Alena Sumová

Publications of Alena Sumová

• Photoperiodic modulation of the hepatic clock by the suprachiasmatic nucleus and feeding regime in mice.

  Authors: Daniela Parkanová, Marta Nováková, Serhiy Sosniyenko, Alena Sumová

  The European journal of neuroscience. 05/2012; 35(9):1446-57.

  Changes in photoperiod modulate the central circadian clock in the suprachiasmatic nucleus (SCN) as well as the peripheral clocks. Consequently, the SCN-driven output rhythms in activity and feedingChanges in photoperiod modulate the central circadian clock in the suprachiasmatic nucleus (SCN) as well as the peripheral clocks. Consequently, the SCN-driven output rhythms in activity and feeding are also modulated by the photoperiod. The aim of the present study was to elucidate whether photoperiodic modulation of the hepatic clock is mediated by changes in feeding or by another SCN-driven pathway. Five days after the change from short photoperiod (SP) to long photoperiod (LP), the profiles of Per2 and Rev-erbα expression in the rostral, middle and caudal regions of the SCN were desynchronized and those in the liver were modulated as in mice fully entrained to LP. The SCN profiles were not affected in mice left under SP and subjected to the 6-h night-time feeding regime for 5 days. In the liver, the profiles were shifted to the same phase, but their waveforms were not modulated compared with those under LP. In mice subjected to the change from SP to LP and fed twice daily during the daytime, the profiles in the SCN were not affected, whereas the waveforms and phases of those in the liver were affected. The data demonstrate that the adjustment of gene expression profiles in the rostral, middle and caudal SCN to the change from SP to LP proceeds within 5 days and is not affected by changes in the feeding regime. The results also suggest that the photoperiod-modulated SCN affects waveforms of gene expression profiles in the liver by food-independent signals.

• Alteration of the circadian clock in children with smith-magenis syndrome.

  Authors: Marta Nováková, Sona Nevsímalová, Iva Príhodová, Martin Sládek, Alena Sumová

  The Journal of clinical endocrinology and metabolism. 12/2011; 97(2):E312-8.

  Context: Smith-Magenis syndrome (SMS) is associated with sleep disturbances and disrupted melatonin production. Objectives: The study aimed to ascertain whether the sleep and melatonin productionContext: Smith-Magenis syndrome (SMS) is associated with sleep disturbances and disrupted melatonin production. Objectives: The study aimed to ascertain whether the sleep and melatonin production anomalies in SMS patients may be due to an alteration of the molecular mechanism of the circadian clock. Subjects and Methods: Five SMS patients (3-17 yr old) and five healthy age-matched control subjects were involved in the study. Saliva and buccal scrub samples were collected every 4 h during a 24-h period. Daily profiles of melatonin were determined in saliva using a direct double-antibody radioimmunoassay. Daily profiles of clock gene mRNA levels (Per1, Per2, and Rev-erbα) were determined in buccal scrub samples by RT-PCR. Results: In controls, melatonin levels were elevated during the nighttime and very low during the daytime. Daily profiles of clock genes, Per1, Per2, and Rev-erbα, mRNA levels in buccal mucosa exhibited significant and mutually synchronized circadian variations (Per1 and Rev-erbα: P < 0.001; Per2: P < 0.05); the mRNA levels were elevated during the daytime and decreased during the nighttime. In SMS patients, melatonin profiles were significantly altered compared with controls, being phase reversed, phase advanced, depressed, or abolished. Only Per1 and Rev-erbα mRNA profiles exhibited significant circadian rhythms (P < 0.05); the Per2 expression exhibited high variability, and the profile was out of phase with the other clock genes. Conclusion: Our findings suggest that the anomalies in melatonin profiles of SMS patients might be due to a disturbance of the molecular circadian clockwork.

• Circadian regulation of electrolyte absorption in the rat colon.

  Authors: M Soták, L Polidarová, J Musílková, M Hock, A Sumová, J Pácha

  American journal of physiology. Gastrointestinal and liver physiology. 09/2011; 301(6):G1066-74.

