Increased GS-II lectin binding and SATB2 downregulation are biological features for sessile serrated lesions and microvesicular hyperplastic polyps.

Sessile serrated lesions (SSLs) and microvesicular hyperplastic polyps (MVHPs) are colorectal lesions displaying gastric differentiation. Griffonia simplicifolia-II (GS-II) is a lectin specific to terminal α/ßGlcNAc residues. Here, we assessed GS-II binding and performed immunostaining for HIK1083 (specific to terminal αGlcNAc residues), MUC5AC, MUC6, and special AT-rich sequence binding protein 2 (SATB2) in SSLs, MVHPs, and tubular adenomas (TAs). We observed MUC5AC positivity in 28 of 30 SSLs, but in only three of 23 TAs. Moreover, 24 of 30 SSLs were MUC6-positive, while none of the 23 TAs were MUC6-positive. None of the 30 SSLs or 23 TAs showed HIK1083 positivity. All 30 SSLs and 26 MVHPs were GS-II-positive, while only seven of 23 were in TAs. GS-II staining was mainly distributed in the Golgi region, but SSLs and MVHPs showed goblet cell distribution, in 20 of 30 and 19 of 26 cases, respectively. All SSLs, MVHPs, and TAs were SATB2-positive, but 21 of 30 SSLs and 12 of 26 MVHPs showed decreased staining intensity relative to adjacent mucosa, a decrease seen in only two of 23 in TAs. These results indicate overall that increased terminal ßGlcNAc and decreased SATB2 expression are characteristics of SSLs and MVHPs.

From forest to pharmacy: Should we be depressed about a sustainable Griffonia simplicifolia (Fabaceae) seed supply chain?

ETHNOPHARMACOLOGICAL RELEVANCE: Griffonia simplicifolia D.C (Baill.) (Fabaceae) seeds are unusually high (6-20% wet weight) in 5-HTP (5-Hydroxytryptophan), a serotonin precursor widely used to treat depression. Consequently, this species is regarded as a herbal "Prozac®". Contemporary use as an anti-depressant contrasts with traditional uses for insecticides, arachnicides, fodder, dyes, mordants and chewing-sticks. G. simplicifolia seeds are wild-harvested for the export trade. Over the past 15 years, use of 5-HTP extracted from G. simplicifolia in cosmetics has added to global demand. Wild populations in West Africa are the sole commercial source of G. simplicifolia seed. AIMS OF THE STUDY: Were to (i) assess the scale of the global trade in G. simplicifolia seeds and (ii) produce a synthesis of the challenges facing sustainable harvest of G. simplicifolia. MATERIALS AND APPROACH: Firstly, we analysed global trade data for G. simplicifolia, taking into account historical trends over the past 40 years. Secondly, we reviewed published studies on the distribution, population biology and harvest impacts of wild G. simplicifolia populations. RESULTS AND CONCLUSION: s: Wild G. simplicifolia populations have been the focus of commercial harvest of their pods (for seeds) for international trade from West Africa for almost 50 years. In the late 1980's, when Ghana exported 75-80 metric tonnes (MT) of G. simplicifolia seed to Europe, this species was already Ghana's main medicinal plant export. Currently, 5 West African countries export G. simplicifolia seeds (Cote d'Ivoire, Ghana, Liberia, Nigeria and Togo). Although in the 1980's, most seed exports were to Europe, today China is the main importer of G. simplicifolia seed. These seeds are value-added for production of 5-HTP extracts, and then re-exported, particularly to North America (c.48% of exports). The low habitat specificity and vigorous re-sprouting of G. simplicifolia after cutting, plus its occurrence in forest reserves and national parks confer some resilience on wild populations. Sustaining future supply chains faces six future challenges, however: (1) Rapid loss of forest habitats; (2) Declining populations of understorey birds and disruption of G. simplicifolia pollination in this bird pollinated species; (3) Negative effects of introduced invasive plant species (Broussonetia papyrifera, Chromolaena odorata) on G. simplicifolia regeneration; (4) Grazing by livestock and use of G. simplicifolia leaves as forage; (5) The long-term impact of industrial scale seed "predation": Over a 9-year period (2005-2013), G. simplicifolia exports from Ghana totalled at least 5550 metric tonnes (or between 9.1 billion to 13.5 billion seeds). This could affect the long-term population dynamics of this species, which produces a low number of seeds per pod (1-4 seeds) and has short distance (ballistic) seed dispersal; and (6) Destructive harvest methods, when plants are cut to harvest get the seed pods. Improved resource management, monitoring, quality control and careful pricing are important if supply chains from wild stocks are to be maintained. If wild populations decline, then 5-HTP biosynthesis may compete with low G. simplicifolia seed yields, leading to loss of income to West African harvesters and traders.

