SLC20A1Is Involved in Urinary Tract and Urorectal Development
SLC20A1Is Involved in Urinary Tract and Urorectal Development
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DOI：
10.3389/fcell.2020.00567
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发表时间：
2020-08-07
影响因子：
5.5
通讯作者：
Reutter, Heiko
中科院分区：
文献类型：
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作者：
Rieke, Johanna Magdalena;Zhang, Rong;Reutter, Heiko
先前对发育中的Xenopus和斑马鱼的研究报道磷酸盐转运蛋白slc 20 a1在前肾中表达。最近发现SLC 20 A1为泄殖腔外翻的单等位候选基因，进一步表明其参与泌尿道和泌尿直肠发育。然而，对SLC 20 A1在泌尿道发育中的功能作用知之甚少。在这里，我们研究了这一点，使用吗啉代寡核苷酸敲低斑马鱼直系同源物lc 20 a1 a。这导致肾囊肿和泄殖腔畸形。此外，在变形，我们表现出功能失调的排尿和后肠开放的缺陷模仿imperanus在人类泄殖腔外翻。此外，我们对未受影响的6周龄人类胚胎进行免疫组织化学检测，并在泌尿道和腹部中线检测到SLC 20 A1，这些结构与泄殖腔外翻的发病机制有关。此外，我们对690例膀胱外翻-尿道上裂综合征（BEEC）患者（包括84例泄殖腔外翻患者）的SLC 20 A1基因进行了重新测序。我们确定了两个额外的单等位基因novovovariants。一个是在一个典型的膀胱外翻的病例-父母三人组中发现的，另一个是在一个受影响的母亲中发现的，她将这种变异传给了她受影响的儿子。为了研究SLC 20 A1变体对细胞的潜在影响，我们在HEK 293细胞中表达了它们。在这里，磷酸盐转运没有受到影响，这表明它不是一种疾病机制。然而，有一个趋势，较低水平的切割半胱天冬酶-3，可能涉及在疾病的细胞凋亡途径。我们的研究结果表明，SLC 20 A1参与泌尿道和泌尿直肠的发育，并暗示SLC 20 A1是BEEC的致病基因。
Previous studies in developingXenopusand zebrafish reported that the phosphate transporterslc20a1ais expressed in pronephric kidneys. The recent identification ofSLC20A1as a monoallelic candidate gene for cloacal exstrophy further suggests its involvement in the urinary tract and urorectal development. However, little is known of the functional role ofSLC20A1in urinary tract development. Here, we investigated this using morpholino oligonucleotide knockdown of the zebrafish orthologslc20a1a. This caused kidney cysts and malformations of the cloaca. Moreover, in morphants we demonstrated dysfunctional voiding and hindgut opening defects mimicking imperforate anus in human cloacal exstrophy. Furthermore, we performed immunohistochemistry of an unaffected 6-week-old human embryo and detectedSLC20A1in the urinary tract and the abdominal midline, structures implicated in the pathogenesis of cloacal exstrophy. Additionally, we resequencedSLC20A1in 690 individuals with bladder exstrophy-epispadias complex (BEEC) including 84 individuals with cloacal exstrophy. We identified two additional monoallelicde novovariants. One was identified in a case-parent trio with classic bladder exstrophy, and one additional novelde novovariant was detected in an affected mother who transmitted this variant to her affected son. To study the potential cellular impact ofSLC20A1variants, we expressed them in HEK293 cells. Here, phosphate transport was not compromised, suggesting that it is not a disease mechanism. However, there was a tendency for lower levels of cleaved caspase-3, perhaps implicating apoptosis pathways in the disease. Our results suggestSLC20A1is involved in urinary tract and urorectal development and implicateSLC20A1as a disease-gene for BEEC.