Sensivity of store-operated calcium entry to antagonists of monotropic receptors

• Autorzy: Gruszczynska-Biegala J. , Sladowska M. , Kuznicki J.
• Języki publikacji: EN

Abstrakty

EN

BACKGROUND AND AIMS: In non-excitable cells Ca2+ enters through Store-Operated Ca2+ Entry (SOCE) pathway, involving STIM1, STIM2 and Orai1 proteins. Upon activation of neurons [Ca2+] is increased in the cytoplasm as a result of Ca2+ influx from the extracellular environment mainly through voltage-operated Ca2+ channels and ionotropic receptor-operated Ca2+ channels (IR). Recently, another possibility has been shown into neurons – Ca2+ influx via SOCE. Our earlier data indicated that both STIMs are involved in Ca2+ homeostasis in neurons, form complexes with endogenous ORAI1 (Gruszczynska-Biegala and Kuznicki 2013, J Neurochem), but played a distinct role in SOCE (Gruszczynska-Biegala et al. 2011, PLoS ONE). The aim of this study is to determine, which receptors react with STIM proteins and are involved in SOCE. The potential STIM partnersin plasma membrane, belong to 3 types of IR (NMDAR, AMPAR and kainate receptors – KR). METHODS: In cultured cortical neurons we recorded single-cell Ca2+ levels using the ratiometric Ca2+ indicator Fura-2AM. SOCE was measured after depletion of intracellular Ca2+ stores by thapsigargin (TG) and subsequent incubation of cells in 2 mM Ca2+ media. To investigate the involvement of IR in TG-induced Ca2+ entry, we applied antagonists of these receptors such as NS-102 (KR), CNQX (AMPAR/KR), NBQX (AMPAR), MK-801 (NMDAR), memantine (NMDAR), and D-AP5 (NMDAR). RESULTS: We found that SOCE was decreased by CNQX, NBQX, D-AP5, memantine but insignificant changes were observed in the presence of MK801 and NS-102. The results showed that NMDA and AMPA receptors are involved in SOCE pathway. The interaction between endogenous STIM1/STIM2 with IR will be checked by co-immunoprecipitation. CONCLUSIONS: The identification of new partners of STIMs will allow us to better understand the mechanisms of SOCE in healthy neurons and during Alzheimer’s disease degeneration. Supported by funds from a National Science Centre (2011/01/D/NZ3/02051, JGB).

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2015
• Tom: 75
• Numer: Supl.
• Opis fizyczny: p.S40

Twórcy

autor

Gruszczynska-Biegala J.

• International Institute of Molecular and Cell Biology, Warsaw, Poland

autor

Sladowska M.

• International Institute of Molecular and Cell Biology, Warsaw, Poland
• Undergraduate Student, Warsaw University of Life Sciences-SGGW, Warsaw, Poland

autor

Kuznicki J.

• Undergraduate Student, Warsaw University of Life Sciences-SGGW, Warsaw, Poland