REGULATION OF NEUROGENESIS BY CALPAINS: RELEVANCE FOR POST-INJURY BRAIN REPAIR

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Organizational Unit

Publications

Calpastatin Overexpression Preserves Cognitive Function Following Seizures, While Maintaining Post-Injury Neurogenesis

Publication . Machado, Vanessa M.; Lourenco, Ana S.; Florindo, Claudia; Fernandes, Raquel; Carvalho, Caetana M.; Araújo, Inês

In the adult mammalian brain, new neurons continue to be produced throughout life in two main regions in the brain, the subgranular zone (SGZ) in the hippocampus and the subventricular zone in the walls of the lateral ventricles. Neural stem cells (NSCs) proliferate in these niches, and migrate as neuroblasts, to further differentiate in locations where new neurons are needed, either in normal or pathological conditions. However, the endogenous attempt of brain repair is not very efficient. Calpains are proteases known to be involved in neuronal damage and in cell proliferation, migration and differentiation of several cell types, though their effects on neurogenesis are not well known. Previous work by our group has shown that the absence of calpastatin (CAST), the endogenous inhibitor of calpains, impairs early stages of neurogenesis. Since the hippocampus is highly associated with learning and memory, we aimed to evaluate whether calpain inhibition would help improve cognitive recovery after lesion and efficiency of post-injury neurogenesis in this region. For that purpose, we used the kainic acid (KA) model of seizure-induced hippocampal lesion and mice overexpressing CAST. Selected cognitive tests were performed on the 3rd and 8th week after KA-induced lesion, and cell proliferation, migration and differentiation in the dentate gyrus (DG) of the hippocampus of adult mice were analyzed using specific markers. Cognitive recovery was evaluated by testing the animals for recognition, spatial and associative learning and memory. Cognitive function was preserved by CAST overexpression following seizures, while modulation of post-injury neurogenesis was similar to wild type (WT) mice. Calpain inhibition could still be potentially able to prevent the impairment in the formation of new neurons, given that the levels of calpain activity could be reduced under a certain threshold and other harmful effects from the pathological environment could also be controlled.

2017Journal article

Open Access

Involvement of calpains in adult neurogenesis: implications for stroke

Publication . Machado, Vanessa M.; Morte, Maria I.; Carreira, Bruno P.; Azevedo, Maria M.; Takano, Jiro; Iwata, Nobuhisa; Saido, Takaomi C.; Asmussen, Hannelore; Horwitz, Alan R.; Carvalho, Caetana M.; Araújo, Inês

Calpains are ubiquitous proteases involved in cell proliferation, adhesion and motility. In the brain, calpains have been associated with neuronal damage in both acute and neurodegenerative disorders, but their physiological function in the nervous system remains elusive. During brain ischemia, there is a large increase in the levels of intracellular calcium, leading to the activation of calpains. Inhibition of these proteases has been shown to reduce neuronal death in a variety of stroke models. On the other hand, after stroke, neural stem cells (NSC) increase their proliferation and newly formed neuroblasts migrate towards the site of injury. However, the process of forming new neurons after injury is not efficient and finding ways to improve it may help with recovery after lesion. Understanding the role of calpains in the process of neurogenesis may therefore open a new window for the treatment of stroke. We investigated the involvement of calpains in NSC proliferation and neuroblast migration in two highly neurogenic regions in the mouse brain, the dentate gyrus (DG) and the subventricular zone (SVZ). We used mice that lack calpastatin, the endogenous calpain inhibitor, and calpains were also modulated directly, using calpeptin, a pharmacological calpain inhibitor. Calpastatin deletion impaired both NSC proliferation and neuroblast migration. Calpain inhibition increased NSC proliferation, migration speed and migration distance in cells from the SVZ. Overall, our work suggests that calpains are important for neurogenesis and encourages further research on their neurogenic role. Prospective therapies targeting calpain activity may improve the formation of new neurons following stroke, in addition to affording neuroprotection.

2015-02Journal article

Open Access

Regulation of neurogenesis by calpains: revelance for post-injury brain repair

Publication . Machado, Vanessa Mendes; Araújo, Inês

No cérebro adulto dos mamíferos, novos neurónios são continuamente adicionados em duas regiões neurogénicas principais: a zona subventricular, nas paredes dos ventrículos laterais, e a zona subgranular, no giro dentado do hipocampo. As células estaminais neurais presentes nestes nichos têm a capacidade de proliferar e dar origem a neuroblastos em migração, que acabam por se diferenciar em novos neurónios. Curiosamente, no evento de uma lesão cerebral, a proliferação das células estaminais neurais aumenta, possivelmente numa tentativa de reparação do dano cerebral. No entanto, esta reparação é limitada pela redução da migração celular ou pela diminuição da sobrevivência dos novos neurónios. Descobrir como aumentar a eficiência da neurogénese após lesão poderá ser portanto benéfico para a reparação cerebral. Por outro lado, o dano cerebral também pode activar diversas protéases, incluindo as calpaínas. As calpaínas são proteases ubíquas, sabendo-se que estão envolvidas na proliferação, migração e diferenciação celular em diversos sistemas. No cérebro, a inibição das calpaínas tem efeito neuroprotector, mas os seus efeitos na neurogénese permanecem ainda pouco conhecidos. A delecção do inibidor endógeno das calpaínas, a calpastatina, já anteriormente mostrou ser capaz de reduzir as primeiras fases da neurogénese in vivo. No presente trabalho, mostramos que a inibição das calpaínas é capaz de reverter este efeito, implicando-as ainda mais no processo. Uma vez que a inibição das calpaínas parece ser um alvo promissor para o aumento da neurogénese após lesão, avaliámos várias fases do processo neurogénico, bem como a recuperação das alterações do comportamento cognitivo após lesão no hipocampo induzida por crises epilépticas, em murganhos com sobrexpressão de calpastatina. Avaliámos ainda os efeitos da delecção das calpaínas 1 e 2 nas primeiras fases da neurogénese no hipocampo. Observámos que o padrão de aumento da neurogénese após lesão foi mantido em murganhos com sobrexpressão de calpastatina, apesar de alguma recuperação cognitiva indicar algum grau de neuroprotecção, que por si só poderá ter mascarado a influência da inibição das calpaínas na neurogénese. Murganhos com falta de calpaínas, no entanto, não apresentaram alterações das primeiras fases da neurogénese no hipocampo, sugerindo que as calpaínas apenas afectam este processo quando a sua actividade está acima de um certo limite, abaixo do qual a neurogénese não é afectada. Isto significa que a inibição das calpaínas poderá potencialmente ser útil para prevenir a redução da formação de novos neurónios após lesão, caso os níveis de actividade das calpaínas possam ser reduzidos abaixo de um certo limite.

2016-10-24Doctoral thesis

Open Access

Funding agency

Fundação para a Ciência e a Tecnologia

Funding Award Number

SFRH/BD/78050/2011