Insulin-like growth factor system components expressed at the conceptus-maternal interface during the establishment of equine pregnancy

In many species, the insulin-like growth factors (IGF1 and IGF2), their receptors and IGF binding proteins play important roles in preparing the endometrium for implantation, and regulating conceptus growth and development. To determine whether the IGF system may contribute to conceptus-maternal interaction during equine pre-implantation development, we evaluated mRNA expression for IGF system components in conceptuses, and endometrium recovered from pregnant and cycling mares, on days 7, 14, 21 and 28 after ovulation. We also investigated expression of IGF1, IGF2 and their receptors 6 and 11 days after transfer of day 8 embryos to synchronous (day 8) or asynchronous (day 3) recipient mares. Expression of IGF1 and IGF2, IGF1R, IGF2R, INSR and IGFBPs 1, 2, 4 and 5 was evident in endometrium and conceptus membranes during days 7-28. Endometrial IGF2, INSR, IGFBP1 and IGFBP2 expression increased between days 7 and 28 of pregnancy. In conceptus membranes, expression of all IGF system components increased with developmental stage. Immunohistochemistry revealed strong expression of IGF1, IGF2 and IGF1R in both endometrium and conceptus membranes, whereas INSR was highly expressed in endometrium but barely detectable in the conceptus. Finally, a negatively asynchronous uterine environment retarded IGF1, IGF2 and INSR expression in the conceptus, whereas in the endometrium only INSR expression was altered by asynchrony. The presence of IGFs, their receptors and IGFBPs in the endometrium and conceptus during early equine pregnancy, and down-regulation in the conceptus following asynchronous embryo transfer, suggest a role in conceptus-maternal communication during the preparation for implantation

Річард Смоллі і знамениті «десять вересневих днів»

У вересні цього року виповнюється 27 років, як було відкрито фулерен — нову сфероподібну форму вуглецю. Ця подія буквально приголомшила вчених, які на той час вважали, що про елементарний вуглець їм відомо практично все. Історія відкриття цієї речовини досить незвичайна. Ще в 1971 р. можливість існування молекули фулерену була передбачена японським ученим Е. Осавою (E. Osawa), за два роки радянські хіміки-теоретики Д.А. Бочвар і О.Г. Гальперн квантово-хімічними розрахунками підтвердили стабільність молекули С60, і лише у 1985 р. Р. Смоллі, Р. Керл та Г. Крото експериментально отримали кластери із 60 атомів вуглецю в стійкій формі, яку вони пояснили структурою молекули у вигляді футбольного м’яча. Натхненнику цього відкриття, видатному вченому, нобелівському лауреату, активному популяризатору нанотехнологій Річарду Смоллі присвячено цей матеріал.В сентябре этого года исполняется 27 лет с момента открытия фуллерена — новой сферообразной формы углерода. Это событие буквально потрясло ученых, которые в то время считали, что об элементарном углероде им известно практически все. История открытия этого вещества довольно необычна. Еще в 1971 г. возможность су ществования молекулы фуллерена была предсказана японским ученым Е. Осавой (E. Osawa), через два года советские химики-теоретики Д.А. Бочвар и Е.Г. Гальперн с помощью квантово-химических расчетов подтвердили стабильность молекулы С60, и только в 1985 г. Р. Смолли, Р. Керл и Г. Крото экспериментально получили кластеры из 60 атомов углерода в устойчивой форме, которую они объяснили структурой молекулы в виде футбольного мяча. Вдохновителю этого открытия, выдающемуся ученому, нобелевскому лауреату, активному популяризатору нанотехнологий Ричарду Смолли посвящен этот материал.27 years since the discovery of fullerene, the new form of carbon, is observed in September of this year. This event has literally shocked scientists, who believed at that time that they know almost everything about the elementary carbon. History of this discovery is rather unusual. Long ago, in 1971 the possibility of the existence of a fullerene molecule was predicted by an young Japanese scientist E. Osawa. Then two Soviet chemists and theorists D.A. Bochvar and E.G. Hal pern confirm the stability of the C60 molecule using quantum chemical calculations, and in 1985 at last R. Smalley, R. Curl and H. Kroto experimentally obtained clusters of 60 carbon atoms in a sustainable form. They explained the structure of this molecule as the structure of a soccer ball. This material is devoted to the inspirer of this discovery, an outstanding scientist, Nobel laureate, active popularizer of nanotechnology — Richard Smalley

Vitrifying expanded equine embryos collapsed by blastocoel aspiration is less damaging than slow-freezing

