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Targeted deletion of Aqp4 promotes the formation of astrocytic gap junctions

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Abstract

Aquaporin-4 (AQP4) is the predominant water channel in the brain and is expressed in high density in astrocytes. By fluxing water along osmotic gradients, AQP4 contributes to brain volume and ion homeostasis. Here we ask whether deletion of Aqp4 leads to upregulation of the gap junctional proteins connexin-43 (Cx43) and connexin-30 (Cx30). These molecules couple adjacent astrocytes to each other and allow water and ions to redistribute within the astrocyte syncytium. Immunogold analysis of parietal cortex and hippocampus showed that the number of gap junctions per capillary profile is increased in AQP4 knockout (AQP4 KO) mice. The most pronounced changes were observed for Cx43 in hippocampus where the number of connexin labeled gap junctions increased by 100% following AQP4 KO. Western blot analysis of whole tissue homogenates showed no change in the amount of Cx43 or Cx30 protein after AQP4 KO. However, AQP4 KO led to a significant increase in the amount of Cx43 in a Triton X-100 insoluble fraction. This fraction is associated with connexin assembly into gap junctional plaques in the plasma membrane. In line with our immunoblot data, RT-qPCR showed no significant increase in Cx43 and Cx30 mRNA levels after AQP4 KO. Our findings suggest that AQP4 KO leads to increased aggregation of Cx43 into gap junctions and provide a putative mechanistic basis for the enhanced tracer coupling in hippocampi of AQP4 KO mice. The increased number of gap junctions in AQP4 deficient mice may explain why Aqp4 deletion has rather modest effects on brain volume and K+ homeostasis.

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Acknowledgements

The authors are grateful to Ms.Bjørg Riber, Karen Marie Gujord and Jorunn Knutsen for the technical assistance and Carina Knudsen for the help with the artwork. Cx30−/−Cx43fl/fl/hGFAP-Cre mice were a kind gift from Dr. Martine Cohen Salmon, École des Neurosciences de Paris Île-de-France (ENP).

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Authors and Affiliations

  1. Division of Anatomy, Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway

    Shirin Katoozi, Nadia Skauli, Soulmaz Rahmani, Laura M. A. Camassa, Henning B. Boldt, Ole P. Ottersen & Mahmood Amiry-Moghaddam

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  1. Shirin Katoozi
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  2. Nadia Skauli
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  3. Soulmaz Rahmani
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  4. Laura M. A. Camassa
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  7. Mahmood Amiry-Moghaddam
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Correspondence to Mahmood Amiry-Moghaddam.

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Katoozi, S., Skauli, N., Rahmani, S. et al. Targeted deletion of Aqp4 promotes the formation of astrocytic gap junctions. Brain Struct Funct 222, 3959–3972 (2017). https://doi.org/10.1007/s00429-017-1448-5

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  • DOI: https://doi.org/10.1007/s00429-017-1448-5

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