later (E 16 + 2). While CS levels increase 100-fold in vivo between E 16 and E 18, the surge of CS is not necessary for expression of PNMT since (a) adrenals removed on E 16 and cultured in the absence of exogenous ACTH fail to increase CS yet still express PNMT and (b) addition of CS (10?5M) to the cultures on E 16 does not alter the time of appearance of the enzyme. CS, on the other hand, increases the amount of PNMT protein and activity 3-fold with respect to control at all time points, without any effect on TH. We conclude that (a) it is the cells already present in the adrenal medulla at E 16 which differentiate to express PNMT; (b) the initial expression of PNMT is not controlled by nerves nor by corticosteroids; and (c) corticosteroids have a selective action on regulating the amount of PNMT, once it is expressed, but not TH enzyme protein. It remains to be determined whether the differentiation of PNMT is elicited by genetic or epigenetic signals. 相似文献
14.
SUBCELLULAR DISTRIBUTION OF DOPAMINE-β-HYDROXYLASE AND INHIBITORS IN THE HIPPOCAMPUS AND CAUDATE NUCLEUS IN SHEEP BRAIN 总被引:1,自引:1,他引:0
—The enzyme dopamine-β-hydroxylase (EC 1.14.17.1) which converts dopamine to noradrenaline was found to be present in substantial amounts in sheep brain hypothalamus and caudate nucleus and was located to the synaptic vesicle fractions in these two brain regions by subcellular fractionation. This dopamine-β-hydroxylase was associated with paniculate matter in these two brain regions since it was resistant to solubilization with butan-1-ol and 0.1% Triton X-100. As highly significant levels of dopamine-β-hydroxylase were present in the caudate nucleus, factors other than a simple lack of this enzyme must operate to maintain the low levels of noradrenaline and high levels of dopamine in the caudate nucleus. Purified adrenal dopamine-β-hydroxylase was substantially inhibited by two factors prepared from sheep brain hypothalamus and caudate nucleus. These were found to be cupric ions and a sulphydryl inhibitor. High levels of the sulphydryl inhibitor of dopamine-β-hydroxylase were found in synaptosomal fractions from sheep brain hypothalamus and caudate nucleus and the levels were comparable in both regions. Upon subfractionation of a synaptosome-containing fraction from the hypothalamus, the inhibitor was located predominantly in the soluble fraction, although there were significant levels in the synaptic vesicle fraction. Therefore, the sulphydryl inhibitor must be considered as a possible regulator of dopamine-β-hydroxylase activity. Free cupric ion concentrations as low as 2·5 μM were found to inhibit purified adrenal dopamine-β-hydroxylase in vitro and the concentration of copper in the soluble tissue component of hypothalamus and caudate nucleus was well above this minimal copper concentration. The percentage content of soluble copper in the caudate nucleus was significantly higher than in the hypothalamus. The importance of the soluble to particulate-bound ratio of copper in brain was shown in studies of the developing rat brain. A rapid increase in the level of copper in brain was found in the first 4 weeks but the level was constant by 2 months of age. The percentage of soluble copper, however, was maximal soon after birth and had declined to a constant figure by 2 months of age. A scheme for the regulation of dopamine-β-hydroxylase activity involving these factors is proposed. 相似文献
15.
Chronic stress is a risk factor for the development of numerous psychopathological conditions in humans including depression. Changes in gene expression of tyrosine-hydroxylase (TH), dopamine-β-hydroxylase (DBH) phenylethanolamine N-methyltransferase (PNMT), β1-, β2- and β3-adrenoceptors in right and left rat atria upon chronic unpredictable mild stress (CMS) were investigated. CMS decreased TH and DBH gene expression levels both in right and left atria and increased PNMT mRNA in left atria. No changes in mRNA levels of β1- and β2-adrenoceptors were recorded, whereas β3-adrenoreceptor mRNA level was significantly elevated in right atria of CMS rats. At the same time, CMS produced a significant increase of β1- and β2-adrenoreceptor mRNA levels in left atria, but did not affect β3-adrenoceptor mRNA level.The results presented here suggest that stress-induced depression expressed differential effects on catecholamine biosynthetic enzymes and β-adrenoceptors at molecular level in right and left atria of adult rat males. Elevated gene expression of PNMT in left atria of rats exposed to CMS can lead to altered physiological response and may play a role in the pathophysiology of cardiovascular function. 相似文献
16.
Micutkova L Rychkova N Sabban EL Krizanova O Kvetnansky R 《Neurochemistry international》2003,43(3):235-242
Enzymes involved in catecholamine synthesis are present in the highest concentration in the adrenal medulla, however they were found also in other, mainly nervous tissues. The aim of our study was to quantify the exact concentration of tyrosine hydroxylase (TH) and dopamine-ss-hydroxylase (DBH) mRNA in rat stellate ganglia under control conditions and at different intervals after exposure to immobilization stress (IMO). In rats immobilized once for 2h, we determined TH and DBH mRNA in different time intervals up to 22 h after the end of the stress stimulus. TH immunoreactive protein levels were also determined in stellate ganglia. TH and DBH mRNA levels were quantified by RT-competitive-PCR.In stellate ganglia, the concentration of TH mRNA was 17+/-1.6 amol/microg of total RNA, which is approximately 30-times lower than in the adrenal medulla. The concentration of DBH mRNA in the stellate ganglia was 2601+/-203 amol/microg of total RNA, which is the concentration similar to adrenal medulla, but is 150-times higher than concentration of TH mRNA in stellate ganglia. After a single 2-h immobilization the highest elevation of TH and DBH mRNA levels was measured 22 h after the termination of the stress stimulus. Repeated immobilization (7 days, 2h daily) did not produce further increase in TH and DBH mRNA levels compared to already elevated levels in adapted control group (immobilized for 6 days, 2h daily and decapitated 22 h later). Levels of TH protein were significantly changed only after the repeated immobilization.This study compared for the first time the precise amounts of TH and DBH mRNA in rat stellate ganglia under control conditions and after immobilization stress, and indicates large differences in their concentration. TH and DBH mRNA concentrations in stellate ganglia are markedly elevated for a prolonged period of time after termination of the stress stimuli. 相似文献
17.
