Neuregulin and erbB Receptors Play a Critical Role in Neuronal Migration

AbstractThe migration of neuronal precursors along radial glial fibers is a critical step in the formation of the nervous system. In this report, we show that neuregulin–erbB receptor signaling plays a crucial role in the migration of cerebellar granule cells along radial glial fibers. Granule cells express neuregulin (NRG), and radial glia cells express erbB4 in the developing cerebellum and in vitro. When the glial erbB receptors are blocked, neurons fail to induce radial glia formation, and their migration along radial glial fibers is impaired. Moreover, soluble NRG is as effective as neuron–glia contact in the induction of radial glia formation. These results suggest that the activation of glial erbB4 by NRG is an early critical step in the neuronal migration program

Laser-Assisted In Vitro Fertilization Facilitates Fertilization of Vitrified-Warmed C57BL/6 Mouse Oocytes with Fresh and Frozen-Thawed Spermatozoa, Producing Live Pups

The utility of cryopreserved mouse gametes for reproduction of transgenic mice depends on development of assisted reproductive technologies, including vitrification of unfertilized mouse oocytes. Due to hardening of the zona pellucida, spermatozoa are often unable to penetrate vitrified-warmed (V-W) oocytes. Laser-assisted in vitro fertilization (LAIVF) facilitates fertilization by allowing easier penetration of spermatozoa through a perforation in the zona. We investigated the efficiency of V-W C57BL/6NTac oocytes drilled by the XYClone laser, compared to fresh oocytes. By using DAP213 for cryoprotection, 83% (1,470/1,762) of vitrified oocytes were recovered after warming and 78% were viable. Four groups were evaluated for two-cell embryo and live offspring efficiency: 1) LAIVF using V-W oocytes, 2) LAIVF using fresh oocytes, 3) conventional IVF using V-W oocytes and 4) conventional IVF using fresh oocytes. First, the groups were tested using fresh C57BL/6NTac spermatozoa (74% motile, 15 million/ml). LAIVF markedly improved the two-cell embryo efficiency using both V-W (76%, 229/298) and fresh oocytes (69%, 135/197), compared to conventional IVF (7%, 12/182; 6%, 14/235, respectively). Then, frozen-thawed C57BL/6NTac spermatozoa (35% motile, 15 million/ml) were used and LAIVF was again found to enhance fertilization efficiency, with two-cell embryo rates of 87% (298/343) using V-W oocytes (P<0.05, compared to fresh spermatozoa), and 73% (195/266) using fresh oocytes. Conventional IVF with frozen-thawed spermatozoa using V-W (6%, 10/168) and fresh (5%, 15/323) oocytes produced few two-cell embryos. Although live offspring efficiency following embryo transfer was greater with conventional IVF (35%, 18/51; LAIVF: 6%, 50/784), advantage was seen with LAIVF in live offspring obtained from total oocytes (5%, 50/1,010; conventional IVF: 2%, 18/908). Our results demonstrated that zona-drilled V-W mouse oocytes can be used for IVF procedures using both fresh and frozen-thawed spermatozoa, producing live pups. The ability to cryopreserve mouse gametes for LAIVF may facilitate management of large-scale transgenic mouse production facilities.National Institutes of Health (U.S.) (NIH National Research Service Award (T32-RR070036))National Institutes of Health (U.S.) (NIH National Cancer Institute Program Project (P01CA10451)

Probiotic Microbes Sustain Youthful Serum Testosterone Levels and Testicular Size in Aging Mice

The decline of circulating testosterone levels in aging men is associated with adverse health effects. During studies of probiotic bacteria and obesity, we discovered that male mice routinely consuming purified lactic acid bacteria originally isolated from human milk had larger testicles and increased serum testosterone levels compared to their age-matched controls. Further investigation using microscopy-assisted histomorphometry of testicular tissue showed that mice consuming Lactobacillus reuteri in their drinking water had significantly increased seminiferous tubule cross-sectional profiles and increased spermatogenesis and Leydig cell numbers per testis when compared with matched diet counterparts This showed that criteria of gonadal aging were reduced after routinely consuming a purified microbe such as L. reuteri. We tested whether these features typical of sustained reproductive fitness may be due to anti-inflammatory properties of L. reuteri, and found that testicular mass and other indicators typical of old age were similarly restored to youthful levels using systemic administration of antibodies blocking pro-inflammatory cytokine interleukin-17A. This indicated that uncontrolled host inflammatory responses contributed to the testicular atrophy phenotype in aged mice. Reduced circulating testosterone levels have been implicated in many adverse effects; dietary L. reuteri or other probiotic supplementation may provide a viable natural approach to prevention of male hypogonadism, absent the controversy and side-effects of traditional therapies, and yield practical options for management of disorders typically associated with normal aging. These novel findings suggest a potential high impact for microbe therapy in public health by imparting hormonal and gonad features of reproductive fitness typical of much younger healthy individuals.National Institutes of Health (U.S.) (Grant P30-ES002109)National Institutes of Health (U.S.) (Grant U01 CA164337)National Institutes of Health (U.S.) (Grant RO1CA108854

