Abstract
We aimed to investigate the spatio-temporal expression of possible CAKUT candidate genes CRKL, AIFM3, and UBASH3A, as well as AIF and BCL2 during human kidney development. Human fetal kidney tissue was stained with antibodies and analyzed by fluorescence microscopy and RT-PCR. Quantification of positive cells was assessed by calculation of area percentage and counting cells in nephron structures. Results showed statistically significant differences in the temporal expression patterns of the examined markers, depending on the investigated developmental stage. Limited but strong expression of CRKL was seen in developing kidneys, with increasing expression up to the period where the majority of nephrons are formed. Results also lead us to conclude that AIFM3 and AIF are important for promoting cell survival, but only AIFM3 is considered a CAKUT candidate gene due to the lack of AIF in nephron developmental structures. Our findings imply great importance of AIFM3 in energy production in nephrogenesis and tubular maturation. UBASH3A raw scores showed greater immunoreactivity in developing structures than mature ones which would point to a meaningful role in nephrogenesis. The fact that mRNA and proteins of CRKL, UBASH3A, and AIFM3 were detected in all phases of kidney development implies their role as renal development control genes.
Highlights
The permanent human kidney or metanephros appears around the 5th developmental week through the reciprocal interactions between the metanephric mesenchyme (MM)and the ureteric bud (UB)
We focus on CRKL, AIFM3, and Ubiquitin associated and SH3 domain containing A (UBASH3A) as possible candidate genes in order to characterize their normal expression pattern during the four phases of kidney development
The expression of CRKL, AIFM3, apoptosis inducing factor (AIF), and UBASH3A was analyzed on fully differentiated glomeruli (G), proximal convoluted tubules (PCTs), distal convoluted tubules (DCTs), immature glomeruli (IG), S-shaped bodies (SSBs), comma-shaped bodies (CSBs), and renal vesicles (RVs) of fetal and postnatal kidney samples by fluorescence microscopy
Summary
The permanent human kidney or metanephros appears around the 5th developmental week through the reciprocal interactions between the metanephric mesenchyme (MM)and the ureteric bud (UB). The permanent human kidney or metanephros appears around the 5th developmental week through the reciprocal interactions between the metanephric mesenchyme (MM). Under the inductive effect of the terminal branches of the UB (ampullae), the MM forms renal vesicles, which gradually differentiate into commaand S-shaped bodies, and later on into more mature nephrons with Bowman’s capsules, glomeruli and nephron tubules that connect to the collecting system. Kidney development can be divided according to the demeanor of the ampullae into four separate phases, in which they variably divide and induce nephrons. During the first phase (Ph1), which takes place from week 5 to around the 14th developmental week, each ampulla induces a single nephron. In phase two (Ph2) (15th to 20th–22nd developmental weeks), the ampullae divide sporadically but one ampulla is able to induce several nephrons which connect with collecting ducts into arcades.
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