Antibodies for Assessing Circadian Clock Proteins in the Rodent Suprachiasmatic Nucleus

Research on the mechanisms underlying circadian rhythmicity and the response of brain and body clocks to environmental and physiological challenges requires assessing levels of circadian clock proteins. Too often, however, it is difficult to acquire antibodies that specifically and reliably label these proteins. Many of these antibodies also lack appropriate validation. The goal of this project was to generate and characterize antibodies against several circadian clock proteins. We examined mice and hamsters at peak and trough times of clock protein expression in the suprachiasmatic nucleus (SCN). In addition, we confirmed specificity by testing the antibodies on mice with targeted disruption of the relevant genes. Our results identify antibodies against PER1, PER2, BMAL1 and CLOCK that are useful for assessing circadian clock proteins in the SCN by immunocytochemistry.

• Publication year

  2012

• Venue

  PLoS ONE

• Publication date

  2012-04-27

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1371/journal.pone.0035938PMID 22558277PMCID 3338757
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Tuning the Period of the Mammalian Circadian Clock: Additive and Independent Effects of CK1εTau and Fbxl3Afh Mutations on Mouse Circadian Behavior and Molecular Pacemaking

  2011cited by this paper
• Effects of Vasoactive Intestinal Peptide Genotype on Circadian Gene Expression in the Suprachiasmatic Nucleus and Peripheral Organs

  2011cited by this paper
• Characterization of orderly spatiotemporal patterns of clock gene activation in mammalian suprachiasmatic nucleus

  2011cited by this paper
• The Acetyltransferase CLOCK Is Dispensable for Circadian Aftereffects in Mice

  2011cited by this paper
• Site-Specific Changes in Brain Extra-SCN Oscillators during Early Pregnancy in the Rat

  2011cited by this paper
• The period of the circadian oscillator is primarily determined by the balance between casein kinase 1 and protein phosphatase 1

  2011cited by this paper
• Impact of aging on diurnal expression patterns of CLOCK and BMAL1 in the mouse brain.

  2010cited by this paper
• Changes in and dorsal to the rat suprachiasmatic nucleus during early pregnancy.

  2010cited by this paper
• Suprachiasmatic nucleus: cell autonomy and network properties.

  2010cited by this paper
• Endogenous Dopamine Regulates the Rhythm of Expression of the Clock Protein PER2 in the Rat Dorsal Striatum via Daily Activation of D2 Dopamine Receptors

  2010cited by this paper
• PER2 rhythms in the amygdala and bed nucleus of the stria terminalis of the diurnal grass rat (Arvicanthis niloticus).

  2010cited by this paper
• Phase preference for the display of activity is associated with the phase of extra-suprachiasmatic nucleus oscillators within and between species.

  2010cited by this paper
• The Comparison between Circadian Oscillators in Mouse Liver and Pituitary Gland Reveals Different Integration of Feeding and Light Schedules

  2010cited by this paper
• Induction of mPer1 expression by GnRH in pituitary gonadotrope cells involves EGR-1.

  2009cited by this paper
• Selective Inhibition of Casein Kinase 1 Epsilon Minimally Alters Circadian Clock Period

  2009cited by this paper
• Circadian rhythms of PERIOD1 expression in the dorsomedial hypothalamic nucleus in the absence of entrained food‐anticipatory activity rhythms in rats

  2009cited by this paper
• Casein Kinase 1 Delta Regulates the Pace of the Mammalian Circadian Clock

  2009cited by this paper
• A Guide to the Perplexed on the Specificity of Antibodies

  2009cited by this paper
• Selective Inhibition of Casein Kinase 1ϵ Minimally Alters Circadian Clock Period

  2009cited by this paper
• Rhythmic expression of clock genes in the ependymal cell layer of the third ventricle of rodents is independent of melatonin signaling

  2008cited by this paper
• Rhythms in expression of PER1 protein in the amygdala and bed nucleus of the stria terminalis of the diurnal grass rat (Arvicanthis niloticus).

  2008cited by this paper
• Daily rhythms in PER1 within and beyond the suprachiasmatic nucleus of female grass rats (Arvicanthis niloticus).

  2008cited by this paper
• PER1-Like Immunoreactivity in Oxytocin Cells of the Hamster Hypothalamo-Neurohypophyseal System

  2007cited by this paper
• Evidence for circadian regulation of activating transcription factor 5 but not tyrosine hydroxylase by the chromaffin cell clock.

  2007cited by this paper
• Temporal and spatial distribution of immunoreactive PER1 and PER2 proteins in the suprachiasmatic nucleus and peri-suprachiasmatic region of the diurnal grass rat (Arvicanthis niloticus).

  2006cited by this paper
• A clock shock: mouse CLOCK is not required for circadian oscillator function.

  2006cited by this paper
• Long-Term Constant Light Induces Constitutive Elevated Expression of mPER2 Protein in the Murine SCN: A Molecular Basis for Aschoff’s Rule?

  2005cited by this paper
• Diurnal variation in CREB phosphorylation and PER1 protein levels in lactotroph cells of melatonin-proficient C3H and melatonin-deficient C57BL mice: similarities and differences

  2005cited by this paper
• Rhythms in clock proteins in the mouse pars tuberalis depend on MT1 melatonin receptor signalling

  2005cited by this paper
• The central and basolateral nuclei of the amygdala exhibit opposite diurnal rhythms of expression of the clock protein Period2.

