Transient transmission of a transgene in mouse offspring following in vivo transfection of male germ cells

2002 ◽  
Vol 62 (4) ◽  
pp. 477-482 ◽  
Author(s):  
Catherine Celebi ◽  
Pierrick Auvray ◽  
Thierry Benvegnu ◽  
Daniel Plusquellec ◽  
Bernard JÉgou ◽  
...  
Keyword(s):  
Germ Cells ◽  
Male Germ Cells ◽  
In Vivo Transfection

scholarly journals The 28 + 28 day design is an effective sampling time for analyzing mutant frequencies in rapidly proliferating tissues of MutaMouse animals

2021 ◽  
Vol 95 (3) ◽  
pp. 1103-1116
Author(s):  
Francesco Marchetti ◽  
Gu Zhou ◽  
Danielle LeBlanc ◽  
Paul A. White ◽  
Andrew Williams ◽  
...  
Keyword(s):  
Germ Cells ◽  
False Negative ◽  
Sampling Time ◽  
Male Germ Cells ◽  
Negative Results ◽  
False Negative Results ◽  
Detection Of Mutations ◽  
The Impact ◽  
Sampling Times

AbstractThe Organisation for Economic Co-Operation and Development Test Guideline 488 (TG 488) uses transgenic rodent models to generate in vivo mutagenesis data for regulatory submission. The recommended design in TG 488, 28 consecutive daily exposures with tissue sampling three days later (28 + 3d), is optimized for rapidly proliferating tissues such as bone marrow (BM). A sampling time of 28 days (28 + 28d) is considered more appropriate for slowly proliferating tissues (e.g., liver) and male germ cells. We evaluated the impact of the sampling time on mutant frequencies (MF) in the BM of MutaMouse males exposed for 28 days to benzo[a]pyrene (BaP), procarbazine (PRC), isopropyl methanesulfonate (iPMS), or triethylenemelamine (TEM) in dose–response studies. BM samples were collected + 3d, + 28d, + 42d or + 70d post exposure and MF quantified using the lacZ assay. All chemicals significantly increased MF with maximum fold increases at 28 + 3d of 162.9, 6.6, 4.7 and 2.8 for BaP, PRC, iPMS and TEM, respectively. MF were relatively stable over the time period investigated, although they were significantly increased only at 28 + 3d and 28 + 28d for TEM. Benchmark dose (BMD) modelling generated overlapping BMD confidence intervals among the four sampling times for each chemical. These results demonstrate that the sampling time does not affect the detection of mutations for strong mutagens. However, for mutagens that produce small increases in MF, sampling times greater than 28 days may produce false-negative results. Thus, the 28 + 28d protocol represents a unifying protocol for simultaneously assessing mutations in rapidly and slowly proliferating somatic tissues and male germ cells.

scholarly journals Generation of male germ cells from induced pluripotent stem cells (iPS cells): an in vitro and in vivo study

2012 ◽  
Vol 14 (4) ◽  
pp. 574-579 ◽  
Author(s):  
Yong Zhu ◽  
Hong-Liang Hu ◽  
Peng Li ◽  
Shi Yang ◽  
Wei Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Induced Pluripotent Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
In Vivo Study ◽  
Male Germ Cells ◽  
Induced Pluripotent

scholarly journals miRNA-dependent poly(A) length control in uncoupling transcription and translation of haploid male germ cells

2021 ◽  
Author(s):  
Chong Tang ◽  
Mei Guo ◽  
Zhuoxing Shi ◽  
Zhuqing Wang ◽  
Chunhai Luo ◽  
...  
Keyword(s):  
Germ Cells ◽  
Regulatory Mechanisms ◽  
Translational Repression ◽  
Dynamic Changes ◽  
Male Germ Cells ◽  
Haploid Male ◽  
Transcriptional Regulatory ◽  
Delayed Translation ◽  
Transcriptional Regulatory Mechanisms

