Author response: Cd59 and inflammation regulate Schwann cell development
Year: 2022
Type: peer-review
Abstract: Article Figures and data Abstract Editor's evaluation eLife digest Introduction Results Discussion Materials and methods Data availability References Decision letter Author response Article and author information Metrics Abstract Efficient neurotransmission is essential for organism survival and is enhanced by myelination. However, the genes that regulate myelin and myelinating glial cell development have not been fully characterized. Data from our lab and others demonstrates that cd59, which encodes for a small GPI-anchored glycoprotein, is highly expressed in developing zebrafish, rodent, and human oligodendrocytes (OLs) and Schwann cells (SCs), and that patients with CD59 dysfunction develop neurological dysfunction during early childhood. Yet, the function of Cd59 in the developing nervous system is currently undefined. In this study, we demonstrate that cd59 is expressed in a subset of developing SCs. Using cd59 mutant zebrafish, we show that developing SCs proliferate excessively and nerves may have reduced myelin volume, altered myelin ultrastructure, and perturbed node of Ranvier assembly. Finally, we demonstrate that complement activity is elevated in cd59 mutants and that inhibiting inflammation restores SC proliferation, myelin volume, and nodes of Ranvier to wildtype levels. Together, this work identifies Cd59 and developmental inflammation as key players in myelinating glial cell development, highlighting the collaboration between glia and the innate immune system to ensure normal neural development. Editor's evaluation This work investigates the function of the small GPI-anchored protein Cd59, a protein known to suppress complement-mediated inflammation, in Schwann cell development and myelination, using zebrafish as a model system. This article will be of interest to developmental biologists, glial biologists, and immunologists as it suggests the interesting and novel findings that Cd59 regulates Schwann cell development, mainly by modulating Schwann cell proliferation. https://doi.org/10.7554/eLife.76640.sa0 Decision letter Reviews on Sciety eLife's review process eLife digest The nervous system of vertebrates is made of up of complex networks of nerve cells that send signals to one another. In addition to these cells, there are a number of supporting cells that help nerves carry out their role. Schwann cells, for example, help nerve cells to transmit information faster by wrapping their long extensions in a fatty membrane called myelin. When myelin is not produced properly, this can disturb the signals between nerve cells, leading to neurological defects. Schwann cells mature simultaneously with nerve cells in the embryo. However, it was not fully understood how Schwann cells generate myelin during development. To investigate, Wiltbank et al. studied the embryos of zebrafish, which, unlike other vertebrates, are transparent and develop outside of the womb. These qualities make it easier to observe how cells in the nervous system grow in real-time using a microscope. First, the team analyzed genetic data collected from the embryo of zebrafish and mice to search for genes that are highly abundant in Schwann cells during development. This led to the discovery of a gene called cd59, which codes for a protein that also interacts with the immune system. To find out whether Schwann cells rely on cd59, Wiltbank et al. deleted the cd59 gene in zebrafish embryos. Without cd59, the Schwann cells produced too many copies of themselves and this, in turn, impaired the appropriate production of myelin. Since the protein encoded by cd59 normally balances inflammation levels, it was possible that losing this gene overactivated the immune system in the zebrafish embryos. In support of this hypothesis, when the cd59 mutant embryos were treated with an anti-inflammatory drug, this corrected Schwann cell overproduction and restored myelin formation. Taken together, these findings reveal how inflammation helps determine the number of Schwann cells produced during development, and that cd59 prevents this process from getting carried away. This suggests that the nervous system and immune system may work together to build the nervous system. In the future, it will be