The Role of Endogenous Carbon Monoxide (CO) on Functioning of Biological Clock in Pig and Wild Boar Hybrid During Long and Short Day Season

Abstract Bacground: The master biological clock (pacemaker) is located in the suprachiasmatic nuclei (SCN) of the preoptic part of the hypothalamus (POA) and makes organisms adjust to the rhythms, both annual and circadian. Intrinsic pacemaker function is based on the 24-hour oscillator of the transcription factor genes Bmal1/Clock or its paralog Npas2 in which an E-box-bound heterodimeric transcription factor drives the expression of Per 1-2, Cry 1-2 and Rev-erb α-β (NR1D1 and NR1D2) genes. The main factor that influences the functioning of this clock rhythm is the light signal reaching the SCN from the retina via the neural pathway, the so-called non-forming image signal. On the other hand, endogenous carbon monoxide (CO) whose formation and availability depends on the amount of light is a chemical signal delivered through the humoral pathway independently of the neural signal. The aim of this experiment was to demonstrate that chemical light carrier CO, regulates the expression of biological clock genes via humoral pathways. Mature males of a wild boar-pig crossbreed, during long and short day season, were used for the study. Autologous blood with experimentally elevated concentrations of endogenous CO (using lamps with white light-emitting diodes) was infused into ophthalmic venous sinus via the right dorsal nasal vein.Results: The results showed that elevated endogenous CO levels through blood irradiation induces changes in genes expressioninvolved in functioning of the major biological clock. Changes in the expression of the transcription factors Bmal1, Clock and Npas2 have a similar pattern in both structures where a very large decrease in gene expression was shown after exposure to elevated endogenous CO levels. The changes in the gene expression of Per 1-2, Cry 1-2, and Rev-erb α-β and Ror β are not the same for both POA and DH structures indicating that both structures respond differently to the received humoral signal.Conclusion: Obtained results indicate that CO is a chemical light molecule whose production in organism depends on the amount of light. An adequate amount of light is an essential factor for the proper functioning of the internal biological clock which is a regulator of many important physiological processes.

Download Full-text

The Role of Endogenous Carbon Monoxide (CO) in the Functioning of Biological Clocks in Pig and Wild Boar Hybrids During Long- and Short-day Seasons

10.21203/rs.3.rs-870647/v1 ◽

2021 ◽

Author(s):

Przemysław GILUN ◽

Barbara Wąsowska ◽

Magdalena Sowa-Kućma ◽

Katarzyna Kozioł ◽

Maria Romerowicz-Misielak ◽

...

Keyword(s):

Gene Expression ◽

Wild Boar ◽

Clock Genes ◽

Autologous Blood ◽

Biological Clock ◽

Biological Clocks ◽

Light Emitting ◽

Short Day ◽

The Right ◽

Induced Changes

Abstract Mature males of a wild boar-pig crossbreed during long- and short-day seasons were used for the study, which demonstrated that the chemical light carrier CO regulates the expression of biological clock genes in the hypothalamus (preoptic area - POA and dorsal part of hypothalamus - DH) via humoral pathways. Autologous blood with experimentally elevated concentrations of endogenous CO (using lamps with white light-emitting diodes) was infused into the ophthalmic venous sinus via the right dorsal nasal vein.The results showed that elevated endogenous CO levels through blood irradiation induced changes in gene expression involved in the functioning of the main biological clock. Changes in the expression of the transcription factors Bmal1, Clock and Npas2 had a similar pattern in both structures, where a very large decrease in gene expression was shown after exposure to elevated endogenous CO levels. The changes in the gene expression of PER 1-2, CRY 1-2, REV-ERB α-β and ROR β are not the same for both POA and DH hypothalamic structures, indicating that both structures respond differently to the received humoral signal.The obtained results indicate that CO is a chemical light molecule whose production in organisms depends on the amount of light. An adequate amount of light is an essential factor for the proper functioning of the main biological clock.

Download Full-text

Cerberus–Nodal–Lefty–Pitx signaling cascade controls left–right asymmetry in amphioxus

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1620519114 ◽

2017 ◽

Vol 114 (14) ◽

pp. 3684-3689 ◽

Cited By ~ 26

Author(s):

Guang Li ◽

Xian Liu ◽

Chaofan Xing ◽

Huayang Zhang ◽

Sebastian M. Shimeld ◽

...

