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At least 163 records · Page 9Linked to original sources
PubMed1997

Expression of preproopiomelanocortin mRNA and preprodynorphin mRNA in brain of spontaneously hypertensive rats.

AIM: To compare the expressions of prepropiomelanocortin (POMC) mRNA and preprodynorphin (PPD) mRNA between 16-wk-old spontaneously hypertensive rats (SHR) and age-matched normotensive Wistar-Kyoto rats (WKY). METHODS: The expression of POMC mRNA and PPD mRNA were detected with nonradioactive in situ hybridization by digoxigenin-labeled RNA probe. RESULTS: POMC mRNA mainly was expressed in arcuate nucleus, compared with WKY, SHR had higher level of POMC mRNA (542). PPD mRNA was found in hippocampus, hypothalamus, central gray, nucleus of the solitary tract (NTS), and thoracic spinal cord (T4-T6). Compared with WKY, PPD mRNA level of SHR decreased in dentate gyrus (2342), NTS (381), and medial preoptic area (467); no difference was observed in arcuate nucleus (263), thoracic spinal cord (750-1800) and CA1, CA2, CA3 of hippocampus (1674, 2014, 2626). CONCLUSION: Increase of POMC mRNA in arcuate nucleus and decrease of PPD mRNA in dentate gyrus of SHR may be associated with the genesis of spontaneous hypertension.

Animals↗
PubMed2000

u-PA and c-MET mRNA expression is co-ordinately enhanced while hepatocyte growth factor mRNA is down-regulated in human hepatocellular carcinoma.

Hepatocyte growth factor/scatter factor (HGF/SF) is one of the most important humoral mediators of liver regeneration. It is potentially related to molecular mechanisms of hepatocarcinogenesis via a paracrine system involving its cellular receptor, c-met. In this study, the expression patterns of HGF and c-met were evidenced by multiplex RT-PCR in different specimens of human hepatic tissues (n = 71). A significant increase of c-met mRNA expression was detected in hepatitis (P = 0.001), cirrhosis (P = 0.006), and hepatocellular carcinoma (HCC) tissue (P = 0.003) compared with normal parenchyma and steatosis. HGF mRNA expression was significantly higher only in hepatitis (P = 0.01). Over-expression of c-met mRNA and under-expression of HGF mRNA were detected in the HCCs compared with the corresponding peri-tumoral tissues. Neither HGF nor c-met expression was related to age, sex, tumor size, grading, presence of pseudocapsula, and proliferative activity of the malignant hepatocytes. A significant inverse correlation was found between c-met mRNA expression level and survival (in months) of patients (P = 0.007), as previously shown for urokinase-type plasminogen activator (u-PA) mRNA (P = 0.027). In addition, c-met mRNA expression was strictly associated with u-PA mRNA level in HCC samples (P = 0.001). These data show that a loss of balance concerning HGF, c-met, and u-PA mRNA expression occurs during hepatocarcinogenesis. Particularly, up-regulation of c-met and u-PA mRNA transcription appears to be coordinately regulated, and their levels of expression are inversely correlated with survival; they must therefore play an important role in the development and progression of human HCC and may also be relevant prognostic markers.

Antigens, Nuclear↗
PubMed2000

[The expression and significance of VEGF mRNA and bFGF mRNA in the malignant neoplasms of ovary].

The expression and significance of VEGF mRNA and bFGF mRNA in the malignant neoplasms of ovary were studied by in situ hybridization technique. The results were that the positive rates of VEGF mRNA and bFGF mRNA were higher in the serous adenocarcinoma and embryonal carcinoma than that in the mucous adenocarcinoma and granulosa cell tumor. The positive rates of VEGF mRNA and bFGF mRNA were lower in the cases of clinical Stage I and without metastasis than that of clinical Stage III-IV and with metastasis. The results suggest that the expressions of VEGF mRNA and bFGF mRNA might be related to the pathological types, clinical stages, and metastasis of the malignant neoplasms of ovary and the patients who have positive expression of VEGF mRNA and bFGF mRNA might have a poor prognosis.