  The intestinal transport of nutrients exhibits distinct diurnal rhythmicity, and the enterocytes harbor a circadian clock. However, temporal regulation of the genes involved in colonic ion transport,The intestinal transport of nutrients exhibits distinct diurnal rhythmicity, and the enterocytes harbor a circadian clock. However, temporal regulation of the genes involved in colonic ion transport, i.e., ion transporters and channels operating in absorption and secretion, remains poorly understood. To address this issue, we assessed the 24-h profiles of expression of genes encoding the sodium pump (subunits Atp1a1 and Atp1b1), channels (α-, β-, and γ-subunits of Enac and Cftr), transporters (Dra, Ae1, Nkcc1, Kcc1, and Nhe3), and the Na(+)/H(+) exchanger (NHE) regulatory factor (Nherf1) in rat colonic mucosa. Furthermore, we investigated temporal changes in the spatial localization of the clock genes Per1, Per2, and Bmal1 and the genes encoding ion transporters and channels along the crypt axis. In rats fed ad libitum, the expression of Atp1a1, γEnac, Dra, Ae1, Nhe3, and Nherf1 showed circadian variation with maximal expression at circadian time 12, i.e., at the beginning of the subjective night. The peak γEnac expression coincided with the rise in plasma aldosterone. Restricted feeding phase advanced the expression of Dra, Ae1, Nherf, and γEnac and decreased expression of Atp1a1. The genes Atp1b1, Cftr, αEnac, βEnac, Nkcc1, and Kcc1 did not show any diurnal variations in mRNA levels. A low-salt diet upregulated the expression of βEnac and γEnac during the subjective night but did not affect expression of αEnac. Similarly, colonic electrogenic Na(+) transport was much higher during the subjective night than the subjective day. These findings indicate that the transporters and channels operating in NaCl absorption undergo diurnal regulation and suggest a role of an intestinal clock in the coordination of colonic NaCl absorption.

• Exposure of pregnant rats to restricted feeding schedule synchronizes the SCN clocks of their fetuses under constant light but not under a light-dark regime.

  Authors: Marta Nováková, Martin Sládek, Alena Sumová

  Journal of biological rhythms. 10/2010; 25(5):350-60.

  The circadian clock in the suprachiasmatic nucleus (SCN) develops gradually during the prenatal and early postnatal period. In the rat, this period lasts from around the 15th day of gestation untilThe circadian clock in the suprachiasmatic nucleus (SCN) develops gradually during the prenatal and early postnatal period. In the rat, this period lasts from around the 15th day of gestation until the 10th day of postnatal development. The circadian system of fetuses and newborn pups is entrained mostly by nonphotic maternal cues during prenatal and early postnatal development. The aim of the present study was to ascertain whether exposure of pregnant rats to a restricted feeding (RF) regime was able to entrain the circadian clock in the SCN of their fetuses during the prenatal period. The potency of RF as an entraining cue was tested under conditions when pregnant rats were entrained to an external light/dark (LD) cycle as well as under conditions when the external timing signal was lacking, i.e., under constant light (LL). The control groups were fed ad libitum and the experimental groups had restricted access to food for 6 h during their resting time throughout pregnancy. Daily profiles of Avp and c-fos gene expression were examined by in situ hybridization in the SCN of 1-day-old pups. The data demonstrated that RF in pregnant rats kept under LD cycle did not notably affect the daily rhythms of c-fos and Avp expression in the SCN of pups. The SCN profiles of Avp and c-fos gene expression did not exhibit circadian rhythms in pups born to mothers maintained in LL and fed ad libitum, likely due to desynchrony among the pups within a litter. However, RF in the pregnant rats kept under LL restored the circadian rhythmicity of c-fos and Avp expression in the SCN of their newborn pups. The results suggest that the fetal SCN clock is dominantly entrained by rhythmic signals from the maternal SCN. However, under conditions when the rhythmic signaling might be lacking, such as LL, regular food intake of the mothers may also play an important role in synchronization of the fetal SCN clock during prenatal ontogenesis.

• Expression and light sensitivity of clock genes Per1 and Per2 and immediate-early gene c-fos within the retina of early postnatal Wistar rats.

  Authors: Kristýna Matejů, Alena Sumová, Zdenka Bendová

  The Journal of comparative neurology. 09/2010; 518(17):3630-44.

  Mammalian retina contains a circadian clock that is composed of components similar to those of the master circadian clock within the suprachiasmatic nuclei of the hypothalamus. The aim of the presentMammalian retina contains a circadian clock that is composed of components similar to those of the master circadian clock within the suprachiasmatic nuclei of the hypothalamus. The aim of the present study was to elucidate whether, when, and where the transcripts of the clock genes Per1 and Per2 and the immediate early gene c-fos are spontaneously expressed and/or induced by light in the newborn rat retina. At postnatal day 1 (P1), P3, P5, and P10, Wistar rat pups were released into constant darkness, and a 30-minute light pulse was administered during the subjective day or during the first or second part of subjective night. Gene expression was determined 30 minutes, 1 hour, 2 hours, and 4 hours after the light pulse by in situ hybridization followed by emulsion autoradiography. Endogenous expression of Per1 was detected in the neuroblastic retina, and Per2 expression was detected in the inner part of the neuroblastic retina from birth. Light pulses induced c-fos expression in ganglion cells from P1. Until P5, the cells were localized in the dorsal part of the retina, but, at P10, they were already distributed across the entire retinal circumference. Light pulses also induced the expression of c-fos and Per1 in the retinal pigment epithelium until P3, but not afterward. Expression of the Per2 gene was not photoresponsive until P10. These data demonstrate that the rat retina is light-sensitive immediately after birth. During early postnatal development, the spatial distribution of spontaneous and light-induced gene expression within the retinal layers changes gradually.

