Ultrastructural analysis of preimplantation lethal yellow (Ay/Ay) mouse embryos

Development ◽  
1978 ◽  
Vol 45 (1) ◽  
pp. 13-24
Author(s):  
Gerald R. Cizadlo ◽  
Nels H. Granholm

Embryos recovered at 62 and 80 h post coitum from reproductive tracts of yellow (Ay/a) and black (a/a) female mice mated to Ay/a males were examined ultrastructurally to define the developmental defect of lethal Ay homozygotes. No abnormalities were observed in five embryos from control matings (♀ a/a × ♂ Ay/a). Of 24 morulae and blastocysts from Ay/a × Ay/a matings, six were observed to possess some morphological aberration. Two of the six abnormal embryos were morulae and contained isolated blastomeres which had developmental features typical of younger embryos; remaining cells of these embryos were normal. The third, an early blastocyst, contained a degenerating trophoblast cell; other cells of this embryo were also abnormal but not in an advanced stage of degeneration. The fourth abnormal embryo (late cleavage stage) was in an advanced stage of degeneration affecting all blastomeres. Finally, the remaining two abnormal morulae had a unique nucleolar morphology and an unusual abundance of intra-cisternal A particles. Presumably, one or more of the six abnormal embryos from Ay/a × Ay/a matings were Ay homozygotes. However, no single ultrastructural alteration characteristic of Ay/Ay embryos was found.


Development ◽  
1980 ◽  
Vol 60 (1) ◽  
pp. 419-428
Author(s):  
Brigid Hogan ◽  
Martha Spiegelman ◽  
Dorothea Bennett

Inner cell masses (ICMs) isolated immunosurgically from mouse blastocysts segregating the homozygous lethal mutants t0/t0 and tw5/tw5 were cultured in vitro. Presumed t0/t0 ICMs fail to grow after three days in culture (equivalent gestational day 7·5) when they consist of an outer layer of endoderm cells surrounding about 30 epiblast cells. Presumed homozygous tw5/tw5 ICMs develop to a more advanced stage in culture and on the seventh day (equivalent gestational day 11·5) consist of an inner core of disorganized ectoderm cells with a small proamniotic cavity, surrounded by multiple layers of endoderm cells.



2003 ◽  
Vol 80 (1) ◽  
pp. 178-183 ◽  
Author(s):  
Jia-Sen Xu ◽  
Samuel Ting-Hon Chan ◽  
Pak-Chung Ho ◽  
William Shu-Biu Yeung


2009 ◽  
Vol 57 (3) ◽  
pp. 399-410 ◽  
Author(s):  
Philip Klambauer ◽  
Zsuzsa Keresztes ◽  
Katalin Kanyó ◽  
Erika Varga ◽  
Rita Kriston ◽  
...  

By decreasing the volume of the cryoprotective solution it is possible to increase dramatically the freezing speed and — at the same time — reduce the toxicity and osmotic side effects of cryoprotectants (CPA). The objective of our study was to vitrify Day-3 cleavage stage mouse embryos (n = 229) with the cryoloop technology using a new composition of vitrification media. Embryos were exposed to a 2-step loading of CPA, ethylene glycol (EG) and propylene glycol (PG), before being placed on the surface of a thin filmy layer formed from the vitrification solution in a small nylon loop, then they were rapidly submerged into liquid nitrogen. After warming, the CPA was diluted out from the embryos by a 3-step procedure. Survival of embryos was based on morphological appearance after thawing and continued development to expanded blastocysts upon subsequent 48-hour culture. Embryos of the two control groups were either treated likewise except that they were not vitrified, or cultured in vitro without any treatment. Our data show that a high percentage of embryos survived (92.7%) vitrification in the mixture of EG and PG combined with cryoloop carrier and developed normally (89.1%) in vitro after thawing. To our knowledge this is the first report of the successful vitrification of cleavage stage mouse embryos using VitroLoop vitrification procedure.



Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 5388-5388
Author(s):  
Eugene E. Zvonkov ◽  
Nelly G. Gabeeva ◽  
Anna K. Morozova ◽  
Olga A. Gavrilina ◽  
Anna A. Sidorova ◽  
...  