The cryotolerance of equine blastocysts larger than 300 μm can be improved by aspirating blastocoele fluid prior to vitrification; however, it is not known whether blastocoele aspiration also enables successful slow-freezing. The aim of this study was therefore to determine whether slow-freezing of expanded equine embryos following blastocoele collapse was more or less damaging than vitrification. Grade 1 blastocysts recovered on day 7 or 8 after ovulation were measured (>300-550 μm, n = 14 and > 550 μm, n = 19) and blastocoele fluid was aspirated prior to slow-freezing in 10% glycerol (n = 14), or vitrification (n = 13) in 16.5% ethylene glycol/16.5% DMSO/0.5 M sucrose. Immediately after thawing or warming, embryos were cultured for 24 h at 38 °C and then graded and measured to assess re-expansion. Control embryos (n = 6) were cultured for 24 h following aspiration of blastocoel fluid, without cryopreservation or exposure to cryoprotectants. Subsequently, embryos were stained to assess live/dead cell proportion (DAPI/TOPRO-3), cytoskeleton quality (Phalloidin) and capsule integrity (WGA). For 300-550 μm embryos, quality grade and re-expansion were impaired after slow-freezing but not affected by vitrification. Slow-freezing embryos >550 μm induced additional cell damage as indicated by a significant increase in dead cell proportion and disruption of the cytoskeleton; neither of these changes were observed in vitrified embryos. Capsule loss was not a significant consequence of either freezing method. In conclusion, slow-freezing of expanded equine blastocysts collapsed by blastocoel aspiration compromises post-thaw embryo quality more than vitrification

In Vitro-Produced Equine Blastocysts Exhibit Greater Dispersal and Intermingling of Inner Cell Mass Cells than In Vivo Embryos

In vitro production (IVP) of equine embryos is increasingly popular in clinical practice but suffers from higher incidences of early embryonic loss and monozygotic twin development than transfer of in vivo derived (IVD) embryos. Early embryo development is classically characterized by two cell fate decisions: (1) first, trophectoderm (TE) cells differentiate from inner cell mass (ICM); (2) second, the ICM segregates into epiblast (EPI) and primitive endoderm (PE). This study examined the influence of embryo type (IVD versus IVP), developmental stage or speed, and culture environment (in vitro versus in vivo) on the expression of the cell lineage markers, CDX-2 (TE), SOX-2 (EPI) and GATA-6 (PE). The numbers and distribution of cells expressing the three lineage markers were evaluated in day 7 IVD early blastocysts ( n = 3) and blastocysts ( n = 3), and in IVP embryos first identified as blastocysts after 7 (fast development, n = 5) or 9 (slow development, n = 9) days. Furthermore, day 7 IVP blastocysts were examined after additional culture for 2 days either in vitro ( n = 5) or in vivo (after transfer into recipient mares, n = 3). In IVD early blastocysts, SOX-2 positive cells were encircled by GATA-6 positive cells in the ICM, with SOX-2 co-expression in some presumed PE cells. In IVD blastocysts, SOX-2 expression was exclusive to the compacted presumptive EPI, while GATA-6 and CDX-2 expression were consistent with PE and TE specification, respectively. In IVP blastocysts, SOX-2 and GATA-6 positive cells were intermingled and relatively dispersed, and co-expression of SOX-2 or GATA-6 was evident in some CDX-2 positive TE cells. IVP blastocysts had lower TE and total cell numbers than IVD blastocysts and displayed larger mean inter-EPI cell distances; these features were more pronounced in slower-developing IVP blastocysts. Transferring IVP blastocysts into recipient mares led to the compaction of SOX-2 positive cells into a presumptive EPI, whereas extended in vitro culture did not. In conclusion, IVP equine embryos have a poorly compacted ICM with intermingled EPI and PE cells; features accentuated in slowly developing embryos but remedied by transfer to a recipient mare

Erratum to: 36th International Symposium on Intensive Care and Emergency Medicine

[This corrects the article DOI: 10.1186/s13054-016-1208-6.]

Expression of leukaemia inhibitory factor at the conceptus-maternal interface during preimplantation development and in the endometrium during the oestrous cycle in the mare

Leukaemia inhibitory factor (LIF) plays a critical role in blastocyst development and implantation in several species. The present study investigated mRNA and protein expression for LIF, as well as the low-affinity LIF receptor (LIFR) and interleukin-6 signal transducer (IL6ST), in equine endometrium, trophoblast and histotroph during early pregnancy and in the endometrium during the oestrous cycle. Endometrial LIF mRNA expression was upregulated after Day 21 of pregnancy, whereas LIF immunoreactivity increased in the endometrium on Day 28. Expression of LIF mRNA in the yolk sac membrane increased from Day 21 of pregnancy, whereas LIF immunoreactivity increased from Day 28 in the trophoblast. LIFR and IL6ST mRNA was expressed in the endometrium during both the oestrous cycle and early pregnancy and, although LIFR and IL6ST protein were localised to the glandular epithelium during the cycle and first 14 days of pregnancy, from Day 21 they were located in the luminal epithelium. Trophoblast expression of LIFR and IL6ST increased as pregnancy proceeded. In conclusion, LIF expression increased at the conceptus-maternal interface during capsule attenuation. Because contemporaneous upregulation of both LIFR and IL6ST was also observed in the trophoblast, we propose that LIF plays an important role in the development of endometrial receptivity for trophoblast growth, apposition and adhesion in mares