Summary The subcellular locilazations of tryrosine hydroxylase (TH), dopamine--hydroxylase (DBH) and phenylethanolamine-N-methyltransferase (PNMT) in the adrenal glands of the frog and rat have been examined by a peroxidase-antiperoxidase (PAP) method. TH was localized in the ground substance of the adrenaline-containing cells and noradrenaline-containing cells, but not in the nucleus or in the mitochondria. TH was also located on the outside of the membrane of the chromaffin granules. DBH was observed only inside the granules. PNMT was found not only in the ground substance but also on the membrane of some adrenaline-containing granules. Cortical lipid cells of the frog adrenals did not show TH-, DBH-, and PNMT-reactions. The negative reactions to TH-, DBH-, and PNMT-antiserum exhibited by the summer cells of the frog adrenals prove that they belong to the cortical cells. 相似文献
18.
The response of adrenomedullary catecholamine synthesizing enzymes to repeated immobilization was studied in hybrid (F1) offspring of 2 inbred rat strains (LEW and F344). Immobilization-induced increases in tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and phenylethanolamine-N-methyl-transferase (PNMT) activities in one of the parental strains (F344) previously were shown to be dependent upon intact adrenal gland innervation but independent of the pituitary gland, while responses in the other parental strain (LEW) were independent of adrenal innervation but dependent upon pituitary function. Factors determining immobilization-induced increases in adrenal enzymes of F1 offspring were enzyme-specific. Increased PNMT activity was pituitary dependent in F1 rats, whereas increased TH and DBH activities after immobilization were dependent upon an intact adrenal gland innervation. These results suggest that the factor(s) regulating PNMT responses are differentiable from those regulating TH and DBH responses. The results also indicate that analysis of PNMT responses to immobilization in backcross populations is feasible, and could indicate whether strain-specific response mechanisms are heritable. 相似文献
19.
An immunohistochemical and immunoelectron microscopic study was used to demonstrate tyrosine hydroxylase (TH) and dopamine -hydroxylase (DBH) immunoreactivities in the rat pancreas. Small TH immunoreactive cells were found in close contact with large TH immunonegative ganglion cells among the exocrine glands and were occasionally found in some islets. Some of these TH immunoreactive cells were also DBH immunopositive. The immunoreaction product was seen diffusely in the cytoplasm and in the granule cores of TH immunoreactive cells. All intra-pancreatic ganglion cells were immunoreactive for DBH, but not for TH. The TH immunoreactive cells were identified as small intensely fluorescent (SIF) cells due to their localization and morphological characteristics and showed no insulin, glucagon, somatostatin or pancreatic polypeptide immunoreactivities. These results indicate that SIF cells may release dopamine or noradrenaline to adequate stimuli while the intra-pancreatic ganglion cells with only DBH may not synthesize catecholamines in a normal biosynthetic pathway. TH immunoreactive nerve bundles without varicosities and fibers with varicosities, associated or unassociated with blood vessels, were found in both the exocrine and endocrine pancreas. Close apposition of TH immunoreactive nerve fibers to the smooth muscle and endothelial cells of the blood vessels was observed. A close apposition between TH immunoreactive nerve fibers and exocrine acinar cells and islet endocrine cells was sometimes found in the pancreas. The immunoreaction product was seen diffusely in the axoplasm and in the granular vesicles of the immunoreactive nerve fibers. Since no TH immunoreactive ganglion cells were present in the rat pancreas, the present study suggests that noradrenergic nerve fibers in the pancreas may be extrinsic in origin, and may exert an effect on the regulation of blood flow and on the secretory acitivity of the acinar cells, duct cells and endocrine cells. 相似文献
20.
Molecular and physiological properties of tyrosine hydroxylase induced by reserpine in rat adrenal gland 总被引:2,自引:0,他引:2
Newly synthesized tyrosine hydroxylase (TH) induced by reserpine was compared with the enzyme in control rats in terms of the molecular and physiological properties. When repeated doses of reserpine were given at daily intervals for three days, the enzyme activity measured in homogenates of the adrenal glands was increased 3-fold. Furthermore, when TH in the adrenal glands from both control and reserpine-treated rats was purified, both total activity of the enzyme and the enzyme protein content purified from reserpine-treated rats were also about 3-fold higher than those of the control rats. The two purified enzymes revealed similar properties; a single subunit with a Mr of 60,000 was observed by SDS polyacrylamide gel electrophoresis, and the Km value for a pterin cofactor, 6-methyl-tetrahydropterin was about 300 microM. In contrast, in situ TH activity measured under physiological conditions at pH 7.2 in adrenal tissue slices was elevated 6-fold by reserpine pretreatment for 3 days, and was stimulated by carbachol (0.1 mM) and elevated K+ (52 mM) in a roughly proportional rather than additive way relative to slices from untreated rats. These results indicate that newly synthesized TH induced by reserpine in rat adrenal gland had similar properties as the enzyme in control rats and that reserpine increased not only the amount of TH molecules but also the in situ activity of TH. Since reserpine also increases the biosynthesis of tetrahydrobiopterin as demonstrated by Viveros and co-workers, this 6-fold increase in in situ TH activity may depend both upon the 3-fold increase in the amount of enzyme molecules and upon the increase of the physiologically available tetrahydrobiopterin in the adrenal gland. 相似文献