Enzymatic modular synthesis and microarray assay of poly-N-acetyllactosamine derivatives.

A facile enzymatic modular assembly strategy for the preparative-scale synthesis of poly-N-acetyllactosamine (poly-LacNAc) glycans with varied lengths and designed sialylation and/or fucosylation patterns is described. These glycans were printed as a microarray to investigate their interactions with a panel of glycan binding proteins (GBPs). Binding affinities revealed that the avidity of GBPs could be largely affected by the length and the patterns of sialylation and fucosylation.

Lectin-Based Method for Deciphering Human Milk IgG Sialylation.

In light of the immunoprotective function of human milk and the incontestable impact of IgG glycosylation on its immune functions, characterization of the sialylation profile of human milk IgG is needed. Lectins as a molecular probe were applied in lectin-IgG-ELISA to analyze the sialylation and galactosylation pattern of skim milk IgG of mothers who delivered at term and prematurely. Well-defined biotinylated lectins were used: Maackia amurensis II (MAA II), Sambucus nigra (SNA), Ricinus communis I (RCA I), and Griffonia simplicifolia II (GSL II) specific to α2,3-Neu5Ac, α2,6-Neu5Ac, Gal(ß1,4)GlcNAc, and agalactosylated glycans, respectively. The sialylation pattern of milk IgG differs qualitatively and quantitatively from maternal plasma IgG and is related to lactation stage and perinatal risk factors. Expression of MAA-, SNA-, and GSL-reactive glycotopes on term milk IgG showed a positive correlation with milk maturation from days 1 to 55. Preterm birth was associated with an increase of MAA-reactive and a decrease of RCA-reactive IgG glycotopes. Moreover, higher SNA- and GSL-reactive and lower RCA-reactive glycoform levels of milk IgG were associated with infection of lactating mothers. Application of a specific and simple method, lectin-IgG-ELISA, reveals the sialylation pattern of milk IgG over milk maturation. However, further investigations are needed in this area.

Chemical Characterization and DNA Fingerprinting of Griffonia simplicifolia Baill.

BACKGROUND: Griffonia simplicifolia Baill. (Caesalpiniaceae) is a medicinal plant whose seeds are widely used in traditional medicine for their high content of 5-hydroxy-l-tryptophan (5-HTP), a direct precursor and enhancer of the activity of the brain hormone serotonin (5-HT). The plant extracts are used in dietary supplements aimed to alleviate serotonin-related disorders. METHODS: In order to characterize the chemical components of G. simplicifolia seeds and their identity, we used a combined methodology by using HPLC-DAD-ESI-MS/MS for the qualitative and quantitative determination of the N-containing compounds, GC-FID and GC-MS for the characterization of the major fatty acids, and DNA fingerprinting based on PCR⁻RFLP for the unequivocal identification of the plant. RESULTS: 5-HTP was the most representative compound, followed by lower percentages of the ß-carboline alkaloid derivative griffonine and other alkaloids. Fatty acids were dominated by the unsaturated fatty acids linoleic acid and oleic acid, followed by the saturated fatty acids stearic and palmitic acids. PCR analysis of the internal transcribed spacer amplified sequence showed a major band at about 758 bp, whereas the PCR⁻RFLP analysis of this sequence using three different restriction enzymes (MspI, HhaI, and HaeIII) generated a specific fingerprinting useful for the plant identification. CONCLUSIONS: The combined chemical and molecular analysis of G. simplicifolia provided an interesting integrated approach for the unequivocal identification of commercial G. simplicifolia seeds.

A Medical Food Formulation of Griffonia simplicifolia/Magnesium for Childhood Periodic Syndrome Therapy: An Open-Label Study on Motion Sickness.