• Different mechanisms of adjustment to a change of the photoperiod in the suprachiasmatic and liver circadian clocks.

  Authors: Serhiy Sosniyenko, Daniela Parkanová, Helena Illnerová, Martin Sládek, Alena Sumová

  American journal of physiology. Regulatory, integrative and comparative physiology. 04/2010; 298(4):R959-71.

  Changes in photoperiod modulate the circadian system, affecting the function of the central clock located in the suprachiasmatic nucleus (SCN) of the hypothalamus. The aim of the present study was toChanges in photoperiod modulate the circadian system, affecting the function of the central clock located in the suprachiasmatic nucleus (SCN) of the hypothalamus. The aim of the present study was to elucidate the dynamics of adjustment to a change of a long photoperiod with 18 h of light to a short photoperiod with 6 h of light of clock gene expression rhythms in the mouse SCN and in the peripheral clock in the liver, as well as of the locomotor activity rhythm. Three, five, and thirteen days after the photoperiod change, daily profiles of Per1, Per2, and Rev-erbalpha expression in the rostral, middle, and caudal parts of the SCN and of Per2 and Rev-erbalpha in the liver were determined by in situ hybridization and real-time RT-PCR, respectively. The clock gene expression rhythms in the different SCN regions, desynchronized under the long photoperiod, attained synchrony gradually following the transition from long to short days, mostly via advancing the expression decline. The photoperiodic modulation of the SCN was due not only to the degree of synchrony among the SCN regions but also to different waveforms of the rhythms in the individual SCN parts. The locomotor activity rhythm adjusted gradually to short days by advancing the activity onset, and the liver rhythms adjusted by advancing the Rev-erbalpha expression rise and Per2 decline. These data indicate different mechanisms of adjustment to a change of the photoperiod in the central SCN clock and the peripheral liver clock.

• Influence of photoperiod duration and light-dark transitions on entrainment of Per1 and Per2 gene and protein expression in subdivisions of the mouse suprachiasmatic nucleus.

  Authors: Serhiy Sosniyenko, Roelof A Hut, Serge Daan, Alena Sumová

  The European journal of neuroscience. 10/2009;

  Abstract The circadian clock located within the suprachiasmatic nuclei (SCN) of the hypothalamus responds to changes in the duration of day length, i.e. photoperiod. Recently, changes in phaseAbstract The circadian clock located within the suprachiasmatic nuclei (SCN) of the hypothalamus responds to changes in the duration of day length, i.e. photoperiod. Recently, changes in phase relationships among the SCN cell subpopulations, especially between the rostral and caudal region, were implicated in the SCN photoperiodic modulation. To date, the effect of abrupt, rectangular, light-to-dark transitions have been studied while in nature organisms experience gradual dawn and twilight transitions. The aim of this study was to compare the effect of a long (18 h of light) and a short (6 h of light) photoperiod with twilight relative to that with rectangular light-to-dark transition on the daily profiles of Per1 and Per2 mRNA (in situ hybridization) and PER1 and PER2 protein (immunohistochemistry) levels within the rostral, middle and caudal regions of the mouse SCN. Under the short but not under the long photoperiod, Per1, Per2 and PER1, PER2 profiles were significantly phase-advanced under the twilight relative to rectangular light-to-dark transition in all SCN regions examined. Under the photoperiods with rectangular light-to-dark transition, Per1 and Per2 mRNA profiles in the caudal SCN were phase-advanced as compared with those in the rostral SCN. The phase differences between the SCN regions were reduced under the long, or completely abolished under the short, photoperiods with twilight. The data indicate that the twilight photoperiod provides stronger synchronization among the individual SCN cell subpopulations than the rectangular one, and the effect is more pronounced under the short than under the long photoperiod.

• Temporal gradient in the clock gene and cell-cycle checkpoint kinase Wee1 expression along the gut.

  Authors: Lenka Polidarová, Matús Soták, Martin Sládek, Jirí Pacha, Alena Sumová

  Chronobiology international. 06/2009; 26(4):607-20.

