Journal: Nature

Article Title: Morphodynamics of human early brain organoid development

doi: 10.1038/s41586-025-09151-3

Figure Lengend Snippet: a ) Photographs of the sample holding chamber with four separated sub-chambers each containing four microwells, for growing and imaging organoids with sixteen different organoids arranged one per microwell n = 16 organoids. b ) Schematic overview of tiled image acquisition and example images showing two timepoints, before and after tiled acquisition and image fusion. Scale Bar is 100 micrometers. Time is in hours. n = 16 organoids. c ) Cross section images (day 7) from 16 simultaneously imaged organoids, generated with cells lines labeled with nuclear membrane (lamin, RFP, magenta), plasma membrane (CAAX, RFP, magenta), actin (GFP, green), tubulin (RFP, magenta), and nuclei (histone, GFP, green) and unlabeled WTC-11. d ) Images showing cross section (z-plane) and orthogonal views (y-plane, x-plane) of one organoid from a timecourse lightsheet imaging experiment shown in c. Scale Bar is 100 micrometers. Time is in hours. e ) Maximum intensity projections from a 2-week imaging acquisition of a mosaic organoid (histone, GFP, green; CAAX, RFP, magenta), unlabeled WTC-11). Scale Bar is 500 micrometers. Time is in hours. n = 16 organoids. f ) Images from a 3-week continuous imaging experiment using a NKX2-1:GFP reporter line. The organoids were given SHH morphogen treatment to induce ventral telencephalic patterning of the organoids. Images are false-colored with the green-fire-blue LUT. Scale Bar is 100 micrometers. Time is in hours. n = 4 organoids.

Article Snippet: The organoids were treated with the average morphogen concentration of SHH (140 ng/ml, Miltenyi, 130-095-727) from day 3–14 together with purmorphamine (0.21 μM, Miltenyi, 130-104-465).

Techniques: Imaging, Generated, Labeling, Membrane, Clinical Proteomics

Journal: microPublication Biology

Article Title: Bone morphogenetic protein 8B (BMP8B) increases the glucose sensitivity of ventromedial hypothalamus (VMH) glucose-inhibited (GI) neurons in female mice.

doi: 10.17912/micropub.biology.001496

Figure Lengend Snippet: A. Representative whole cell current clamps traces from a glucose inhibited (GI) neuron in the ventrolateral ventromedial hypothalamus (vlVMH) in response to a glucose decrease from 2.5 to 0.1 mM glucose in the presence and absence of 1 nM bone morphogenetic protein (BMP) 8B. Top trace: This neuron depolarized and increased resistance in 0.1 mM glucose but returned to baseline before glucose was returned to 2.5 mM indicating that it belongs to the subcategory of adapting (Ad) GI neurons. Bottom trace: This depolarization and increased resistance was blocked in the presence of BMP8B. B. Representative voltage responses to a constant -20 pA hyperpolarizing current pulse in 2.5- and 0.1-mM glucose in the absence (top) and presence (bottom) of BMP8B. In the absence of BMP8B the voltage response increased in 0.1 mM glucose; however, BMP8B blocked this response. C. Bar graph of the percentage change in resistance as glucose was lowered from 2.5 to 0.1 mM glucose in the absence (con) and presence (BMP) of BMP8B. BMP8B significantly decreased the percentage change in resistance. Paired t-test. *p = 0.01. White square: AdGI neurons; black dot: sustained (s) GI neurons. D. Bar graph of the percentage of nNOS expressing cells that also express the BMP receptor BMPR1a in males and females. E. Immunohistochemical images of the VMH from males (left) and females (right) showing colocalization of neuronal nitric oxide synthase (nNOS, green) and BMPR1a (red). Dapi nuclear stain in blue. White arrows indicate cells expressing both nNOS and BMPR1a. F. Hypothetical model for the effect of BMP8B on VMH GI neurons. Glucose inhibits VMH nNOS-GI neurons by inhibiting AMPK. BMP8B also exerts it's metabolic effects by inhibiting VMH AMPK. Thus, BMP8B may regulate blood glucose in females due to increased expression of BMPR1a on nNOS expressing VMH GI neurons compared to males. * p<0.01, independent students t-test.

Article Snippet: Materials: Bone Morphogenic Protein 8B (BMP8B) human (Millipore Sigma, SRP 6322, Burlington, MA), Antibodies: primary rabbit anti-NOS1 (1:200, Cell Signaling Technology C7D7,Danvers, MA), secondary Alexa FluorTM 488 (1:500, Donkey anti-Rabbit; A21206, Invitrogen, Waltham, MA); primary mouse Anti-BMPR1A (1:250, Santa Cruz Biotechnology, sc293175, Dallas, TX), secondary Alexa FluorTM 594 (1:500, Donkey anti -Mouse; A21203, Invitrogen, Waltham, MA) Cell Counting: Images were obtained using the Nikon A1R Confocal Laser Microscope System.

Techniques: Expressing, Immunohistochemical staining, Staining