Laminin chain expression suggests that laminin-10 is a major isoform in the mouse hippocampus and is degraded by the tissue plasminogen activator/plasmin protease cascade during excitotoxic injury

doi:10.1016/S0306-4522(02)00704-2

Neuroscience

Volume 116, Issue 2, 31 January 2003, Pages 359-371

https://doi.org/10.1016/S0306-4522(02)00704-2 Get rights and content

Abstract

Laminins are important components of the extracellular matrix, and participate in neuronal development, survival and regeneration. The tissue plasminogen activator/plasmin extracellular protease cascade and downstream laminin degradation are implicated in excitotoxin-induced neuronal degeneration. To determine which specific laminin chains are involved, we investigated the expression of laminins in the hippocampus, and the cell types expressing them. Reverse transcription-PCR demonstrated that the messenger RNAs for all laminin chains could be detected in the hippocampus. To determine the localization of laminin chain expression, immunostaining was used. This method showed that α5, β1 and γ1 are most highly expressed in the neuronal cell layers. Immunoblotting confirmed the hippocampal expression of the chains α5, β1 and γ1, and RNA in situ hybridization showed a neuronal expression pattern of α5, β1 and γ1. At early time points following intrahippocampal injection of kainate, α5, β1 and γ1 chain immunoreactivities were lost. In addition, tissue plasminogen activator-deficient mice, which are resistant to kainate-induced neuronal death, show no significant change in laminins α5, β1 and γ1 after intrahippocampal kainate injection.

Taken together, these results suggest that laminin-10 (α5-β1-γ1) comprises a major neuronal laminin in the mouse hippocampus, and is degraded before neuronal death during excitotoxic injury by the tissue plasminogen activator/plasmin protease cascade. By identifying a neuronal laminin (laminin-10) that participates in neuronal degeneration after excitotoxic injury, this study clarifies the molecular definition of the extracellular matrix in the hippocampus and further defines a pathway for mechanisms of neuronal death.

Section snippets

Reverse transcription-polymerase chain reaction

Using chain-specific primers for the different mouse laminin genes (Niimi et al., 1997), reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect expression of 11 different laminin chains. Hippocampi from approximately 10 adult C57BL6/J inbred mice (Taconic, Germantown, NY, USA, or Jackson Laboratories, Bar Harbor, ME, USA) were dissected and messenger RNA (mRNA) was isolated using a Poly(A)Pure kit (Ambion, Austin, TX, USA), and reverse transcribed using random

Laminin RNA expression in the mouse hippocampus

Previous work has shown that after excitotoxin injection in the CNS, laminin is degraded via a plasmin-mediated mechanism, in a manner that correlates spatially and temporally with neuronal death (Chen and Strickland, 1997). Since those experiments were performed using a pan-laminin-1 antibody, the data only supported the involvement of the laminin chains that form laminin-1, namely either α1, β1 or γ1. In order to specifically determine which laminin chains are degraded in this process, we

Discussion

Whereas most tissues, such as the kidney and intestine, possess a very organized basement membrane, the organization of the ECM in the CNS is unusual (Ruoslahti, 1996). Laminins appear to be important components of the ECM in the CNS, and are critical for neuronal survival, as loss of a laminin foundation can predispose neurons to excitotoxic death (Chen and Strickland, 1997). Here we present evidence that multiple laminin chains are expressed in the mouse hippocampus. Although by RT-PCR most

Acknowledgements

This work was supported by grants from the National Institutes of Health (S.E.T., S.S.) and the Medical Scientist Training Program (J.A.I.). We are grateful to Dr Katerina Akassoglou for critical reading of the manuscript and all the members of the Strickland and Tsirka laboratories for their input and support. We also thank Drs J. R. Sanes, and J. H. Miner (Washington University, St. Louis, MO, USA), and L. Sorokin (University of Erlangen-Nuremberg, Erlangen, Germany) for their generous gift

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Original contributionLaminin chain expression suggests that laminin-10 is a major isoform in the mouse hippocampus and is degraded by the tissue plasminogen activator/plasmin protease cascade during excitotoxic injury

Abstract

Section snippets

Reverse transcription-polymerase chain reaction

Laminin RNA expression in the mouse hippocampus

Discussion

Acknowledgements

Matrix Biol

Matrix Biol

Cell

J Biol Chem

Dev Biol

Curr Opin Cell Biol

J Biol Chem

J Biol Chem

J Biol Chem

Neuroscience

Neuron

Brain Res

J Biol Chem

Brain Res Mol Brain Res

J Biol Chem

Matrix Biol

Brain Res Rev

Trends Cell Biol

J Biol Chem

FEBS Lett

Matrix Biol

Int J Biochem Cell Biol

J Biol Chem

J Biol Chem

Brain Res

Dev Biol

Brain Res

Cell

J Biol Chem

J Neurol Sci

Selective immunoreactivities of kidney basement membranes to monoclonal antibodies against lamininlocalization of the end of the long arm and the short arms to discrete microdomains

J Cell Biol

The role of laminins in basement membrane function

J Anat

Physiological consequences of loss of plasminogen activator gene function in mice

Nature

Expression and activity-dependent changes of a novel limbic-serine protease gene in the hippocampus

J Neurosci

Original contribution
Laminin chain expression suggests that laminin-10 is a major isoform in the mouse hippocampus and is degraded by the tissue plasminogen activator/plasmin protease cascade during excitotoxic injury