Motion sickness (MS) is a disabling condition dominated by disagreement between visually perceived movement and the vestibular system's sense of movement, with symptoms like dizziness, fatigue, and nausea, and other autonomic disabling symptoms. Preparations of Griffonia simplicifolia, containing high concentrations of 5-HTP, might be effective for serotonin-related disorders, including MS. Therefore, the aim of the present study is to assess the efficacy and safety of the G. simplicifolia/magnesium complex in a pediatric population with MS. The Griffonia/magnesium complex (50 and 200 mg, respectively) was orally administered as a prophylactic therapy for MS twice a day for 3 months to group A, and no therapy for MS was administered to group B. The MS clinical signs were recorded by parents or, where possible, directly from children by a specific module, which included validated questions for the diagnoses that were administered to all subjects and parents of both groups. Two study groups were matched for age (P=.224), sex (P=.801), and z-score body-mass index (P=.173). At T0, all recruited subjects in both groups complained about MS. After 3 months (T1), group A showed an MS prevalence of 36%, significantly lower than MS prevalence in group B (73%) (P<.001). The findings of the present study suggest the role of the Griffonia/magnesium complex as a potential treatment with middle-term efficacy even for MS.

A new ß-carboline alkaloid from the seeds of Griffonia simplicifolia.

AIM: To study the chemical constituents from the seeds of Griffonia simplicifolia Baill. METHODS: The chemical constituents were isolated and purified by a combination of chromatographic materials including silica gel, Sephadex LH-20, and preparative TLC. Their structures were elucidated on the basis of MS and NMR data analysis. The cytotoxic activities were evaluated against HepG2 cancer cell line using the MTT colorimetric method. RESULTS: A new ß-carboline alkaloid, griffonine (1), together with seven known alkaloids, hyrtioerectine B (2), 3-carboxy-6-hydroxy-ß-carboline (3), hyrtiosulawesine (4), 5-hydroxyindole-3-carbaldehyde (5), 5-hydroxy-3-(2-hydroxyethyl) indole (6), trigonelline (7), and 5-hydroxytryptamine (8) were isolated and identified. Alkaloids 1, 2 and 4 showed growth inhibitory effects on the HepG2 cell line with IC50 values of 23.5, 9.6 and 19.3 µmol·L(-1), respectively. CONCLUSION: Alkaloid 1 is new and was named griffonine. Alkaloids 2-7 were isolated from this plant for the first time. Alkaloids 1, 2 and 4 were potentially cytotoxic.

A new β-carboline alkaloid from the seeds of Griffonia simplicifolia / 中国天然药物

AIM@#To study the chemical constituents from the seeds of Griffonia simplicifolia Baill.@*METHODS@#The chemical constituents were isolated and purified by a combination of chromatographic materials including silica gel, Sephadex LH-20, and preparative TLC. Their structures were elucidated on the basis of MS and NMR data analysis. The cytotoxic activities were evaluated against HepG2 cancer cell line using the MTT colorimetric method.@*RESULTS@#A new β-carboline alkaloid, griffonine (1), together with seven known alkaloids, hyrtioerectine B (2), 3-carboxy-6-hydroxy-β-carboline (3), hyrtiosulawesine (4), 5-hydroxyindole-3-carbaldehyde (5), 5-hydroxy-3-(2-hydroxyethyl) indole (6), trigonelline (7), and 5-hydroxytryptamine (8) were isolated and identified. Alkaloids 1, 2 and 4 showed growth inhibitory effects on the HepG2 cell line with IC50 values of 23.5, 9.6 and 19.3 μmol·L(-1), respectively.@*CONCLUSION@#Alkaloid 1 is new and was named griffonine. Alkaloids 2-7 were isolated from this plant for the first time. Alkaloids 1, 2 and 4 were potentially cytotoxic.

Nutraceutical preparations in childhood migraine prophylaxis: effects on headache outcomes including disability and behaviour.

Migraine is common in children, but few specific drugs are available. We performed an open-label comparison of effects of two nutraceutical preparations (ginkgolide B vs. Griffonia simplicifolia extract) on outcomes in 374 school-age children (mean 10.7 years) with migraine without aura. Half of them received ginkgolide B; and half, Griffonia simplicifolia. Both preparations were given orally twice a day for 6 months. Patients kept a headache diary. Outcomes at the beginning and end of treatment were compared. Both preparations reduced all outcome measures after 6 months of treatment. However, reductions in headache frequency, duration and intensity, PedMIDAS score and behavioural reactions to headache were significantly greater in the ginkgolide B group. Both nutraceutical treatments appear promising in paediatric migraine without aura, particularly because of their lack of side effects. However, the ginkgolide B preparation was significantly more effective in the medium-term (6 months).

