scholarly journals Transfer at the blastocyst stage of embryos derived from testicular round spermatid injection

2002 ◽  
Vol 17 (3) ◽  
pp. 741-743 ◽  
Author(s):  
B. Urman ◽  
C. Alatas ◽  
S. Aksoy ◽  
R. Mercan ◽  
A. Nuhoglu ◽  
...  
Keyword(s):  
Round Spermatid ◽  
Blastocyst Stage ◽  
Round Spermatid Injection

Tricostatin A-treated round spermatid enhances preimplantation embryo developmental competency following round spermatid injection in mice

Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Sara Hosseini ◽  
Mohammad Salehi
Keyword(s):  
Preimplantation Embryo ◽  
Trichostatin A ◽  
Round Spermatid ◽  
Fertilization Rate ◽  
Blastocyst Stage ◽  
Expression Level ◽  
Fluid Medium ◽  
Preimplantation Embryo Development ◽  
Significant Difference ◽  
Round Spermatid Injection

Summary It has been documented that the inefficacy of round spermatid injection (ROSI) might be caused by abnormal epigenetic modifications. Therefore, this study aimed to evaluate the effect of trichostatin A (TSA) as an epigenetic modifier of preimplantation embryo development in activated ROSI oocytes. Matured oocytes were collected from superovulated female mice. Testes were placed in human tubal fluid medium and masses were then cut into small pieces to disperse spermatogenic cells. Round spermatids were treated with TSA and subsequently injected into oocytes. The expression level of the development-related genes including Oct4, Sox2, Nanog, Dnmt and Hdac transcripts were evaluated using qRT-PCR. Immunohistochemistry was performed to confirm the presence of Oct-4 protein at the blastocyst stage. There was no statistically significant difference in fertilization rate following ROSI/+TSA compared with the non-treated ROSI and intracytoplasmic sperm injection (ICSI) groups. Importantly, TSA treatment increased blastocyst formation from 38% in non-treated ROSI to 68%. The relative expression level of developmentally related genes increased and Dnmt transcripts decreased in ROSI/+TSA-derived embryos, similar to the expression levels observed in the ICSI-derived embryos. In conclusion, our results indicate that spermatid treatment with TSA prior to ROSI would increase the success rate of development to the blastocyst stage and proportion of pluripotent cells.

Fate of microinjected spermatid mitochondria in the mouse oocyte and embryo

Zygote ◽  
1998 ◽  
Vol 6 (3) ◽  
pp. 213-222 ◽  
Author(s):  
James M. Cummins ◽  
Teruhiko Wakayama ◽  
Ryuzo Yanagimachi
Keyword(s):  
Significant Risk ◽  
Primary Spermatocyte ◽  
Round Spermatid ◽  
Blastocyst Stage ◽  
Infertile Men ◽  
Oocyte Cytoplasm ◽  
Almost All ◽  
Cytoskeletal Elements ◽  
Round Spermatid Injection ◽  
Cell Transition

Mouse round spermatids labelled with MitoTracker were microinjected into Sr2+-activated mouse oocytes. The labelled mitochondria were tracked up to the morula/blastocyst stage using fluorescence microscopy. The overall incidence of embryos with labelled mitochondria fell from 80% in the 1-cell zygote to 25% in 2-cell, 9% in 4-cell and ~1% in 8-cell or later stages. Thus it appears that almost all round spermatid mitochondria finally disappear from embryos during the 4-cell to 8-cell transition, as happens for mature spermatozoa (Cummins et al.Zygote 1997, 5: 301–8). The spermatid mitochondria remained tightly bound together during this process. In contrast, labelled primary spermatocyte and cumulus mitochondria dispersed rapidly throughout the oocyte cytoplasm within 3 h. We hypothesise that spermatid mitochondria may be bound together by cytoskeletal elements produced in the early haploid spermatid. These elements, together with terminal differentiation of the sperm mitochondria, may be central to the processes by which the embryo ‘recognises’ the sperm mitochondria and inhibits inheritance of paternal mitochondrial DNA. These results suggest that round spermatid injection for infertile men will not pose a significant risk to offspring by transmitting abnormal mitochondrial genomes.

Is Round Spermatid Injection (ROSI) a Therapy for Male Infertility?: ROSI in the Rhesus Monkey is Unsuccessful

2000 ◽  
Vol 74 (3) ◽  
pp. S68 ◽  
Author(s):  
L Hewitson ◽  
C Martinovich ◽  
C Simerly ◽  
T Dominko ◽  
G Schatten
Keyword(s):  
Male Infertility ◽  
Rhesus Monkey ◽  
Round Spermatid ◽  
Round Spermatid Injection

scholarly journals Mice generated after round spermatid injection into haploid two-cell blastomeres

Cell Research ◽  
2011 ◽  
Vol 21 (5) ◽  
pp. 854-858 ◽  
Author(s):  
Hui Yang ◽  
Linyu Shi ◽  
Charlie Degui Chen ◽  
Jinsong Li
Keyword(s):  
Round Spermatid ◽  
Round Spermatid Injection

Round spermatids stained with MitoTracker can be used to produce offspring more simply