Abstract Background. Aggressive lymphomas accounts for about 80% of primary gastrointestinal non-Hodgkin`s lymphomas (PGIAL). In adults the most common type is diffuse large B-cell lymphoma (DLBCL). Burkitt lymphoma (BL) is rare and mainly affects children. R-CHOP chemotherapy can induce favorable result for localized-stage. But the presence of adverse factors (AF) and advanced stage decrease the efficacy of this therapy: 3-year progression-free survival (PFS) and overall survival (OS) are about 50% and 60% respectively. The optimal treatment strategy for this pts still remains unknown. Aim. Efficacy and safety assessment of the modified chemotherapy protocol NHL-BFM-90 (m NHL-BFM-90 and LB-M-04) in the treatment of the PGIAL with advanced stage and AF. Patients. 74 previously untreated pts with PGIAL underwent mNHL-BFM-90 or LB-M-04 treatment between January 2002 and December 2015; out of them, 45 pts - primary gastric lymphoma (PGL), 29 pts - primary intestinal lymphoma (PIL); median age 39 years (range 14-72); age ≥60 years 10 pts (13,5%); M\F=44\30; stage >I 58 pts (78,3%); B-symptoms 30 pts (40,5%); Bulky disease 28 pts (37,8%). Patients characteristics in groups presented in Table 1. In the PIL group compared with the PGL was predominance of male (M\F=20\9 versus 21\24), stage II-IV (89,6% versus 71%), high level of LDH (72% versus 49%), Bulky disease (55% versus 27%). In pts with high Ki-67 (>40%) FISH test on t(8;14) was performed. Burkitt`s lymphoma diagnosed in 5 pts (11%) with PGL and 9 pts (31%) with PIL. In the PIL group more than half (58,6%) pts received surgical treatment before chemotherapy, and only 2 pts (4,4%) - in the PGL group. Of the 74 pts, 60 (81%) were diagnosed with DLBCL, 14 (19%) - BL. All pts with DLBCL received treatment according to the mNHL-BFM-90 program (2 courses A and 2 courses B) that was modified in the following way: doxorubicin (50mg\m2) was added on the third day of course A. All patients with BL received treatment according to the LB-M-04 program (2 courses A and 2 courses C) that was modified in the following way: doxorubicin (50mg\m2) was added on the third day of course A, methotrexate was administered on the 1st day of course C at a dose 1500mg/m2 for 12 hours. No one received consolidation radiotherapy from both groups. Results. In DLBCL group the overall response rate (ORR) was 95%. Complete remission (CR) was achieved in 38 from 40 pts (95%) in PGL, and 17 from 20 pts (85%) PIL. With a median follow-up of 74 months (range, 1-156) disease-free and overall survival of 60 pts with DLBCL constituted 86.7% and 91,7%, respectively. In BL group all pts achieved CR and alive with no signs of progression with a median follow-up of 110 months (range, 62-154). Hematologic toxicity of grade 3 and 4 was observed in 80% of pts. Severe complications became the reason for subsequent switch to CHOP therapy after 2 courses in 6 pts with PGL DLBCL. There was no treatment-related mortality. Conclusions. The mNHL-BFM-90 and LB-M-04 demonstrated acceptable toxicity and high efficacy in patients with PGIAL. Burkett lymphoma is not rare in adults with PGIAL and detection of t(8;14) can improve treatment outcomes. Table 1 Characteristics of the patients with aggressive primary gastrointestinal non-Hodgkin`s lymphomas. Table 1. Characteristics of the patients with aggressive primary gastrointestinal non-Hodgkin`s lymphomas. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.





Development ◽  
1983 ◽  
Vol 74 (1) ◽  
pp. 79-96
Author(s):  
Joanne T. Fujii ◽  
Gail R. Martin

