Qazi Waheed-Ullah, Anna Wilsdon, Aseel Abbad, Sophie Rochette, Frances Bu'Lock, Marc-Phillip Hitz, Gregor Dombrowsky, Friederike Cuello, J. David Brook, Siobhan Loughna
{"title":"缺失蛋白激酶 PRKD1 对小鼠胚胎心脏发育的影响","authors":"Qazi Waheed-Ullah, Anna Wilsdon, Aseel Abbad, Sophie Rochette, Frances Bu'Lock, Marc-Phillip Hitz, Gregor Dombrowsky, Friederike Cuello, J. David Brook, Siobhan Loughna","doi":"10.1111/joa.14033","DOIUrl":null,"url":null,"abstract":"<p>Congenital heart disease (CHD) is the most common congenital anomaly, with an overall incidence of approximately 1% in the United Kingdom. Exome sequencing in large CHD cohorts has been performed to provide insights into the genetic aetiology of CHD. This includes a study of 1891 probands by our group in collaboration with others, which identified three novel genes—<i>CDK13, PRKD1</i>, and <i>CHD4</i>, in patients with syndromic CHD. <i>PRKD1</i> encodes a serine/threonine protein kinase, which is important in a variety of fundamental cellular functions. Individuals with a heterozygous mutation in <i>PRKD1</i> may have facial dysmorphism, ectodermal dysplasia and may have CHDs such as pulmonary stenosis, atrioventricular septal defects, coarctation of the aorta and bicuspid aortic valve. To obtain a greater appreciation for the role that this essential protein kinase plays in cardiogenesis and CHD, we have analysed a <i>Prkd1</i> transgenic mouse model (<i>Prkd1</i><sup><i>em1</i></sup>) carrying deletion of exon 2, causing loss of function. High-resolution episcopic microscopy affords detailed morphological 3D analysis of the developing heart and provides evidence for an essential role of <i>Prkd1</i> in both normal cardiac development and CHD. We show that homozygous deletion of <i>Prkd1</i> is associated with complex forms of CHD such as atrioventricular septal defects, and bicuspid aortic and pulmonary valves, and is lethal. Even in heterozygotes, cardiac differences occur. However, given that 97% of <i>Prkd1</i> heterozygous mice display normal heart development, it is likely that one normal allele is sufficient, with the defects seen most likely to represent sporadic events. Moreover, mRNA and protein expression levels were investigated by RT-qPCR and western immunoblotting, respectively. A significant reduction in <i>Prkd1</i> mRNA levels was seen in homozygotes, but not heterozygotes, compared to WT littermates. While a trend towards lower PRKD1 protein expression was seen in the heterozygotes, the difference was only significant in the homozygotes. There was no compensation by the related <i>Prkd2</i> and <i>Prkd3</i> at transcript level, as evidenced by RT-qPCR. Overall, we demonstrate a vital role of <i>Prkd1</i> in heart development and the aetiology of CHD.</p>","PeriodicalId":14971,"journal":{"name":"Journal of Anatomy","volume":null,"pages":null},"PeriodicalIF":1.8000,"publicationDate":"2024-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/joa.14033","citationCount":"0","resultStr":"{\"title\":\"Effect of deletion of the protein kinase PRKD1 on development of the mouse embryonic heart\",\"authors\":\"Qazi Waheed-Ullah, Anna Wilsdon, Aseel Abbad, Sophie Rochette, Frances Bu'Lock, Marc-Phillip Hitz, Gregor Dombrowsky, Friederike Cuello, J. David Brook, Siobhan Loughna\",\"doi\":\"10.1111/joa.14033\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Congenital heart disease (CHD) is the most common congenital anomaly, with an overall incidence of approximately 1% in the United Kingdom. Exome sequencing in large CHD cohorts has been performed to provide insights into the genetic aetiology of CHD. This includes a study of 1891 probands by our group in collaboration with others, which identified three novel genes—<i>CDK13, PRKD1</i>, and <i>CHD4</i>, in patients with syndromic CHD. <i>PRKD1</i> encodes a serine/threonine protein kinase, which is important in a variety of fundamental cellular functions. Individuals with a heterozygous mutation in <i>PRKD1</i> may have facial dysmorphism, ectodermal dysplasia and may have CHDs such as pulmonary stenosis, atrioventricular septal defects, coarctation of the aorta and bicuspid aortic valve. To obtain a greater appreciation for the role that this essential protein kinase plays in cardiogenesis and CHD, we have analysed a <i>Prkd1</i> transgenic mouse model (<i>Prkd1</i><sup><i>em1</i></sup>) carrying deletion of exon 2, causing loss of function. High-resolution episcopic microscopy affords detailed morphological 3D analysis of the developing heart and provides evidence for an essential role of <i>Prkd1</i> in both normal cardiac development and CHD. We show that homozygous deletion of <i>Prkd1</i> is associated with complex forms of CHD such as atrioventricular septal defects, and bicuspid aortic and pulmonary valves, and is lethal. Even in heterozygotes, cardiac differences occur. However, given that 97% of <i>Prkd1</i> heterozygous mice display normal heart development, it is likely that one normal allele is sufficient, with the defects seen most likely to represent sporadic events. Moreover, mRNA and protein expression levels were investigated by RT-qPCR and western immunoblotting, respectively. A significant reduction in <i>Prkd1</i> mRNA levels was seen in homozygotes, but not heterozygotes, compared to WT littermates. While a trend towards lower PRKD1 protein expression was seen in the heterozygotes, the difference was only significant in the homozygotes. There was no compensation by the related <i>Prkd2</i> and <i>Prkd3</i> at transcript level, as evidenced by RT-qPCR. Overall, we demonstrate a vital role of <i>Prkd1</i> in heart development and the aetiology of CHD.