VPAC1 and VPAC2 Receptor Heterozygosity Confers Distinct Biological Properties to BV2 Microglial Cells.

Song, XYR; Jansen, MI; Marzagalli, R; Musumeci, G; D'Agata, V; Castorina, A

VPAC1 and VPAC2 Receptor Heterozygosity Confers Distinct Biological Properties to BV2 Microglial Cells.

Song, XYR Jansen, MI Marzagalli, R Musumeci, G D'Agata, V Castorina, A

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Publisher:: MDPI
Publication Type:: Journal Article
Citation:: Cells, 2025, 14, (11), pp. 769
Issue Date:: 2025-05-23

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Field	Value	Language
dc.contributor.author	Song, XYR
dc.contributor.author	Jansen, MI
dc.contributor.author	Marzagalli, R
dc.contributor.author	Musumeci, G
dc.contributor.author	D'Agata, V
dc.contributor.author	Castorina, A https://orcid.org/0000-0001-7037-759X
dc.date.accessioned	2026-01-08T01:03:19Z
dc.date.available	2025-05-22
dc.date.available	2026-01-08T01:03:19Z
dc.date.issued	2025-05-23
dc.identifier.citation	Cells, 2025, 14, (11), pp. 769
dc.identifier.issn	2073-4409
dc.identifier.issn	2073-4409
dc.identifier.uri	http://hdl.handle.net/10453/191486
dc.description.abstract	Microglial cells, the resident immune cells of the central nervous system (CNS), are essential for maintaining CNS homeostasis. Dysregulation of microglial function is implicated in the pathogenesis of various neurodegenerative diseases. Vasoactive intestinal polypeptide receptors 1 and 2 (VPAC1 and VPAC2) are G-protein-coupled receptors (GPCRs) expressed by microglia, with their primary ligands being pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP). However, the specific roles of VPAC-type receptors in microglial regulation remain poorly understood. In this study, we generated VPAC1+/- and VPAC2+/- BV2 microglial cell lines using CRISPR-Cas9 gene editing and conducted a series of biological and molecular assays to elucidate the functions of these receptors. Our findings demonstrated that both mutant cell lines exhibited a polarized phenotype and increased migratory activity. VPAC1+/- cells showed enhanced survivability and baseline activation of the unfolded protein response (UPR), a protective mechanism triggered by endoplasmic reticulum (ER) stress, whereas this response appeared impaired in VPAC2+/- cells. In contrast, under lipopolysaccharide (LPS)-induced inflammatory conditions, UPR activation was impaired in VPAC1+/- cells but restored in VPAC2+/- cells, resulting in improved survival of VPAC2+/- cells, whereas VPAC1+/- cells exhibited reduced resilience. Overall, our findings suggest that VPAC1 and VPAC2 receptors play distinct yet complementary roles in BV2 microglia. VPAC2 is critical for regulating survival, ER stress responses, and polarization under basal conditions, while VPAC1 is essential for adaptive responses to inflammatory stimuli such as LPS. These insights advance our understanding of microglial receptor signaling and may inform therapeutic strategies targeting microglial dysfunction in neurodegenerative diseases.
dc.format	Electronic
dc.language	eng
dc.publisher	MDPI
dc.relation.ispartof	Cells
dc.relation.isbasedon	10.3390/cells14110769
dc.rights	info:eu-repo/semantics/openAccess
dc.subject.classification	31 Biological sciences
dc.subject.classification	32 Biomedical and clinical sciences
dc.subject.mesh	Microglia
dc.subject.mesh	Animals
dc.subject.mesh	Receptors, Vasoactive Intestinal Peptide, Type II
dc.subject.mesh	Mice
dc.subject.mesh	Receptors, Vasoactive Intestinal Polypeptide, Type I
dc.subject.mesh	Cell Line
dc.subject.mesh	Lipopolysaccharides
dc.subject.mesh	Cell Movement
dc.subject.mesh	Unfolded Protein Response
dc.subject.mesh	Endoplasmic Reticulum Stress
dc.subject.mesh	Microglia
dc.subject.mesh	Cell Line
dc.subject.mesh	Animals
dc.subject.mesh	Mice
dc.subject.mesh	Lipopolysaccharides
dc.subject.mesh	Cell Movement
dc.subject.mesh	Receptors, Vasoactive Intestinal Polypeptide, Type I
dc.subject.mesh	Receptors, Vasoactive Intestinal Peptide, Type II
dc.subject.mesh	Unfolded Protein Response
dc.subject.mesh	Endoplasmic Reticulum Stress
dc.subject.mesh	Microglia
dc.subject.mesh	Animals
dc.subject.mesh	Receptors, Vasoactive Intestinal Peptide, Type II
dc.subject.mesh	Mice
dc.subject.mesh	Receptors, Vasoactive Intestinal Polypeptide, Type I
dc.subject.mesh	Cell Line
dc.subject.mesh	Lipopolysaccharides
dc.subject.mesh	Cell Movement
dc.subject.mesh	Unfolded Protein Response
dc.subject.mesh	Endoplasmic Reticulum Stress
dc.title	VPAC1 and VPAC2 Receptor Heterozygosity Confers Distinct Biological Properties to BV2 Microglial Cells.
dc.type	Journal Article
utslib.citation.volume	14
utslib.location.activity	Switzerland
pubs.organisational-group	University of Technology Sydney
pubs.organisational-group	University of Technology Sydney/Faculty of Science
pubs.organisational-group	University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	open_access	*
dc.rights.license	This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0). To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/
dc.date.updated	2026-01-08T01:03:15Z
pubs.issue	11
pubs.publication-status	Published online
pubs.volume	14
utslib.citation.issue	11

Abstract:

Microglial cells, the resident immune cells of the central nervous system (CNS), are essential for maintaining CNS homeostasis. Dysregulation of microglial function is implicated in the pathogenesis of various neurodegenerative diseases. Vasoactive intestinal polypeptide receptors 1 and 2 (VPAC1 and VPAC2) are G-protein-coupled receptors (GPCRs) expressed by microglia, with their primary ligands being pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal peptide (VIP). However, the specific roles of VPAC-type receptors in microglial regulation remain poorly understood. In this study, we generated VPAC1+/- and VPAC2+/- BV2 microglial cell lines using CRISPR-Cas9 gene editing and conducted a series of biological and molecular assays to elucidate the functions of these receptors. Our findings demonstrated that both mutant cell lines exhibited a polarized phenotype and increased migratory activity. VPAC1+/- cells showed enhanced survivability and baseline activation of the unfolded protein response (UPR), a protective mechanism triggered by endoplasmic reticulum (ER) stress, whereas this response appeared impaired in VPAC2+/- cells. In contrast, under lipopolysaccharide (LPS)-induced inflammatory conditions, UPR activation was impaired in VPAC1+/- cells but restored in VPAC2+/- cells, resulting in improved survival of VPAC2+/- cells, whereas VPAC1+/- cells exhibited reduced resilience. Overall, our findings suggest that VPAC1 and VPAC2 receptors play distinct yet complementary roles in BV2 microglia. VPAC2 is critical for regulating survival, ER stress responses, and polarization under basal conditions, while VPAC1 is essential for adaptive responses to inflammatory stimuli such as LPS. These insights advance our understanding of microglial receptor signaling and may inform therapeutic strategies targeting microglial dysfunction in neurodegenerative diseases.

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http://hdl.handle.net/10453/191486