Relationship between the absorption of 5-hydroxytryptophan from an integrated diet, by means of Griffonia simplicifolia extract, and the effect on satiety in overweight females after oral spray administration.

The management of overweight may include the use of dietary supplements targeted to counter the feeling of hunger. A randomized, double-blind, placebo-controlled trial has been performed in 20 overweight females. These subjects were randomly assigned to supplement their diet with either an extract from Griffonia Simplicifolia (10 subjects) or a placebo (10 matched subjects) for 4-weeks, in conjunction with a personalised reduced calorie diet. The main aim of this study was to evaluate the efficacy, by the assessment of 24-h urinary 5-hydroxyindoleacetic acid levels (5-HIAA), of 1-month administration of a dietary supplement containing 5-hydroxytryptophan (5-HTP) from botanical extracts in healthy, overweight females. Secondary endpoints were the assessment of sensation of appetite (by Haber score), body composition, and severity of binge eating. The supplemented group had a significant increase of 24-h urinary 5-HIAA levels (p<0.001), and a decrease in Haber score (p<0.001) while the placebo group did not show significant changes. With regard to changes in body composition, statistically significant differences between the treatment groups were found for the mean change in BMI, suprailiac skinfold thicknesses, arm circumference and hip circumference. Other parameters were found to be similar in the treated and in the placebo groups. In conclusion, this study shows that the 5-hydroxytryptophan present in the Griffonia extract, administered via spray to the oral cavity, is adequately absorbed, as confirmed by the increase in 24-h urinary 5-HIAA, and that the supplementation of the diet of overweight women with 5-hydroxytryptophan increases the feeling of satiety associated with a decrease in BMI.

Nestin expressing progenitor cells during establishment of the neural retina and its vasculature / 대한해부학회지

In order to test if nestin is a useful marker for various types of progenitor cells, we explored nestin expression in the retina during development. Nestin expression was co-evaluated with bromodeoxyuridine (BrdU) labeling and Griffonia simplicifolia isolectin B4 (GSIB4) histochemistry. Nestin immunoreactivity appears in cell soma of dividing neural progenitor cells and their leading processes in retinas from embryonic day (E) 13 to E20, in accordance with a BrdU-labeled pattern. At postnatal day (P) 5, it is restricted to the end feet of Muller cells. BrdU-labeled nuclei were mainly in the inner part of the inner nuclear layer in postnatal neonates. The retinal vessels demarcated with GSIB4-positive endothelial cells were first distributed in the nerve fiber layer from P3. Afterward the vascular branches sprouted and penetrated deeply into the retina. The endothelial cells positive for GSIB4 and the pericytes in the microvessels were additionally immunoreactive for nestin. Interestingly, the presumed migrating microglial cells showing only GSIB4 reactivity preceded the microvessels throughout the neuroblast layer during vascular sprouting and extension. These findings may suggest that nestin expression represents the proliferation and movement potential of the neural progenitor cells as well as the progenitor cells of the endothelial cell and the pericyte during retinal development. Thus, Muller glial cells might be potential neural progenitor cells of the retina, and the retinal microvasculature established by both the endothelial and the pericyte progenitor cells via vasculogenesis along microglia migrating routes sustains its angiogenic potential.

Influence of Griffonia simplicifolia on male sexual behavior in rats: behavioral and neurochemical study.

The seeds of Griffonia simplicifolia Baill. are rich in 5-HTP (5-hydroxytryptophan), a direct precursor of the neurotransmitter serotonin. In the present study we investigated the influence of the plant extract on male sexual behavior. The seed extract was orally administered to Sprague-Dawley male rats at three dose levels (25, 50 and 100 mg/kg) both acutely and subchronically (daily for 9 days). Mating test with receptive female rats was performed 60 min after the acute treatment or the last dose when repetitively administered. Mount, intromission and ejaculation latencies and post-ejaculatory interval were recorded. Food intake and body weight were measured over the 9-day period of treatment. Microdialysis technique was used to detect the extracellular levels of serotonin (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in rat brain following the acute administration of the extract dosed at 100mg/kg. The acute treatment significantly increased mount latency (at any dosage), intromission and ejaculation latencies (at 100 mg/kg) and post-ejaculatory interval (at 50 and 100 mg/kg). On the contrary the subchronic treatment failed to exert a significant influence on copulatory behavior. The daily administration of the extract dosed at 50 and 100 mg/kg for 9 days significantly reduced food intake and body weight. Finally in the microdialysis experiments we found a dramatic increase in 5-HT and its metabolite 5-HIAA.