Adolescent↗
PubMed2004

[Effect of nandrolone phenylpropionate on hepatic albumin mRNA and granulational alpha 1(I) procollagen mRNA in burned rats].

OBJECTIVE: To assess the mechanism for the effect of nandrolone phenypropionate (NP) on hepatic albumin mRNA and granulational alpha 1(I) procollagen mRNA in burned rats with the aim to underpin the clinical application of anabolic steroids. METHODS: Thirty-two Wistar rats with a deep second-degree cutaneous burn of 20% total body surface area were randomly divided into two groups to receive either 5 mg/kg NP (NP group) or normal saline as placebo(control group) every other day. The expression copy quantities of albumin-mRNA in liver tissue and alpha 1 (I) procollagen mRNA in granulation wound were measured by quantitative fluorescent RT-PCR respectively on the post-burned days 4, 7, 14 and 21. RESULTS: The expression levels of albumin-mRNA and alpha 1 (I) procollagen mRNA in NP group were much higher than those in control group. The 7th and 14th days were the periods in which the albumin-mRNA and alpha 1 (I) procollagen mRNA expression had been increasing obviously (P < 0.01). CONCLUSION: Nandrolone phenylpropionate could effectively up-regulate the expression of albumin-mRNA in liver tissue and the alpha 1 (I) procollagen mRNA in granulation wound.

Albumins↗
PubMed1992

Analysis of cytokine mRNA expression in the central nervous system of mice with experimental autoimmune encephalomyelitis reveals that IL-10 mRNA expression correlates with recovery.

Experimental autoimmune encephalomyelitis (EAE) serves as an important animal model for understanding the events that lead to immune-mediated inflammation and tissue destruction within the central nervous system. We have utilized a murine adoptive transfer model of EAE and semiquantitative reverse transcriptase-polymerase chain reaction analysis to examine cytokine mRNA expression within the central nervous system in relation to the onset and resolution of paralysis associated with EAE. Spinal cord samples, obtained from mice as they progressed through discrete clinical stages of EAE, were examined for the expression of six cytokine genes (IL-1 alpha, IL-2, IL-4, IL-6, IL-10, and IFN-gamma). Distinct patterns of cytokine gene expression were observed during the acute, recovery, and chronic phases of EAE. The acute phase of disease was characterized by rapid increases in the levels of mRNA for IL-2, IL-4, IL-6, IFN-gamma, and IL-1 alpha. In fact, peak expression of several cytokine mRNA (e.g., IL-2, IL-4, IL-6, and IFN-gamma) occurred before the peak in clinical severity. In contrast, IL-1 alpha mRNA levels were elevated throughout the initial disease course. IL-10 mRNA demonstrated only modest increases during the acute phase of EAE. Stabilization of the clinical symptoms was characterized by rapid declines in the mRNA levels of IL-2, IL-4, IL-6, and IFN-gamma. The decreases in these four cytokine mRNA levels occurred concomitant with a dramatic rise in IL-10 mRNA. Finally, of the six cytokine mRNA examined, only IL-1 alpha, IFN-gamma, and IL-10 mRNA remained elevated during the early chronic stage. These results suggest that local cytokine production varies significantly during the course of EAE and that increases in discrete sets of cytokines are associated with the acute response and the recovery/chronic phase of disease.

Animals↗
PubMed1992

Regulation of usage of membrane and secreted 3' termini of alpha mRNA differs from mu mRNA.