Zygote ◽  
2005 ◽  
Vol 13 (1) ◽  
pp. 55-61 ◽  
Author(s):  
Takafusa Hikichi ◽  
Satoshi Kishigami ◽  
Nguyen Van Thuan ◽  
Hiroshi Ohta ◽  
Eiji Mizutani ◽  
...  
Keyword(s):  
Embryo Development ◽  
Intracytoplasmic Sperm Injection ◽  
Round Spermatid ◽  
Testicular Cells ◽  
Infertility Treatments ◽  
Developmental Capacity ◽  
20 Nm ◽  
Harmful Effects ◽  
Round Spermatid Injection ◽  
Selection Of

Although both intracytoplasmic sperm injection (ICSI) and round spermatid injection (ROSI) are used in infertility treatments, the rate of offspring achieved with ROSI is low compared with that achieved with ICSI. The difficulty in correctly selecting round spermatids from testicular cells is one of the causes of this phenomenon. We easily selected live round spermatids from testicular cells stained with 20 nM MitoTracker, which visualizes mitochondria without killing the cell. Using this method, we divided round spermatids into three groups based on the polarization of their mitochondria, and performed ROSI. The rate of successful offspring achieved with MitoTracker-stained ROSI was the same in all groups. This indicates that changes in the polarization of mitochondria in round spermatids are not directly related to the developmental capacity of subsequently fertilized embryos. Because this staining has no harmful effects on embryo development, the selection of spermatids by MitoTracker under a fluorescence microscope should be useful in research into and the treatment of infertility.

scholarly journals The employment of strontium to activate mouse oocytes: effects on spermatid-injection outcome

Reproduction ◽  
2006 ◽  
Vol 131 (2) ◽  
pp. 259-267 ◽  
Author(s):  
Jean Loren ◽  
Orly Lacham-Kaplan
Keyword(s):  
Birth Outcomes ◽  
Round Spermatid ◽  
Oocyte Activation ◽  
Embryo Viability ◽  
Blastocyst Development ◽  
Incubation Periods ◽  
Artificial Oocyte Activation ◽  
The Impact ◽  
Post Implantation ◽  
Round Spermatid Injection

The present research investigated the effects of various strontium concentrations, in combination with different incubation periods, on mouse parthenogentic oocyte activation and blastocyst development. The results for blastocyst development showed a trend indicating that 10 mM strontium for 3 h was the optimal strontium protocol. Ethanol, an agent that incites oocyte activation via a monotonic rise in calcium, was employed as a control. The outcome of blastocyst formation arising from parthenogenic ethanol activation was significantly less (P < 0.001) than that achieved by the optimal strontium protocol. To assess the impact of strontium oocyte activation on embryo viability following fertilization with immature germ cells, the protocol of 10 mM strontium for 3 h was applied to oocytes injected with round spermatids and then compared with other protocols. The results indicate that following round-spermatid injection the benefits derived from strontium artificial oocyte activation are evident during both pre- and post-implantation development. However, in order to adjust the protocol to the most effective round-spermatid injection in relation to the oocyte cell cycle, injection was done 1.5 h after strontium activation followed by another 1.5 h activation in strontium. The implementation of round-spermatid injection in combination with this oocyte-activation protocol led to live-birth outcomes not significantly different to those outcomes obtained by mature spermatozoa.

Harmful or Not: Trichostatin A treatment of embryos generated by ICSI or ROSI

2006 ◽  
Vol 1 (3) ◽  
pp. 376-385 ◽  
Author(s):  
Satoshi Kishigami ◽  
Hiroshi Ohta ◽  
Eiji Mizutani ◽  
Sayaka Wakayama ◽  
Hong-Thuy Bui ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Trichostatin A ◽  
Reproductive Technologies ◽  
Round Spermatid ◽  
Developmental Arrest ◽  
Offspring Production ◽  
Deacetylase Inhibitor ◽  
Cloning Method ◽  
Round Spermatid Injection ◽  
Parthenogenetic Embryos

AbstractTrichostatin A (TSA), a histone deacetylase inhibitor, is a known teratogen causing malformations such as vertebral fusions when applied during the postimplantation period; TSA also causes developmental arrest when applied during the preimplantation period. Regardless of these hindrances, we have succeeded in the establishment of an efficient somatic cloning method for the mouse where reconstructed embryos are treated with TSA. To elucidate this apparent discrepancy, we treated fertilized mouse embryos generated either by intracytoplasmic sperm injection (ICSI) or round spermatid injection (ROSI) with 50 nM TSA for 20 h after fertilization as well as parthenogenetic embryos and found that TSA treatment inhibited the preimplantation development of ICSI embryos but not ROSI or parthenogenetic embryos. And, although we often observed hypomorphism following TSA treatment in embryos grown to full term produced by both ICSI (av. of body weight: 1.7 g vs. 1.5 g) and ROSI (1.6 g vs. 1.2 g), TSA treatment reduced the offspring production rate for ICSI from 57% to 34% but not for ROSI from 30% to 36%. Thus, these data indicate that the effects, harmful or not, of TSA treatment on embryonic development depend on their nuclear derivations. Also, the resulting hypomorphism after TSA treatment is a caveat for this procedure in current Assisted Reproductive Technologies.

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