interesting to investigate whether cd59 acts in a similar way during the development of the human nervous system. Introduction Myelin is a highly specialized, lipid-rich membrane that insulates axons in the nervous system to enhance neurotransmission (Ritchie, 1982) while preserving axon health (Stadelmann et al., 2019). Myelin is produced and maintained by myelinating glial cells such as motor exit point glia (Fontenas and Kucenas, 2021; Fontenas and Kucenas, 2018; Smith et al., 2014) and myelinating Schwann cells (SCs; Figure 1A; Ackerman and Monk, 2016; BEN GEREN, 1954; Jessen and Mirsky, 2005) in the peripheral nervous system (PNS) as well as oligodendrocytes (OLs) in the central nervous system (CNS) (Figure 1B; Ackerman and Monk, 2016; Maturana, 1960; Nave and Werner, 2014; Peters, 1960a; Peters, 1960b). Normal myelin and myelinating glial cell development facilitate an efficient and effective nervous system, ensuring precise motor, sensory, and cognitive function (Almeida and Lyons, 2017; Berger et al., 2006; Stadelmann et al., 2019; Wei et al., 2019). When this process is impaired, as seen with inherited disorders that cause abnormal myelination, patients present with neurological dysfunction that often leads to severe physical and intellectual disabilities (Berger et al., 2006; Stadelmann et al., 2019; van der Knaap and Bugiani, 2017; Wei et al., 2019). Figure 1 with 1 supplement see all Download asset Open asset cd59 is expressed in myelinating glial cells during nervous system development. (A) Timeline of Schwann cell (SC) (orange) development (top panel). SC developmental stages for zebrafish (hours post fertilization [hpf]) and mice (embryonic day [E]) are indicated in the bottom panel. (B) Timeline of oligodendrocyte (OL) (blue) development. OL developmental stages for zebrafish (hpf) and mice (E and postnatal day) are indicated in the bottom panel. (C) Scatter plot of cd59 expression (TPM) in oligodendrocyte lineage cells (OLCs), SCs, and neurons (N) at 72 hpf (left; mean ± SEM: OLC: 2145.1 ± 1215.1; SC: 40.1 ± 16.3; N: 240.5 ± 173.3; dot = replicate) as well as SCs at 36 and 72 hpf (right; mean ± SEM: 36 hpf: 0.0 ± 0.0, 72 hpf: 40.2 ± 16.3; dot = replicate). (D) Schematic of the relative cd59/Cd59a expression in developing SCs (orange) and OLs (blue) determined from RNAseq analysis in Figure 1—figure supplement 1. Developmental stage numbers correspond with stages indicated in (A) and (B). Artwork created by Ashtyn T. Wiltbank with Illustrator (Adobe) based on previous schematics and electron micrographs published in Ackerman and Monk, 2016; Cunningham and Monk, 2018; Jessen and Mirsky, 2005. Figure 1—source data 1 Source data for cd59 bulk, RNAseq expression depicted in Figure 1C. Data contributed to scatter plot of cd59 expression (TPM) in oligodendrocyte lineage cells (OLCs), Schwann cells (SCs), and neurons (N) at 72 hours post fertilization (hpf) (left; mean ± SEM: OLC: 2145.1 ± 1215.1, SC: 40.1 ± 16.3; N: 240.5 ± 173.3; data point = replicate) as well as SCs at 36 and 72 hpf (right; mean ± SEM: 36 hpf: 0.0 ± 0.0, 72 hpf: 40.2 ± 16.3; data point = replicate). https://cdn.elifesciences.org/articles/76640/elife-76640-fig1-data1-v2.xlsx Download elife-76640-fig1-data1-v2.xlsx Despite the importance of myelin, we still lack a complete understanding of myelin and myelinating glial cell development. With the recent boon of transcriptomic and proteomic analyses, many genes and proteins have been highlighted as differentially expressed during myelinating glial cell development, yet their precise functions remain unknown. For example, cd59, a gene that encodes for a small, glycosylphosphotidylinositol (GPI)-anchored glycoprotein, is a particularly interesting candidate for exploration. This gene features in several RNA sequencing (RNAseq) and proteomic analyses of zebrafish and rodent myelinating glial cells (Gerber et al., 2021; Howard et al., 2021; Marisca et al., 2020; Marques et al., 2018; Marques et al., 2016; Piller et al., 2021; Saunders et al., 2019; Siems et al., 2021; Wolbert et al., 2020; Zhu et al., 2019), all of which demonstrate high expression of cd59 in developing oligodendrocyte lineage cells (OLCs) and SCs. cd59 continues to be expressed in juvenile and adult zebrafish (Saunders et al., 2019; Siems et al., 2021; Sun et al., 2013) and becomes the fourth