Previous studies demonstrated that the addition of transforming growth factor-beta (TGF-beta) to LPS-stimulated B cell cultures induced cells to express membrane IgA and to mature to IgA-secreting cells without a parallel change in usage of 3' termini by alpha mRNA. In these cultures, the secreted form of alpha mRNA was predominant even before expression of membrane IgA could be detected. In the present study, we demonstrate that the preferential usage of the secreted terminus of alpha mRNA in these cultures is not caused by transcription termination and reflects a difference in the regulation of choice of 3' terminus for alpha and mu mRNA. The addition of TGF-beta to LPS-stimulated cultures causes an increase in the steady state level of alpha mRNA using the secreted 3' terminus. In contrast, TGF-beta decreases the steady state level of mu mRNA and inhibits usage of the 3' terminus for the secreted form of mu, suggesting that the choice of 3' terminus for alpha and mu mRNA is regulated differently in LPS-stimulated cultures. To determine whether the difference in usage of 3' termini by alpha and mu mRNA was a property of the culture system or whether it reflected a difference in regulation, C alpha was transfected into cell lines representing different stages of B cell development. The secreted form of alpha mRNA predominates regardless of the ratio of membrane to secreted forms of the endogenous C mu gene. A similar dichotomy in 3' terminus usage occurred in a stable C alpha transfectant of the BCL1 lymphoma, suggesting that trans-acting factors are not limiting. Furthermore, as was the case with normal B cells, the predominance of the secreted form of the transfected C alpha genes was not due to transcription termination. These data demonstrate that usage of 3' terminus in alpha and mu mRNA is regulated differently.

Animals↗
PubMed2006

[Study on the correlation between the expressions of nm23H1 mRNA, VEGF-C mRNA and lymphatic metastases, survival rate with gastric cancer].

The correlations between the expression of nm23H1 mRNA,VEGF-C mRNA, VEGFR-3 and neoangiogenesis, hyperplasia of micro-lymphatic, tumor metastases in gastric cancer were studied, the results supplied an experimental foundation for clinical treatment and prognosis. All the specimens were marked by VEGFR-3 antibody, the expressions of nm23H1 mRNA, VEGF-C mRNA were detected in 78 cases of gastric cancer in situ hybridization with EnVision and Leica-Qwin computer image analysis system. The Weidner's highest vessel density counting method was used to analyse micro-lymphatics count (MLC) of the specimens, and the patients' viability after operation was investigated. The positive expression of nm23H1 mRNA in gastric cancer was 69.23% (54 cases). There was negative correlation between the positive expression of nm23H1 mRNA and lymph node metastasizing, TNM staging, MLC (P < 0.01 or P < 0.05). The expression of VEGF-C mRNA was 46.15% (36 cases). It was positive related to the lymph node metastasis, TNM stages, MLC in gastric cancer (P < 0.01 or P < 0.05), and was significantly different compared with adjacent nontumorous tissue (P < 0.01 or 0.05). The expression of nm23H1 in stage I and II gastric cancer was high, while in stage III and IV the expression was lower or even none. The MLC (8.37 +/- 2.29/mm2) in gastric cancer was higher than that in adjacent nontumorous tissue (4.82 +/- 3.48/mm2), which has statistical significance (P < 0.05). There was negative correlation between the expression of nm23H1 mRNA and VEGFR-3 (p < 0.05, r = 0.8479), but positive correlation between VEGF-C mRNA and VEGFR-3 (P < 0.05, r = 0.8362). The MLC (10.82 +/- 2.51/mm2) in those who died in five years (48 cases) was higher than that (6.53 +/- 2.09/mm2) in those who were still alive, the difference has statistical significance (P < 0.05). In different tumor pathology grading, different differentiation, the nm23H1 positive expression is significantly different (P < 0.05). Higher the nm23H1 expressed, lower the tumor lymph metastasized, but the viability rate was higher, so the nm23H1 gene was thought to have the effect of suppressing gastric cancer occurring and lymph metastasizing. Higher the VEGF-C mRNA expressed, higher the tumor lymph metastasized, but the viability rate was lower. There is close correlation between the VEGFR-3 expression and gastric cancer lymph metastasizing. The higher MLC level indicated neoangiogenesis in gastric cancer. VEGF-C promote neoangiogenesis induced by tumor and play an important role in lymph metastasizing.

Adult↗
PubMed1988

Relationship between the levels of calbindin synthesis and calbindin mRNA in chick intestine. Quantitation of calbindin mRNA.