Keyword(s):

Gene Expression ◽

Transcription Factor ◽

Feedback Loops ◽

Signaling Cascade ◽

Nodal Signaling ◽

Transcription Activator ◽

Gene Expression Cassette ◽

Left Right Asymmetry ◽

The Right ◽

Target Transcription Factor

Many bilaterally symmetrical animals develop genetically programmed left–right asymmetries. In vertebrates, this process is under the control of Nodal signaling, which is restricted to the left side by Nodal antagonists Cerberus and Lefty. Amphioxus, the earliest diverging chordate lineage, has profound left–right asymmetry as a larva. We show that Cerberus, Nodal, Lefty, and their target transcription factor Pitx are sequentially activated in amphioxus embryos. We then address their function by transcription activator-like effector nucleases (TALEN)-based knockout and heat-shock promoter (HSP)-driven overexpression. Knockout of Cerberus leads to ectopic right-sided expression of Nodal, Lefty, and Pitx, whereas overexpression of Cerberus represses their left-sided expression. Overexpression of Nodal in turn represses Cerberus and activates Lefty and Pitx ectopically on the right side. We also show Lefty represses Nodal, whereas Pitx activates Nodal. These data combine in a model in which Cerberus determines whether the left-sided gene expression cassette is activated or repressed. These regulatory steps are essential for normal left–right asymmetry to develop, as when they are disrupted embryos may instead form two phenotypic left sides or two phenotypic right sides. Our study shows the regulatory cassette controlling left–right asymmetry was in place in the ancestor of amphioxus and vertebrates. This includes the Nodal inhibitors Cerberus and Lefty, both of which operate in feedback loops with Nodal and combine to establish asymmetric Pitx expression. Cerberus and Lefty are missing from most invertebrate lineages, marking this mechanism as an innovation in the lineage leading to modern chordates.

Download Full-text

Changing planetary rotation rescues the biological clock mutantlhy cca1ofArabidopsis thaliana

10.1101/034629 ◽

2015 ◽

Author(s):

Andrew J. Millar ◽

Jamie T. Carrington ◽

Wei Ven Tee ◽

Sarah K. Hodge

Keyword(s):

Gene Expression ◽

Circadian Clock ◽

Double Mutant ◽

Clock Genes ◽

Biological Clock ◽

Laboratory Model ◽

Wild Type ◽

Planetary Rotation ◽

Short Period ◽

Clock Mechanism

Background: Pervasive, 24-hour rhythms from the biological clock affect diverse biological processes in metabolism and behaviour, including the human cell division cycle and sleep-wake cycle, nightly transpiration and energy balance in plants, and seasonal breeding in both plants and animals. The clock mechanism in the laboratory model plant species Arabidopsis thaliana is complex, in part due to the multiple interlocking, negative feedback loops that link the clock genes. Clock gene mutants are powerful tools to manipulate and understand the clock mechanism and its effects on physiology. The LATE ELONGATED HYPOCOTYL and CIRCADIAN CLOCK ASSOCIATED 1 genes encode dawn-expressed, Myb-related repressor proteins that delay the expression of other clock genes until late in the day. Double mutant plants (lhy cca1) have low-amplitude, short-period rhythms that have been used in multiple studies of the plant circadian clock. Results: We used in vivo imaging of several luciferase (LUC) reporter genes to test how the rhythmic gene expression of wild-type and lhy cca1 mutant plants responded to light:dark cycles. Red, blue and red+blue light were similarly able to entrain these gene expression rhythms. The timing of expression rhythms in double mutant plants showed little or no response to the duration of light under 24h light:dark cycles (dusk sensitivity), in contrast to the wild type. As the period of the mutant clock is about 18h, we tested light:dark cycles of different duration (T cycles), simulating altered rotation of planet Earth. lhy cca1 double mutants regained as much dusk sensitivity in 20h T cycles as the wild type in 24h cycles, though the phase of the rhythm in the mutants was much earlier than wild type. The severe, triple lhy cca1 gi mutants also regained dusk sensitivity in 20h cycles. The double mutant showed some dusk sensitivity under 28h cycles. lhy cca1 double mutants under 28h cycles with short photoperiods, however, had the same apparent phase as wild-type plants. Conclusion: Simulating altered planetary rotation with light:dark cycles can reveal normal circadian performance in clock mutants that have been described as arrhythmic under standard conditions. The features rescued here comprise a dynamic behaviour (apparent phase under 28h cycles) and a dynamic property (dusk sensitivity under 20h cycles). These conditional clock phenotypes indicate that parts of the clock mechanism continue to function independently of LHY and CCA1, despite the major role of these genes in wild-type plants under standard conditions. Accessibility: Most results here will be published only in this format, citable by the DOI. Data and analysis are publicly accessible on the BioDare resource (www.biodare.ed.ac.uk), as detailed in the links below. Transgenic lines are linked to Stock Centre IDs below (Table 7).