  Circadian clocks were recently discovered in the rat and mouse colon as well as mouse stomach and jejunum. The aim of this study was to determine whether clocks in the upper part of the gut areCircadian clocks were recently discovered in the rat and mouse colon as well as mouse stomach and jejunum. The aim of this study was to determine whether clocks in the upper part of the gut are synchronized with those in the lower part, or whether there is a difference in their circadian phases. Moreover, the profiles of core clock-gene expression were compared with the profiles of the clock-driven Wee1 gene expression in the upper and lower parts of the gut. Adult rats were transferred to constant darkness on the day of sampling. 24 h expression profiles of the clock genes Per1, Per2, Rev-erbalpha, and Bmal1 and the cell-cycle regulator Wee1 were examined by a reverse transcriptase-polymerase chain reaction within the epithelium of the rat duodenum, ileum, jejunum, and colon. In contrast to the duodenum, the rhythms in expression of all genes but Rev-erbalpha and Bmal1 in the colon exhibited non-sinusoidal profiles. Therefore, a detailed analysis of the gene expression every 1 h within the 12 h interval corresponding to the previous lights-on was performed. The data demonstrate that rhythmic profiles of the clock gene Per1, Per2, Bmal1, Rev-erbalpha, and clock-driven Wee1 expression within the epithelium from different parts of the rat gut exhibited a difference in phasing, such that the upper part of the gut, as represented by the duodenum, was phase-advanced to the lower part, as represented by the distal colon. Our data demonstrate that the circadian clocks within each part of the gut are mutually synchronized with a phase delay in the cranio-caudal axis. Moreover, they support the view that the individual circadian clocks may control the timing of cell cycle within different regions of the gut.

• Development of the light sensitivity of the clock genes Period1 and Period2, and immediate-early gene c-fos within the rat suprachiasmatic nucleus.

  Authors: Kristýna Matejů, Zdena Bendová, Rehab El-Hennamy, Martin Sládek, Serhiy Sosniyenko, Alena Sumová

  The European journal of neuroscience. 03/2009; 29(3):490-501.

  The molecular mechanism underlying circadian rhythmicity within the suprachiasmatic nuclei (SCN) of the hypothalamus has two light-sensitive components, namely the clock genes Per1 and Per2. Besides,The molecular mechanism underlying circadian rhythmicity within the suprachiasmatic nuclei (SCN) of the hypothalamus has two light-sensitive components, namely the clock genes Per1 and Per2. Besides, light induces the immediate-early gene c-fos. In adult rats, expression of all three genes is induced by light administered during the subjective night but not subjective day. The aim of the present study was to ascertain when and where within the SCN the photic sensitivity of Per1, Per2 and c-fos develops during early postnatal ontogenesis. The specific aim was to find out when the circadian clock starts to gate photic sensitivity. The effect of a light pulse administered during either the subjective day or the first or second part of the subjective night on gene expression within the rat SCN was determined at postnatal days (P) 1, 3, 5 and 10. Per1, Per2 and c-fos mRNA levels were assessed 30 min, 1 and 2 h after the start of each light pulse by in situ hybridization histochemistry. Expression of Per1 and c-fos was light responsive from P1, and the responses began to be gated by the circadian clock at P3 and P10, respectively. Expression of Per2 was only slightly light responsive at P3, and the response was not fully gated until P5. These data demonstrate that the light sensitivity of the circadian clock develops gradually during postnatal ontogenesis before the circadian clock starts to control the response. The photoinduction of the clock gene Per2 develops later than that of Per1.

• Maternal control of the fetal and neonatal rat suprachiasmatic nucleus.

  Authors: Rehab El-Hennamy, Kristyna Mateju, Zdena Bendová, Serhiy Sosniyenko, Alena Sumová

  Journal of biological rhythms. 11/2008; 23(5):435-44.

  The molecular clockwork underlying the generation of circadian rhythmicity within the suprachiasmatic nucleus (SCN) develops gradually during ontogenesis. The authors' previous work has shown thatThe molecular clockwork underlying the generation of circadian rhythmicity within the suprachiasmatic nucleus (SCN) develops gradually during ontogenesis. The authors' previous work has shown that rhythms in clock gene expression in the rat SCN are not detectable at embryonic day (E) 19, start to form at E20 and develop further via increasing amplitude until postnatal day (P) 10. The aim of the present work was to elucidate whether and how swiftly the immature fetal and neonatal molecular SCN clocks can be reset by maternal cues. Pregnant rats maintained under a light-dark (LD) regimen with 12 h of light and 12 h of darkness were exposed to a 6-h delay of the dark period and released into constant darkness at different stages of the fetal SCN development. Adult rats maintained under the same LD regimen were exposed to an identical shifting procedure. Daily rhythms in spontaneous c-fos, Avp, Per1, and Per2 expression were examined within the adult and newborn SCN by in situ hybridization. Exposure of adult rats to the shifting procedure induced a significant phase delay of locomotor activity within 3 days after the phase shift as well as a delay in the rhythms of c-fos and Avp expression within 3 days and Per1 and Per2 expression within 5 days. Exposure of pregnant rats to the shifting procedure at E18, but not at E20, delayed the rhythm in c-fos and Avp expression in the SCN of newborn pups at P0-1. The shifting procedure at E20 did, however, induce a phase delay of Per1 and Per2 expression rhythms at P3 and P6. Hence, 5 days were necessary for phase-shifting the pups' SCN clock by maternal cues, be it the interval between E18 and P0-1 or the interval between E20 and P3, while only 3 days were necessary for phase-shifting the maternal SCN by photic cues. These results demonstrate that the SCN clock is capable of significant phase shifts at fetal developmental stages when no or very faint molecular oscillations can be detected.