most abundant CNS myelin protein in adult zebrafish (Siems et al., 2021). Phylogenetic analysis demonstrates that Cd59 is conserved within vertebrate evolution (Siems et al., 2021), and accordingly, CD59 protein is also found in developing and adult human myelin (Erne et al., 2002; Koski et al., 2002; Scolding et al., 1998; Zajicek et al., 1995). Collectively, these data indicate that cd59 is a key component of the genetic program that orchestrates myelinating glial cell development. However, despite this knowledge of Cd59 expression, little is known about the function of Cd59 in the developing nervous system. Outside of the nervous system, Cd59 is best known as a complement inhibitory protein. Complement is a molecular cascade within the innate immune system that aids in cell lysis of invading pathogens or aberrant cells in the body (Merle et al., 2015). This process lacks specificity and requires inhibitory proteins, such as Cd59, to protect healthy cells from lytic death (Davies et al., 1989). Cd59, specifically, acts at the end of the complement cascade where it binds to complement proteins 8 and 9 (C8 and C9, respectively) to prevent the polymerization of C9 and the subsequent formation of lytic pores (also known as membrane attack complexes [MACs] or terminal complement complexes [TCCs]) in healthy cell membranes (Meri et al., 1990; Ninomiya and Sims, 1992; Rollins et al., 1991). This interaction is important throughout the adult body, including the nervous system, where CD59a is neuroprotective in models of multiple sclerosis (Mead et al., 2004) and neuromyelitis optica (Yao and Verkman, 2017). Beyond complement-inhibitory functions, Cd59 can also facilitate vesicle signaling in insulin-producing pancreatic cells (Golec et al., 2019; Krus et al., 2014), suppress cell proliferation in T cells responding to a viral infection (Longhi et al., 2005) or smooth muscle cells in models of atherosclerosis (Li et al., 2013), and orchestrate proximal-distal cell identity during limb regeneration (Echeverri and Tanaka, 2005). Like complement inhibition, these processes are critical to normal nervous system function (Almeida et al., 2021; Baron and Hoekstra, 2010; Reiter and Bongarzone, 2020; White and Krämer-Albers, 2014). However, it is unclear exactly what role Cd59 is playing in myelinating glial cell development. Though the precise function is unknown, it is clear that CD59 does impact human nervous system development. Patients with CD59 dysfunction, such as those with congenital CD59 deficiency (Haliloglu et al., 2015; Höchsmann and Schrezenmeier, 2015; Karbian et al., 2018; Solmaz et al., 2020) or germline paroxysmal nocturnal hemoglobinuria (Johnston et al., 2012), present with polyneuropathies during infancy and continue to have nervous system dysfunction throughout their lives. Intriguingly, these neurological symptoms persist even with complement inhibition, the most common treatment for congenital CD59 deficiency (Höchsmann and Schrezenmeier, 2015). Together, these observations further indicate that CD59 has an additional role in the developing nervous system and requires further investigation. Here, utilizing the zebrafish model, we examined the role of Cd59 in the developing nervous system. In this study, we found that cd59 is highly expressed in a subset of developing SCs in addition to mature OLs and myelinating and nonmyelinating SCs. We chose to focus on SCs and found that cd59 mutant zebrafish have excessive SC proliferation. These mutants also have impaired myelin and node of Ranvier development. Finally, we demonstrate that complement activity increases in cd59 mutants and that unregulated inflammation contributes to SC overproliferation and mutant larvae may also have aberrant myelin and node of Ranvier formation. Overall, this data demonstrates that developmental inflammation stimulates SC proliferation and that this process is balanced by Cd59 to ensure normal SC and myelinated nerve development. Results Cd59 is expressed in myelinating glial cells during nervous system development Myelinating glial cell development is a complex process (Figure 1A and B) orchestrated by a genetic program that is not yet fully elucidated. To identify unexplored genes that may impact myelinating glial cell or myelin development, we evaluated bulk and single-cell (sc) RNAseq