DNA reveals long-distance partial migratory behavior in a cryptic owl lineage

10.1186/s40657-015-0035-9Avian Research612

Viewing early life without labels: optical approaches for imaging the early embryo

OnlinePublEmbryo quality is an important determinant of successful implantation and a resultant live birth. Current clinical approaches for evaluating embryo quality rely on subjective morphology assessments or an invasive biopsy for genetic testing. However, both approaches can be inherently inaccurate and crucially, fail to improve the live birth rate following the transfer of in vitro produced embryos. Optical imaging offers a potential non-invasive and accurate avenue for assessing embryo viability. Recent advances in various label-free optical imaging approaches have garnered increased interest in the field of reproductive biology due to their ability to rapidly capture images at high resolution, delivering both morphological and molecular information. This burgeoning field holds immense potential for further development, with profound implications for clinical translation. Here, our review aims to: (1) describe the principles of various imaging systems, distinguishing between approaches that capture morphological and molecular information, (2) highlight the recent application of these technologies in the field of reproductive biology, and (3) assess their respective merits and limitations concerning the capacity to evaluate embryo quality. Additionally, the review summarizes challenges in the translation of optical imaging systems into routine clinical practice, providing recommendations for their future development. Finally, we identify suitable imaging approaches for interrogating the mechanisms underpinning successful embryo development.Darren J.X. Chow, Tiffany C.Y. Tan, Avinash Upadhya, Megan Lim, Kishan Dholakia, and Kylie R. Dunnin

Novel genome and genome-wide SNPs reveal early fragmentation effects in an edge-tolerant songbird population across an urbanized tropical metropolis

10.1038/s41598-018-31074-5Scientific Reports811280

A comprehensive assessment of diversity loss in a well-documented tropical insect fauna: almost half of Singapore’s butterfly species extirpated in 160 years

Insects as a group are suffering rapid declines in many parts of the world but are also poorly studied relative to vertebrate taxa. Comprehensive assessments of insect declines must account for both detected and undetected species. We studied extirpations among butterflies, a particularly well-known insect group, in the highly developed and biologically well-surveyed island city-state of Singapore. Building on existing butterfly species lists, we collated museum and naturalist records over the last two centuries and used statistical models to estimate the total extirpation rate since the first major collections in 1854. In addition, we compiled a set of traits for each butterfly species and explored how they relate to species discovery and extirpation. With a database of 413 native species, 132 (32%) of which are recorded as extirpated in Singapore, we used a statistical model to infer that, in addition, 104 unknown species (95% CI 60–162) were likely extirpated before they were ever discovered, suggesting a total extirpation rate of 46% (41–51%). In the trait analyses, we found that butterfly species that were discovered later were weakly associated with rarer larval host plants and smaller wingspans, while species that persisted for longer were weakly associated with higher larval host plant abundance and lower forest-dependence. This exercise is one of the first to offer a holistic estimate of extirpations for a group of insects by accounting for undetected extirpations. It suggests that extirpations among insects, specifically in the tropics, may be higher than naïve estimates based only on known records.Meryl Theng, Wan F.A.Jusoh, Anuj Jain, Blanca Huertas, David J.X.Tan, Hui Zhen Tan ... et al

Vitrification within a nanoliter volume: oocyte and embryo cryopreservation within a 3D photopolymerized device

Corrected by: Correction to: Vitrification within a nanoliter volume: oocyte and embryo cryopreservation within a 3D photopolymerized device, in Vol. 39(10):2435 2022. The correct wavelength of the laser used to excite an intracellular fluorophore should be “405 nm” and not “305 nm”. The original article has been corrected.Purpose: Vitrification permits long-term banking of oocytes and embryos. It is a technically challenging procedure requiring direct handling and movement of cells between potentially cytotoxic cryoprotectant solutions. Variation in adherence to timing, and ability to trace cells during the procedure, affects survival post-warming. We hypothesized that minimizing direct handling will simplify the procedure and improve traceability. To address this, we present a novel photopolymerized device that houses the sample during vitrification. Methods: The fabricated device consisted of two components: the Pod and Garage. Single mouse oocytes or embryos were housed in a Pod, with multiple Pods docked into a Garage. The suitability of the device for cryogenic application was assessed by repeated vitrification and warming cycles. Oocytes or early blastocyst-stage embryos were vitrified either using standard practice or within Pods and a Garage and compared to non-vitrified control groups. Post-warming, we assessed survival rate, oocyte developmental potential (fertilization and subsequent development) and metabolism (autofluorescence). Results: Vitrification within the device occurred within ~ 3 nL of cryoprotectant: this volume being ~ 1000-fold lower than standard vitrification. Compared to standard practice, vitrification and warming within our device showed no differences in viability, developmental competency, or metabolism for oocytes and embryos. The device housed the sample during processing, which improved traceability and minimized handling. Interestingly, vitrification-warming itself, altered oocyte and embryo metabolism. Conclusion: The Pod and Garage system minimized the volume of cryoprotectant at vitrification—by ~ 1000-fold—improved traceability and reduced direct handling of the sample. This is a major step in simplifying the procedure.Suliman H. Yagoub, Megan Lim, Tiffany C. Y. Tan, Darren J. X. Chow, Kishan Dholakia, Brant C. Gibson, Jeremy G. Thompson, Kylie R. Dunnin

Optical imaging of cleavage stage bovine embryos using hyperspectral and confocal approaches reveals metabolic differences between on-time and fast-developing embryos

The assessment of embryo quality aims to enhance subsequent pregnancy and live birth outcomes. Metabolic analysis of embryos has immense potential in this regard. As a step towards this goal, here we assess the metabolism of bovine embryos using label-free optical imaging. We compared embryos defined as either on-time or fast-developing, as fast dividing embryos are more likely to develop to the blastocyst stage. Specifically, bovine embryos at 48 (Day 2) and 96 (Day 4) hours post fertilization were fixed and separated based on morphological assessment: on-time (Day 2: 2 cell; Day 4: 5-7 cell) or fast-developing (Day 2: 3-7 cell; Day 4: 8-16 cell). Embryos with different developmental rates on Day 2 and Day 4 were correlated with metabolic activity and DNA damage. Confocal microscopy was used to assess metabolic activity by quantification of cellular autofluorescence specific for the endogenous fluorophores NAD(P)H and FAD with a subsequent calculation of the optical redox ratio. Separately, hyperspectral microscopy was employed to assess a broader range of endogenous fluorophores. DNA damage was determined using γH2AX immunohistochemistry. Hyperspectral imaging showed significantly lower abundance of endogenous fluorophores in fast-developing compared to on-time embryos on Day 2, indicating a lower metabolic activity. On Day 4 of development there was no difference in the abundance of FAD between on-time and fast-developing embryos. There was, however, significantly higher levels of NAD(P)H in fast-developing embryos leading to a significantly lower optical redox ratio when compared to on-time embryos. Collectively, these results demonstrate that fast-developing embryos present a 'quiet' metabolic pattern on Day 2 and Day 4 of development, compared to on-time embryos. There was no difference in the level of DNA damage between on-time and fast-developing embryos on either day of development. To our knowledge, this is the first collective use of confocal and hyperspectral imaging in cleavage-stage bovine embryos in the absence of fluorescent tags.Clara AnaSantos Monteiro, Darren J.X.Chow, Gabriela Ramos Leal, Tiffany CY.Tan, Ana Maria Reis Ferreira, Jeremy G.Thompson, Kylie R.Dunnin