Anxiolytic-like effect of Griffonia simplicifolia Baill. seed extract in rats.

The seeds of Griffonia simplicifolia Baill., a tropical shrub native to West Africa, are rich in 5-hydroxy-l-tryptophan (5-HTP), a direct precursor in the synthesis of serotonin (5-HT). In spite of the modern therapeutic application of Griffonia simplicifolia seed extract in mood disorders, no scientific evidence has been provided till now. For this reason the aim of our study was to investigate the effect of Griffonia simplicifolia seed extract on anxiety behavior. Griffonia simplicifolia seed extract, dosed at 1, 5, 10 and 25 mg/kg, was orally administered in rats which were submitted to the dark-light test and open field test, 60 min after the treatment. In the dark-light test, the administration of the extract at the doses of 10 and 25 mg/kg was able to significantly increase the time spent in the light compartment (P<0.05). In the open field test, the extract dosed at 5, 10 and 25 mg/kg induced an anti-tigmotactic effect, as indicated by a significant increase of time spent in the central area of the open field (P<0.01). In conclusion these findings indicate that Griffonia simplicifolia seed extract exerts anxiolytic-like effect in rats and suggest its potential usefulness for the treatment of anxiety in humans.

Regulation of CD1, Ki-67, PCNA mRNA expression, and Akt activation in estrogen-responsive human breast adenocarcinoma cell line, MCF-7 cells, by griffonianone C, an isoflavone derived from Millettia griffoniana.

CONTEXT: Millettia griffoniana Baill. (Fabaceae), which contains isoflavonoids like griffonianone C (Griff C), is commonly used in the folk medicine in Cameroon to treat various ailments. Possible health benefits of Griff C which include alleviation of menopausal symptoms, limitation of bone resorption, and lowering of the risks of cancer and cardiovascular diseases attracted our interest. OBJECTIVE: The effects of Griff C on the regulation of the expression of proliferation markers such as proliferating cell nuclear antigen (PCNA), cyclin D1 (CD1) and Ki-67 are investigated here. Its role in apoptosis or cell survival, through the phosphatidylinositol 3 kinase-Akt (PI3K-Akt) signaling pathway is further studied. MATERIALS AND METHODS: Semiquantitative real-time PCR was performed to analyze the effects of Griff C on gene expression in MCF-7 cells. Western blot analysis was used to assess the role of Griff C on the expression of phosphorylated Akt in MCF-7 cells. RESULTS: Griff C induced a 4.84-fold increase in the expression of Ki-67 mRNA at the concentration of 10(-8) M and a 3.90-fold increase of CD1 mRNA at 10(-7) M. Griff C slightly increased the phosphorylation of Akt at its serine 473 residue. Akt phosphorylation was inhibited by the PI3K inhibitor, LY294002, but not by the specific estrogen receptor antagonist, fulvestrant. DISCUSSION AND CONCLUSION: These findings suggest that Griff C can modulate proliferation of MCF-7 cells. Our results also suggest that Griff C can affect the PI3K-related signaling pathway. Thus, Griff C may exert part of its low proliferative and antiapoptotic effects by a nongenomic mode of action.

Isolectin B4binding in populations of rat trigeminal ganglion cells.

We have recently classified dissociated trigeminal ganglion cells into nine types using electrophysiological current signatures. In the present study, we investigated the relationship between isolectin B(4) (IB(4)) binding and the cell types in rat trigeminal ganglion cells. We found that IB(4) was bound to all type 2 cells and more than 70% of cell types 1 and 13; however, it was bound to less than 20% of cell types 7 and 8 and did not bind at all to cell types 3-5 and 9. Thus, each trigeminal ganglion cell type showed high homogeneity in IB(4) binding. These results correspond to reported IB(4) binding profiles in the matched dorsal root ganglion cell types, except for types 5 and 7.

Construction of N-glycan microarrays by using modular synthesis and on-chip nanoscale enzymatic glycosylation.