Distribution and Evolution of Supraglacial Lakes in Greenland during the 2016–2018 Melt Seasons

In recent decades, the melting of the Greenland Ice Sheet (GrIS) has become one of the major causes of global sea-level rise. Supraglacial lakes (SGLs) are typical hydrological features produced on the surface of the GrIS during the melt seasons. The existence and evolution of SGLs play an important role in the melting process of the ice sheet surface. To understand the distribution and recent changes of SGLs in Greenland, this study developed a random forest (RF) algorithm incorporating the texture and morphological features to automatically identify SGLs based on the Google Earth Engine (GEE) platform. Sentinel-2 imagery was used to map the SGLs inventory in Greenland during the 2016–2018 melt seasons and to explore the spatial and temporal variability characteristics of SGLs. Our results show changes in SGLs from 2016 to 2018, with the total area decreasing by ~1152.22 km2 and the number increasing by 1134; SGLs are mainly distributed in western Greenland (SW, CW, NW) and northeastern Greenland (NE), where the NE region has the largest number of observed SGLs and the largest SGL was with the surface area of 16.60 km2 (2016). SGLs were found to be most active in the area with the elevation of 800–1600 m and the slope of 0–5°, and showed a phenomenon of retreating to lower elevation areas and developing to steeper slope areas. Our work provided a method for rapid inventory of SGLs. This study will help monitor the mass balance of the GrIS and predict future rapid ice loss from Greenland

Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair

Searching for factors to improve knockin efficiency for therapeutic applications, biotechnology, and generation of non-human primate models of disease, we found that the strand exchange protein RAD51 can significantly increase Cas9-mediated homozygous knockin in mouse embryos through an interhomolog repair (IHR) mechanism. IHR is a hallmark of meiosis but only occurs at low frequencies in somatic cells, and its occurrence in zygotes is controversial. Using multiple approaches, we provide evidence for an endogenous IHR mechanism in the early embryo that can be enhanced by RAD51. This process can be harnessed to generate homozygotes from wild-type zygotes using exogenous donors and to convert heterozygous alleles into homozygous alleles without exogenous templates. Furthermore, we identify additional IHR-promoting factors and describe features of IHR events. Together, our findings show conclusive evidence for IHR in mouse embryos and describe an efficient method for enhanced gene conversion

Live offspring efficiency of laser-assisted IVF (LAIVF) and conventional IVF (Conv IVF) using vitrified-warmed (V-W) and fresh oocytes.

<p>Resorption is abbreviated as resorp, and implantation is abbreviated as implant.</p>a<p>Means ± SEM of % viable E17 embryos and % live pups were calculated from % viable E17 embryos or % live pups [# viable E17 embryos or # live pups/# two-cell embryos transferred (transf) *100] of each replicate; 2–4 replicates were included in each group where SEM is indicated.</p>b<p>One replicate included number of live pups obtained from 2 embryo transfer surgery recipients co-housed together post-operatively, with a combined total of 56 transferred two-cell embryos.</p>c<p>Mean ± SEM of % live offspring was calculated from % viable E17 embryos and live pups (# viable E17 embryos and live pups/total # two-cell embryos transferred *100) of each replicate; 3–7 replicates were included in each group where SEM is indicated.</p><p>NA indicates data not available; too few two-cell embryos were obtained via conventional IVF to evaluate live pup efficiency.</p

Summary of two-cell embryo and live offspring efficiencies of current and prior studies evaluating various techniques to improve penetration of the zona pellucida (ZP) of vitrified-warmed (V-W) mouse oocytes by spermatozoa.

<p>Laser-assisted IVF is abbreviated as LAIVF, and vitrification is abbreviated as vit.</p>a<p>Kunming mice are outbred, and C57BL/6 mice are inbred.</p>b<p>Percent two-cell embryos was calculated as # two-cell embryos/# total cells post-recovery and prior to treatment *100; data presented differently in the original manuscript were reconfigured to match the format of data presentation in the current study.</p>c<p>Percent live offspring was calculated as # live offspring/# two-cell embryos transferred to all recipients (pregnant and nonpregnant) *100; data presented differently in the original manuscript were reconfigured to match the format of data presentation in the current study.</p><p>NA indicates data not available.</p