An RNA-excess filter hybridization assay was established to measure the absolute amount of calbindin mRNA in chick tissues. The tissue with the highest level of mRNA is intestine, followed by kidney and cerebellum; the mRNA was not detected in liver and skin. Calbindin mRNA in intestine and kidney is vitamin D-dependent. The maximum concentration of calbindin and its mRNA found after dosing vitamin D-deficient chicks with dihydroxyvitamin D3 (1,25-(OH)2D3) is less than 5% of that found with vitamin D dosing. Secondary 1,25-(OH)2D3 stimulation produced greatly increased amounts of both calbindin mRNA and the protein, at least reaching levels similar to those found after vitamin D dosing. In this last case, each mucosal cell contains about 2000 calbindin mRNA molecules which are translated at a rate sufficient to account for the levels of calbindin found. Calbindin mRNA is translated most rapidly in the very short time periods after its release into the cytoplasm. 1,25-(OH)2D3 has two effects on calbindin mRNA formation: first, to permit the expression of the calbindin gene and a second effect, of slower onset but more persistent, which increases either the rate of calbindin gene transcription or the stability of calbindin mRNA.

Animals↗
PubMed1996

Detection of breast cancer micrometastases in axillary lymph nodes by means of reverse transcriptase-polymerase chain reaction. Comparison between MUC1 mRNA and keratin 19 mRNA amplification.

Usefulness of MUC1 mRNA and keratin 19 mRNA as a target of reverse-transcriptase polymerase chain reaction (RT-PCR) was compared in the detection of breast cancer micrometastases in axillary lymph nodes. RT-PCR amplification of MUC1 mRNA and keratin 19 mRNA was conducted using total RNA samples. RT-PCR products were stained with ethidium bromide and analyzed by agarose gel electrophoresis. Expression of both MUC1 mRNA and keratin 19 mRNA was detected by RT-PCR in a breast cancer cell line (MRK) and in all the 23 primary breast cancers but not in the control lymph nodes obtained from patients with benign diseases. A serial dilution study of MRK cells against normal lymph node cells has shown that detection sensitivity of MUC1 RT-PCR and keratin 19 RT-PCR were 1/10(5) and 1/10(6) (cancer/lymph node cells), respectively. Sixty-three axillary lymph nodes were obtained from 23 patients with primary breast cancer, and metastases in each lymph node were investigated by histological examination (hematoxylin and eosin sections) and RT-PCR method. In all 10 lymph nodes, which were histologically metastasis-positive, both MUC1 mRNA and keratin mRNA were detected by RT-PCR. Of the 53 histologically negative lymph nodes, 3 (6%) and 5 (9%) lymph nodes were found to express MUC1 mRNA and keratin 19 mRNA, respectively, indicating the presence of micrometastases which could be detected by RT-PCR but not by histological examination. These results demonstrate the usefulness of both MUC1 RT-PCR and keratin 19 RT-PCR in the detection of breast cancer micrometastases in lymph nodes, and also indicate the superiority of keratin 19 RT-PCR over MUC1 RT-PCR because of its higher detection sensitivity.

Axilla↗
PubMed1995

The polyribosomal protein bound to the 3' end of histone mRNA can function in histone pre-mRNA processing.

Cell cycle-regulated histone mRNAs end in a conserved 26-nt sequence that can form a stem-loop with a six-base stem and a four-base loop. The 3' end of histone mRNA has distinct functions in the nucleus and in the cytoplasm. In the nucleus it functions in pre-mRNA processing and transport, whereas in the cytoplasm it functions in translation and regulation of histone mRNA stability. The stem-loop binding protein (SLBP), present in both nuclei and polyribosomes, is likely the trans-acting factor that binds to the 3' end of mature histone mRNA and mediates its function. A nuclear extract that efficiently processes histone pre-mRNA was prepared from mouse myeloma cells. The factor(s) that bind to the 3' end of histone mRNA can be depleted from this extract using a biotinylated oligonucleotide containing the conserved stem-loop sequence. Using this depleted extract which is deficient in histone pre-mRNA processing, we show that SLBP found in polyribosomes can restore processing, suggesting that SLBP associates with histone pre-mRNA in the nucleus, participates in processing, and then accompanies the mature mRNA to the cytoplasm.

Animals↗
PubMed2001

Regional distribution and cellular localization of 5-HT2C receptor mRNA in monkey brain: comparison with [3H]mesulergine binding sites and choline acetyltransferase mRNA.

