PIWI-interacting RNAs (piRNAs) support the germline by suppressing retrotransposons. Studies of the pathway in mice have strongly shaped the view that mammalian piRNAs are essential for male but not for female fertility. Here, we report that the role of the piRNA pathway substantially differs in golden hamsters (Mesocricetus auratus), the piRNA pathway setup of which more closely resembles that of other mammals, including humans. The loss of the Mov10l1 RNA helicase-an essential piRNA biogenesis factor-leads to striking phenotypes in both sexes. In contrast to mice, female Mov10l1-/- hamsters are sterile because their oocytes do not sustain zygotic development. Furthermore, Mov10l1-/- male hamsters have impaired establishment of spermatogonia accompanied by transcriptome dysregulation and an expression surge of a young retrotransposon subfamily. Our results show that the mammalian piRNA pathway has essential roles in both sexes and its adaptive nature allows it to manage emerging genomic threats and acquire new critical roles in the germline.

• MeSH
• křečci praví MeSH
• křeček rodu Mesocricetus metabolismus   MeSH
• malá interferující RNA genetika   MeSH
• oocyty metabolismus   patologie   MeSH
• retroelementy fyziologie   MeSH
• RNA-helikasy genetika   MeSH
• spermatogeneze genetika   fyziologie   MeSH
• spermatogonie metabolismus   patologie   MeSH
• testis metabolismus   MeSH
• umlčování genů fyziologie   MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• malá interferující RNA MeSH
• retroelementy MeSH
• RNA-helikasy MeSH

Germline genome defense evolves to recognize and suppress retrotransposons. One of defensive mechanisms is the PIWI-associated RNA (piRNA) pathway, which employs small RNAs for sequence-specific repression. The loss of the piRNA pathway in mice causes male sterility while females remain fertile. Unlike spermatogenic cells, mouse oocytes posses also RNA interference (RNAi), another small RNA pathway capable of retrotransposon suppression. To examine whether RNAi compensates the loss of the piRNA pathway, we produced a new RNAi pathway mutant DicerSOM and crossed it with a catalytically-dead mutant of Mili, an essential piRNA gene. Normal follicular and oocyte development in double mutants showed that RNAi does not suppress a strong ovarian piRNA knock-out phenotype. However, we observed redundant and non-redundant targeting of specific retrotransposon families illustrating stochasticity of recognition and targeting of invading retrotransposons. Intracisternal A Particle retrotransposon was mainly targeted by the piRNA pathway, MaLR and RLTR10 retrotransposons were targeted mainly by RNAi. Double mutants showed accumulations of LINE-1 retrotransposon transcripts. However, we did not find strong evidence for transcriptional activation and mobilization of retrotransposition competent LINE-1 elements suggesting that while both defense pathways are simultaneously expendable for ovarian oocyte development, yet another transcriptional silencing mechanism prevents mobilization of LINE-1 elements.

• MeSH
• Argonaut proteiny genetika   MeSH
• DEAD-box RNA-helikasy genetika   MeSH
• malá interferující RNA genetika   MeSH
• mutace MeSH
• myši MeSH
• oocyty chemie   růst a vývoj   MeSH
• retroelementy * MeSH
• ribonukleasa III genetika   MeSH
• RNA interference * MeSH
• signální transdukce MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Argonaut proteiny MeSH
• DEAD-box RNA-helikasy MeSH
• Dicer1 protein, mouse MeSH Prohlížeč
• malá interferující RNA MeSH
• Piwil2 protein, mouse MeSH Prohlížeč
• retroelementy * MeSH
• ribonukleasa III MeSH

Small non-coding RNAs control normal development and differentiation in the embryo. These regulatory molecules play a key role in the development of human diseases and are used often today for researching new treatments for different pathologies. In this study, CaCo2 colorectal adenocarcinoma cells were initially epigenetically reprogrammed and transformed into CD4+ cells with nano-sized complexes of amphiphilic poly-(N-vinylpyrrolidone) (PVP) with miRNA-152 and piRNA-30074. The transformation of cells was confirmed by morphological and genetic changes in the dynamic of reprogramming. CD4+ lymphocytes marker was detected using immunofluorescence. Amphiphilic poly-(N-vinylpyrrolidone)/small non-coding RNAs complexes were investigated for transfection efficiency and duration of transfection of CaCo2 colorectal adenocarcinoma cells using fluorescence.

