Alpha-SNAP (M105I) mutation promotes neuronal differentiation of neural stem/progenitor cells through overactivation of AMPK
Background: The M105I point mutation in α-SNAP (Soluble N-ethylmaleimide-sensitive factor attachment protein-alpha) leads in mice to a complex phenotype known as hyh (hydrocephalus with hop gait), characterized by cortical malformation and hydrocephalus, among other neuropathological features. Studi...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-04-01
|
| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2023.1061777/full |
| _version_ | 1827969359354527744 |
|---|---|
| author | Felipe A. Bustamante-Barrientos Maxs Méndez-Ruette Maxs Méndez-Ruette Maxs Méndez-Ruette Luis Molina Tania Koning Pamela Ehrenfeld Pamela Ehrenfeld Carlos B. González Ursula Wyneken Ursula Wyneken Ursula Wyneken Roberto Henzi Roberto Henzi Luis Federico Bátiz Luis Federico Bátiz Luis Federico Bátiz |
| author_facet | Felipe A. Bustamante-Barrientos Maxs Méndez-Ruette Maxs Méndez-Ruette Maxs Méndez-Ruette Luis Molina Tania Koning Pamela Ehrenfeld Pamela Ehrenfeld Carlos B. González Ursula Wyneken Ursula Wyneken Ursula Wyneken Roberto Henzi Roberto Henzi Luis Federico Bátiz Luis Federico Bátiz Luis Federico Bátiz |
| author_sort | Felipe A. Bustamante-Barrientos |
| collection | DOAJ |
| description | Background: The M105I point mutation in α-SNAP (Soluble N-ethylmaleimide-sensitive factor attachment protein-alpha) leads in mice to a complex phenotype known as hyh (hydrocephalus with hop gait), characterized by cortical malformation and hydrocephalus, among other neuropathological features. Studies performed by our laboratory and others support that the hyh phenotype is triggered by a primary alteration in embryonic neural stem/progenitor cells (NSPCs) that leads to a disruption of the ventricular and subventricular zones (VZ/SVZ) during the neurogenic period. Besides the canonical role of α-SNAP in SNARE-mediated intracellular membrane fusion dynamics, it also negatively modulates AMP-activated protein kinase (AMPK) activity. AMPK is a conserved metabolic sensor associated with the proliferation/differentiation balance in NSPCs.Methods: Brain samples from hyh mutant mice (hydrocephalus with hop gait) (B6C3Fe-a/a-Napahyh/J) were analyzed by light microscopy, immunofluorescence, and Western blot at different developmental stages. In addition, NSPCs derived from WT and hyh mutant mice were cultured as neurospheres for in vitro characterization and pharmacological assays. BrdU labeling was used to assess proliferative activity in situ and in vitro. Pharmacological modulation of AMPK was performed using Compound C (AMPK inhibitor) and AICAR (AMPK activator).Results: α-SNAP was preferentially expressed in the brain, showing variations in the levels of α-SNAP protein in different brain regions and developmental stages. NSPCs from hyh mice (hyh-NSPCs) displayed reduced levels of α-SNAP and increased levels of phosphorylated AMPKα (pAMPKαThr172), which were associated with a reduction in their proliferative activity and a preferential commitment with the neuronal lineage. Interestingly, pharmacological inhibition of AMPK in hyh-NSPCs increased proliferative activity and completely abolished the increased generation of neurons. Conversely, AICAR-mediated activation of AMPK in WT-NSPCs reduced proliferation and boosted neuronal differentiation.Discussion: Our findings support that α-SNAP regulates AMPK signaling in NSPCs, further modulating their neurogenic capacity. The naturally occurring M105I mutation of α-SNAP provokes an AMPK overactivation in NSPCs, thus connecting the α-SNAP/AMPK axis with the etiopathogenesis and neuropathology of the hyh phenotype. |
| first_indexed | 2024-04-09T18:36:35Z |
| format | Article |
| id | doaj.art-abb45c1e04ef4c9d84988bcc2e9f5e4a |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-09T18:36:35Z |
