Biology subjects

Gonzalez, B. J.

Publications and source records attributed to Gonzalez, B. J..

3 recordsLinked to original sources

bioRxiv2026

Spatio-temporal 3D Mapping of Mouse Cerebellar Vascularization during Embryonic Development

Despite major advances in the study of cerebellar neurogenesis, cerebellar angiogenesis during embryogenesis remains poorly described. Recent advances in tissue clearing, light-sheet microscopy, and artificial intelligence have increasingly enabled detailed 3D modelling of cerebellar vasculature at early developmental stages. Here, vascular networks in mouse embryos from E11 to birth (P0) were labelled with podocalyxin, SMA, and PECAM-1 antibodies together with the nuclear marker TO-PRO-3 iodide, cleared, imaged by light-sheet microscopy, and finally modelled and quantitatively analyzed using Imaris and VesselVio. Our mapping reveals that the three main paired cerebellar arteries--the superior (SCA), anterior inferior (AICA), and posterior inferior (PICA) cerebellar arteries--emerge sequentially between E11 and E13 and display significant topographical variability comparable to that observed in humans. Morphometric analysis demonstrates distinct developmental dynamics, with SCA growth proportional to cerebellar expansion, whereas the AICA and PICA exhibit accelerated extension during later embryonic stages. Interestingly, the PICA does not reach the cerebellum before birth, highlighting the question of its contribution to embryonic cerebellar vascularization. The intrinsic vascular network evolves from a rudimentary bilayer at E11 into a highly branched architecture organized around radial penetrating vessels, giving rise to collaterals that progressively colonized the cerebellar parenchyma during foliation and lobulation. These vascular changes temporally coincided with the successive stages of cerebellar neurogenesis, supporting an interplay between vascular and neuronal development. Together, our findings provide the first spatio-temporal three-dimensional atlas of cerebellar vascularization during mouse embryogenesis, establishing a reference framework for investigating cerebellar angiogenesis in developmental and pathological conditions. HighlightsO_LIThis work is the first 3D modelling of the cerebellar vasculature in mouse embryo. C_LIO_LISCA, AICA, PICA develop through distinct spatial and temporal growth programs. C_LIO_LIPICA does not contribute to cerebellar vascularization before birth. C_LIO_LIThe intra-cerebellar vascularization evolves at E11 from a simple vessel bilayer. C_LIO_LIBetween E13 and P0, radial vessels form collaterals colonizing cerebellum. C_LIO_LIThe vascular changes temporally coincided with cerebellar neurogenesis. C_LI

neuroscience↗

bioRxiv2025

Endothelial NMDA receptor involvement in retinal neurovascular damage following prenatal alcohol exposure in mouse model

Prenatal alcohol exposure (PAE) induces neurodevelopmental damage leading to fetal alcohol spectrum disorders (FASD) by altering both brain and ocular development. Recent data showed that PAE impairs brain cortical and retinal vasculature leading to defective positioning of interneurons. In the retina, PAE disturbs vascular development and the association of calretinin neurons with vessels. The NMDA receptor (NMDAR) is a major target of alcohol in the brain, and both ligand binding to NMDARs and the expression of NMDAR subunits are altered in FASD. Given that NMDAR is also expressed in endothelial cells and that glutamate stimulation of endothelial NMDAR (eNMDAR) regulates cortical interneuron positioning along blood vessels, we hypothesize that eNMDAR is critical for retinal vascular development and mediates PAE-induced defects. Using an in vivo model of FASD and transgenic mice lacking, specifically in endothelial cells, the GluN1 subunit of the NMDAR, this study aimed to characterize the neurovascular phenotype of the developing retina. The results show that eNMDAR knockout delays the formation of the superficial vascular plexus and prevents the alterations in vascular organization and neuronal density induced by PAE, particularly cells positioned closer to the retinal vasculature, namely ganglion, amacrine, and horizontal cells. Moreover, eNMDAR deletion led to an increased number of calretinin-positive interneurons in contact with perforating vessels and prevents the decrease induced by PAE. Together, these findings demonstrate that eNMDARs are essential for normal retinal neurovascular development and mediate, at least in part, the detrimental effects of ethanol exposure in FASD. Significance statementUsing a murine model of Fetal Alcohol Spectrum Disorder (FASD) and transgenic mice lacking the GluN1 subunit of the NMDA receptor specifically in endothelial cells (eNMDAR), this study demonstrates that eNMDAR plays a critical role in mediating prenatal alcohol exposure (PAE)-induced neurovascular abnormalities in the retina. Loss of eNMDAR alters the progression of the superficial vascular plexus and prevents the vascular impairments typically observed following PAE. In addition, eNMDAR deletion protects against PAE-induced neuronal damage, particularly affecting retinal ganglion cells, calbindin-positive, and calretinin-positive interneurons. Notably, this study identifies, for the first time, a role for endothelial NMDAR in regulating neurovascular interactions between retinal vessels and calretinin-positive neurons, highlighting this receptor as a key molecular mediator of ethanol-induced retinal damage.

neuroscience↗

bioRxiv2021

Human stem cell model of HNF1A deficiency shows uncoupled insulin to C-peptide secretion with accumulation of abnormal insulin granules

Mutations in HNF1A cause Maturity Onset Diabetes of the Young type 3 (MODY3), the most prevalent form of monogenic diabetes. We generated stem cell-derived pancreatic endocrine cells from human embryonic stem cells (hESCs) with induced hypomorphic mutations in HNF1A. Using these cells, we show that HNF1A orchestrates a transcriptional program required for distinct aspects of {beta}-cell fate and function. During islet cell differentiation, HNF1A deficiency biases islet endocrine cells towards an -cell fate associated with PAX4 down-regulation. HNF1A- deficient {beta}-cells display impaired basal and glucose stimulated-insulin secretion in association with a reduction in CACNA1A and intracellular calcium levels, and impaired insulin granule exocytosis in association with SYT13 down-regulation. Knockout of PAX4, CACNA1A and SYT13 reproduce the relevant phenotypes. Reduction of insulin secretion is associated with accumulation of enlarged secretory granules, and altered stoichiometry of secreted insulin to C-peptide. In HNF1A deficient {beta}-cells, glibenclamide, a sulfonylurea drug used in the treatment of MODY3 patients, increases intracellular calcium to levels beyond those achieved by glucose, and restores C-peptide and insulin secretion to a normal stoichiometric ratio. To study HNF1A deficiency in the context of a human disease model, we also generated stem cell-derived pancreatic endocrine cells from two MODY3 patients induced pluripotent stem cells (iPSCs). While insulin secretion defects are constitutive in cells with complete HNF1A loss of function, {beta}-cells heterozygous for hypomorphic HNF1A mutations are initially normal, but lose the ability to secrete insulin and acquire abnormal stoichiometric secretion ratios. Importantly, the defects observed in these stem cell models are also seen in circulating proportions of insulin:C-peptide in nine MODY3 patients. One sentence of summaryDeficiency of the transcription factor HNF1A biases islet endocrine cell fate towards -cells, impairs intracellular calcium homeostasis and insulin exocytosis, alters the stoichiometry of insulin to C-peptide release, and leads to an accumulation of abnormal insulin secretory granules in {beta}-cells.

cell biology↗