The distribution of serotonin 5-HT(2C) receptor mRNA in monkey brain was studied by in situ hybridization and compared with the distribution of [3H]mesulergine binding sites as visualized by receptor autoradiography. 5-HT(2C) receptor transcripts showed a widespread and heterogeneous distribution. The strongest hybridization signal was detected in choroid plexus. In neocortex, 5-HT(2C) mRNA was detected in layer V of all cortical regions examined except in the calcarine sulcus, which was devoid of signal. Several structures within the striatum and basal forebrain were strongly labeled: nucleus accumbens, ventral aspects of anterior caudate and putamen, septal nuclei, diagonal band, ventral striatum, and extended amygdala. Several thalamic, midbrain, and brainstem nuclei also contained 5-HT(2C) mRNA. Comparison of the distributions of 5-HT(2C) mRNA and specific [3H]mesulergine binding sites showed a good agreement in the majority of brain regions, suggesting a predominant somatodendritic localization of 5-HT(2C) receptors. A possible localization to axon terminals of 5-HT(2C) receptors is suggested by the disagreement observed in some regions such as septal nuclei and horizontal limb of the diagonal band (presence of mRNA with apparent absence of binding sites) and interpeduncular nucleus (presence of binding sites with apparent absence of mRNA). Comparison of 5-HT(2C) receptor and choline acetyltransferase mRNA distributions indicate that some regions where cholinergic cells are located are also enriched in cells containing 5-HT(2C) mRNA. Although the present methodology does not allow strict colocalization of both mRNA species to the same cells, the codistribution observed in several regions provides a possible anatomical substrate for the described modulation of acetylcholine release by 5-HT(2C) receptors.

Animals↗
PubMed2001

Dispersion of cyclin B mRNA aggregation is coupled with translational activation of the mRNA during zebrafish oocyte maturation.

Cyclin B mRNA stored in immature zebrafish oocytes is translationally activated upon the stimulation of 17alpha,20beta-dihydroxy-4-pregnen-3-one (17alpha,20beta-DP), an event prerequisite for initiating oocyte maturation in this species. We investigated localization of cyclin B mRNA in zebrafish oocytes. Cyclin B mRNA was found to be exclusively localized as an aggregation along the cytoplasm at the animal pole of full-grown immature oocytes. When oocytes were treated with 17alpha,20beta-DP, a meshwork of microfilaments in the oocyte cortex disappeared and the aggregation of cyclin B mRNA dispersed just prior to the initiation of cyclin B synthesis and germinal vesicle breakdown (GVBD). Cytochalasin B, but not nocodazole or taxol, deformed the aggregation of cyclin B mRNA, indicating the involvement of microfilaments in organizing this form. Like 17alpha,20beta-DP, cytochalasin B (10 microg/ml) induced both complete dispersion of the aggregation and translational activation of cyclin B mRNA, forcing the oocytes to undergo GVBD without 17alpha,20beta-DP. Conversely, disturbance of the aggregation of cyclin B mRNA with a low concentration (1 microg/ml) of cytochalasin B inhibited 17alpha,20beta-DP-induced GVBD. These results suggest that the direct change in cyclin B mRNA from the aggregated form to the dispersed form is responsible for translational activation of the mRNA during zebrafish oocyte maturation.

Animals↗
PubMed1976

Direct demonstration of a complementarity between mRNA and double-stranded sequences of pre-mRNA.

The total poly(A)-containing mRNA from mouse liver or Ehrlich ascites carcinoma cells was annealed with denatured ds RNA prepared from heavy nuclear 3H-labeled pre-mRNA of the same tissue. The hybrids formed were detected by binding of complexes to poly(U)-Sepharose columns through the poly(A) of mRNA. With this technique, about 30% of labeled ds RNA was bound to poly(U)-Sepharose after annealing it with an mRNA excess. The proportion of hybrid material detected by RNase treatment was two to three times lower than that obtained by poly(U)-Sepharose binding. The length of the RNase-stable acid precipitable hybrid material consisted of heterogeneous sequences of 10-100 nucleotides long when cytoplasmic, and 10-60 nucleotides long when polysomal mRNA was used in the hybridization reaction. The results obtained show that at least some of the mRNA molecules contain sequences complementary to one of the branches of the pre-mRNA hairpins. These results are compatible with the idea that the hairpin-like sequences in pre-mRNA are localized between mRNA and the non-informative part of the precursor molecule.