  2005cited by this paper
• Resetting the brain clock: time course and localization of mPER1 and mPER2 protein expression in suprachiasmatic nuclei during phase shifts

  2004cited by this paper
• Expression of mCLOCK and Other Circadian Clock‐Relevant Proteins in the Mouse Suprachiasmatic Nuclei

  2003cited by this paper
• Light does not degrade the constitutively expressed BMAL1 protein in the mouse suprachiasmatic nucleus

  2003cited by this paper
• A short half-life GFP mouse model for analysis of suprachiasmatic nucleus organization.

  2003cited by this paper
• The Biological Clock Nucleus: A Multiphasic Oscillator Network Regulated by Light

  2003cited by this paper
• Coordination of circadian timing in mammals

  2002cited by this paper
• Rhythmic gene expression in pituitary depends on heterologous sensitization by the neurohormone melatonin

  2002cited by this paper
• Differential functions of mPer1, mPer2, and mPer3 in the SCN circadian clock.

  2001cited by this paper
• Posttranslational mechanisms regulate the mammalian circadian clock.

  2001cited by this paper
• Mop3 is an essential component of the master circadian pacemaker in mammals.

  2000cited by this paper
• Analysis of clock proteins in mouse SCN demonstrates phylogenetic divergence of the circadian clockwork and resetting mechanisms.

  2000influential reference
• Interacting molecular loops in the mammalian circadian clock.

  2000cited by this paper
• Light and Glutamate-Induced Degradation of the Circadian Oscillating Protein BMAL1 during the Mammalian Clock Resetting

  2000cited by this paper
• Differential Regulation of mPER1 and mTIM Proteins in the Mouse Suprachiasmatic Nuclei: New Insights into a Core Clock Mechanism

  1999influential reference

• Analytical insights into the behavior of finite amplitude waves in plasma fluid dynamics

  2024cites this paper
• Chronobiotics: classifications of existing circadian clock modulators, future perspectives.

  2024cites this paper
• Neuroendocrine effects of the duper mutation in Syrian hamsters: a role for Cryptochrome 1

  2024cites this paper
• Does Patient-Applied Testosterone Replacement Therapy Pose Risk for Blood Pressure Elevation? Circadian Medicine Perspectives.

  2022cites this paper
• Anatomical Methods to Study the Suprachiasmatic Nucleus.

  2022cites this paper
• Intense light elicited alveolar type 2 specific circadian PER2 protects from bacterial lung injury via BPIFB1.

  2022cites this paper
• Development of human cartilage circadian rhythm in a stem cell-chondrogenesis model

  2022cites this paper
• Rhythmic Regulation of DNA Methylation Factors and Core-Clock Genes in Brain Structures Activated by Cocaine or Sucrose: Potential Role of Chromatin Remodeling

  2021influential citation
• A role for the adenosine ADORA2B receptor in midazolam induced cognitive dysfunction.

  2020cites this paper
• Removing Short Wavelengths From Polychromatic White Light Attenuates Circadian Phase Resetting in Rats

  2019cites this paper
• Suprachiasmatic function in a circadian period mutant: Duper alters light‐induced activation of vasoactive intestinal peptide cells and PERIOD1 immunostaining

  2018cites this paper
• Functionally Complete Excision of Conditional Alleles in the Mouse Suprachiasmatic Nucleus by Vgat-ires-Cre

  2018cites this paper
• The Period 2 Enhancer Nobiletin as Novel Therapy in Murine Models of Circadian Disruption Resembling Delirium*

  2018cites this paper
• Effects of a free-choice high-fat high-sugar diet on brain PER2 and BMAL1 protein expression in mice.

  2017cites this paper
• Differential localization of PER1 and PER2 in the brain master circadian clock

  2017cites this paper
• Impact of Ataxin-2 knock out on circadian locomotor behavior and PER immunoreaction in the SCN of mice

  2017cites this paper
• Loss of CLOCK Results in Dysfunction of Brain Circuits Underlying Focal Epilepsy.

  2017cites this paper
• Behavioral, Neurobiological, and Genetic Analysis of the Circadian Mutant Duper

  2015cites this paper
• The Circadian Clock Gene Period1 Connects the Molecular Clock to Neural Activity in the Suprachiasmatic Nucleus

  2015influential citation
• Clock genes control cortical critical period timing.

  2015cites this paper
• Cell type-specific transcriptomics of hypothalamic energy-sensing neuron responses to weight-loss

  2015cites this paper
• Dim Light at Night Disrupts Molecular Circadian Rhythms and Affects Metabolism

  2014influential citation
• Sleep disorders and depression: brief review of the literature, case report, and nonpharmacologic interventions for depression

  2013cites this paper
• Dim Light at Night Disrupts Molecular Circadian Rhythms and Increases Body Weight

  2013influential citation
• Light at Night Alters Daily Patterns of Cortisol and Clock Proteins in Female Siberian Hamsters

  2013cites this paper
• Physiological consequences of circadian disruption by nighttime light exposure

  2013cites this paper
• Sleep disorders and depression: brief review of the literature, case report, and nonpharmacologic interventions for depression

  2013cites this paper
• Sleep disorders and depression: brief review of the literature, case report, and nonpharmacologic interventions for depression

  2013cites this paper
• Long-Lasting Effects of Sepsis on Circadian Rhythms in the Mouse

  2012cites this paper
• Heterogeneous Expression of the Core Circadian Clock Proteins among Neuronal Cell Types in Mouse Retina

  2012influential citation