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The Effect of Hapln4 Link Protein Deficiency on Extracellular Space Diffusion Parameters and Perineuronal Nets in the Auditory System During Aging

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Abstract

Hapln4 is a link protein which stabilizes the binding between lecticans and hyaluronan in perineuronal nets (PNNs) in specific brain regions, including the medial nucleus of the trapezoid body (MNTB). The aim of this study was: (1) to reveal possible age-related alterations in the extracellular matrix composition in the MNTB and inferior colliculus, which was devoid of Hapln4 and served as a negative control, (2) to determine the impact of the Hapln4 deletion on the values of the ECS diffusion parameters in young and aged animals and (3) to verify that PNNs moderate age-related changes in the ECS diffusion, and that Hapln4-brevican complex is indispensable for the correct protective function of the PNNs. To achieve this, we evaluated the ECS diffusion parameters using the real-time iontophoretic method in the selected region in young adult (3 to 6-months-old) and aged (12 to 18-months-old) wild type and Hapln4 knock-out (KO) mice. The results were correlated with an immunohistochemical analysis of the ECM composition and astrocyte morphology. We report that the ECM composition is altered in the aged MNTB and aging is a critical point, revealing the effect of Hapln4 deficiency on the ECS diffusion. All of our findings support the hypothesis that the ECM changes in the MNTB of aged KO animals affect the ECS parameters indirectly, via morphological changes of astrocytes, which are in direct contact with synapses and can be influenced by the ongoing synaptic transmission altered by shifts in the ECM composition.

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Funding

This study was supported by the Czech Science Foundation (GACR) (Grant No. 16-10214S to LV) and a Grant-in-Aid for Scientific Research on Innovative Areas (Grant No. 19H04754 to TO) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan. We wish to thank Fran Zatrepalkova for her useful comments and suggestions, Sophie Chowdhury for her contribution to diffusion measurements and Lenka Kohoutova for her excellent technical services. We also wish to thank Eva Sykova, who helped to initiate the collaboration of the Czech and Japanese teams.

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Authors and Affiliations

  1. Department of Neuroscience, 2nd Faculty of Medicine, Charles University, V Úvalu 84, 150 06, Prague 5, Czech Republic

    Petra Sucha, Martina Chmelova, Monika Kamenicka, Marcel Bochin & Lydia Vargova

  2. Department of Cellular Neurophysiology, Institute of Experimental Medicine AS CR, v.v.i., Prague, Czech Republic

    Petra Sucha, Martina Chmelova, Monika Kamenicka, Marcel Bochin & Lydia Vargova

  3. Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

    Toshitaka Oohashi

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  1. Petra Sucha
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Correspondence to Lydia Vargova.

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All experiments were performed in accordance with the European Communities Council Directive 24 November 1986 (86/609/EEC), animal care guidelines approved by the Institute of Experimental Medicine, Academy of Science of the Czech Republic Animal Care Committee on the 17th April 2009; approval number 12/2013 and in accordance with the Policy on the Care and Use of the Laboratory Animals of Okayama University (Approval Number: OKU-2016045), approved by the Animal Care and Use Committee of Okayama University.

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Sucha, P., Chmelova, M., Kamenicka, M. et al. The Effect of Hapln4 Link Protein Deficiency on Extracellular Space Diffusion Parameters and Perineuronal Nets in the Auditory System During Aging. Neurochem Res 45, 68–82 (2020). https://doi.org/10.1007/s11064-019-02894-2

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