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. 2019 Mar 28;7:35. doi: 10.3389/fcell.2019.00035

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Copyright © 2019 Huss, Saias, Hamamah, Singh, Wang, Dave, Kim, Eberwine and Lansford.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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PGCs and their surrounding cells in the germinal crescent express integrin beta 1 subunit protein on their cell surfaces. Whole-mount immunofluorescence for fibronectin and laminin receptors (integrin beta 1 subunit, abJG-22) and PGC-specific Dazl at two separate locations within an HH4 [Tg(hUbC:Membrane-eGFP)] quail embryo germinal crescent. (A) Central germinal crescent (see panel C for location). Left to right: Integrin beta 1 subunit (Itgb1, abJG-22, red), Dazl (PGC marker, green), membrane-GFP (blue), Itgb1 and membrane-GFP merge, Itgb1, membrane-GFP, Dazl merge (bounding box shown at higher magnification in E). White arrows indicate one PGC imaged in cross section with a clearly defined perimeter of Itgb1. 20x 0.8x zoom. (B) Lateral germinal crescent (see panel C for location). Left to right: Integrin beta 1 subunit (Itgb1, abJG-22, red), Dazl (PGC marker, green), membrane-GFP (blue), Itgb1 and membrane-GFP merge, Itgb1, membrane-GFP, DAZL merge (bounding box shown at higher magnification in D). White arrows indicate one PGC with a clearly defined perimeter of Itgb1 that was imaged at an oblique angle. 20x 1.0x zoom. (C) Tiled 10x confocal Z stack maximum intensity projections of the whole-mount HH4 embryo. Bounding boxes indicate location of images in (A,B). Itgb1 (red), Dazl (green). Scale bar C = 500μm. (D,E) Images are digital zooms of bounding boxes in (B,A) Merge. White arrows in (D,E) highlight particular PGCs with integrin beta 1 subunit receptor labeled puncta on the cell surface which was imaged in oblique (D) or in cross-section (E) by the confocal optical slice. Scale bar A, B, D, E = 25 μm.