Neuroanatomical and behavioral deficits in mice haploinsufficient for Pericentriolar material 1 (Pcm1)

doi:10.1016/j.neures.2015.02.002

Neuroscience Research

Volume 98, September 2015, Pages 45-49

https://doi.org/10.1016/j.neures.2015.02.002 Get rights and content

Highlights

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First study of the effects of Pcm1 haploinsufficiency on brain structure and behavior.
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Pcm1^+/− mice had a slightly smaller brain compared to wild-type mice.
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Pcm1^+/− mice were impaired in the social novelty test, but not in novel object recognition.

Abstract

The pericentriolar material (PCM) is composed of proteins responsible for microtubule nucleation/anchoring at the centrosome, some of which have been associated with genetic susceptibility to schizophrenia. Here, we show that mice haploinsufficient for Pericentriolar material 1 (Pcm1^+/−), which encodes a component of the PCM found to bear rare loss of function mutations in patients with psychiatric illness, manifest neuroanatomical phenotypes and behavioral abnormalities. Using ex vivo magnetic resonance imaging of the Pcm1^+/− brain, we detect reduced whole brain volume. Pcm1 mutant mice show impairment in social interaction, specifically in the social novelty phase, but not in the sociability phase of the three-chamber social interaction test. In contrast, Pcm1^+/− mice show normal preference for a novel object, suggesting specific impairment in response to novel social stimulus. In addition, Pcm1^+/− mice display significantly reduced rearing activity in the open field. Pcm1^+/− mice behave normally in the elevated plus maze, rotarod, prepulse inhibition, and progressive ratio tests. Together, our results suggest that haploinsufficiency at the Pcm1 locus can induce a range of neuroanatomical and behavioral phenotypes that support the candidacy of this locus in neuropsychiatric disorders.

Introduction

Schizophrenia is a common disorder of largely obscure etiology. Genetic analyses have highlighted the influence of susceptibility alleles for schizophrenia, which, in turn, raised the expectation that the cloning of such loci will illuminate the key pathways for the understanding of the disease pathology.

One candidate, PCM1 (Pericentriolar material 1), is attractive in molecular psychiatry for two reasons. First, we and others have linked the chromosomal location 8p22 and specifically PCM1 to schizophrenia in European populations. Family and trio samples showed significant transmission disequilibrium between a marker in the PCM1 locus and schizophrenia. A case–control sample also found significant association between PCM1 markers and schizophrenia (Gurling et al., 2006). This was followed by identification of specific non-synonymous mutations in coding and regulatory regions of PCM1 in schizophrenia (Kamiya et al., 2008, Datta et al., 2010). Of note, a meta-analysis in Japanese failed to find a significant association (Hashimoto et al., 2011). Second, we have reported previously that transient depletion of Pcm1 by in utero gene transfer can affect the neuroarchitecture of the developing cortex (Kamiya et al., 2008). PCM1 is recruited to the centrosome by Disrupted in Schizophrenia-1 (DISC1) and Bardet–Biedl syndrome 4 proteins (Kamiya et al., 2008, Narayan et al., 2013). PCM1 also interacts with Huntingtin, which is critical for regulation of ciliogenesis (Keryer et al., 2011).

Given these observations, we targeted the Pcm1 locus through homologous recombination. Here we show that mice haploinsufficient for Pcm1 show a significant reduction in brain volume. Pcm1^+/− mice were impaired in response to a novel social stimulus, but not to a novel object. In addition, they displayed reduced rearing in the open field test, whereas motor function appeared unimpaired.

Section snippets

Mouse colony

Mice deficient in Pcm1 were generated in a previous study from our group (Brodar, 2015). In brief, the Pcm1 gene was targeted through insertion of a gene-trap cassette between exon 4 and 5. The depletion of Pcm1 protein is demonstrated in the present manuscript (Fig. 1A). Mice heterozygous for Pcm1 were backcrossed to C57BL/6N nine times. All the experiments were performed with heterozygous males and their wild-type (WT) littermates. All animal experiments were carried out in accordance with

Results

To achieve a homogeneous genetic background, Pcm1^+/− mice were backcrossed nine generations to a C57BL/6N (Charles River) background. Pcm1^−/− animals were produced at a low frequency from Pcm1^+/− mating, suggesting early lethality. Gross histological analyses of Pcm1^+/− and wild-type littermates showed no overt pathology; therefore this genotype was used for subsequent experiments to address key significant questions in molecular psychiatry.

