Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Jun 17;35(24):9007-16.
doi: 10.1523/JNEUROSCI.4024-14.2015.

Pharmacological Inhibition of the Psychiatric Risk Factor FKBP51 Has Anxiolytic Properties

Jakob Hartmann  1 , Klaus V Wagner  2 , Steffen Gaali  2 , Alexander Kirschner  2 , Christian Kozany  2 , Gerd Rühter  3 , Nina Dedic  2 , Alexander S Häusl  2 , Lianne Hoeijmakers  2 , Sören Westerholz  2 , Christian Namendorf  2 , Tamara Gerlach  2 , Manfred Uhr  2 , Alon Chen  2 , Jan M Deussing  2 , Florian Holsboer  2 , Felix Hausch  1 , Mathias V Schmidt  1
Affiliations

Pharmacological Inhibition of the Psychiatric Risk Factor FKBP51 Has Anxiolytic Properties

Jakob Hartmann et al. J Neurosci. .

Abstract

Anxiety-related psychiatric disorders represent one of the largest health burdens worldwide. Single nucleotide polymorphisms of the FK506 binding protein 51 (FKBP51) gene have been repeatedly associated with anxiety-related disorders and stress sensitivity. Given the intimate relationship of stress and anxiety, we hypothesized that amygdala FKBP51 may mediate anxiety-related behaviors. Mimicking the stress effect by specifically overexpressing FKBP51 in the basolateral amygdala (BLA) or central amygdala resulted in increased anxiety-related behavior, respectively. In contrast, application of a highly selective FKBP51 point mutant antagonist, following FKBP51(mut) BLA-overexpression, reduced the anxiogenic phenotype. We subsequently tested a novel FKBP51 antagonist, SAFit2, in wild-type mice via BLA microinjections, which reduced anxiety-related behavior. Remarkably, the same effect was observed following peripheral administration of SAFit2. To our knowledge, this is the first in vivo study using a specific FKBP51 antagonist, thereby unraveling the role of FKBP51 and its potential as a novel drug target for the improved treatment of anxiety-related disorders.