</p>\",\"PeriodicalId\":14971,\"journal\":{\"name\":\"Journal of Anatomy\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.8000,\"publicationDate\":\"2024-02-28\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1111/joa.14033\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Anatomy\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1111/joa.14033\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"ANATOMY & MORPHOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Anatomy","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/joa.14033","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"ANATOMY & MORPHOLOGY","Score":null,"Total":0}
Effect of deletion of the protein kinase PRKD1 on development of the mouse embryonic heart
Congenital heart disease (CHD) is the most common congenital anomaly, with an overall incidence of approximately 1% in the United Kingdom. Exome sequencing in large CHD cohorts has been performed to provide insights into the genetic aetiology of CHD. This includes a study of 1891 probands by our group in collaboration with others, which identified three novel genes—CDK13, PRKD1, and CHD4, in patients with syndromic CHD. PRKD1 encodes a serine/threonine protein kinase, which is important in a variety of fundamental cellular functions. Individuals with a heterozygous mutation in PRKD1 may have facial dysmorphism, ectodermal dysplasia and may have CHDs such as pulmonary stenosis, atrioventricular septal defects, coarctation of the aorta and bicuspid aortic valve. To obtain a greater appreciation for the role that this essential protein kinase plays in cardiogenesis and CHD, we have analysed a Prkd1 transgenic mouse model (Prkd1em1) carrying deletion of exon 2, causing loss of function. High-resolution episcopic microscopy affords detailed morphological 3D analysis of the developing heart and provides evidence for an essential role of Prkd1 in both normal cardiac development and CHD. We show that homozygous deletion of Prkd1 is associated with complex forms of CHD such as atrioventricular septal defects, and bicuspid aortic and pulmonary valves, and is lethal. Even in heterozygotes, cardiac differences occur. However, given that 97% of Prkd1 heterozygous mice display normal heart development, it is likely that one normal allele is sufficient, with the defects seen most likely to represent sporadic events. Moreover, mRNA and protein expression levels were investigated by RT-qPCR and western immunoblotting, respectively. A significant reduction in Prkd1 mRNA levels was seen in homozygotes, but not heterozygotes, compared to WT littermates. While a trend towards lower PRKD1 protein expression was seen in the heterozygotes, the difference was only significant in the homozygotes. There was no compensation by the related Prkd2 and Prkd3 at transcript level, as evidenced by RT-qPCR. Overall, we demonstrate a vital role of Prkd1 in heart development and the aetiology of CHD.
期刊介绍:
Journal of Anatomy is an international peer-reviewed journal sponsored by the Anatomical Society. The journal publishes original papers, invited review articles and book reviews. Its main focus is to understand anatomy through an analysis of structure, function, development and evolution. Priority will be given to studies of that clearly articulate their relevance to the anatomical community. Focal areas include: experimental studies, contributions based on molecular and cell biology and on the application of modern imaging techniques and papers with novel methods or synthetic perspective on an anatomical system.
Studies that are essentially descriptive anatomy are appropriate only if they communicate clearly a broader functional or evolutionary significance. You must clearly state the broader implications of your work in the abstract.
We particularly welcome submissions in the following areas:
Cell biology and tissue architecture
Comparative functional morphology
Developmental biology
Evolutionary developmental biology
Evolutionary morphology
Functional human anatomy
Integrative vertebrate paleontology
Methodological innovations in anatomical research
Musculoskeletal system
Neuroanatomy and neurodegeneration
Significant advances in anatomical education.