Three‐dimensional microscopy and image fusion reconstruction analysis of the thyroid gland during morphogenesis

Abstract

Rui-jia Zhang, Liu Yang and Feng Sun are co-first authors. Huai-dong Song and Mei Dong are cocorresponding authors.

Disruption of the foxe1 gene in zebrafish reveals conserved functions in development of the craniofacial skeleton and the thyroid

“…The zebrafish thyroid is not organized as a discrete gland, but as individual follicles scattered along the ventral aorta ( Alt et al, 2006 ). These follicles appear from 72 to 96 hpf and can be visualized using thyroid hormone (T4) antibody staining ( Zhang et al, 2021 ). Figure 3 shows two sequential sections of the same individual stained with Foxe1-and T4-antibodies.…”

“…(C) Representative images of wild type and mutant thyroid follicles. Larvae were imaged from the ventral side and T4-positive follicle surfaces were rendered from z-stack images using Imaris 9.0 as previously described ( Zhang et al, 2021 ). Scale bar 200 µm.…”

“…Whole mount immunohistochemistry was performed exactly as described previously ( Hammond-Weinberger and ZeRuth 2020 ) using T4 antibody (MP Biomedicals rabbit anti-T4) diluted at 1:1,000 for whole embryos and Foxe1 Boster Bio (DZ41149) at 1:500 on scales. Z-stack images of the T4 staining were taken on an SPX8 confocal microscope at magnification ×20 and were processed by surface rendering using Imaris 9.0 software to calculate volumes as previously described ( Zhang et al, 2021 ).…”

“…The zebrafish thyroid is not organized as a discrete gland, but as individual follicles scattered along the ventral aorta (Alt et al, 2006). These follicles appear from 72 to 96 hpf and can be visualized using thyroid hormone (T4) antibody staining (Zhang et al, 2021). Figure 3 shows two sequential sections of (J-Q) Detailed protein localization at 96 hpf showed Foxe1-positive cells in the brain (J), the ceratobranchials (K), in the oral epithelium and in a linear cluster in the subpharyngeal area (white arrow) (L), the eyes (M), on the ethmoid plate (N), on the lining of the ceratohyal (O), in the mouth opening (P) and to some extent in the fin tips (Q), upper image) and in the notochordal sheath ((Q), lower image).…”