Dissecting the role of plasmalogen lipids in synapse function

This research aims to reveal fundamental mechanisms of neurodegenerative disorders by studying the consequences of disrupted lipid homeostasis in neurons by means of functional synapse assays and protein expression analyses. In neurodegeneration, the physiology of synapses is altered. Here, plasmalogen lipids are abundant and their peculiar conical shape makes them ideal in supporting vesicle fusion. In addition, plasmalogen levels decrease with pathological progression in Alzheimer’s disease. Since current evidence is correlative, we aim to provide tools to directly test whether plasmalogens support synaptic transmission for normal neuronal cell function. Specifically, we seek to prove that plasmalogens are required for synapse function and later explore their potential for neuro-regenerative supplementation therapies.

Methods:

My work will include western blot and ICC/IF detection of markers of mature neurons (NeuN, b3-tubulin) alongside established synaptic and vesicular markers (Synaptophysin1, PSD95, SV2, VAChT), and an assay for neurotransmitter release (Ach). Wet-lab assays will be performed on otherwise untreated differentiated cells alongside differentiated cells genetically silenced by using siRNA for plasmalogen biosynthetic enzymes FAR1 and PEDS1. Specifically, we will modulate plasmalogen levels in differentiated human SH-SY5Y cells as they embody a convenient model for developing assays. This work will be complemented with lipidomic analyses and will be soon translated to relevant iPSC-derived neurons.

Statistics:

Based on our preliminary data and previous literature, we expect the data from lipidomic and synaptic assays to be normally distributed. Comparison of multiple groups (n=6) at one timepoint (e.g., normal vs plasmalogen-deficient) will use one-way ANOVA with Bonferroni correction (GraphPad Software).

Content

Author and article information

Journal

Title: ScienceOpen Posters

Publisher: ScienceOpen

Publication date (Electronic preprint): 16 April 2023

Affiliations

[1 ] Swansea University Medical School;

Author notes

[* ]Email: sadilovaa@ 123456gmail.com .

Author information

Anna Sadilova https://orcid.org/0009-0006-3109-1576

Roberto Angelini https://orcid.org/0000-0001-5136-5921

Article

DOI: 10.14293/P2199-8442.1.SOP-.PJDZ5Y.v1

SO-VID: 3e2e1c57-6ef4-49ca-84b2-bdc7d3b9ae74

License:

This work has been published open access under Creative Commons Attribution License CC BY 4.0 , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Conditions, terms of use and publishing policy can be found at www.scienceopen.com .

History

Date received : 16 April 2023

References

Shipley Mackenzie M., Mangold Colleen A., Szpara Moriah L.. Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line. Journal of Visualized Experiments. (108)2016. MyJove Corporation. [Cross Ref]
Senanayake Vijitha, Goodenowe Dayan B.. Plasmalogen deficiency and neuropathology in Alzheimer's disease: Causation or coincidence? Alzheimer's & Dementia: Translational Research & Clinical Interventions. Vol. 5(1):524–532. 2019. Wiley. [Cross Ref]
Rouser George, Yamamoto Akira. Curvilinear regression course of human brain lipid composition changes with age. Lipids. Vol. 3(3):284–287. 1968. Wiley. [Cross Ref]
Han Xianlin, Holtzman David M., McKeel Daniel W.. Plasmalogen deficiency in early Alzheimer's disease subjects and in animal models: molecular characterization using electrospray ionization mass spectrometry. Journal of Neurochemistry. Vol. 77(4):1168–1180. 2001. Wiley. [Cross Ref]
Verstraelen Peter, Garcia-Diaz Barriga Gerardo, Verschuuren Marlies, Asselbergh Bob, Nuydens Rony, Larsen Peter H., Timmermans Jean-Pierre, De Vos Winnok H.. Systematic Quantification of Synapses in Primary Neuronal Culture. iScience. Vol. 23(9)2020. Elsevier BV. [Cross Ref]
Dravid Anusha, Raos Brad, Svirskis Darren, O’Carroll Simon J.. Optimised techniques for high-throughput screening of differentiated SH-SY5Y cells and application for neurite outgrowth assays. Scientific Reports. Vol. 11(1)2021. Springer Science and Business Media LLC. [Cross Ref]
Shipley Mackenzie M., Mangold Colleen A., Szpara Moriah L.. Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line. Journal of Visualized Experiments. (108)2016. MyJove Corporation. [Cross Ref]
Ippolito Dominic M., Eroglu Cagla. Quantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse Number. Journal of Visualized Experiments. (45)2010. MyJove Corporation. [Cross Ref]
Rouser George, Yamamoto Akira. Curvilinear regression course of human brain lipid composition changes with age. Lipids. Vol. 3(3):284–287. 1968. Wiley. [Cross Ref]
Bozelli José Carlos, Epand Richard M.. Plasmalogen Replacement Therapy. Membranes. Vol. 11(11)2021. MDPI AG. [Cross Ref]
Senanayake Vijitha, Goodenowe Dayan B.. Plasmalogen deficiency and neuropathology in Alzheimer's disease: Causation or coincidence? Alzheimer's & Dementia: Translational Research & Clinical Interventions. Vol. 5(1):524–532. 2019. Wiley. [Cross Ref]
Gu Jinxin, Chen Lixue, Sun Ran, Wang Jie-Li, Wang Juntao, Lin Yingjun, Lei Shuwen, Zhang Yang, Lv Dan, Jiang Faqin, Deng Yuru, Collman James P., Fu Lei. Plasmalogens Eliminate Aging-Associated Synaptic Defects and Microglia-Mediated Neuroinflammation in Mice. Frontiers in Molecular Biosciences. Vol. 9:2022. Frontiers Media SA. [Cross Ref]
Verstraelen Peter, Garcia-Diaz Barriga Gerardo, Verschuuren Marlies, Asselbergh Bob, Nuydens Rony, Larsen Peter H., Timmermans Jean-Pierre, De Vos Winnok H.. Systematic Quantification of Synapses in Primary Neuronal Culture. iScience. Vol. 23(9)2020. Elsevier BV. [Cross Ref]
Longair Mark H., Baker Dean A., Armstrong J. Douglas. Simple Neurite Tracer: open source software for reconstruction, visualization and analysis of neuronal processes. Bioinformatics. Vol. 27(17):2453–2454. 2011. Oxford University Press (OUP). [Cross Ref]
Wang Yizhi, Wang Congchao, Ranefall Petter, Broussard Gerard Joey, Wang Yinxue, Shi Guilai, Lyu Boyu, Wu Chiung-Ting, Wang Yue, Tian Lin, Yu Guoqiang. SynQuant: an automatic tool to quantify synapses from microscopy images. Bioinformatics. Vol. 36(5):1599–1606. 2020. Oxford University Press (OUP). [Cross Ref]

