Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1371/journal.pone.0246289 http://hdl.handle.net/11449/210687 |
Resumo: | Background Here, we have demonstrated that gestational low-protein (LP) intake offspring present lower birth weight, reduced nephron numbers, renal salt excretion, arterial hypertension, and renal failure development compared to regular protein (NP) intake rats in adulthood. We evaluated the expression of various miRNAs and predicted target genes in the kidney in gestational 17-days LP (DG-17) fetal metanephros to identify molecular pathways involved in the proliferation and differentiation of renal embryonic or fetal cells. Methods Pregnant Wistar rats were classified into two groups based on protein supply during pregnancy: NP (regular protein diet, 17%) or LP diet (6%). Renal miRNA sequencing (miRNA-Seq) performed on the MiSeq platform, RT-qPCR of predicted target genes, immunohistochemistry, and morphological analysis of 17-DG NP and LP offspring were performed using previously described methods. Results A total of 44 miRNAs, of which 19 were up and 25 downregulated, were identified in 17-DG LP fetuses compared to age-matched NP offspring. We selected 7 miRNAs involved in proliferation, differentiation, and cellular apoptosis. Our findings revealed reduced cell number and Six-2 and c-Myc immunoreactivity in metanephros cap (CM) and ureter bud (UB) in 17-DG LP fetuses. Ki-67 immunoreactivity in CM was 48% lesser in LP compared to age-matched NP fetuses. Conversely, in LP CM and UB, beta-catenin was 154%, and 85% increased, respectively. Furthermore, mTOR immunoreactivity was higher in LP CM (139%) and UB (104%) compared to that in NP offspring. TGF beta-1 positive cells in the UB increased by approximately 30% in the LP offspring. Moreover, ZEB1 metanephros-stained cells increased by 30% in the LP offspring. ZEB2 immunofluorescence, although present in the entire metanephros, was similar in both experimental groups. Conclusions Maternal protein restriction changes the expression of miRNAs, mRNAs, and proteins involved in proliferation, differentiation, and apoptosis during renal development. Renal ontogenic dysfunction, caused by maternal protein restriction, promotes reduced reciprocal interaction between CM and UB; consequently, a programmed and expressive decrease in nephron number occurs in the fetus. |
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Repositório Institucional da UNESP |
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Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetusBackground Here, we have demonstrated that gestational low-protein (LP) intake offspring present lower birth weight, reduced nephron numbers, renal salt excretion, arterial hypertension, and renal failure development compared to regular protein (NP) intake rats in adulthood. We evaluated the expression of various miRNAs and predicted target genes in the kidney in gestational 17-days LP (DG-17) fetal metanephros to identify molecular pathways involved in the proliferation and differentiation of renal embryonic or fetal cells. Methods Pregnant Wistar rats were classified into two groups based on protein supply during pregnancy: NP (regular protein diet, 17%) or LP diet (6%). Renal miRNA sequencing (miRNA-Seq) performed on the MiSeq platform, RT-qPCR of predicted target genes, immunohistochemistry, and morphological analysis of 17-DG NP and LP offspring were performed using previously described methods. Results A total of 44 miRNAs, of which 19 were up and 25 downregulated, were identified in 17-DG LP fetuses compared to age-matched NP offspring. We selected 7 miRNAs involved in proliferation, differentiation, and cellular apoptosis. Our findings revealed reduced cell number and Six-2 and c-Myc immunoreactivity in metanephros cap (CM) and ureter bud (UB) in 17-DG LP fetuses. Ki-67 immunoreactivity in CM was 48% lesser in LP compared to age-matched NP fetuses. Conversely, in LP CM and UB, beta-catenin was 154%, and 85% increased, respectively. Furthermore, mTOR immunoreactivity was higher in LP CM (139%) and UB (104%) compared to that in NP offspring. TGF beta-1 positive cells in the UB increased by approximately 30% in the LP offspring. Moreover, ZEB1 metanephros-stained cells increased by 30% in the LP offspring. ZEB2 immunofluorescence, although present in the entire metanephros, was similar in both experimental groups. Conclusions Maternal protein restriction changes the expression of miRNAs, mRNAs, and proteins involved in proliferation, differentiation, and apoptosis during renal development. Renal ontogenic dysfunction, caused by maternal protein restriction, promotes reduced reciprocal interaction between CM and UB; consequently, a programmed and expressive decrease in nephron number occurs in the fetus.