datasets that assessed myelinating glial cells during nervous system development (Gerber et al., 2021; Howard et al., 2021; Marisca et al., 2020; Marques et al., 2018; Marques et al., 2016; Piller et al., 2021; Saunders et al., 2019; Wolbert et al., 2020; Zhu et al., 2019). Across multiple datasets, cd59 (zebrafish) or Cd59a (mouse), which encodes for a small, GPI-anchored glycoprotein that has no known function in the developing nervous system, is expressed in SCs and OLCs (Figure 1C, Figure 1—figure supplement 1). When looking at SCs, data from developing zebrafish and mice indicated that cd59/Cd59a, respectively, is minimally expressed in neural crest cells (NCCs; the multipotent progenitors that give rise to SCs; Figure 1—figure supplement 1A; Howard et al., 2021; Zhu et al., 2019) but increases in expression in SC precursors (SCPs; Figure 1—figure supplement 1B.1,B.2; Gerber et al., 2021), immature SCs (iSCs; Figure 1—figure supplement 1B.1,B.2; Gerber et al., 2021), pro-myelinating SCs (pmSCs; Figure 1—figure supplement 1B.1,B.2; Gerber et al., 2021), and mature myelinating SCs (MSCs) and non-myelinating SCs (MSCs and NMSCs; Figure 1C, Figure 1—figure supplement 1B.3,C.1,C.2; Gerber et al., 2021; Piller et al., 2021; Saunders et al., 2019; Wolbert et al., 2020). This data indicates that Cd59 may be important during early stages of SC development as well as in mature MSCs and NMSCs (Figure 1D). In contrast, bulk RNAseq of zebrafish oligodendrocyte progenitor cells (OPCs) and OLs indicated that cd59 is expressed near the onset of myelination (72 hours post fertilization [hpf]; Figure 1C; Piller et al., 2021). scRNAseq (Marisca et al., 2020; Marques et al., 2018; Marques et al., 2016) and in situ hybridization (ISH; Siems et al., 2021) of zebrafish and mouse OLCs showed that cd59/CD59a is mostly expressed in mature OLs and not OPCs (Figure 1—figure supplement 1D). CD59 protein is also present within newly formed human myelin sheaths as is evident in immunostaining of third trimester fetal brains (Zajicek et al., 1995). Based on these findings, it is likely that Cd59 does not play a role in early stages of OLC development but could influence mature, myelinating OL function (Figure 1D). Intrigued by this elevated expression of cd59 in developing myelinating glial cells, we verified that cd59 RNA was expressed in OLCs and SCs with chromogenic ISH (CISH) and fluorescent ISH (FISH) at 3 days post fertilization (dpf) (Figure 2A–C). Morphology, location, and co-expression with sox10:megfp, which is a marker for both OLCs and SCs, showed that cd59 RNA is expressed in both cell types at 3 dpf (Figure 2A–C). Beyond expression in the heart and pancreas, cd59 expression is largely confined to developing myelinating glial cells at this stage. Figure 2 with 2 supplements see all Download asset Open asset cd59 is expressed in Schwann cells (SCs) and oligodendrocytes (OLs). (A) Bright-field images of whole-mount chromogenic in situ hybridization (CISH) showing cd59 expression (purple) in the heart, pancreas, dorsal and ventral oligodendrocyte lineage cells (OLCs) (dOLCs and vOLCs, respectively; filled arrows), and posterior lateral line nerve (pLLN) SCs (empty arrows) at 3 days post fertilization (dpf). Schematic (right panel) indicates location of spinal cord, notochord, and pLLN in transverse section. (B) Fluorescent in situ hybridization (FISH) (RNAscope; ACD) showing cd59 expression (cyan) in sox10:megfp-positive, pLLN SCs (cd59-positive orange cells indicated by empty arrows), and spinal cord OLs (cd59-positive orange cells indicated by filled arrows) at 3 dpf in transverse sections (top row). Representative image (top row) displays one SC on each pLLN (left and right empty arrows) as well as multiple OLs in the spinal cord (filled arrows). Images were acquired with confocal imaging. (C) Imaris renderings of the confocal images shown in (B), including the full image (left panel). From the full image (left panel, C), a single SC (indicated with the open magenta arrow and enlarged in the middle panel, C’) and a single OLC (indicated with the filled magenta arrow and enlarged in the right panel, C’’) illustrate the cd59 puncta localized within the myelinating cells. (D) Mosaic labeling showing a cd59:tagrfp-positive OL in the spinal cord (left) and two SCs on the pLLN (right) at 3 