| publishDate | 2023-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-abb45c1e04ef4c9d84988bcc2e9f5e4a2023-04-11T10:06:36ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2023-04-011110.3389/fcell.2023.10617771061777Alpha-SNAP (M105I) mutation promotes neuronal differentiation of neural stem/progenitor cells through overactivation of AMPKFelipe A. Bustamante-Barrientos0Maxs Méndez-Ruette1Maxs Méndez-Ruette2Maxs Méndez-Ruette3Luis Molina4Tania Koning5Pamela Ehrenfeld6Pamela Ehrenfeld7Carlos B. González8Ursula Wyneken9Ursula Wyneken10Ursula Wyneken11Roberto Henzi12Roberto Henzi13Luis Federico Bátiz14Luis Federico Bátiz15Luis Federico Bátiz16Immunology Program, Centro de Investigación e Innovación Biomédica (CiiB), Universidad de los Andes, Santiago, ChileNeuroscience Program, Centro de Investigación e Innovación Biomédica (CiiB), Universidad de los Andes, Santiago, ChilePhD Program in Biomedicine, Facultad de Medicina, Universidad de los Andes, Santiago, ChileIMPACT, Center of Interventional Medicine for Precision and Advanced Cellular Therapy, Santiago, ChileFacultad de Medicina y Ciencia, Universidad San Sebastián, Puerto Montt, ChileInstituto de Inmunología, Facultad de Medicina, Universidad Austral de Chile, Valdivia, ChileLaboratory of Cellular Pathology, Institute of Anatomy, Histology and Pathology, Faculty of Medicine, Universidad Austral de Chile, Valdivia, ChileCenter for Interdisciplinary Studies on Nervous System (CISNe), Universidad Austral de Chile, Valdivia, ChileInstituto de Fisiología, Facultad de Medicina, Universidad Austral de Chile, Valdivia, ChileNeuroscience Program, Centro de Investigación e Innovación Biomédica (CiiB), Universidad de los Andes, Santiago, ChileIMPACT, Center of Interventional Medicine for Precision and Advanced Cellular Therapy, Santiago, Chile0School of Medicine, Facultad de Medicina, Universidad de Los Andes, Santiago, ChileNeuroscience Program, Centro de Investigación e Innovación Biomédica (CiiB), Universidad de los Andes, Santiago, Chile1Laboratorio de Reproducción Animal, Escuela de Medicina Veterinaria, Facultad de Recursos Naturales, Universidad Católica de Temuco, Temuco, ChileNeuroscience Program, Centro de Investigación e Innovación Biomédica (CiiB), Universidad de los Andes, Santiago, ChileIMPACT, Center of Interventional Medicine for Precision and Advanced Cellular Therapy, Santiago, Chile0School of Medicine, Facultad de Medicina, Universidad de Los Andes, Santiago, ChileBackground: The M105I point mutation in α-SNAP (Soluble N-ethylmaleimide-sensitive factor attachment protein-alpha) leads in mice to a complex phenotype known as hyh (hydrocephalus with hop gait), characterized by cortical malformation and hydrocephalus, among other neuropathological features. Studies performed by our laboratory and others support that the hyh phenotype is triggered by a primary alteration in embryonic neural stem/progenitor cells (NSPCs) that leads to a disruption of the ventricular and subventricular zones (VZ/SVZ) during the neurogenic period. Besides the canonical role of α-SNAP in SNARE-mediated intracellular membrane fusion dynamics, it also negatively modulates AMP-activated protein kinase (AMPK) activity. AMPK is a conserved metabolic sensor associated with the proliferation/differentiation balance in NSPCs.Methods: Brain samples from hyh mutant mice (hydrocephalus with hop gait) (B6C3Fe-a/a-Napahyh/J) were analyzed by light microscopy, immunofluorescence, and Western blot at different developmental stages. In addition, NSPCs derived from WT and hyh mutant mice were cultured as neurospheres for in vitro characterization and pharmacological assays. BrdU labeling was used to assess proliferative activity in situ and in vitro. Pharmacological modulation of AMPK was performed using Compound C (AMPK inhibitor) and AICAR (AMPK activator).Results: α-SNAP was preferentially expressed in the brain, showing variations in the levels of α-SNAP protein in different brain regions and developmental stages. NSPCs from