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Mice lacking microRNAs in Pax8-expressing cells develop hypothyroidism and end-stage renal failure.

Bartram MP, Amendola E, Benzing T, Schermer B, de Vita G, Müller RU - BMC Mol. Biol. (2016)

Bottom Line: The Dgcr8/Drosha complex processes pri-miRNAs in the nucleus before they are exported as pre-miRNAs for further maturation by Dicer in the cytoplasm.Pax8Cre-mediated conditional loss of DiGeorge syndrome critical region 8 (Dgcr8), an essential component of the nuclear machinery that is required for microRNA biogenesis, resulted in severe hypothyroidism, massively reduced body weight and ultimately led to renal failure and death of the animals.These data provide further insight into the importance of miRNAs in organ homeostasis using a Dicer independent model.

View Article: PubMed Central - PubMed

Affiliation: Department II of Internal Medicine and Center for Molecular Medicine, University of Cologne, Kerpener Str. 62, 50937, Cologne, Germany.

ABSTRACT

Background: Non-coding RNAs have gained increasing attention during the last decade. The first large group of non-coding RNAs to be characterized systematically starting at the beginning of the 21st century were small oligonucleotides--the so-called microRNAs (miRNAs). By now we have learnt that microRNAs are indispensable for most biological processes including organogenesis and maintenance of organ structure and function. The role of microRNAs has been studied extensively in the development of a number of organs, so far most studies focussed on e.g. the heart or the brain whilst the role of microRNAs in the development and maintenance of complex epithelial organs is less well understood. Furthermore most analyses regarding microRNA function in epithelial organs employed conditional knockout mouse models of the RNAse III Dicer to abrogate microRNA biogenesis. However, there is increasing evidence for Dicer to have multiple functions independent from microRNA maturation. Therefore Dicer independent models are needed to gain further insight into the complex biology of miRNA dependent processes.

Results: Here we analyze the contribution of microRNA-dependent transcriptional control in Pax8-expressing epithelial cells. Pax8 is a transcription factor that is crucial to the development of epithelial organs. The miRNA machinery was disrupted by crossing conditional DiGeorge syndrome critical region 8 (Dgcr8) fl/fl mice to Pax8Cre mice. The Dgcr8/Drosha complex processes pri-miRNAs in the nucleus before they are exported as pre-miRNAs for further maturation by Dicer in the cytoplasm. Dgcr8 fl/fl; Pax8Cre+ knockout mice died prematurely, developed massive hypothyroidism and end stage renal disease due to a loss of miRNAs in Pax8 expressing tissue.

Conclusion: Pax8Cre-mediated conditional loss of DiGeorge syndrome critical region 8 (Dgcr8), an essential component of the nuclear machinery that is required for microRNA biogenesis, resulted in severe hypothyroidism, massively reduced body weight and ultimately led to renal failure and death of the animals. These data provide further insight into the importance of miRNAs in organ homeostasis using a Dicer independent model.

No MeSH data available.


Related in: MedlinePlus

Analysis of differentiation markers in Dgcr8–Pax8Cre knockout thyroid glands. Immunohistochemistry of thyroid differentiation markers show a strong reduction of Tg expression. Pax8, Nkx2.1 and Foxe1 expression levels are unaffected in the residual follicular cells, as well as the expression of their target Nis. No alteration in calcitonin cells was observed. (40× magnification for Tg, Pax8, calcitonin, Nkx2.1 and Foxe1 bar 50 μm; 100× magnification for Nis, bar 10 µm, age of the mice 4–8 weeks)
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Fig2: Analysis of differentiation markers in Dgcr8–Pax8Cre knockout thyroid glands. Immunohistochemistry of thyroid differentiation markers show a strong reduction of Tg expression. Pax8, Nkx2.1 and Foxe1 expression levels are unaffected in the residual follicular cells, as well as the expression of their target Nis. No alteration in calcitonin cells was observed. (40× magnification for Tg, Pax8, calcitonin, Nkx2.1 and Foxe1 bar 50 μm; 100× magnification for Nis, bar 10 µm, age of the mice 4–8 weeks)

Mentions: Molecular analysis of differentiation markers on knockout thyroid glands showed that the expression of the early differentiation markers Pax8, Nkx2.1 and Foxe1 is retained in the residual follicular cells, as well as the expression of their target Nis. Conversely, the expression of another late differentiation marker—Tg—resulted strongly reduced in the residual follicles. No alterations in calcitonin secreting cells were observed (Fig. 2). Thus, Dgcr8 knockout mice show functional defects partly similar to those from Dicer knockout mice [3, 4]. The main difference is that Nis is still expressed in Dgcr8 knockout mice, while being completely absent in Dicer knockout animals.Fig. 2


Mice lacking microRNAs in Pax8-expressing cells develop hypothyroidism and end-stage renal failure.