dpf. (E) Immunofluorescence (IF) showing cd59:tagrfp expression (cyan) in Sox10-positive SCs (orange) along the pLLN at 3 and 7 dpf (left and right panels, respectively). Double-positive cells are indicated with white arrows. White dashed lines outline the pLLN. Sox10-positive pigment cells outside of the dashed lines were not included in the analysis. (F) Scatter plot of percent cd59:tagrfp-positive SCs on the pLLN from 3 to 7 dpf (mean ± SEM: 3 dpf: 9.4 ± 1.5; 4 dpf: 10.0 ± 1.0; 5 dpf: 7.8 ± 1.2; 6 dpf: 4.4 ± 0.6; 7 dpf: 5.8± 0 .6; p-values: 3 vs. 6 dpf: p=0.0126, 4 vs. 6 dpf: p=0.0095, 4 vs. 7 dpf: Pp0.0477; dot = 1 fish). Data collected from somites 11–13 (~320 µm) and normalized to units per 100 µm. These data were compared with a one-way ANOVA with Tukey’s post-hoc test using GraphPad Prism. All fluorescent images were acquired with confocal imaging. Scale bars: (A) lateral view, 100 µm; transverse section, 25 µm; (B, D) 10 µm; (E) 25 µm. Artwork created by Ashtyn T. Wiltbank with Illustrator (Adobe). Figure 2—source data 1 Source data for the quantification of percent cd59:tagrfp-positive cells per 100 µm depicted in Figure 2E. Data contributed to scatter plot of percent cd59:tagrfp-positive Schwann cells (SCs) on the posterior lateral line nerve (pLLN) from 3 to 7 days post fertilization (dpf) (mean ± SEM: 3 dpf: 9.4 ± 1.5; 4 dpf: 10.0 ± 1.0; 5 dpf: 7.8 ± 1.2; 6 dpf: 4.4 ± 0.6; 7 dpf: 5.8 ± 0.6; p-values: 3 vs. 6 dpf: p=0.0126, 4 vs. 6 dpf: p=0.0095, 4 vs. 7 dpf: p=0.0477; data point = 1 fish). These data were compared with a one-way ANOVA with Tukey’s post-hoc test using GraphPad Prism. Data collected from somites 11–13 (~320 μm) and normalized to units per 100 μm. https://cdn.elifesciences.org/articles/76640/elife-76640-fig2-data1-v2.xlsx Download elife-76640-fig2-data1-v2.xlsx To study these cells in vivo, we created a transcriptional reporter zebrafish line for cd59 (herein referred to as cd59:tagrfp). This line utilizes 5 kb of the cd59 promoter as well as the first exon and intron, the latter containing an enhancer that is important for cd59 expression (Tone et al., 1999), to drive cytoplasmic expression of TagRFP in cd59-expressing cells. With in vivo, confocal imaging, mosaic labeling with our cd59:tagrfp construct revealed labeling of OLs and SCs with myelin-like processes (Figure 2D), indicating that cd59 is expressed in mature myelinating glial cells. Bulk and scRNAseq data indicated that cd59 is expressed during early stages of SC development whereas cd59 expression was restricted to mature OLs. To verify this expression pattern, we looked at cd59 RNA expression at earlier stages in the SC using and we not observe cd59 expression in we see it in the and of the spinal cord at this stage (Figure supplement 1A and 36 and when to develop and we expression of cd59 with in cells along the posterior lateral line nerve (pLLN) in the (Figure supplement and 72 we cd59 expression within SCs along the pLLN (Figure supplement and These data demonstrate that cd59 is expressed in developing SCs, the expression seen in the RNAseq datasets we analyzed (Figure 1—figure supplement 1). From our ISH we that cd59 was not expressed in SCs with spinal motor nerves not cd59 (Figure 2A–C). cd59 expression was largely confined to SCs along the pLLN (Figure 2A–C). To investigate this expression we our cd59:tagrfp This line in labeling of cd59-expressing pLLN SCs, which we using with a cd59 in cd59:tagrfp-positive SCs (Figure supplement To verify that cd59:tagrfp-positive SCs expressed SC including 10 and myelin protein a we examined pLLN SCs in and gene larvae these with in confocal imaging. cd59:tagrfp expression was in and SCs at 72 and 7 dpf (Figure supplement that cd59:tagrfp-positive SCs other known SC To determine whether cd59 was expressed in all pLLN SCs, we (IF) to cd59:tagrfp-positive SCs with an which all SCs along the pLLN. when looking at SCs from 3 to 7 cd59:tagrfp was expressed in a subset of pLLN SCs of 4.4 ± to 9.4 ± of pLLN SCs; Figure and We also that cd59 expression was expressed in a subset of OLs (Figure and was in Taken together, these data demonstrate that cd59 expression is in developing myelinating glial cells. of cd59 mutant zebrafish with findings demonstrate that cd59 is expressed in a subset of developing SCs. However, it is unclear how Cd59 SC development. In