First we confirmed by immunoblot that the mice were

Discussion

The significance of the present study is to report brain and psychiatry-associated phenotypes (gross anatomical changes and unique set of behavioral alteration) in Pcm1^+/− mice that our group originally developed (Brodar, 2015).

The two most prominent behavioral phenotypes of Pcm1^+/− mice were reduced rearing in the open field and reduced social interaction in the three-chamber social interaction test. Rearing is a motor function aimed at the visual scanning of the (novel) environment. Reduced

Funding

This research was funded by MH-084018, MH-094268 Silvo O. Conte Center, MH-069853, MH-085226, MH-088753, MH-092443, MH-105660, as well as grants from Stanley, S-R, RUSK, NARSAD, and MSCRF.

Conflict of interest

There are no known conflicts of interest for any author.

Acknowledgements

We thank Drs. Akiharu Kubo and Sachiko Tsukita for the Pcm1 antibody and Ms. Yukiko Lema for organizing the figures.

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PCM1, which is a DISC1- interacting protein, is involved in the maintenance of centrosome integrity and regulation of microtubule cytoskeleton and dopamine transmission (Eastwood et al., 2009; Kamiya et al., 2008; Rampino et al., 2014). Suppression of PCM1 affects neuronal migration and plays a role in cortical development and genetic susceptibility to schizophrenia (Gurling et al., 2006b), and Pcm1+/− mice display deficits in social interaction and show significant reduction in brain volume (Zoubovsky et al., 2015). The striking alterations in the expressions of GPCRs in the four psychiatric disorders, particularly in SCZ (75% of cilia GPCRs) indicate a crucial role for the cilia GPCRs in the pathogenesis of these disorders.
Primary cilia function as cells' antennas to detect and transduce external stimuli and play crucial roles in cell signaling and communication. The vast majority of cilia genes that are causally linked with ciliopathies are also associated with neurological deficits, such as cognitive impairments. Yet, the roles of cilia dysfunctions in the pathogenesis of psychiatric disorders have not been studied. Our aim is to identify patterns of cilia gene dysregulation in the four major psychiatric disorders: schizophrenia (SCZ), autism spectrum disorder (ASD), bipolar disorder (BP), and major depressive disorder (MDD). For this purpose, we acquired differentially expressed genes (DEGs) from the largest and most recent publicly available databases. We found that 42%, 24%, 17%, and 15% of brain-expressed cilia genes were significantly differentially expressed in SCZ, ASD, BP, and MDD, respectively. Several genes exhibited cross-disorder overlap, suggesting that typical cilia signaling pathways' dysfunctions determine susceptibility to more than one psychiatric disorder or may partially underlie their pathophysiology. Our study revealed that genes encoding proteins of almost all sub-cilia structural and functional compartments were dysregulated in the four psychiatric disorders. Strikingly, the genes of 75% of cilia GPCRs and 50% of the transition zone proteins were differentially expressed in SCZ.
The present study is the first to draw associations between cilia and major psychiatric disorders, and is the first step toward understanding the role that cilia components play in their pathophysiological processes, which may lead to novel therapeutic targets for these disorders.
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PCM1 granules are distinct from pericentrin-containing granules (Kubo and Tsukita, 2003). Pcm1+/− mice manifest neuroanatomical phenotypes with behavioral abnormalities and show significant reduction in brain volume (Zoubovsky et al., 2015). Presence of a retina phenotype was not investigated.
Photoreceptors are polarized neurons, with very specific subcellular compartmentalization and unique requirements for protein expression and trafficking. Each photoreceptor contains an outer segment, the site of photon capture that initiates vision, an inner segment that houses the biosynthetic machinery and a synaptic terminal for signal transmission to downstream neurons. Outer segments and inner segments are connected by a connecting cilium (CC), the equivalent of a transition zone (TZ) of primary cilia. The connecting cilium is part of the basal body/axoneme backbone that stabilizes the outer segment. This report will update the reader on late developments in photoreceptor ciliogenesis and transition zone formation, specifically in mouse photoreceptors, focusing on early events in photoreceptor ciliogenesis. The connecting cilium, an elongated and narrow structure through which all outer segment proteins and membrane components must traffic, functions as a gate that controls access to the outer segment. Here we will review genes and their protein products essential for basal body maturation and for CC/TZ genesis, sorted by phenotype. Emphasis is given to naturally occurring mouse mutants and gene knockouts that interfere with CC/TZ formation and ciliogenesis.