AbstractAs one of the post-transcriptional regulatory mechanisms, transcription and translation’s uncoupling plays an essential role in development and adulthood physiology. However, it remains elusive how thousands of mRNAs get translationally silenced while stability is maintained for up to hours or even days before translation. In addition to oocytes and neurons, developing spermatids have significant uncoupling of transcription and translation for delayed translation. Therefore, spermiogenesis represents an excellent in vivo model for investigating the mechanism underlying uncoupled transcription and translation. Through full-length poly(A) deep sequencing, we discovered dynamic changes in poly(A) length through deadenylation and re-polyadenylation. Deadenylation appeared to be mediated by microRNAs (miRNAs), and transcripts with shorter poly(A) tails tend to be sequestered into ribonucleoproteins (RNPs) for translational repression and stabilization. In contrast, re-polyadenylation allows for translocation of the translationally repressed transcripts from RNPs to polysomes for translation. Overall, our data suggest that miRNA-dependent poly(A) length control represents a novel mechanism underlying uncoupled translation and transcription in haploid male germ cells.

scholarly journals Partial Sperm beta1 Integrin Subunit Deletion Proves Its Involvement in Mouse Gamete Adhesion/Fusion

2020 ◽  
Vol 21 (22) ◽  
pp. 8494
Author(s):  
Virginie Barraud-Lange ◽  
Côme Ialy-Radio ◽  
Céline Chalas ◽  
Isabelle Holtzmann ◽  
Jean-Philippe Wolf ◽  
...  
Keyword(s):  
Western Blot ◽  
Germ Cells ◽  
Blot Analysis ◽  
Integrin Subunit ◽  
Perivitelline Space ◽  
Male Germ Cells ◽  
Beta1 Integrin ◽  
First Time

We have previously shown, using antibodies, that the sperm alpha6beta1 integrin is involved in mouse gamete fusion in vitro. Here we report the conditional knockdown of the sperm Itgb1 gene. It induced a drastic failure of sperm fusogenic ability with sperm accumulation in the perivitelline space of in vitro inseminated oocytes deleted or not for the Itgb1 gene. These data demonstrate that sperm, but not oocyte, beta1 integrin subunit is involved in gamete adhesion/fusion. Curiously, knockdown males were fertile in vivo probably because of the incomplete Cre-mediated deletion of the sperm Itgb1 floxed gene. Indeed, this was shown by Western blot analysis and confirmed by both the viability and litter size of pups obtained by mating partially sperm Itgb1 deleted males with females producing completely deleted Itgb1 oocytes. Because of the total peri-implantation lethality of Itgb1 deletion in mice, we assume that sperm that escaped the Itgb1 excision seemed to be preferentially used to fertilize in vivo. Here, we showed for the first time that the deletion, even partial, of the sperm Itgb1 gene makes the sperm unable to normally fertilize oocytes. However, to elucidate the question of the essentiality of its role during fertilization, further investigations using a mouse expressing a recombinase more effective in male germ cells are necessary.

A new method for producing transgenic birds via direct in vivo transfection of primordial germ cells

2013 ◽  
Vol 22 (6) ◽  
pp. 1257-1264 ◽  
Author(s):  
Scott G. Tyack ◽  
Kristie A. Jenkins ◽  
Terri E. O’Neil ◽  
Terry G. Wise ◽  
Kirsten R. Morris ◽  
...  
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
New Method ◽  
In Vivo Transfection

scholarly journals Spermatogonial Stem Cells for In Vitro Spermatogenesis and In Vivo Restoration of Fertility

Cells ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 745 ◽  
Author(s):  
Fahar Ibtisham ◽  
Ali Honaramooz
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Recent Progress ◽  
Male Germ Cells ◽  
Culture Systems ◽  
In Vitro Spermatogenesis ◽  
Haploid Male ◽  
Restoration Of Fertility

Spermatogonial stem cells (SSCs) are the only adult stem cells capable of passing genes onto the next generation. SSCs also have the potential to provide important knowledge about stem cells in general and to offer critical in vitro and in vivo applications in assisted reproductive technologies. After century-long research, proof-of-principle culture systems have been introduced to support the in vitro differentiation of SSCs from rodent models into haploid male germ cells. Despite recent progress in organotypic testicular tissue culture and two-dimensional or three-dimensional cell culture systems, to achieve complete in vitro spermatogenesis (IVS) using non-rodent species remains challenging. Successful in vitro production of human haploid male germ cells will foster hopes of preserving the fertility potential of prepubertal cancer patients who frequently face infertility due to the gonadotoxic side-effects of cancer treatment. Moreover, the development of optimal systems for IVS would allow designing experiments that are otherwise difficult or impossible to be performed directly in vivo, such as genetic manipulation of germ cells or correction of genetic disorders. This review outlines the recent progress in the use of SSCs for IVS and potential in vivo applications for the restoration of fertility.