other cells, Cd59 has many functions, including cell lysis (Davies et al., signaling (Golec et al., 2019; Krus et al., 2014), cell proliferation et al., et al., et al., et al., and proximal-distal cell identity (Echeverri and Tanaka, 2005). is little known about Cd59 in the developing nervous system, To the function of Cd59 in SC development, we cd59 mutant zebrafish with the end of the first exon Figure we an with a that the exon 2 a (herein referred to as mutant Figure and sequencing analysis of mutant revealed that multiple were produced compared to wildtype cd59, which produced a single (Figure supplement 1A and of the mutant indicated that an early was in the a protein compared to the wildtype Figure supplement and the proteins the and (Figure supplement and the mutant Cd59 protein was to be We also an additional cd59 mutant which was produced with the RNA This mutant a 6 near the end of exon 2 that not the (Figure supplement and D) and produced a single mutant (Figure supplement of the mutant the of two in the which a protein compared to wildtype Cd59 Figure supplement and Though the protein was not we that with the prevent protein to the and protein Collectively, mutant larvae have perturbed RNA that in formation and protein and mutant larvae have a these both mutants were to have impaired Cd59 Figure 3 with 1 supplement see all Download asset Open asset of cd59 mutant zebrafish with (A) Schematic of zebrafish cd59 The cd59 encodes the protein with the indicated in The is indicated in The dashed lines indicate the and the (B) for wildtype cd59 and mutant showing a at the between exon 2 (blue) and 2 of the cd59 (C) Schematic of Cd59 protein made in wildtype panel) and mutant bottom panel). (D) Bright-field images of wildtype and mutant larvae at 72 hours post fertilization (hpf) showing no as a of cd59 (E) in situ hybridization (CISH) showing oligodendrocytes (OLs) in the spinal cord (filled arrows) and Schwann cells (SCs) on the posterior lateral line nerve (pLLN) (empty arrows) at 72 hpf in transverse (F) showing OLs in the spinal cord (filled arrows) and SCs on the pLLN (empty arrows) at 72 hpf in transverse showing SCs on the pLLN (empty arrows) and neurons in the (filled arrows) at 72 hpf in transverse Fluorescent in situ hybridization (FISH) (RNAscope; ACD) showing cd59 expression (cyan) in SCs (orange) along the pLLN at 72 Representative images each a transverse of µm) of single SC on the pLLN. Imaris renderings show cd59 puncta that are localized within each Scatter plot of the number of cd59 RNA puncta in pLLN SCs (mean ± SEM: ± ± dot = 1 = 7 per These data were compared with using GraphPad Prism. and images were acquired with confocal imaging. Scale bars: (A) 25 µm; µm. Artwork created by Ashtyn T. Wiltbank with Illustrator (Adobe). Figure data 1 Source data for the quantification of the number of cd59 puncta in Schwann cells (SCs) at 72 hours post fertilization (hpf) depicted in Figure Data contributed to scatter plot of the number of cd59 RNA puncta in posterior lateral line nerve (pLLN) SCs (mean ± SEM: ± ± data point = 1 = 7 per These data were compared with using GraphPad Prism. Download To our evaluation of these we to developmental between cd59 mutant larvae and their wildtype and were and produced embryos with no or (Figure Figure supplement we to SCs were present in mutant which were in the of our Using we wildtype and mutant larvae with RNA (also known as early response or and at 72 wildtype and mutant larvae and SCs along the pLLN (Figure indicating that Cd59 was not for SC or expression of genes for SC and myelination. that SCs were we were how cd59 expression was in mutant SCs. to our mutant embryos produced RNA with (Figure supplement RNA with can a process by which normal and mutant gene expression is RNA et al., et al., and et al., 2014). To determine mutant RNA we to wildtype and mutant larvae with a cd59 (Figure and of cd59 RNA puncta showed that cd59 expression was reduced in mutant SCs compared to wildtype SCs (Figure indicating that mutant RNA was and could in a of protein. to the lack of a for Cd59, we were to determine these in RNA
Source: eLife
Institution: University of Virginia
Language: English
Subfield: Developmental Neuroscience
Field: Neuroscience
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $2,500
OpenAlex ID: W4283373949