Identification of porcine polycystic kidney disease 1 (PKD1) gene: Molecular cloning, expression profile, and implication in disease model

He, Jin; Wang, Qingsong; Ye, Jianhua; Hu, Xiaoxiang; Li, Ning

doi:10.1016/j.gene.2011.08.027

Cited by 8 publications

(8 citation statements)

References 88 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As in mouse models that disrupted either PKD1 or PKD2 , prenatal and perinatal deaths were common phenomena, which demonstrates that both genes are required for normal placental development 24 . Because PKD1 is ubiquitously expressed in various organs 18 , qRT-PCR was performed using RNA from ear biopsies of these piglets. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Membrane protein was extracted from the kidneys using the Membrane and Cytosol Protein Extraction Kit (Beyotime, Nantong, China). Eighty micrograms of membrane protein per lane was separated, transferred, and blotted according to a previously established protocol 18 . The antibodies used to detect PC1 were 7e12 (gift from Dr. Christopher Ward) with 1:500 dilution.…”

Section: Methodsmentioning

confidence: 99%

“…In previous studies, we cloned and characterized the pig PKD1 and PKD2 genes and demonstrated a remarkable similarity between the human and pig orthologs at molecular level 18 , 19 . Furthermore, a porcine PKD2 transgenic model was generated, but there were no cystic manifestations by one year of age in that model 20 .…”

Section: Introductionmentioning

confidence: 95%

See 2 more Smart Citations

PKD1 Mono-Allelic Knockout Is Sufficient to Trigger Renal Cystogenesis in a Mini-Pig Model

He¹,

Li²,

Fang³

et al. 2015

Int. J. Biol. Sci.

Self Cite

View full text Add to dashboard Cite

PKD1 and PKD2 mutations could lead to autosomal dominant polycystic kidney disease (ADPKD), which afflicts millions of people worldwide. Due to the marked differences in the lifespan, size, anatomy, and physiology from humans, rodent ADPKD models cannot fully mimic the disease. To obtain a large animal model that recapitulates the disease, we constructed a mini-pig model by mono-allelic knockout (KO) of PKD1 using zinc finger nuclease. The mono-allelic KO pigs had lower PKD1 expression than their wild-type littermates at both the transcriptional and translational levels. After approximately six months, renal cysts appeared and grew progressively in the KO pigs. Histological analysis showed that renal cysts were scatteredly distributed in the mutant pig kidneys and were lined by either cuboidal or flattened epithelial cells. Contrast-enhanced computed tomography confirmed that all of the mutant pigs had renal and hepatic cysts, when they were 11-month-old. Immunohistochemical analysis revealed that most of the cysts were derived from the proximal tubules and collecting ducts. Therefore, the PKD1 mono-allelic knockout is sufficient to trigger renal cystogenesis, and this pig model may provide a platform for future study of renal cyst formation.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

PKD1 Mono-Allelic Knockout Is Sufficient to Trigger Renal Cystogenesis in a Mini-Pig Model

He¹,

Li²,

Fang³

et al. 2015

Int. J. Biol. Sci.

Self Cite

View full text Add to dashboard Cite

show abstract

“…The PCR reactions (35 cycles of 94 °C for 30 s, 60 °C for 30 s, and 72 °C for 30 s or 1 min) were then performed in 25 μl volume with 2.5 U Taq (Takara), 1 μl cDNA, 400 nM each primer, 200 mM MgCl 2 , and 200 μM dNTPs. Subsequently, to obtain a more accurate expression results, quantitative real-time PCR (qRT-PCR) was performed in triplicate for each sample from all the three pigs using a similar protocol to a previous report 27 . The qRT-PCR results were analysed using the 2 −ΔΔCt method 28 .…”