Keywords: FKBP51; PTSD; amygdala; antidepressants; anxiety.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Overexpression of FKBP51 in the BLA induces anxiety-related behavior. A, Schematic representation of the viral injection into the BLA. Gray dots represent the location of the injection tip. B, Representative autoradiographs of viral FKBP51 mRNA expression in the BLA 7 weeks after injection. C, Quantification of FKBP51 mRNA levels in the BLA. Infection with the viral construct induced a strong increase. D, Representative immunohistochemistry images of viral FKBP51 expression (Ha-tag Immuno, green; DAPI, blue) in the BLA region 7 weeks after injection. E, F, FKBP51 overexpression in the BLA leads to significantly reduced open arm time and open arm entries in the EPM. G, 51OEBLA mice show a trend toward increased latency to enter the lit compartment in the DaLi. H, General locomotion as assessed in the OF is not affected by FKBP51 overexpression in the BLA. I–L, 51OEBLA mice show reduced stress-coping behavior in the FST, reflected in decreased time struggling, increased time floating and decreased latency to first floating, whereas time swimming remained unaffected; n = 12–14 per group; #p < 0.05. Data are expressed as mean + SEM. Scale bars, 250 μm.
Figure 2.
Figure 2.
Overexpression of FKBP51 in the CeA. A, Schematic representation of the viral injection into the CeA. Gray dots represent the location of the injection tip. B, Representative autoradiographs of viral FKBP51 mRNA expression in the CeA 7 weeks after injection .C, Quantification of FKBP51 mRNA levels in the CeA. Infection with the viral construct induced a strong increase. D, Representative immunohistochemistry images of viral FKBP51 expression (Ha-tag Immuno, green; DAPI, blue) in the CeA region 7 weeks after injection. E, F, FKBP51 overexpression in the CeA leads to significantly reduced open arm time and a trend toward decreased open arm entries in the EPM. G, H, 51OECeA mice show no alterations in the DaLi. I–L, General locomotion as assessed in the OF and stress-coping behavior in the FST are not affected by FKBP51 overexpression in the CeA; n = 9–10; #p < 0.05. Data are expressed as mean + SEM. Scale bars, 250 μm.
Figure 3.
Figure 3.
Overexpression of FKBP51 in the dHc. A, Schematic representation of the viral injection sites into the CA1 and DG of the dHc. B, Representative immunohistochemistry images of viral FKBP51 expression (Ha-tag Immuno, green; DAPI, blue) in the dHc 7 weeks after injection. C, Representative autoradiographs of viral FKBP51 mRNA expression in the dHc 7 weeks after injection. D, Quantification of FKBP51 mRNA levels in the CA1, CA3, and DG of the dHc. Infection with the viral construct induced a strong increase. E–H, No differences between 51OEdHc and Empty groups were found in the EPM and DaLi; n = 12–13, #p < 0.05. Data are expressed as mean + SEM. Scale bars, 500 μm.
Figure 4.
Figure 4.
The FKBP51F67V antagonist Ligand2 reverses the anxiogenic phenotype induced by overexpression of FKBP51F67V in the amygdala. A, The anti-neuritotrophic effect of mutant FKBP51F67V, but not of FKBP51wt, can be fully blocked by Ligand2. N2a neuroblastoma cells were transfected with a plasmid encoding myr-Venus together with plasmids encoding FKBP51wt, FKBP51F67V or an empty plasmid (control). Cells were treated with or without 20 μm Ligand2 for 24 h, followed by fixation, fluorescence imaging, and morphological analysis using ImageJ. Data represent mean + SEM of 25–40 analyzed cells, post hoc tukey: #p < 0.001. B, Ligand2 binds to FKBP51F67V, but not to wild-type FKBP51 or FKBP52. Chemical structure of the FKBP51F67V-selective antagonist Ligand2 and binding affinities for FKBPs determined in a biochemical fluorescence polarization assay using 3 nm of the fluorescent tracers C40-(Fluoresceinyl-Gly)-Rapamycin and 2 nm of purified human FKBP51. C, D, Ligand2 pharmacokinetics. The compound was administered intraperitoneally at the indicated dose and the plasma and brain concentrations were determined after 1 h. Ligand2 accumulates in the brain in a dose-dependent manner (n = 4). E, Schematic representation of the viral injection into the BLA. Gray dots represent the location of the injection tip. F, Representative immunohistochemistry images of viral FKBP51F67V expression (Ha-tag Immuno, green; DAPI, blue) in the BLA 7 weeks after injection. G, Representative autoradiographs of viral FKBP51F67V mRNA expression in the BLA 7 weeks after injection. H, Quantification of FKBP51 mRNA levels in the BLA. Infection with the viral construct induced a strong increase. I, FKBP51F67V overexpression in the BLA leads to significantly reduced open arm time, independent of treatment group in the EPM. J, K, Vehicle treated 51OEF67V-BLA mice spend significantly less time and travel less distance in the lit compartment of the DaLi compared with vehicle treated control animals. This effect is abolished in the Ligand2 treated groups. L, General locomotion in the OF does not differ between the experimental groups. M, Vehicle treated 51OEF67V-BLA mice spend significantly less time struggling in the FST compared with vehicle treated control animals. This effect is abolished in the Ligand2 treated groups. N–P, No differences were found for the parameters time swimming, floating and latency to first floating between the groups in the FST; n = 11–13; *p < 0.05 (ANOVA genotype effect); #p < 0.05 (Tukey post hoc test); Q–T, Ligand2 did not show any overt behavioral side effects in the EPM and DaLi in naive mice (n = 8). Data are expressed as mean + SEM. Scale bars, 250 μm.
Figure 5.
Figure 5.
Inhibition of wild-type FKBP51 by SAFit2 induces anxiolytic behavior in a time-dependent manner. A–D, Mice tested 1 h after peripheral SAFit2 injection (i.p.) shown no alterations in anxiety-related behavior in the EPM or DaLi. E, Mice show a significant increase in the open arm time of the EPM 16 h after peripheral administration of SAFit2. F, G, SAFit2 leads to significantly increased entries to the lit compartment and significantly more time spent in the lit compartment of the DaLi 16 h after peripheral application. H, I, Mice treated with SAFit2 showed a significantly decreased latency to enter and a significantly increased distance traveled in the lit compartment of the DaLi 16 h after the intraperitoneal injection; n = 10–11. J, Schematic representation of the cannula placement. X represents the tip of the guiding cannula. K, mice treated with SAFit2 via BLA microinjections show a significant increase in the number of lit compartment entries in the DaLi. L, M, SAFit2 BLA microinjections lead toward a trend to increased time spent in the lit compartment and a significantly decreased latency to enter the lit compartment of the DaLi; n = 14, #p < 0.05. Data are expressed as mean + SEM.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Appel K, Schwahn C, Mahler J, Schulz A, Spitzer C, Fenske K, Stender J, Barnow S, John U, Teumer A, Biffar R, Nauck M, Völzke H, Freyberger HJ, Grabe HJ. Moderation of adult depression by a polymorphism in the FKBP5 gene and childhood physical abuse in the general population. Neuropsychopharmacology. 2011;36:1982–1991. doi: 10.1038/npp.2011.81. - DOI - PMC - PubMed
    1. Attwood BK, Bourgognon JM, Patel S, Mucha M, Schiavon E, Skrzypiec AE, Young KW, Shiosaka S, Korostynski M, Piechota M, Przewlocki R, Pawlak R. Neuropsin cleaves EphB2 in the amygdala to control anxiety. Nature. 2011;473:372–375. doi: 10.1038/nature09938. - DOI - PMC - PubMed
    1. Binder EB. The role of FKBP5, a co-chaperone of the glucocorticoid receptor in the pathogenesis and therapy of affective and anxiety disorders. Psychoneuroendocrinology. 2009;34:S186–S195. doi: 10.1016/j.psyneuen.2009.05.021. - DOI - PubMed
    1. Binder EB, Holsboer F. Pharmacogenomics and antidepressant drugs. Ann Med. 2006;38:82–94. doi: 10.1080/07853890600551045. - DOI - PubMed
    1. Binder EB, Salyakina D, Lichtner P, Wochnik GM, Ising M, Pütz B, Papiol S, Seaman S, Lucae S, Kohli MA, Nickel T, Künzel HE, Fuchs B, Majer M, Pfennig A, Kern N, Brunner J, Modell S, Baghai T, Deiml T, et al. Polymorphisms in FKBP5 are associated with increased recurrence of depressive episodes and rapid response to antidepressant treatment. Nat Genet. 2004;36:1319–1325. doi: 10.1038/ng1479. - DOI - PubMed

Publication types