An effective chemoenzymatic strategy is reported that has allowed the construction, for the first time, of a focused microarray of synthetic N-glycans. Based on modular approaches, a variety of N-glycan core structures have been chemically synthesized and covalently immobilized on a glass surface. The printed structures were then enzymatically diversified by the action of three different glycosyltransferases in nanodroplets placed on top of individual spots of the microarray by a printing robot. Conversion was followed by lectin binding specific for the terminal sugars. This enzymatic extension of surface-bound ligands in nanodroplets reduces the amount of precious glycosyltransferases needed by seven orders of magnitude relative to reactions carried out in the solution phase. Moreover, only those ligands that have been shown to be substrates to a specific glycosyltransferase can be individually chosen for elongation on the array. The methodology described here, combining focused modular synthesis and nanoscale on-chip enzymatic elongation, could open the way for the much needed rapid construction of large synthetic glycan arrays.

Griffonia simplicifolia negatively affects sexual behavior in female rats.

At present Griffonia simplicifolia is used in food supplement aimed to treat mood disorders as well as to reduce food intake and body weight. The plant has gained increasing interest for its high content in 5-hydroxy-L-tryptophan (5-HTP) particularly in the seed. The present study was designed to evaluate the influence of a seed extract of the plant, dosed at 25, 50 and 100 mg/kg, on the sexual behavior of ovariectomized hormone-primed rats after acute and subchronic treatment. The single administration of G. simplicifolia significantly reduced lordosis response and increased rejection behavior in female rats treated with the highest dose while it did not influence proceptive behaviors. On the other hand the subchronic administration of the extract significantly reduced proceptivity but not receptivity, and increased rejection behavior. All the tested dosages were able to markedly decrease food intake and body weight after a 9-day treatment. Taken together the present results, possibly ascribed to increased levels of 5-hydroxytryptamine (5-HT) in the brain, suggest a cautious administration of the plant extract owing to its negative influence on female sexual behavior.

O-glycosylation expression in fibroadenoma.

Fibroadenomas are human benign breast tumors characterized by proliferation of epithelial and stroma cells of the terminal ductal unit. Expression of O-glycans seems to contribute to the proliferation and transformation events. With this in mind, we evaluated the expression of glycans in fibroadenoma tissue through immunohistochemistry with antibodies against mucin epitopes (Anti CA15-3 and MUC1), as well as with lectins specific for glycans linked to proteins or lipids, and we compared findings with healthy breast specimens. Our results show positive expression of CA15-3 and MUC1 in fibroadenoma tissue, mainly in duct and stroma cells, whereas, in normal samples, staining was observed in duct cells. The lectin from Glycine max recognized equally well duct and stroma cells; this was the only lectin showing co-localization with anti-CA15-3 in healthy and tumor tissues. Dolichos biflorus, Artocarpus integrifolia, and Griffonia simplicifolia lectins recognized duct cells in control healthy tissues as well as in fibroadenoma tissue. The lectin from Amaranthus leucocarpus recognized only duct cells in control samples, whereas, in fibroadenoma tissue, it recognized duct and some stromal cells, suggesting that O-glycans-type mucin linked to proteins and mucin participate in the development of fibroadenomas.

Semaphorin3a regulates endothelial cell number and podocyte differentiation during glomerular development.

Semaphorin3a (Sema3a), a chemorepellant guidance protein, plays crucial roles in neural, cardiac and peripheral vascular patterning. Sema3a is expressed in the developing nephron, mature podocytes and collecting tubules. Sema3a acts as a negative regulator of ureteric bud branching, but its function in glomerular development has not been examined. Here we tested the hypothesis that Sema3a regulates glomerular vascular development using loss- and gain-of-function mouse models. Sema3a deletion resulted in defects in renal vascular patterning, excess endothelial cells within glomerular capillaries, effaced podocytes with extremely wide foot processes and albuminuria. Podocyte Sema3a overexpression during organogenesis resulted in glomerular hypoplasia, characterized by glomerular endothelial cell apoptosis, delayed and abnormal podocyte foot process development, a complete absence of slit diaphragms and congenital proteinuria. Nephrin, WT1 and VEGFR2 were downregulated in Sema3a-overexpressing kidneys. We conclude that Sema3a is an essential negative regulator of endothelial cell survival in developing glomeruli and plays a crucial role in podocyte differentiation in vivo. Hence, a tight regulation of Sema3a dosage is required for the establishment of a normal glomerular filtration barrier.