Embryonal carcinoma cells were aggregated with cleavage stage mouse embryos, cultured briefly, and transferred as morulae to the uteri of pseudopregnant females. When midgestation foetuses were examined, many were morphologically abnormal. The severity of this abnormal development was correlated with the extent of contribution by embryonal carcinoma cells to the foetuses as indicated by GPI (glucose phosphate isomerase) analysis. This was true for all three of the cell lines studied, NG-2, PSA-1, and LT1-2D. The clear correlation between increasingly abnormal development and more extensive participation by the embryonal carcinoma cells was not observed in control experiments in which embryos of different stages of development were aggregated together. The data therefore suggest that the embryonal carcinoma cells studied here are unable to support normal development in the absence of a substantial number of host embryonic cells. It remains unclear whether this is a consequence of the karyotypic abnormalities of the cells tested, or whether it reflects a characteristic limitation in the ability of embryonal carcinoma cells to independently direct normal development. When aggregates were allowed to develop to term and the extent of chimaerism was examined in the live-born animals, it was found to be sporadic and limited. This is consistent with the results indicating that large contributions by embryonal carcinoma cells are not compatible with normal development at midgestation. The chimaerism observed in the live-born animals was comparable in both frequency and in tissue distribution to that generally obtained in other studies using either the aggregation or blastocyst injection techniques.



Development ◽  
1977 ◽  
Vol 38 (1) ◽  
pp. 77-92
Author(s):  
S. A. Iles ◽  
E. P. Evans

Karyotype and capacity for differentiation were determined in four transplantable teratomas, and their embryoid bodies, derived from C3H mouse embryos. An apparently normal karyotype was retained by one tumour and one subline that were able to differentiate into a wide range of tissues, but some chromosomal alterations were found in the two tumours and one subline that showed almost identical restrictions in their capacity for differentiation. Trisomy for chromosome 11 was shared by all three restricted tumours; two of the tumours had similar length changes in the same two chromosome (1 and 14) while the third was generally trisomic for four other chromosomes.



2020 ◽  
Vol 11 ◽  
Author(s):  
Lon J. Van Winkle

The osmolality of mouse oviductal fluid ranges from about 300 mOsmol/kg in the ampulla 0–3 h post coitus (h p.c.) to more than 350 mOsmol/kg in the isthmus 34–36 h p.c. Thus, it has been surprising to find that development of one-cell and cleavage-stage mouse embryos arrests in vitro in media exceeding 300 mOsmol/kg, and they develop best in unphysiological, hypotonic media. The glycine concentration in oviductal fluid can, however, rescue development in hypertonic media, so physiological conditions in vivo and in vitro likely work together to foster embryo well-being. Glycine acts on one-cell and cleavage-stage mouse embryos through the glycine-gated chloride channel, GLRA4, and uptake via the glycine neurotransmitter transporter, GLYT1. Since these processes lead to further signaling in neurons, the presence and function of such signaling in preimplantation embryos also should be investigated. The more we know about the interactions of physiological processes and conditions in vivo, the better we would be able to reproduce them in vitro. Such improvements in assisted reproductive technology (ART) could improve patient outcomes for IVF and potentially help prevent unwanted developmental abnormalities in early embryos, which might include undesirable epigenetic DNA and histone modifications. These epigenetic modifications may lead to transgenerational adult disorders such as metabolic syndrome and related conditions.



Author(s):  
Joachim T. Haug ◽  
Andreas Maas ◽  
Dieter Waloszek

ABSTRACTA detailed account of the morphology and ontogeny of the late Middle Cambrian crustacean †Henningsmoenicaris scutula is presented. Ten successive ontogenetic stages could be recognised in the material collected from various localities in Sweden. Morphogenetic changes include the development of a pair of stalked lateral eyes and the increase in the number and size of appendages and their setal armature. Notably, early stages lack ‘proximal endites’ on all post-antennular appendages; such a spine-bearing endite has previously been thought to appear simultaneously on these limbs. In †H. scutula a single functional endite appears on the third limb in an advanced stage; an additional endite appears on the second limb and, subsequently, further endites appear on more posterior limbs. Furthermore, a single specimen of †Sandtorpia vestrogothiensis gen. et sp. nov. is described. Based on this new information and data of other ‘Orsten’ taxa, particularly those assigned already to the early evolutionary lineage of Crustacea, a small-scale computer-based phylogenetic analysis was performed. This resolved the basal branchings of Crustacea s. l. as follows: †Oelandocarididae (=†Oelandocaris oelandica+†H. scutula+†S. vestrogothiensis)+(†Cambropachycopidae (=†Goticaris longispinosa+†Cambropachycope clarksoni)+ (†Martinssonia elongata+Labrophora (=†Phosphatocopina+Eucrustacea))). Plotting ontogenetic data on the phylogram and comparing the ground pattern at every node led to the detection of three peramorphic heterochronic events in the evolutionary lineage towards Eucrustacea.



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