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RFGF21 induces neurite outgrowth in GnRH neurons in vitro, stimulates GnRH secretion ex vivo, and reaches GnRH neurons by fenestrated vessels in vivo rFGF21.

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Presentation on theme: "RFGF21 induces neurite outgrowth in GnRH neurons in vitro, stimulates GnRH secretion ex vivo, and reaches GnRH neurons by fenestrated vessels in vivo rFGF21."— Presentation transcript:

1 rFGF21 induces neurite outgrowth in GnRH neurons in vitro, stimulates GnRH secretion ex vivo, and reaches GnRH neurons by fenestrated vessels in vivo rFGF21 induces neurite outgrowth in GnRH neurons in vitro, stimulates GnRH secretion ex vivo, and reaches GnRH neurons by fenestrated vessels in vivo ARepresentative images of GT1‐7 immortalized GnRH neurite traces after 48 h of culture in control medium (SFM) alone or with 100 nM rFGF21 (n = 4 per group). Scale bar 30 μm.BAverage of the longest neurite length of GT1‐7 cells in aforementioned conditions, unpaired t‐test (n = 4 per group).CNeurite length distribution histogram showing significant increase of cells with longer neurites (> 80, 60–80 μm) and reduced number with shorter neurites (20–40 μm) after rFGF21 treatment, unpaired t‐test (n = 4 per group).DIsolation protocol of GnRH neurons from GnRH::GFP mice by fluorescence‐activated cell sorting (FACS) and Klb mRNA expression quantification in GnRH‐positive (POS) and GnRH‐negative (NEG) hypothalamic FACS‐isolated cells (n = 3 per group), unpaired t‐test. POA, preoptic area of the hypothalamus.EGnRH peptide content in conditioned medium from ME explant cultures treated with increasing doses of rFGF21 (n = 5 per group) for 20 min. Data were analyzed by two‐way ANOVA followed by Sidak's multiple comparisons test.FRepresentative photomicrographs of the preoptic region showing GnRH neurons (GFP, green) and blood vessels from the BBB (empty arrowhead) and the OVLT (arrowhead) labeled by fluorescent rFGF21 (5 nmol/animal, white staining). Note the presence of white staining in GFP‐expressing neuronal processes, dendrites, and cell body (arrows, inset 1′ and 2′). Nuclear contrast staining: DAPI; 3V, third ventricle; OVLT, organum vasculosum of the lamina terminalis; oc, optic chiasma. Scale bar: 150 μm for inset 1′, 40 μm for inset 2′, and 300 μm for other panels.GRepresentative photomicrographs showing blood vessels from BBB (empty arrowhead) and in ME (arrowhead) labeled by fluorescent rFGF21 (5 nmol/animal, white staining) in the tuberal region of the hypothalamus. Note that fluorescent rFGF21 administered intravenously diffuses into ME tissue (white staining, asterisk) where GnRH neuroendocrine axons are distributed (green, arrows). Nuclear contrast staining: DAPI; ME, median eminence; ARH, arcuate nucleus of the hypothalamus. Scale bar: 50 μm for inset 1′ and 130 μm for other panels.Data information: Values shown are mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001. Cheng Xu et al. EMBO Mol Med. 2017;9: © as stated in the article, figure or figure legend

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