scholarly journals Pentacle gold–copper alloy nanocrystals: a new system for entering male germ cells in vitro and in vivo

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Yu Lin ◽  
Rong He ◽  
Liping Sun ◽  
Yushan Yang ◽  
Wenqing Li ◽  
...  
Keyword(s):  
Germ Cells ◽  
Copper Alloy ◽  
Male Germ Cells ◽  
Alloy Nanocrystals ◽  
New System

scholarly journals Structure and expression of murine mgcRacGAP: its developmental regulation suggests a role for the Rac/MgcRacGAP signalling pathway in neurogenesis

1999 ◽  
Vol 343 (1) ◽  
pp. 225-230 ◽  
Author(s):  
Chantal ARAR ◽  
Marie-Odile OTT ◽  
Aminata TOURÉ ◽  
Gérard GACON
Keyword(s):  
Germ Cells ◽  
Developmental Regulation ◽  
Ventricular Zone ◽  
Developmentally Regulated ◽  
Gene Encoding ◽  
Male Germ Cells ◽  
Preferential Expression ◽  
Wide Range ◽  
Developing Neurons

Rho-family GTPases regulate a wide range of biological functions including cell migration, cell adhesion and cell growth. Recently, results from studies in vivo in Drosophila, mouse and humans have demonstrated the involvement of these GTPases in mechanisms controlling neuronal differentiation and the development of the central nervous system (CNS). However, the signalling pathways underlying these functions and the proteins directly regulating RhoGTPases in developing neurons are poorly defined. Here we report the structure and expression pattern of the murine orthologue of mgcRacGAP, a human gene encoding a RacGTPase partner expressed in male germ cells [Touré, Dorseuil, Morin, Timmons, Jegou, Reibel and Gacon (1998) J. Biol. Chem. 273, 6019-6023]. In contrast with that from humans, murine mgcRacGAP encodes two distinct transcripts. Both are developmentally regulated. A 2.2 kb transcript is strongly expressed in mature testis and is up-regulated with spermatogenesis. A 3 kb RNA is predominant in the embryo and is expressed primarily in the CNS during the neurogenic phase, decreasing after birth. In situ hybridization analysis in embryonic-day 14.5 mouse embryos demonstrates a preferential expression of mgcRacGAP in the proliferative ventricular zone of the cortex. In addition to the expression of mgcRacGAP in male germ cells already reported in humans and suggesting an involvement in spermatogenesis, we characterize an embryonic transcript whose expression is closely correlated with neurogenesis. This result addresses the question of the role of Rac/MgcRacGAP pathway in neuronal proliferation.

scholarly journals Gonadal sex reversal of the developing marsupial ovary in vivo and in vitro

Development ◽  
1996 ◽  
Vol 122 (12) ◽  
pp. 4057-4063 ◽  
Author(s):  
D.J. Whitworth ◽  
G. Shaw ◽  
M.B. Renfree
Keyword(s):  
Germ Cells ◽  
Normal Development ◽  
Tammar Wallaby ◽  
Mitotic Division ◽  
Sex Reversal ◽  
Direct Result ◽  
Male Germ Cells ◽  
Well Differentiated

Undifferentiated tammar wallaby ovaries were transplanted under the skin of male pouch young during the period of mitotic division of the XX germ cells. After 25 days, all the germ cells had disappeared and the ovaries contained seminiferous-like cords. Similarly, undifferentiated ovaries cultured for 4 days with recombinant human Mullerian-inhibiting substance (rhMIS) also contained well-differentiated seminiferous-like cords and few or no surviving germ cells. The majority of controls cultured without rhMIS developed as normal ovaries. However, in a few control ovaries seminiferous-like cords developed in those regions of the ovaries that were partially necrotic and contained few germ cells. These results strongly suggest that sex-reversal of the tammar ovary is the direct result of loss of mitotic germ cells, rather than an effect of MIS on female somatic cells. MIS is apparently toxic to these female germ cells in mitosis, but not to male germ cells in mitosis. Thus, in normal development in the tammar, the presence of XX germ cells in the ovary inhibits the formation of seminiferous cords so that the gonad develops as an ovary.

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