Animals↗
PubMed1995

Effect of fasting and growth hormone (GH) administration on GH receptor (GHR) messenger ribonucleic acid (mRNA) and GH-binding protein (GHBP) mRNA levels in male rats.

To elucidate whether GHR and GHBP are coordinately regulated or not, we studied the effect of fasting with or without GH administration on the GHR and GHBP mRNAs in the liver as well as in extrahepatic tissues in rats. Tissues were collected from 7-week-old male rats by decapitation 1,3, and 7 days after the start of fasting. Liver GHR mRNA levels were not affected 1 day after the start of fasting but progressively decreased for the subsequent 3 and 7 days of fasting as compared with those in control rats fed ad libitum. In contrast, liver GHBP mRNA levels significantly rose after 1 day fasting, returned to the control level after 3 days and further reduced after 7 days of fasting. Changes in GHBP mRNA level after fasting were different among the tissues. A transient increase in GHBP mRNA levels was observed in muscle and heart as well as liver, while the GHBP mRNA levels in fat tissues did not change throughout 7 days of fasting. Next, bovine GH(bGH) was administered ip to the fasted rats and control fed rats for either 1 day(100 micrograms [corrected], tid) or 5 days(150 micrograms [corrected], daily). In fed rats, liver GHR mRNA level was significantly increased by 1 day bGH treatment, but after 5 days treatment with bGH it was not different from the level in saline-injected control. Accordingly, net increment of plasma IGF-I was 296.0 ng/ml with 1 day bGH treatment and 234.2 ng/ml with bGH administration for 5 days. In fasted rats, liver GHR mRNA level did not changed after 1 day treatment with bGH, but markedly decreased 5 days after bGH administration. Net increment of plasma IGF-I was slightly reduced to 284 ng/ml with 1 day treatment with bGH, and markedly decreased to 37.0 with bGH administration for 5 days. The effect of GH administration on liver GHBP mRNA level was virtually absent in either fasting or fed state. These findings suggest that GHR and GHBP mRNAs in the liver are expressed in different ways and that expression of GHBP mRNA is differently regulated among tissues.

Animals↗
PubMed1989

Differences in the effects of membrane depolarization on levels of preprosomatostatin mRNA and tyrosine hydroxylase mRNA in rat sympathetic neurons in vivo and in culture.

Regulation of preprosomatostatin mRNA and tyrosine hydroxylase mRNA were examined in sympathetic neurons of the rat superior cervical ganglion (SCG). Surgical denervation of the adult SCG increased ganglion levels of preprosomatostatin (SS) mRNA more than 11-fold, and levels of the mRNA remained elevated 14 days after surgery. By contrast, denervation decreased levels of tyrosine hydroxylase (TH) mRNA. Potassium- or veratridine-induced membrane depolarization of cultured neonatal sympathetic neurons decreased levels of SS mRNA but elevated levels of TH mRNA. Sodium channel blockade with tetrodotoxin prevented the effects of veratridine on SS and TH mRNAs. In toto these observations suggest that transsynaptic nerve impulse activity and sympathetic neuron membrane depolarization decrease SS synthesis but increase TH synthesis at the mRNA level. Thus nerve impulse activity may alter the relative levels of different transmitters co-expressed in the same neuronal population by inhibiting levels of some species of mRNA while simultaneously stimulating levels of others.

Animals↗
PubMed1995

Photoperiodic exposure and time of day modulate the expression of arginine vasopressin mRNA and vasoactive intestinal peptide mRNA in the suprachiasmatic nuclei of Siberian hamsters.

