Diurnal rhythms in neurexins transcripts and inhibitory/excitatory synapse scaffold proteins in the biological clock

PLoS One. 2012;7(5):e37894. doi: 10.1371/journal.pone.0037894. Epub 2012 May 25.

Abstract

The neurexin genes (NRXN1/2/3) encode two families (α and β) of highly polymorphic presynaptic proteins that are involved in excitatory/inhibitory synaptic balance. Recent studies indicate that neuronal activation and memory formation affect NRXN1/2/3α expression and alternative splicing at splice sites 3 and 4 (SS#3/SS#4). Neurons in the biological clock residing in the suprachiasmatic nuclei of the hypothalamus (SCN) act as self-sustained oscillators, generating rhythms in gene expression and electrical activity, to entrain circadian bodily rhythms to the 24 hours day/night cycles. Cell autonomous oscillations in NRXN1/2/3α expression and SS#3/SS#4 exons splicing and their links to rhythms in excitatory/inhibitory synaptic balance in the circadian clock were explored. NRXN1/2/3α expression and SS#3/SS#4 splicing, levels of neurexin-2α and the synaptic scaffolding proteins PSD-95 and gephyrin (representing excitatory and inhibitory synapses, respectively) were studied in mRNA and protein extracts obtained from SCN of C3H/J mice at different times of the 24 hours day/night cycle. Further studies explored the circadian oscillations in these components and causality relationships in immortalized rat SCN2.2 cells. Diurnal rhythms in mNRXN1α and mNRXN2α transcription, SS#3/SS#4 exon-inclusion and PSD-95 gephyrin and neurexin-2α levels were found in the SCN in vivo. No such rhythms were found with mNRXN3α. SCN2.2 cells also exhibited autonomous circadian rhythms in rNRXN1/2 expression SS#3/SS#4 exon inclusion and PSD-95, gephyrin and neurexin-2α levels. rNRXN3α and rNRXN1/2β were not expressed. Causal relationships were demonstrated, by use of specific siRNAs, between rNRXN2α SS#3 exon included transcripts and gephyrin levels in the SCN2.2 cells. These results show for the first time dynamic, cell autonomous, diurnal rhythms in expression and splicing of NRXN1/2 and subsequent effects on the expression of neurexin-2α and postsynaptic scaffolding proteins in SCN across the 24-h cycle. NRXNs gene transcripts may have a role in coupling the circadian clock to diurnal rhythms in excitatory/inhibitory synaptic balance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing
  • Animals
  • Biological Clocks / drug effects
  • Biological Clocks / genetics*
  • Calcium-Binding Proteins
  • Carrier Proteins / metabolism
  • Cell Line
  • Central Nervous System Depressants / pharmacology
  • Circadian Rhythm / drug effects
  • Circadian Rhythm / genetics*
  • Disks Large Homolog 4 Protein
  • Guanylate Kinases / metabolism
  • Male
  • Melatonin / pharmacology
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Inbred C3H
  • Neural Cell Adhesion Molecules / genetics*
  • Neural Cell Adhesion Molecules / metabolism
  • Period Circadian Proteins / genetics
  • Protein Transport
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Suprachiasmatic Nucleus / drug effects
  • Suprachiasmatic Nucleus / metabolism
  • Synapses / metabolism*
  • Transcription, Genetic*

Substances

  • Calcium-Binding Proteins
  • Carrier Proteins
  • Central Nervous System Depressants
  • Disks Large Homolog 4 Protein
  • Dlg4 protein, mouse
  • Membrane Proteins
  • Neural Cell Adhesion Molecules
  • Nrxn1 protein, mouse
  • Period Circadian Proteins
  • RNA, Messenger
  • gephyrin
  • Guanylate Kinases
  • Melatonin