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Estadual Campinas, Fetal Programming & Hydroelectrolyte Metab Lab, Nucleus Med & Expt Surg, Dept Internal Med,Fac Med Sci, Campinas, SP, BrazilSao Paulo State Univ, Biosci Inst, Dept Struct & Funct Biol, Botucatu, SP, BrazilSao Paulo State Univ, Biosci Inst, Dept Struct & Funct Biol, Botucatu, SP, BrazilFAPESP: 2013/12486-5FAPESP: 2014/50938-8CNPq: 465699/2014-6Public Library ScienceUniversidade Estadual de Campinas (UNICAMP)Universidade Estadual Paulista (Unesp)Sene, Leticia de BarrosScarano, Wellerson Rodrigo [UNESP]Zapparoli, AdrianaRocha Gontijo, Jose AntonioBoer, Patricia Aline2021-06-26T02:53:53Z2021-06-26T02:53:53Z2021-02-05info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article24http://dx.doi.org/10.1371/journal.pone.0246289Plos One. San Francisco: Public Library Science, v. 16, n. 2, 24 p., 2021.1932-6203http://hdl.handle.net/11449/21068710.1371/journal.pone.0246289WOS:000617991900051Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPlos Oneinfo:eu-repo/semantics/openAccess2021-10-23T22:13:49Zoai:repositorio.unesp.br:11449/210687Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T22:13:49Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus |
title |
Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus |
spellingShingle |
Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus Sene, Leticia de Barros |
title_short |
Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus |
title_full |
Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus |
title_fullStr |
Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus |
title_full_unstemmed |
Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus |
title_sort |
Impact of gestational low-protein intake on embryonic kidney microRNA expression and in nephron progenitor cells of the male fetus |
author |
Sene, Leticia de Barros |
author_facet |
Sene, Leticia de Barros Scarano, Wellerson Rodrigo [UNESP] Zapparoli, Adriana Rocha Gontijo, Jose Antonio Boer, Patricia Aline |
author_role |
author |
author2 |
Scarano, Wellerson Rodrigo [UNESP] Zapparoli, Adriana Rocha Gontijo, Jose Antonio Boer, Patricia Aline |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual de Campinas (UNICAMP) Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Sene, Leticia de Barros Scarano, Wellerson Rodrigo [UNESP] Zapparoli, Adriana Rocha Gontijo, Jose Antonio Boer, Patricia Aline |
description |
Background Here, we have demonstrated that gestational low-protein (LP) intake offspring present lower birth weight, reduced nephron numbers, renal salt excretion, arterial hypertension, and renal failure development compared to regular protein (NP) intake rats in adulthood. We evaluated the expression of various miRNAs and predicted target genes in the kidney in gestational 17-days LP (DG-17) fetal metanephros to identify molecular pathways involved in the proliferation and differentiation of renal embryonic or fetal cells. Methods Pregnant Wistar rats were classified into two groups based on protein supply during pregnancy: NP (regular protein diet, 17%) or LP diet (6%). Renal miRNA sequencing (miRNA-Seq) performed on the MiSeq platform, RT-qPCR of predicted target genes, immunohistochemistry, and morphological analysis of 17-DG NP and LP offspring were performed using previously described methods. Results A total of 44 miRNAs, of which 19 were up and 25 downregulated, were identified in 17-DG LP fetuses compared to age-matched NP offspring. We selected 7 miRNAs involved in proliferation, differentiation, and cellular apoptosis. Our findings revealed reduced cell number and Six-2 and c-Myc immunoreactivity in metanephros cap (CM) and ureter bud (UB) in 17-DG LP fetuses. Ki-67 immunoreactivity in CM was 48% lesser in LP compared to age-matched NP fetuses. Conversely, in LP CM and UB, beta-catenin was 154%, and 85% increased, respectively. Furthermore, mTOR immunoreactivity was higher in LP CM (139%) and UB (104%) compared to that in NP offspring. TGF beta-1 positive cells in the UB increased by approximately 30% in the LP offspring. Moreover, ZEB1 metanephros-stained cells increased by 30% in the LP offspring. ZEB2 immunofluorescence, although present in the entire metanephros, was similar in both experimental groups. Conclusions Maternal protein restriction changes the expression of miRNAs, mRNAs, and proteins involved in proliferation, differentiation, and apoptosis during renal development. Renal ontogenic dysfunction, caused by maternal protein restriction, promotes reduced reciprocal interaction between CM and UB; consequently, a programmed and expressive decrease in nephron number occurs in the fetus. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-06-26T02:53:53Z 2021-06-26T02:53:53Z 2021-02-05 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1371/journal.pone.0246289 Plos One. San Francisco: Public Library Science, v. 16, n. 2, 24 p., 2021. 1932-6203 http://hdl.handle.net/11449/210687 10.1371/journal.pone.0246289 WOS:000617991900051 |
url |
http://dx.doi.org/10.1371/journal.pone.0246289 http://hdl.handle.net/11449/210687 |
identifier_str_mv |
Plos One. San Francisco: Public Library Science, v. 16, n. 2, 24 p., 2021. 1932-6203 10.1371/journal.pone.0246289 WOS:000617991900051 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Plos One |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
24 |
dc.publisher.none.fl_str_mv |
Public Library Science |
publisher.none.fl_str_mv |
Public Library Science |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1803650234862010368 |