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Chemokines play important roles in the central nervous system, including mediating neuroinflammation and guiding the intracortical migration of interneurons during development. Alteration in parvalbumin-positive interneurons is a key neuropathological hallmark of multiple mental conditions. We recently reported a significant reduction in the expression of CXCL12 in olfactory neurons from sporadic cases with schizophrenia compared with matched controls, suggesting a role for CXCR4/CXCL12 signaling in mental conditions. Thus, we depleted the chemokine receptor Cxcr4 from mice using the parvalbumin-2A-Cre line. The conditional knockout mice exhibited a unique behavioral phenotype involving increased stereotypy. Stereotypy is observed in many psychiatric conditions, including schizophrenia, autism, and dementia. Thus, the Cxcr4 conditional knockout mice may serve as a model for this symptomatic feature.
Disruption of social cognition in the sub-chronic PCP rat model of schizophrenia: Possible involvement of the endocannabinoid system
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In contrast to their intact sociability, PCP-treated animals showed a selective impairment of social novelty preference that was not olfactory-based (no alteration in their ability to discriminate social over neutral odors) nor linked to anxiety [no alteration in motor functions and anxiety-like behaviors during the first 5 min of the habituation phase; confirming previously published data (McLean et al., 2010; Seillier and Giuffrida, 2011)]. Interestingly, this selective impairment of social novelty preference has been reported across a broad range of animal models of schizophrenia (Kaminitz et al., 2014; O׳Tuathaigh et al., 2007; Shimamoto et al., 2014; Zoubovsky et al., 2015, but O׳Tuathaigh et al., 2010a). Eagle et al. (2013) proposed that this selective impairment might be related to the differences between sensation-seeking versus novelty-seeking behaviors, the latter requiring to recall and recognize a familiar versus a novel object.
Previous studies have shown that social withdrawal in the phencyclidine (PCP) rat model of schizophrenia results from deficient endocannabinoid-induced activation of CB₁ receptors. To understand the underlying cognitive mechanisms of the social deficit in PCP-treated rats, we examined the impact of pharmacological manipulation of the endocannabinoid system on sociability (i.e. social approach) and social novelty preference (which relies on social recognition).
Control rats showed a clear preference for a “social” cage (i.e. unfamiliar stimulus rat placed under a wire mesh cage) versus an “empty” cage, and spent more time exploring a “novel” cage (i.e. new stimulus rat) versus a “familiar” cage. In contrast, rats receiving PCP (5 mg/kg, b.i.d. for 7 days, followed by a 7 day-washout period) showed intact sociability, but lacked social novelty preference. This PCP-induced deficit was due to increased activity at CB₁ receptors as it was reversed by systemic administration of the CB₁ antagonist AM251 (1 mg/kg). In agreement with this hypothesis, the cannabinoid agonist CP55,940 (0.003–0.03 mg/kg) dose-dependently suppressed social novelty preference in control animals without affecting sociability.
Taken together, these data suggest that PCP-treated rats have a deficit in social cognition, possibly induced by increased stimulation of CB₁ receptors. This deficit, however, is distinct from the social withdrawal previously observed in these animals, as the latter is due to deficient, rather than increased, CB₁ stimulation.
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¹: Present address: Department of Psychology, Michigan State University, East Lansing, MI 48824, United States.

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Neuroanatomical and behavioral deficits in mice haploinsufficient for Pericentriolar material 1 (Pcm1)

Highlights

Abstract

Introduction

Section snippets

Mouse colony

Results

Discussion

Funding

Conflict of interest

Acknowledgements

Schizophr. Res.

Prog. Brain Res.

Dev. Cogn. Neurosci.

Trends Cogn. Sci.

Linking neurodevelopmental and synaptic theories of mental illness through DISC1

Nat. Rev. Neurosci.

PCM1 is necessary for the maintenance of focal ciliary integrity and dopamine signaling in the postnatal brain

A threonine to isoleucine missense mutation in the pericentriolar material 1 gene is strongly associated with schizophrenia

Mol. Psychiatry