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Sex-Dependent Behavioral Functions of the Purkinje Cell-Specific Gαi/o Binding Protein, Pcp2(L7)

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Abstract

We previously reported motor and non-motor enhancements in a mouse mutant with an inactivated Purkinje cell-specific gene, Pcp2(L7), that encodes a GoLoco domain-containing modulator of Gi/o protein-coupled receptors. Effects included elevated learning asymptote with repeated rotarod training, increased acquisition rate in tone-cued fear conditioning (FC), and subtle male-specific changes in both acoustic startle habituation and pre-pulse inhibition. We have further analyzed this mutant strain this time with a focus on male–female differences, and here we report a sex-dependent anxiety-like phenotype: male mutants are less anxious, and female mutants are more anxious, than same-sex wild types. Similarly, the fear responses measured during the tone in FC acquisition are decreased in male mutants and increased in female mutants relative to same-sex wild types. Overall, the dynamics of both acquisition and extinction of FC is affected in mutants but memory was not affected. In the social realm, compositional analysis of sociability and preference for social novelty data supports that both L7 genotype and sex contribute to these behaviors. These results provide direct evidence of emotional functions of the cerebellum due to the unambiguous cerebellar specificity of Pcp2(L7) expression and the lack of any confounding motor defects in the mutant. We attempt to synthesize these new data with what is previously known both about Pcp2(L7) and about the effects of sex and sex hormones on anxiety and fear behaviors: specifically, L7 is a bidirectional and sex-dependent damper that regulates the amplitude and/or rate of sensorimotor responses, potentially acting as a mood stabilizer.

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Acknowledgments

This research was supported by NIH grant RO1-NS37504 to JO. Additional support was provided from NIH grant P30-NS045758. We thank Dr. Derick Lindquist for his critical reading of the manuscript.

Conflicts of Interest

The authors declare they have no conflicts of interest and are aware of no individuals or entity that would benefit from the studies described herein.

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Authors and Affiliations

  1. Department of Neuroscience, The Ohio State University Wexner Medical Center, Columbus, OH, 43210, USA

    James C. Walton, Randy J. Nelson & John Oberdick

  2. Neuroscience Graduate Studies Program, The Ohio State University, Columbus, OH, 43210, USA

    James C. Walton

  3. Anatomisches Institut, Anatomie & Zellbiologie, Rheinische Friedrich-Wilhelms-Universität, Nussallee 10, 53115, Bonn, Germany

    Karl Schilling

  4. Department of Neuroscience, The Ohio State University Wexner Medical Center, 226 Rightmire Hall, 1060 Carmack Rd, Columbus, OH, 43210, USA

    John Oberdick

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Correspondence to Karl Schilling or John Oberdick.

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Figure S1

Lack of significant genotype contributions in long-term fear memory extinction when freezing is measured during the inter-trial interval. The normalized percent freezing decreases with repeated CS-alone trials when freezing is measured during the interval between trials. Note that there is significant extinction (asterisk, trial 1 vs. trial 8), and that there is no observable anticipation in the first few trials as there is when freezing is measured during the tone (see Fig. 2). *p < 0.05, calculated from un-normalized data (JPEG 253 kb)

Figure S2

Increased motor learning on the rotarod in sexually mature males. Mutant males (left panel) have a significantly different learning curve shape than wild-type males primarily due to an increased learning asymptote; *p < 0.05 for genotype × trial interaction. This is a subset of data we reported earlier [18]. In that study, we showed a significant individual effect of genotype in adolescent mutants (two cohorts, 4–5 weeks old), and a significant individual effect of genotype in males alone when the data from adolescent and mature cohorts were pooled (four cohorts total) (JPEG 289 kb)

Figure S3

Compositional analysis of preference for social novelty data. When both genotypes are considered together, there is a significant sex effect as determined by lack of overlap of 95% confidence ellipses (JPEG 200 kb)

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Walton, J.C., Schilling, K., Nelson, R.J. et al. Sex-Dependent Behavioral Functions of the Purkinje Cell-Specific Gαi/o Binding Protein, Pcp2(L7). Cerebellum 11, 982–1001 (2012). https://doi.org/10.1007/s12311-012-0368-4

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