• Insight into the circadian clock within rat colonic epithelial cells.

  Authors: Martin Sládek, Markéta Rybová, Zuzana Jindráková, Zdena Zemanová, Lenka Polidarová, Libor Mrnka, John O'Neill, Jirí Pácha, Alena Sumová

  Gastroenterology. 10/2007; 133(4):1240-9.

  BACKGROUND & AIMS: The gastrointestinal tract exhibits diurnal rhythms in many physiologic functions. These rhythms are driven by food intake but are also preserved during food deprivation,BACKGROUND & AIMS: The gastrointestinal tract exhibits diurnal rhythms in many physiologic functions. These rhythms are driven by food intake but are also preserved during food deprivation, suggesting the presence of endogenous circadian rhythmicity. The aim of the study was to provide insight into the circadian core clock mechanism within the rat colon. Moreover, the potency of a restricted feeding regime to shift the circadian clock in the colon was tested. The question of whether the colonic clock drives circadian expression in NHE3, an electroneutral Na(+)/H(+) exchanger, was also addressed. METHODS: Daily profiles in expression of clock genes Per1, Per2, Cry1, Bmal1, Clock, and Rev-erbalpha, and the NHE3 transporter were examined by reverse transcriptase-polymerase chain reaction and their mRNA levels, as well as PER1 and BMAL1 protein levels, were localized in the colonic epithelium by in situ hybridization and immunocytochemistry, respectively. RESULTS: Expression of Per1, Per2, Cry1, Bmal1, Clock, Rev-erbalpha, and NHE3, as well as PER1 and BMAL1 protein levels, exhibited circadian rhythmicity in the colon. The rhythms were in phase with those in the liver but phase-delayed relative to the master clock in the suprachiasmatic nucleus. Restricted feeding entrained the clock in the colon, because rhythms in clock genes as well as in NHE3 expression were phase-advanced similarly to the clock in the liver. CONCLUSIONS: The rat colon harbors a circadian clock. The colonic clock is likely to drive rhythmic NHE3 expression. Restricted feeding resets the colonic clock similarly to the clock in the liver.

• Dynamics of the adjustment of clock gene expression in the rat suprachiasmatic nucleus to an asymmetrical change from a long to a short photoperiod.

  Authors: Alena Sumová, Zuzana Kováciková, Helena Illnerová

  Journal of biological rhythms. 07/2007; 22(3):259-67.

  The molecular clockwork of the rat suprachiasmatic nucleus, the site of the circadian clock, is affected by the photoperiod (Sumová et al., 2003). The aim of the present study was to partly elucidateThe molecular clockwork of the rat suprachiasmatic nucleus, the site of the circadian clock, is affected by the photoperiod (Sumová et al., 2003). The aim of the present study was to partly elucidate the dynamics of the adjustment of the clockwork to a change from a long to a short photoperiod accomplished by an asymmetrical prolongation of the dark period into the morning hours. Rats maintained under a regime with 16 h of light and 8 h of darkness per day (LD 16:8) were transferred to LD 8:16, and after 2, 3, and 13 days, daily profiles of Per1, Per2, Bmal1, and Cry1 mRNA were assessed by in situ hybridization. The rhythms of Per1, Per2, and Bmal1 expression adjusted to the change from a long to a short photoperiod with larger phase delays of the morning Per mRNA rise and Bmal1 mRNA decline than of the evening and nighttime Per mRNA decline and Bmal1 mRNA rise. The rhythm of Cry1 expression adjusted to the change by parallel delays of the Cry1 mRNA rise and decline. Adjustment of the Cry1 mRNA rhythm to short days was almost accomplished within 13 days, whereas adjustment of the Per1 and Bmal1 mRNA rhythms took longer. Different dynamics of the adjustment of rhythms in clock gene expression to a change from a long to a short photoperiod suggests complex resetting effects of the photoperiod change.

• Postnatal ontogenesis of the circadian clock within the rat liver.