Section: Rna Preparation and Gene Cloningmentioning

confidence: 99%

Evolutionary and Molecular Characterization of liver-enriched gene 1

Dang

Wang

Liu

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

Liver-enriched gene 1 (Leg1) is a newly identified gene with little available functional information. To evolutionarily and molecularly characterize Leg1 genes, a phylogenetic study was first conducted, which indicated that Leg1 is a conserved gene that exists from bacteria to mammals. During the evolution of mammals, Leg1s underwent tandem duplications, which gave rise to Leg1a, Leg1b, and Leg1c clades. Analysis of the pig genome showed the presence of all three paralogs of pig Leg1 genes (pLeg1s), whereas only Leg1a could be found in the human (hLeg1a) or mouse (mLeg1a) genomes. purifying force acts on the evolution of Leg1 genes, likely subjecting them to functional constraint. Molecularly, pLeg1a and its coded protein, pig LEG1a (pLEG1a), displayed high similarities to its human and mouse homologs in terms of gene organization, expression patterns, and structures. Hence, pLeg1a, hLeg1a, and mLeg1a might preserve similar functions. Additionally, expression analysis of the three Leg1as suggested that eutherian Leg1as might have different functions from those of zebrafish and platypus due to subfunctionalization. Therefore, pLeg1a might provide essential information about eutherian Leg1a. Moreover, a preliminary functional study using RNA-seq suggested that pLeg1a is involved in the lipid homeostasis. In conclusion, our study provides some basic information on the aspects of evolution and molecular function, which could be applied for further validation of Leg1 using pig models. Leg1 (liver-enriched gene 1, or C6orf58 homolog) is a newly identified gene with very little available functional information 1,2. It is characterized by the presence of Domain of Unknown Function 781 (DUF781 or LEG1 domain) in its encoded protein 1,2. Leg1 was first identified in a zebrafish (Danio rerio) microarray study, in which it was named on the basis of its abundance in the liver 3. Functional experiments later demonstrated that Leg1 is involved in liver development, as knock-down of Leg1 in embryos results in small liver phenotype due to blocked liver expansion 1. Another functional study of the Leg1 gene was performed in the platypus (Ornithorhynchus anatinus), in which the human C6orf58 paralog MLP encodes monotreme lactation protein (MLP). MLP protein is a secreted protein that is enriched in milk, where it can exert antibacterial activity. Thus, it is presumed that MLP is related to the innate immunity of monotremes during the nipple-less delivery of milk to the hatchlings 2. Proteomic studies in eutherian species revealed that the Leg1-encoded N-glycosylated LEG1 protein is mainly present in saliva and seminal plasma 4,5. However, no further functional studies were carried out in eutherian animals. Expression profiling analyses of mouse (Mus musculus) and human (Homo sapiens) Leg1s (mLeg1 and hLeg1) reported in the Expression Atlas (www.ebi.ac.uk/gxa/home) showed that the gene is not expressed in the liver or mammary glands, in contrast to studies in monotremes and fishes, implying that eutherian Leg1s might have different ...

show abstract

“…When the ADPKD is concerned, people from different area may have nuances [28], for instance, Bunyong Phakdeekitcharoen studied the mutations of the replicated region of PKD1 in Asian population. So the codon usage is a reference for identification of porcine polycystic kidney disease [29,30].…”

Section: Figure3 Rscu Of the Pkd1 Coding Genementioning

confidence: 99%

Codon Usage of Autosomal Dominant Polycystic Kidney Disease Genes PKD1 and PKD2

2017

ARC Journal of Urology

View full text Add to dashboard Cite

Identification of porcine polycystic kidney disease 1 (PKD1) gene: Molecular cloning, expression profile, and implication in disease model

Cited by 8 publications

References 88 publications

PKD1 Mono-Allelic Knockout Is Sufficient to Trigger Renal Cystogenesis in a Mini-Pig Model

PKD1 Mono-Allelic Knockout Is Sufficient to Trigger Renal Cystogenesis in a Mini-Pig Model

Evolutionary and Molecular Characterization of liver-enriched gene 1

Codon Usage of Autosomal Dominant Polycystic Kidney Disease Genes PKD1 and PKD2

Contact Info

Product

Resources

About