title: Urokinase-type plasminogen activator modulates mammalian circadian clock phase regulation in tissue-type plasminogen activator knockout mice.
abstract: Glutamate phase shifts the circadian clock in the mammalian suprachiasmatic nucleus (SCN) by activating NMDA receptors. Tissue-type plasminogen activator (tPA) gates phase shifts by activating plasmin to generate m(ature) BDNF, which binds TrkB receptors allowing clock phase shifts. Here, we investigate phase shifting in tPA knockout (tPA-/-; B6.129S2-Plattm1Mlg/J) mice, and identify urokinase-type plasminogen activator (uPA) as an additional circadian clock regulator. Behavioral activity rhythms in tPA-/-mice entrain to a light-dark (LD) cycle and phase shift in response to nocturnal light pulses with no apparent loss in sensitivity. When the LD cycle is inverted, tPA-/-mice take significantly longer to entrain than C57BL/6J wild-type (WT) mice. SCN brain slices from tPA-/-mice exhibit entrained neuronal activity rhythms and phase shift in response to nocturnal glutamate with no change in dose-dependency. Pre-treating slices with the tPA/uPA inhibitor, plasminogen activator inhibitor-1 (PAI-1), inhibits glutamate-induced phase delays in tPA-/-slices. Selective inhibition of uPA with UK122 prevents glutamate-induced phase resetting in tPA-/-but not WT SCN slices. tPA expression is higher at night than the day in WT SCN, while uPA expression remains constant in WT and tPA-/-slices. Casein-plasminogen zymography reveals that neither tPA nor uPA total proteolytic activity is under circadian control in WT or tPA-/-SCN. Finally, tPA-/-SCN tissue has lower mBDNF levels than WT tissue, while UK122 does not affect mBDNF levels in either strain. Together, these results suggest that either tPA or uPA can support photic/glutamatergic phase shifts of the SCN circadian clock, possibly acting through distinct mechanisms.
authors.affiliation: Department of Biochemistry and Cellular and Molecular Biology, NeuroNET Research Center, M407 Walters Life Sciences, University of Tennessee, Knoxville, TN, 37996-0001, USA.
authors.forename: Joanna M
authors.initials: JM
authors.lastname: Cooper
authors.affiliation: Department of Biological Sciences, Kent State University, Kent, OH, USA.
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Rastogi
authors.affiliation: Department of Biological Sciences, Kent State University, Kent, OH, USA.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Krizo
authors.affiliation: Department of Biological Sciences, Kent State University, Kent, OH, USA.
authors.forename: Eric M
authors.initials: EM
authors.lastname: Mintz
authors.affiliation: Department of Biochemistry and Cellular and Molecular Biology, NeuroNET Research Center, M407 Walters Life Sciences, University of Tennessee, Knoxville, TN, 37996-0001, USA.
authors.forename: Rebecca A
authors.identifier: 0000-0003-1636-7272
authors.initials: RA
authors.lastname: Prosser
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D057906:Circadian Clocks; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015849:Plasminogen Inactivators / Q000494:pharmacology; D059748:Proteolysis; D013493:Suprachiasmatic Nucleus / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D010959:Tissue Plasminogen Activator / Q000235:genetics* / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D015849:Plasminogen Inactivators; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator
keywords: circadian rhythms; suprachiasmatic nucleus; tissue-type plasminogen activator; urokinase-type plasminogen activator