  Authors: Martin Sládek, Zuzana Jindráková, Zdenka Bendová, Alena Sumová

  American journal of physiology. Regulatory, integrative and comparative physiology. 04/2007; 292(3):R1224-9.

  In mammals, the circadian oscillator within the suprachiasmatic nuclei (SCN) entrains circadian clocks in numerous peripheral tissues. Central and peripheral clocks share a molecular core clockIn mammals, the circadian oscillator within the suprachiasmatic nuclei (SCN) entrains circadian clocks in numerous peripheral tissues. Central and peripheral clocks share a molecular core clock mechanism governing daily time measurement. In the rat SCN, the molecular clockwork develops gradually during postnatal ontogenesis. The aim of the present work was to elucidate when during ontogenesis the expression of clock genes in the rat liver starts to be rhythmic. Daily profiles of mRNA expression of clock genes Per1, Per2, Cry1, Clock, Rev-Erbalpha, and Bmal1 were analyzed in the liver of fetuses at embryonic day 20 (E20) or pups at postnatal age 2 (P2), P10, P20, P30, and in adults by real-time RT-PCR. At E20, only a high-amplitude rhythm in Rev-Erbalpha and a low-amplitude variation in Cry1 but no clear circadian rhythms in expression of other clock genes were detectable. At P2, a high-amplitude rhythm in Rev-Erbalpha and a low-amplitude variation in Bmal1 but no rhythms in expression of other genes were detected. At P10, significant rhythms only in Per1 and Rev-Erbalpha expression were present. At P20, clear circadian rhythms in the expression of Per1, Per2, Rev-Erbalpha, and Bmal1, but not yet of Cry1 and Clock, were detected. At P30, all clock genes were expressed rhythmically. The phase of the rhythms shifted between all studied developmental periods until the adult stage was achieved. The data indicate that the development of the molecular clockwork in the rat liver proceeds gradually and is roughly completed by 30 days after birth.

• Setting the biological time in central and peripheral clocks during ontogenesis.

  Authors: Alena Sumová, Zdenka Bendová, Martin Sládek, Rehab El-Hennamy, Kristýna Laurinová, Zuzana Jindráková, Helena Illnerová

  FEBS letters. 06/2006; 580(12):2836-42.

  In mammals, the principal circadian clock within the suprachiasmatic nucleus (SCN) entrains the phase of clocks in numerous peripheral tissues and controls the rhythmicity in various body functions.In mammals, the principal circadian clock within the suprachiasmatic nucleus (SCN) entrains the phase of clocks in numerous peripheral tissues and controls the rhythmicity in various body functions. During ontogenesis, the molecular mechanism responsible for generating circadian rhythmicity develops gradually from the prenatal to the postnatal period. In the beginning, the maternal signals set the phase of the newly developing fetal and early postnatal clocks, whereas the external light-dark cycle starts to entrain the clocks only later. This minireview discusses the complexity of signaling pathways from mothers and the outside world to the fetal and newborn animals' circadian clocks.

• Expression of clock and clock-driven genes in the rat suprachiasmatic nucleus during late fetal and early postnatal development.

  Authors: Zuzana Kováciková, Martin Sládek, Zdenka Bendová, Helena Illnerová, Alena Sumová

  Journal of biological rhythms. 05/2006; 21(2):140-8.

  The SCN as a site of the circadian clock itself exhibits rhythmicity. A molecular clockwork responsible for the rhythmicity consists of clock genes and their negative and positiveThe SCN as a site of the circadian clock itself exhibits rhythmicity. A molecular clockwork responsible for the rhythmicity consists of clock genes and their negative and positive transcriptional-translational feedback loops. The authors' previous work showed that rhythms in clock gene expression in the rat SCN were not yet detectable at embryonic day (E) 19 but were already present at postnatal day (P) 3. The aim of the present study was to elucidate when during the interval E19-P3 the rhythms start to develop in clock gene expression and in clock-controlled, namely in arginine-vasopressin (AVP), gene expression. Daily profiles of Per1, Per2, Cry1, Bmal1, and Clock mRNA and of AVP heteronuclear (hn) RNA as an indicator of AVP gene transcription were assessed in the SCN of fetuses at E20 and of newborn rats at P1 and P2 by the in situ hybridization method. At E20, formation of a rhythm in Per1 expression was indicated, but no rhythms in expression of other clock genes or of the AVP gene were detected. At P1, rhythms in Per1, Bmal1, and AVP and a forming rhythm in Per2 but no rhythm in Cry1 expression were present in the SCN. The Per1 mRNA rhythm was, however, only slightly pronounced. The Bmal1 mRNA rhythm, although pronounced, exhibited still an atypical shape. Only the AVP hnRNA rhythm resembled that of adult rats. At P2, marked rhythms of Per1, Per2, and Bmal1 and a forming rhythm of Cry1, but not of Clock, expression were present. The data suggest that rhythms in clock gene expression for the most part develop postnatally and that other mechanisms besides the core clockwork might be involved in the generation of the rhythmic AVP gene expression in the rat SCN during early ontogenesis.