hyh mice (hyh-NSPCs) displayed reduced levels of α-SNAP and increased levels of phosphorylated AMPKα (pAMPKαThr172), which were associated with a reduction in their proliferative activity and a preferential commitment with the neuronal lineage. Interestingly, pharmacological inhibition of AMPK in hyh-NSPCs increased proliferative activity and completely abolished the increased generation of neurons. Conversely, AICAR-mediated activation of AMPK in WT-NSPCs reduced proliferation and boosted neuronal differentiation.Discussion: Our findings support that α-SNAP regulates AMPK signaling in NSPCs, further modulating their neurogenic capacity. The naturally occurring M105I mutation of α-SNAP provokes an AMPK overactivation in NSPCs, thus connecting the α-SNAP/AMPK axis with the etiopathogenesis and neuropathology of the hyh phenotype.https://www.frontiersin.org/articles/10.3389/fcell.2023.1061777/fullneurogenesishydrocephalus with hop gaitbrain developmentcell metabolismcell fateAMPK phosphatase |
| spellingShingle | Felipe A. Bustamante-Barrientos Maxs Méndez-Ruette Maxs Méndez-Ruette Maxs Méndez-Ruette Luis Molina Tania Koning Pamela Ehrenfeld Pamela Ehrenfeld Carlos B. González Ursula Wyneken Ursula Wyneken Ursula Wyneken Roberto Henzi Roberto Henzi Luis Federico Bátiz Luis Federico Bátiz Luis Federico Bátiz Alpha-SNAP (M105I) mutation promotes neuronal differentiation of neural stem/progenitor cells through overactivation of AMPK Frontiers in Cell and Developmental Biology neurogenesis hydrocephalus with hop gait brain development cell metabolism cell fate AMPK phosphatase |
| title | Alpha-SNAP (M105I) mutation promotes neuronal differentiation of neural stem/progenitor cells through overactivation of AMPK |
| title_full | Alpha-SNAP (M105I) mutation promotes neuronal differentiation of neural stem/progenitor cells through overactivation of AMPK |
| title_fullStr | Alpha-SNAP (M105I) mutation promotes neuronal differentiation of neural stem/progenitor cells through overactivation of AMPK |
| title_full_unstemmed | Alpha-SNAP (M105I) mutation promotes neuronal differentiation of neural stem/progenitor cells through overactivation of AMPK |
| title_short | Alpha-SNAP (M105I) mutation promotes neuronal differentiation of neural stem/progenitor cells through overactivation of AMPK |
| title_sort | alpha snap m105i mutation promotes neuronal differentiation of neural stem progenitor cells through overactivation of ampk |
| topic | neurogenesis hydrocephalus with hop gait brain development cell metabolism cell fate AMPK phosphatase |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2023.1061777/full |
| work_keys_str_mv | AT felipeabustamantebarrientos alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT maxsmendezruette alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT maxsmendezruette alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT maxsmendezruette alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT luismolina alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT taniakoning alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT pamelaehrenfeld alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT pamelaehrenfeld alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT carlosbgonzalez alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT ursulawyneken alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT ursulawyneken alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT ursulawyneken alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT robertohenzi alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT robertohenzi alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT luisfedericobatiz alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT luisfedericobatiz alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk AT luisfedericobatiz alphasnapm105imutationpromotesneuronaldifferentiationofneuralstemprogenitorcellsthroughoveractivationofampk |