Comments

Comment on this article

[1] Shipley Mackenzie M., Mangold Colleen A., Szpara Moriah L.. Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line. Journal of Visualized Experiments. (108)2016. MyJove Corporation. [Cross Ref]

[2] Senanayake Vijitha, Goodenowe Dayan B.. Plasmalogen deficiency and neuropathology in Alzheimer's disease: Causation or coincidence? Alzheimer's & Dementia: Translational Research & Clinical Interventions. Vol. 5(1):524–532. 2019. Wiley. [Cross Ref]

[3] Rouser George, Yamamoto Akira. Curvilinear regression course of human brain lipid composition changes with age. Lipids. Vol. 3(3):284–287. 1968. Wiley. [Cross Ref]

[4] Han Xianlin, Holtzman David M., McKeel Daniel W.. Plasmalogen deficiency in early Alzheimer's disease subjects and in animal models: molecular characterization using electrospray ionization mass spectrometry. Journal of Neurochemistry. Vol. 77(4):1168–1180. 2001. Wiley. [Cross Ref]

[5] Verstraelen Peter, Garcia-Diaz Barriga Gerardo, Verschuuren Marlies, Asselbergh Bob, Nuydens Rony, Larsen Peter H., Timmermans Jean-Pierre, De Vos Winnok H.. Systematic Quantification of Synapses in Primary Neuronal Culture. iScience. Vol. 23(9)2020. Elsevier BV. [Cross Ref]

[6] Dravid Anusha, Raos Brad, Svirskis Darren, O’Carroll Simon J.. Optimised techniques for high-throughput screening of differentiated SH-SY5Y cells and application for neurite outgrowth assays. Scientific Reports. Vol. 11(1)2021. Springer Science and Business Media LLC. [Cross Ref]

[7] Shipley Mackenzie M., Mangold Colleen A., Szpara Moriah L.. Differentiation of the SH-SY5Y Human Neuroblastoma Cell Line. Journal of Visualized Experiments. (108)2016. MyJove Corporation. [Cross Ref]

[8] Ippolito Dominic M., Eroglu Cagla. Quantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse Number. Journal of Visualized Experiments. (45)2010. MyJove Corporation. [Cross Ref]

[9] Rouser George, Yamamoto Akira. Curvilinear regression course of human brain lipid composition changes with age. Lipids. Vol. 3(3):284–287. 1968. Wiley. [Cross Ref]

[10] Bozelli José Carlos, Epand Richard M.. Plasmalogen Replacement Therapy. Membranes. Vol. 11(11)2021. MDPI AG. [Cross Ref]

[11] Senanayake Vijitha, Goodenowe Dayan B.. Plasmalogen deficiency and neuropathology in Alzheimer's disease: Causation or coincidence? Alzheimer's & Dementia: Translational Research & Clinical Interventions. Vol. 5(1):524–532. 2019. Wiley. [Cross Ref]

[12] Gu Jinxin, Chen Lixue, Sun Ran, Wang Jie-Li, Wang Juntao, Lin Yingjun, Lei Shuwen, Zhang Yang, Lv Dan, Jiang Faqin, Deng Yuru, Collman James P., Fu Lei. Plasmalogens Eliminate Aging-Associated Synaptic Defects and Microglia-Mediated Neuroinflammation in Mice. Frontiers in Molecular Biosciences. Vol. 9:2022. Frontiers Media SA. [Cross Ref]

[13] Verstraelen Peter, Garcia-Diaz Barriga Gerardo, Verschuuren Marlies, Asselbergh Bob, Nuydens Rony, Larsen Peter H., Timmermans Jean-Pierre, De Vos Winnok H.. Systematic Quantification of Synapses in Primary Neuronal Culture. iScience. Vol. 23(9)2020. Elsevier BV. [Cross Ref]

[14] Longair Mark H., Baker Dean A., Armstrong J. Douglas. Simple Neurite Tracer: open source software for reconstruction, visualization and analysis of neuronal processes. Bioinformatics. Vol. 27(17):2453–2454. 2011. Oxford University Press (OUP). [Cross Ref]

[15] Wang Yizhi, Wang Congchao, Ranefall Petter, Broussard Gerard Joey, Wang Yinxue, Shi Guilai, Lyu Boyu, Wu Chiung-Ting, Wang Yue, Tian Lin, Yu Guoqiang. SynQuant: an automatic tool to quantify synapses from microscopy images. Bioinformatics. Vol. 36(5):1599–1606. 2020. Oxford University Press (OUP). [Cross Ref]