• The rat circadian clockwork and its photoperiodic entrainment during development.

  Authors: Alena Sumová, Zdenka Bendová, Martin Sládek, Zuzana Kováciková, Rehab El-Hennamy, Kristýna Laurinová, Helena Illnerová

  Chronobiology international. 02/2006; 23(1-2):237-43.

  The mammalian circadian pacemaker is located in the suprachiasmatic nucleus (SCN), which is composed of dorsomedial (dm) and ventrolateral (vl) regions. The molecular clockwork responsible for theThe mammalian circadian pacemaker is located in the suprachiasmatic nucleus (SCN), which is composed of dorsomedial (dm) and ventrolateral (vl) regions. The molecular clockwork responsible for the SCN rhythmicity consists of clock genes and their transcriptional-translational feedback loops. The rat SCN rhythmicity and clockwork are affected by the photoperiod. The aim of this study was to elucidate development of the rat SCN rhythmicity, namely of the rhythmicity of the dm- and vl-SCN and of expression of clock genes and to ascertain when the photoperiod starts to affect the SCN rhythmicity. Rhythmicity of the dm-SCN, measured as the rhythm in spontaneous c-FOS production, developed earlier than that of the vl-SCN, which was measured as the rhythm in c-FOS photoinduction. However, photoperiodic affection of the rhythmicity occurred earlier in the vl-SCN than in the dm-SCN. From the 4 clock genes (Per1, Per2, Cry1 and Bmal1) studied, the expression of Bmal1 and Per1 was rhythmic already in 1-day-old rats; at this age, the Per2 mRNA rhythm just started to form and no rhythm in Cry1 expression was detected. After the second postnatal day, all 4 genes were expressed in a rhythmic manner. Thereafter, the rhythms matured gradually via increasing amplitude. Per1 and Per2 mRNA rhythms started to be affected by the photoperiod at the 10th postnatal day. The data suggest that the rhythms in clock genes expression in the rat SCN develop mostly postnatally. The molecular clockwork may start to be photoperiod-dependent around the 10th postnatal day.

• [Development of the mammalian circadian system]

  Authors: Kristýna Laurinová, Alena Sumová

  Ceskoslovenská fysiologie / Ústrední ústav biologický. 02/2006; 55(4):148-54.

  Many behavioral, physiological and molecular processes exhibit diurnal rhythms. Endogenous rhythms with period close to 24 hours are called circadian rhythms. Light entrains circadian rhythms to a 24Many behavioral, physiological and molecular processes exhibit diurnal rhythms. Endogenous rhythms with period close to 24 hours are called circadian rhythms. Light entrains circadian rhythms to a 24 period of solar day. Circadian system consists of pacemaker, which is in mammals located in the suprachiasmatic nuclei of hypothalamus (SCN), its input and output pathways and peripheral clocks in numerous tissues. The generation of circadian rhytmicity is based on interactive transcription-translational feedback loops in SCN. These feedback loops consist of so called clock genes and their protein products which positively or negatively regulate their own transcription. Studies in rodent embros and neonates demonstrate that individual components of circadian system matures gradually during prenatal and postnatal period Mechanism of light entrainment of the circadian system develops postnatally. During early postnatal period, the developing circadian system is synchronized mainly by maternal cues.

• Effect of photic stimuli disturbing overt circadian rhythms on the dorsomedial and ventrolateral SCN rhythmicity.

  Authors: Alena Sumová, Helena Illnerová

  Brain research. 07/2005; 1048(1-2):161-9.

