Expression of zaqp8a.1 in zebrafish embryos was analyzed by whole mount in situ hybridization. Lateral view with anterior to the left (c-j). (a, b) The expression of zaqp8a.1 was not detected at the one cell stage or at 3 ss. For selection of 3 ss embryos precisely, embryos were co-stained with myod1 (b). (c-d) The expression of zaqp8a.1 was first detected in the posterior region of LPM at 12 ss and then anteriorly expanded at 18 ss (arrowhead). (e-j) The expression of zaqp8a.1 was detected in blood vessels. (e) In DA (arrowhead) and ISV (arrow), expression of zaqp8a.1 was seen at 24 hpf, but it greatly decreased in DA and ISV at 72 hpf (h) and 4 dpf (i), respectively. (f) Expression of zaqp8a.1 in DLAV (asterisks) was firstly seen from 36 hpf, and persisted until 7 dpf (j).

Spatiotemporal expression pattern of zaqp8a.1 in the vascular system and gastrointestinal tract. Lateral view with anterior to the left (a, c, e); ventral view with anterior to the left (b). (a) Expression of zaqp8a.1 in DA (arrowhead), ISV (asterisk), and DLAV (arrow). The zaqp8a.1 transcripts in DA and ISV gradually reduced at the anterior region of the trunk. (b) Expression of zaqp8a.1 in the hypobranchial artery (arrowhead). (c) The zaqp8a.1 transcripts were detected in the pectoral artery at 72 hpf (arrow). (d) The expression of zaqp8a.1 in the embryonic heart tube from 28 hpf Tg(cmlc2:EGFP) transgenic zebrafish embryos was detected by whole mount in situ hybridization and immunofluorescence staining. (e) The zaqp8a.1 is expressed in the heart (arrow head) and gastrointestinal tract, such as liver (L) and intestinal tract (I) at 4 to 7 dpf.

Spatiotemporal expression pattern of zaqp8a.2 at an early developmental stage. Lateral view with anterior to the left (d, g, i, and k); dorsal view with anterior to the left (e and f); ventral view with anterior to the left (h, j and l). (a-d) Expression of zaqp8a.2 was not detectable from one cell to 48 hpf. (e-l) Between 72 hpf and 7 dpf, zaqp8a.2 was only expressed in the intestinal tract (arrowhead).

Pronephros-specific expression pattern of zaqp8b was analyzed by whole mount in situ hybridization. Dorsal view to the left (b-l). Staining without myod1 RNA probe (a, d-l) or co-staining with myod1 probe (b, c). (b) In 9 ss embryos, zaqp8b began to express in a subset of cells in IM (arrowhead). (c-g) Bilateral expression of zaqp8b in pronephros (arrowhead). (h-k) Expression of zaqp8b in the PCT (arrow head) from 72 hpf to 6 dpf. (l) The zaqp8b transcripts in the PCT almost completely disappeared at 7 dpf.