In hamsters, changes in ambient photoperiod lead to alterations in the circadian rhythm of pineal melatonin secretion and subsequent changes in reproductive function. The present study examined whether photoperiod also alters 24-h rhythms in neuropeptide mRNA levels in the SCN of Siberian hamsters. In situ hybridization and quantitative autoradiography were used to assess messenger RNA levels for vasopressin (AVP) and vasoactive intestinal peptide (VIP) in the SCN of hamsters sacrificed at six times of day following exposure to long (16 h light/day) or short (10 h light/day) photoperiod for 2 weeks. Both AVP mRNA and VIP mRNA in the SCN were significantly affected by time of day and photoperiodic exposure. The 24-h profiles of AVP mRNA and VIP mRNA showed different relationships to the light: dark cycle, suggesting that these profiles are differentially regulated. In general, short photoperiod tended to suppress AVP mRNA and VIP mRNA in the SCN; this effect on AVP mRNA was significant at two times of day. These results complement and extend previous findings of 24-h h profiles in neuropeptide mRNA expression in the rat SCN by showing that these 24-h profiles are also characteristic of the Siberian hamster SCN and that they can be modulated by photoperiod.

Animals↗
PubMed1989

Histamine activates proenkephalin A mRNA but not phenylethanolamine N-methyltransferase mRNA expression in cultured bovine adrenal chromaffin cells.

The effects of histamine on the regulation of proenkephalin A (ProEnk A) and phenylethanolamine N-methyltransferase (PNMT) mRNA expression were examined in cultures of bovine adrenal chromaffin cells. Prolonged incubation with histamine resulted in a concentration-dependent increase in the levels of ProEnk A mRNA with little effect on the levels of PNMT mRNA. The activation of ProEnK A mRNA by histamine followed a slow time course, reaching 2-3 fold basal levels after 48 h incubation. This activation was antagonized by the H1-antagonist mepyramine but not by the H2-antagonist cimetidine indicating involvement of H1-histamine receptors. The histamine-induced activation of ProEnK mRNA was blocked by the RNA synthesis inhibitor actinomycin D, suggesting that the novo synthesis of ProEnkA mRNA is a requirement for activation. In the presence of the calcium channel blocker D600, the histamine-induced increase in ProEnk A mRNA was greatly reduced, though not abolished. Prolonged incubation with histamine also caused a substantial release of catecholamines and opioid peptides from these cells. These results suggest that the synthesis and release of opioid peptides is controlled by histamine via H1-receptors. The differential effects of histamine on ProEnk A mRNA and PNMT mRNA expression suggest that different regulatory mechanisms are called upon to regulate the synthesis of opioid peptides and adrenaline in response to stimulation of the chromaffin cells.

Adrenal Glands↗
PubMed2006

Relation between mRNA expression and sequence information in Desulfovibrio vulgaris: combinatorial contributions of upstream regulatory motifs and coding sequence features to variations in mRNA abundance.

The context-dependent expression of genes is the core for biological activities, and significant attention has been given to identification of various factors contributing to gene expression at genomic scale. However, so far this type of analysis has been focused either on relation between mRNA expression and non-coding sequence features such as upstream regulatory motifs or on correlation between mRNA abundance and non-random features in coding sequences (e.g., codon usage and amino acid usage). In this study multiple regression analyses of the mRNA abundance and all sequence information in Desulfovibrio vulgaris were performed, with the goal to investigate how much coding and non-coding sequence features contribute to the variations in mRNA expression, and in what manner they act together. Using the AlignACE program, 442 over-represented motifs were identified from the upstream 100bp region of 293 genes located in the known regulons. Regression of mRNA expression data against the measures of coding and non-coding sequence features indicated that 54.1% of the variations in mRNA abundance can be explained by the presence of upstream motifs, while coding sequences alone contribute to 29.7% of the variations in mRNA abundance. Interestingly, most of contribution from coding sequences is overlapping with that from upstream motifs; thereby a total of 60.3% of the variations in mRNA abundance can be explained when coding and non-coding information was included. This result demonstrates that upstream regulatory motifs and coding sequence information contribute to the overall mRNA expression in a combinatorial rather than an additive manner.

Base Sequence↗