Bartram MP, Amendola E, Benzing T, Schermer B, de Vita G, Müller RU - BMC Mol. Biol. (2016)

Analysis of differentiation markers in Dgcr8–Pax8Cre knockout thyroid glands. Immunohistochemistry of thyroid differentiation markers show a strong reduction of Tg expression. Pax8, Nkx2.1 and Foxe1 expression levels are unaffected in the residual follicular cells, as well as the expression of their target Nis. No alteration in calcitonin cells was observed. (40× magnification for Tg, Pax8, calcitonin, Nkx2.1 and Foxe1 bar 50 μm; 100× magnification for Nis, bar 10 µm, age of the mice 4–8 weeks)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4835897&req=5

Fig2: Analysis of differentiation markers in Dgcr8–Pax8Cre knockout thyroid glands. Immunohistochemistry of thyroid differentiation markers show a strong reduction of Tg expression. Pax8, Nkx2.1 and Foxe1 expression levels are unaffected in the residual follicular cells, as well as the expression of their target Nis. No alteration in calcitonin cells was observed. (40× magnification for Tg, Pax8, calcitonin, Nkx2.1 and Foxe1 bar 50 μm; 100× magnification for Nis, bar 10 µm, age of the mice 4–8 weeks)
Mentions: Molecular analysis of differentiation markers on knockout thyroid glands showed that the expression of the early differentiation markers Pax8, Nkx2.1 and Foxe1 is retained in the residual follicular cells, as well as the expression of their target Nis. Conversely, the expression of another late differentiation marker—Tg—resulted strongly reduced in the residual follicles. No alterations in calcitonin secreting cells were observed (Fig. 2). Thus, Dgcr8 knockout mice show functional defects partly similar to those from Dicer knockout mice [3, 4]. The main difference is that Nis is still expressed in Dgcr8 knockout mice, while being completely absent in Dicer knockout animals.Fig. 2

Bottom Line: The Dgcr8/Drosha complex processes pri-miRNAs in the nucleus before they are exported as pre-miRNAs for further maturation by Dicer in the cytoplasm.Pax8Cre-mediated conditional loss of DiGeorge syndrome critical region 8 (Dgcr8), an essential component of the nuclear machinery that is required for microRNA biogenesis, resulted in severe hypothyroidism, massively reduced body weight and ultimately led to renal failure and death of the animals.These data provide further insight into the importance of miRNAs in organ homeostasis using a Dicer independent model.

View Article: PubMed Central - PubMed

Affiliation: Department II of Internal Medicine and Center for Molecular Medicine, University of Cologne, Kerpener Str. 62, 50937, Cologne, Germany.

ABSTRACT

Background: Non-coding RNAs have gained increasing attention during the last decade. The first large group of non-coding RNAs to be characterized systematically starting at the beginning of the 21st century were small oligonucleotides--the so-called microRNAs (miRNAs). By now we have learnt that microRNAs are indispensable for most biological processes including organogenesis and maintenance of organ structure and function. The role of microRNAs has been studied extensively in the development of a number of organs, so far most studies focussed on e.g. the heart or the brain whilst the role of microRNAs in the development and maintenance of complex epithelial organs is less well understood. Furthermore most analyses regarding microRNA function in epithelial organs employed conditional knockout mouse models of the RNAse III Dicer to abrogate microRNA biogenesis. However, there is increasing evidence for Dicer to have multiple functions independent from microRNA maturation. Therefore Dicer independent models are needed to gain further insight into the complex biology of miRNA dependent processes.

Results: Here we analyze the contribution of microRNA-dependent transcriptional control in Pax8-expressing epithelial cells. Pax8 is a transcription factor that is crucial to the development of epithelial organs. The miRNA machinery was disrupted by crossing conditional DiGeorge syndrome critical region 8 (Dgcr8) fl/fl mice to Pax8Cre mice. The Dgcr8/Drosha complex processes pri-miRNAs in the nucleus before they are exported as pre-miRNAs for further maturation by Dicer in the cytoplasm. Dgcr8 fl/fl; Pax8Cre+ knockout mice died prematurely, developed massive hypothyroidism and end stage renal disease due to a loss of miRNAs in Pax8 expressing tissue.

Conclusion: Pax8Cre-mediated conditional loss of DiGeorge syndrome critical region 8 (Dgcr8), an essential component of the nuclear machinery that is required for microRNA biogenesis, resulted in severe hypothyroidism, massively reduced body weight and ultimately led to renal failure and death of the animals. These data provide further insight into the importance of miRNAs in organ homeostasis using a Dicer independent model.

No MeSH data available.


Related in: MedlinePlus