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1.
Regul Pept ; 186: 49-56, 2013 Sep 10.
Article in English | MEDLINE | ID: mdl-23850797

ABSTRACT

Endogenous daily rhythms are generated by the hierarchically organized circadian system predominantly synchronized by the external light (L): dark (D) cycle. During recent years several humoral signals have been found to influence the generation and manifestation of daily rhythm. Since most studies have been performed under in vitro conditions, the mechanisms employed under in vivo conditions need to be investigated. Our study focused on angiotensin II (angII)-mediated regulation of Per2 expression in the suprachiasmatic nuclei (SCN) and heart and spontaneous locomotor activity in Wistar rats under synchronized conditions. Angiotensin II was infused (100ng/kg/min) via subcutaneously implanted osmotic minipumps for 7 or 28days. Samples were taken in 4-h intervals during a 24hcycle and after a light pulse applied in the first and second part of the dark phase. Gene expression was measured using real time PCR. Locomotor activity was monitored using an infrared camera with a remote control installed in the animal facility. Seven days of angII infusion caused an increase in blood pressure and heart/body weight index and 28days of angII infusion also increased water intake in comparison with controls. We observed a distinct daily rhythm in Per2 expression in the SCN and heart of control rats and infused rats. Seven days of angII infusion did not influence Per2 expression in the heart. 28days of angII treatment caused significant phase advance and a decrease in nighttime expression of Per2 and influenced expression of clock controlled genes Rev-erb alpha and Dbp in the heart compared to the control. Four weeks of angII infusion decreased the responsiveness of Per2 expression in the SCN to a light pulse at the end of the dark phase of the 24hcycle. Expression of mRNA coding angiotensin-converting enzyme (ACE) and angiotensin-converting enzyme 2 (ACE2) showed a daily rhythm in the heart of control rats. Four weeks of angII infusion caused a decrease in amplitude of rhythmic expression of Ace, the disappearance of rhythm and an increase in Ace2 expression. The Ace/Ace2 ratio showed a rhythmic pattern in the heart of control rats with peak levels during the dark phase. Angiotensin II infusion decreased the mean Ace/Ace2 mRNA ratio in the heart. We observed a significant daily rhythm in expression of brain natriuretic peptide (BNP) in the heart of control rats. In hypertensive rats mean value of Bnp expression increased. Locomotor activity showed a distinct daily rhythm in both groups. Angiotensin II time dependently decreased ratio of locomotor activity in active versus passive phase of 24hcycle. To conclude, 28days of subcutaneous infusion of angII modulates the functioning of the central and peripheral circadian system measured at the level of Per2 expression and locomotor activity.


Subject(s)
Angiotensin II/physiology , Myocardium/metabolism , Period Circadian Proteins/metabolism , Suprachiasmatic Nucleus/metabolism , ARNTL Transcription Factors/genetics , ARNTL Transcription Factors/metabolism , Angiotensin II/pharmacology , Angiotensin-Converting Enzyme 2 , Animals , Blood Pressure/drug effects , CLOCK Proteins/genetics , CLOCK Proteins/metabolism , Cardiomegaly/chemically induced , Circadian Rhythm , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Gene Expression , Motor Activity , Myocardium/pathology , Nuclear Receptor Subfamily 1, Group D, Member 1/genetics , Nuclear Receptor Subfamily 1, Group D, Member 1/metabolism , Peptidyl-Dipeptidase A/genetics , Peptidyl-Dipeptidase A/metabolism , Period Circadian Proteins/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Wistar , Transcription Factors/genetics , Transcription Factors/metabolism
2.
Mol Cell Biochem ; 348(1-2): 53-60, 2011 Feb.
Article in English | MEDLINE | ID: mdl-21076970

ABSTRACT

Under synchronized conditions daily rhythms run in precise phase relationships. Long lasting shift-work disturbs circadian rhythms and causes metabolism dysfunction. As a result of frequent shifts of the light (L):dark (D) cycle the circadian system has to adjust to a new regimen repeatedly, and organism can never achieve complete adjustment of all circadian rhythms. Nuclear receptor PPARα is supposed to be a functional interface between circadian clock and metabolism, and its interconnection with rev-erbα and pdk4 was proven. The aim of this study was to elucidate responsiveness of the circadian system to the LD cycle mimicking the rotating shift-work with 8-h phase delay every second day. Expression of key clock genes and clock controlled metabolic genes rev-erbα, pparα, and pdk4 was analyzed in the liver and heart of rats by real time PCR. Control Wistar rats were exposed to the regular LD cycle 12:12. The second group was exposed to the LD regimen mimicking shift-work with 8-h phase delays during period of 10 weeks. Sampling was performed in 4-h intervals during 24-h cycle. Clock gene expression in the heart and liver of shifted rats was rhythmic and phase delayed by 8-9 h compared to control. Expression of metabolic genes was influenced more in the liver than in the heart. Results indicate that frequent shifts of LD cycle may interfere with control of lipid metabolism.