Download Full-text

Melatonin alleviated the immune response and improved the salivary gland function in primary Sjögren’s syndrome

10.21203/rs.3.rs-46098/v1 ◽

2020 ◽

Author(s):

Yi Liu ◽

Xiuhong Weng ◽

Shaoling Yu ◽

Yumei Ding ◽

Bo Cheng

Keyword(s):

Gene Expression ◽

Salivary Gland ◽

Salivary Glands ◽

Clock Gene ◽

Clock Genes ◽

Biological Clock ◽

Lymphocyte Infiltration ◽

Clock Gene Expression ◽

Melatonin Administration ◽

Gland Function

Abstract Background Excessive inflammatory reactions participate in primary Sjögren’s syndrome (pSS) progression. In addition, biological clock genes have been detected in the salivary glands, which indicates that clock genes regulate the growth and development of the salivary glands as well as the quality and quantity of saliva secretion. Melatonin is an amine hormone secreted by the pineal gland that has many physiological functions, such as regulating immunity and correcting disorder in the biological clock rhythm. The purpose of this study was to clarify the correlation between pSS and the biological clock rhythm and explore the possibility of applying melatonin to treat pSS. Methods Melatonin (10 mg/kg/d or 15 mg/kg/d) or vehicle was administered to NOD/Ltj mice by intraperitoneal injection for 4 weeks. Clock gene expression levels in labial gland biopsy specimens from pSS patients and submandibular gland specimens from mice were measured by Western blotting (WB) and RT-PCR. The salivary flow rate of mice was measured at 12, 14, and 16 weeks. The severity of lymphocyte infiltration in the salivary glands was analysed by haematoxylin and eosin (H&E) staining. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining were used to detect the expression levels of related inflammatory factors in mice. The percentages of Th17, Th2, and Treg cells were analysed by flow cytometry. Results There was a distinct expression profile for clock genes in pSS patients compared with controls. Continuous melatonin administration improved salivary gland function in NOD/Ltj mice, with decreased lymphocyte infiltration in the submandibular glands and reduced related inflammatory factor expression in the serum and salivary glands. Melatonin treatment skewed T cells towards the Treg and Th2 subsets while suppressing Th17 responses. Additionally, melatonin administration regulated clock gene expression in NOD/Ltj mice. Conclusion pSS pathogenesis and progression are correlated with abnormal circadian gene expression. Melatonin improves the hypofunction of the salivary glands and inhibits the inflammatory development of pSS in NOD/Ltj mice. This study provides a theoretical basis and potential approach for the clinical prevention and treatment of pSS.

Download Full-text

The brain structure and neurosecretory cells of noctuid moth Anadevidia peponis: Noctuidae

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100159722 ◽

1990 ◽

Vol 48 (3) ◽

pp. 436-437

Author(s):

M. Sato ◽

Y. Ogawa ◽

M. Sasaki ◽

T. Matsuo

Keyword(s):

Biological Clock ◽

Virgin Female ◽

Basic Structure ◽

Fixed Time ◽

Neurosecretory Cells ◽

Nerve Cells ◽

Noctuid Moth ◽

The Right ◽

20 Nm ◽

The Brain

A virgin female of the noctuid moth, a kind of noctuidae that eats cucumis, etc. performs calling at a fixed time of each day, depending on the length of a day. The photoreceptors that induce this calling are located around the neurosecretory cells (NSC) in the central portion of the protocerebrum. Besides, it is considered that the female’s biological clock is located also in the cerebral lobe. In order to elucidate the calling and the function of the biological clock, it is necessary to clarify the basic structure of the brain. The observation results of 12 or 30 day-old noctuid moths showed that their brains are basically composed of an outer and an inner portion-neural lamella (about 2.5 μm) of collagen fibril and perineurium cells. Furthermore, nerve cells surround the cerebral lobes, in which NSCs, mushroom bodies, and central nerve cells, etc. are observed. The NSCs are large-sized (20 to 30 μm dia.) cells, which are located in the pons intercerebralis of the head section and at the rear of the mushroom body (two each on the right and left). Furthermore, the cells were classified into two types: one having many free ribosoms 15 to 20 nm in dia. and the other having granules 150 to 350 nm in dia. (Fig. 1).

Download Full-text