• Klíčová slova
• AGO2, argonaute 2, Amphiphilic poly-(N-vinylpyrrolidone), BACH1, BTB domain and CNC homolog 1, CD, cluster of differentiation, CaCo2 colorectal adenocarcinoma, DICER1, ribonuclease III, DNMT1, DNA methyltransferase 1, DTT, dithyothreitol, ERK1/2, extracellular signal regulated kinase ½, FGF2, fibroblast growth factor 2, GITR3A, glucocorticoid-induced TNFR-related protein, H3K9me3, tri-methyl lysine 9 of histone H3, HILI, human piwi, HMOX1, heme oxygenase 1, HOXA10, homebox A10, ICOS1B, inducible T-cell co-stimulator, IL, interleukin, KIR1DL2, CD158b, expressed on natural killer cells and a subset of T cells, MKI-67, marker of proliferation ki-67, OCT4, octamer-binding transcription factor 4, PIWIL1, piwi-like protein 1, PNVP, poly-(N-vinylpyrrolidone), Polymer carriers, RB1, retinoblastoma 1, Reprogramming, SncRNAs, small non-coding RNAs, TE, transposon elements, TGFBR2, transforming growth factor beta receptor 2, TNFRS6B, TNF receptor superfamily 6B, TSS, transcriptional start sites, VMAF, musculoaponeurotic fibrosarcoma, Wnt-1, wingless type MMTV integration site family, member 1, iPS, induced pluripotent stem cells, mTOR, mechanistic target of rapamycin, miR, micro-RNA, miRNA-152, piR, piwi-interacting RNA, P-element induced wimpy testis interacting RNA, piRNA-30074,
• Publikační typ
• časopisecké články MeSH

Rat hypodactyly (hd) mutation is characterized by abnormal spermatogenesis and sperm decapitation, limb malformation (missing digits II and III) and growth retardation. We have previously reported centrobin (centrosome BRCA2-interacting protein) truncation at the C-terminus in the hd mutant. Here, we report data from a transgenic rescue experiment carried out to determine a role of centrobin in pathogenesis of hd. The transgenic construct, consisting of full-length-coding cDNA linked to a ubiquitous strong promoter/enhancer combination, was inserted to chromosome 16 into a LINE repeat. No known gene is present in the vicinity of the insertion site. Transgenic centrobin was expressed in all tissues tested, including testis. Transgenic animals show normal body weight and limb morphology as well as average weight of testis and epididymis. Yet, abnormal spermatogenesis and sperm decapitation persisted in the transgenic animals. Western blotting showed the coexistence of full-length and truncated or partially degraded centrobin in sperm of the rescued transgenic animals. Immunocytochemistry showed a buildup of centrobin and ODF2 (outer dense fiber 2) at the sperm decapitation site in the hd mutant and rescued transgenic rats. Additional findings included bulge-like formations and thread-like focal dissociations along the sperm flagellum and the organization of multiple whorls of truncated sperm flagella in the epididymal lumen. We conclude that centrobin is essential for normal patterning of the limb autopod. Centrobin may be required for stabilizing the attachment of the sperm head to flagellum and for maintaining the structural integrity of the sperm flagellum. We postulate that the presence of truncated centrobin, coexisting with full-length centrobin, together with incorrect timing of transgenic centrobin expression may hamper the rescue of fertility in hd male rats.

• MeSH
• epididymis patologie   MeSH
• exprese genu MeSH
• fertilita genetika   MeSH
• homeodoménové proteiny genetika   MeSH
• krysa rodu Rattus MeSH
• mutace * MeSH
• myši MeSH
• potkani transgenní MeSH
• proteiny buněčného cyklu genetika   metabolismus   MeSH
• proteiny teplotního šoku metabolismus   MeSH
• spermie růst a vývoj   metabolismus   MeSH
• testis patologie   MeSH
• transport proteinů MeSH
• velikost orgánu genetika   MeSH
• vrozené deformity končetin genetika   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• Cntrob protein, rat MeSH Prohlížeč
• homeobox protein HOXA13 MeSH Prohlížeč
• homeodoménové proteiny MeSH
• Odf2 protein, rat MeSH Prohlížeč
• proteiny buněčného cyklu MeSH
• proteiny teplotního šoku MeSH

Small RNA molecules regulating gene expression received a status of omnipresent master regulators of eukaryotic lives with almost supernatural powers. Mammals hold at least three mechanisms employing small RNA molecules for regulating gene expression. One of these mechanisms, the microRNA (miRNA) pathway, involves currently over a thousand of genome-encoded different miRNAs that are claimed to extend their control over more than a half of a genome. Here, I discuss how and why mouse oocytes and early embryos ignore the regulatory power of miRNAs, adding another surprising feature to the field of small RNAs.

• MeSH
• embryo savčí metabolismus   MeSH
• mikro RNA genetika   MeSH
• myši MeSH
• oocyty metabolismus   MeSH
• RNA interference * MeSH
• RNA messenger skladovaná MeSH
• vývojová regulace genové exprese * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• mikro RNA MeSH
• RNA messenger skladovaná MeSH