Subject(s)
ARNTL Transcription Factors/genetics , Circadian Rhythm/genetics , Liver/metabolism , Myocardium/metabolism , Nuclear Receptor Subfamily 1, Group D, Member 1/genetics , PPAR alpha/genetics , Period Circadian Proteins/genetics , Photoperiod , Protein Serine-Threonine Kinases/genetics , Animals , Gene Expression Regulation , Male , Pyruvate Dehydrogenase Acetyl-Transferring Kinase , RNA, Messenger/metabolism , Rats , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Time Factors
3.
J Hypertens Suppl ; 27(6): S21-6, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19633447

ABSTRACT

OBJECTIVES: Plasma melatonin concentrations in non-dipping patients show a blunted daily rhythm. Melatonin has a capacity to improve disturbances in biological rhythms. Hypertensive TGR(mRen2)27 (TGR) rats with an upregulated renin-angiotensin system and inverted blood pressure profile were used to elucidate whether melatonin is able to influence the control of blood pressure. DESIGN: Melatonin was administered in drinking water to normotensive Sprague-Dawley (SD) and hypertensive TGR rats during the dark phase of the light: dark cycle 12: 12 for 4 weeks. METHODS: The effect of melatonin on blood pressure was monitored, and the expression of clock genes per2 and bmal1 and melatonin receptor MT1 in the suprachiasmatic nucleus (SCN) and the heart was measured by real time polymerase chain reaction during a 24-h cycle. RESULTS AND CONCLUSION: The administration of melatonin did not influence clock gene expression in the SCN but its effect on clock gene expression in the heart was phase dependent in both SD and TGR rats. Melatonin administration did not decrease the expression of melatonin receptors in the SCN and the heart. Melatonin did not decrease blood pressure in TGR rats but influenced the peripheral oscillator in the heart independently of the SCN. A modified function of molecular circadian oscillators in the heart can interfere with anticipation and disturb the adaptation of this organ to pressure overload.


Subject(s)
Gene Expression Regulation , Hypertension/metabolism , Melatonin/administration & dosage , Myocardium/metabolism , Renin/genetics , Suprachiasmatic Nucleus/metabolism , Trans-Activators/biosynthesis , Animals , Blood Pressure , CLOCK Proteins , Circadian Rhythm , Male , Melatonin/blood , Rats , Rats, Sprague-Dawley , Renin-Angiotensin System
4.
Article in English | MEDLINE | ID: mdl-19447189

ABSTRACT

The circadian system drives postnatally rhythmic processes in the peripheral tissues. To extend information about embryonic stages, ontogeny of the circadian oscillations in chicken (Gallus gallus) heart and liver was analyzed. Nineteen day old embryos and 4-day old chicks were synchronized to a light:dark cycle and then the effects of light pulses applied during the dark and subjective light phases of the 24 h cycle were tested. Expression of Per2 and Bmal1 was measured by RT-PCR. The effects of light pulses on plasma glucose and melatonin levels were also analyzed. The expression of Per2 and Bmal1 was rhythmic in the heart and liver of 4-day old chicks, however, more pronounced rhythmic expression was observed in the liver. The light pulse induced a significant increase in Per2 expression in the liver. Plasma glucose was up- and melatonin down-regulated after the light pulse postnatally. Expression of Per2 did not show a rhythmic pattern and the light pulse did not cause changes in Per2 expression in the liver and heart of the embryos. The absence of the rhythmic expression of the clock genes in the embryonic heart and liver may reflect missing rhythmic cues since early after hatching peripheral oscillators function in a mature-like manner.


Subject(s)
Circadian Rhythm/genetics , Heart/embryology , Liver/embryology , ARNTL Transcription Factors , Animals , Basic Helix-Loop-Helix Transcription Factors/biosynthesis , Blood Glucose/metabolism , Chick Embryo , Chickens/genetics , Down-Regulation , Heart/physiology , Light , Liver/metabolism , Melatonin/blood , Nuclear Proteins/biosynthesis , Transcription Factors/biosynthesis , Up-Regulation
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