  To ascertain how photic stimuli disturbing overt circadian rhythms affect the endogenous rhythmicity of the suprachiasmatic nucleus (SCN), rats were subjected to constant light (LL) or to a 9-h lightTo ascertain how photic stimuli disturbing overt circadian rhythms affect the endogenous rhythmicity of the suprachiasmatic nucleus (SCN), rats were subjected to constant light (LL) or to a 9-h light pulse encompassing midnight, and rhythms of abundance of the c-Fos-immunoreactive (c-Fos-ir) and the PER1-immunoreactive (PER1-ir) cells were studied during the first 1-2 cycles following release into LL or darkness (DD) within the whole SCN as well as in its ventrolateral (vl) and the dorsomedial (dm) part. LL seemingly abolished the c-Fos rhythm in the whole SCN, while the rhythm persisted in the vl- and dm-SCN. In the dm-SCN, the rhythm of c-Fos-ir was phase-delayed by about 4 h in LL, whereas the rhythm of PER1-ir was affected just slightly. In the vl-SCN, the rhythm of c-Fos photo-induction might be delayed by 5-6 h as compared with the reported rhythm [A. Sumova and H. Illnerova, Am. J. Physiol. 274 (1998) R857-R863], whereas the PER1 profile appeared to be out of phase with that in DD. After a 9-h light pulse encompassing midnight, the rhythm of PER1-ir in the dm-SCN changed just slightly, whereas the PER1 rhythm in the vl-SCN was abolished and there was just an indication of extension of elevated PER1-ir. Altogether, the data indicate that photic stimuli disturbing circadian rhythms affect more dramatically the vl- than the dm-SCN rhythmicity within the first cycles and that in the dm-SCN shifting of the c-Fos rhythm proceeds more rapidly than that of the Per1 rhythm.

• Insight into molecular core clock mechanism of embryonic and early postnatal rat suprachiasmatic nucleus.

  Authors: Martin Sládek, Alena Sumová, Zuzana Kováciková, Zdenka Bendová, Kristyna Laurinová, Helena Illnerová

  Proceedings of the National Academy of Sciences of the United States of America. 05/2004; 101(16):6231-6.

  Rhythmicity of the rat suprachiasmatic nucleus (SCN), a site of the circadian clock, develops prenatally. A molecular clockwork responsible for the rhythmicity consists of clock genes and theirRhythmicity of the rat suprachiasmatic nucleus (SCN), a site of the circadian clock, develops prenatally. A molecular clockwork responsible for the rhythmicity consists of clock genes and their negative and positive transcriptional-translational feedback loops. The aim of the present study was to discover the development of the clockwork during ontogenesis. Daily profiles of Per1, Per2, Cry1, Bmal1, and Clock mRNA in the SCN of fetuses at the embryonic day (E)19 and of newborn rats at the postnatal day (P)3 and P10 were assessed by the in situ hybridization method. In addition, daily profiles of PER1, PER2, and CRY1 proteins at E19 were assessed by immunohistochemistry. As early as at E19, all the studied clock genes were already expressed in the SCN. However, no SCN rhythm in their expression was detected; Per1, Cry1, and Clock mRNA levels were low, whereas Bmal1 mRNA levels were high and Per2 mRNA levels were medium. Moreover, no rhythms of PER1, PER2, and CRY1 were detectable, as no immunoreactive cells were present at E19. At P3, rhythms in Per1, Per2, Cry1, and Bmal1, but not in Clock mRNA, were expressed in the SCN. The rhythm matured gradually; at P10, the amplitude of Per1, Per2, and Bmal1 mRNA rhythms was more pronounced than at P3. Altogether, the data show a gradual development of both the positive and negative elements of the molecular clockwork, from no detectable rhythmicity at E19 to highly developed rhythms at P10.

• Development of circadian rhythmicity and photoperiodic response in subdivisions of the rat suprachiasmatic nucleus.

  Authors: Zdenka Bendová, Alena Sumová, Helena Illnerová

  Brain research. Developmental brain research. 02/2004; 148(1):105-12.

  To ascertain whether the circadian rhythmicity of the ventrolateral (vl) suprachiasmatic nucleus (SCN) develops concurrently with that of the dorsomedial (dm) SCN and when the rhythmicity starts toTo ascertain whether the circadian rhythmicity of the ventrolateral (vl) suprachiasmatic nucleus (SCN) develops concurrently with that of the dorsomedial (dm) SCN and when the rhythmicity starts to respond to day length, i.e., to the photoperiod, rats with their offspring were maintained under either a long photoperiod with 16 h of light and 8 h of darkness per day (LD 16:8) or under a short, LD 8:16 photoperiod. The rhythms of spontaneous c-Fos immunoreactivity in the dm-SCN and of the light-induced c-Fos immunoreactivity in the vl-SCN were studied in the pups. In 3- and 10-day-old rats, the dm-SCN rhythm in spontaneous c-Fos immunoreactivty was already well expressed but a response to a photoperiod similar to that in adult rats has not yet been developed. The vl-SCN gate for insensitivity of c-Fos production to light at certain times was detected in 10-day but not yet in 3-day-old rats: in the latter, light exposure at any daytime induced high c-Fos immunoreactivity. In the 10-day-old pups, similarly as with adult rats, the gate was shorter under LD 8:16 than under LD 16:8, but the difference in the gate duration between the short and the long photoperiod did not yet attain that of adult animals. The data indicate that the circadian rhythmicity may develop sooner in the dm-SCN, than in the vl-SCN, whereas the photoperiodic response may develop sooner in the vl-SCN.