Discovery of abundant tsRNAs in mature sperm

In year 2012, by analyzing mature sperm small RNAs by RNA-seq in mice, we serendipitously found that the mature sperm contain a unique subset of tRNA–derived small RNAs (tsRNAs), mainly from 5′ transfer RNA halves and ranging in size from 29-34 nucleotides. tsRNAs show a drastic increase at late- spermatogenesis and post- spermatogenesis during epididymis maturation, suggesting regulated tRNA cleavage or/and selective concentrating mechanisms at these stages. tsRNAs are enriched in the sperm head and thus could be delivered into oocytes during fertilization. Like tRNAs, tsRNAs can be highly conserved leading us to propose that tsRNAs might serve as an ancient paternal element with evolutionarily conserved functions. (Cell Res 2012; Highlighted in Cell Res 2013)

Hidden information in sperm small RNAs: tRNA-derived small RNA (tsRNA)

Discovery of abundant tsRNAs in serum, across vertebrate species

After identifying tsRNAs as a new type of sperm small RNAs, we further discovered that tsRNAs are also abundantly and conservatively present in the serum of a wide range of vertebrates (from fish to human), and that serum tsRNAs are sensitive to pathological conditions (e.g. active infection) in mice, monkey and human beings. Importantly, we found that tsRNAs extracted from serum are more stable than chemically synthetic tsRNAs of the same sequence, suggesting RNA modifications in tsRNAs would contribute to their stabilization in the serum, thus represent another layer of information. (J Mol Cell Biol 2014,as Cover Story)
Daddy's diet haunt over generations, contributed by
sperm RNAs and sperm RNA modifications

Sperm tsRNAs, and RNA modifications, contribute to epigenetic inheritance of paternally acquired traits

Parallel with the discovery of tsRNAs, we are fascinated about the increasing evidence that certain acquired traits during paternal exposure can be “memorized” in the sperm and inherited by the offspring, leading to a resurrection of the ideas of "Lamarckian inheritance”. However, the underlining epigenetic mechanisms remain unclear. Recently, we found changes in both expression profiles and RNA modifications in sperm tsRNAs of mice with a paternal high-fat diet (HFD). By injecting various RNAs into normal zygotes, we found that sperm tsRNAs, along with their RNA modifications, represent a carrier of paternal epigenetic information that contributes to intergenerational inheritance of diet-induced metabolic disorder (Science 2016; Highlighted in Science, Nat Rev Genet, Cell Metab, Cell Res, FASEB J, Biol Reprod, Our discovery coincided with Oliver Rando lab's story on sperm tsRNAs in a low-protein diet model (Science 2016). We are now working to understand how sperm tsRNAs, along with their RNA modifications and responsible enzymes, affect embryo development mediate phenotypic in the offspring (Nat Rev Genet 2016; Trends Mol Med 2016). 
Mammalian pre-implantation embryos provide a unique opportunity to study how a single cell diverges from a totipotent state into different fates within limited rounds of cell division. There are two longstanding questions about this process: When does the first blastomere-to-blastomere asymmetry emerge and how does this relate to the development of distinct cell fates? It remains debatable whether the first bifurcation of cell fate emerges randomly at the morula stage, or if it has been initiated at earlier stages before morphological differences arise. Combining single-cell RNA-seq analysis and mathematical modeling, we recently showed that the very first symmetry-breaking process involves both chance separation and defined transcriptional circuits (Development 2015; Highlighted in Development 2015), a new framework for our future detailed investigations.

Single-cell analysis: bridging randomness and determinism

From our single-embryo transcriptome analysis, small biases at molecular level will inevitably emerge at the 2-cell embryo stage, following a binomial distribution due to the cleavage division. At this stage, the blastomere-to-blastomere distribution seems random but during subsequent zygotic transcriptional activation, a “bistable pattern” emerges in some genes. Several lineage specifiers show a strong bias between different blastomeres thus providing potential for further increased asymmetry subsequently (Development 2015). These observations suggest a scenario of how order is created from a seemingly random process through the differential triggering of existing master regulators by the emergence of their small bias. This is an inevitable process due to the imperfection of cleavage division.

Symmetry-breaking in mammalian early embryo: when and how?

Old dog, new tricks? novel function of Aquaporins beyond simple permeability

Discovery of Aquaporin-3 for rapid sperm osmoadaptation

In the journey from the male to female reproductive tract, mammalian sperm experiences a natural osmotic decrease (e.g., in mouse, from ~415 mOsm in the cauda epididymis to ~310 mOsm in the uterine cavity). On one hand, the hypotonic stress upon ejaculation is beneficial for the onset of mouse sperm motility (an evolutionary trait from fish sperm). However, this is a double-edged sword because the hypotonic stress could also cause potential harm to sperm function by inducing un-wanted swelling. To counteract this negative impact, mammalian sperm have acquired mechanisms for rapid transmembrane water movement to efficiently regulate cell volume. However, the specific sperm proteins responsible for this rapid osmoadaptation remain elusive. Using a knockout model, we discovered that Aquaporin-3 (AQP3) is an essential membrane protein for sperm regulatory volume decrease (RVD) upon physiological hypotonicity, balancing the “trade-off” between hypotonic induced sperm motility and cell swelling, thereby optimizing postcopulatory sperm behavior (Cell Res 2011).
However, we found that AQP3's role in sperm osmoadaptation cannot be fully explained by considering AQPs as inert pores simply for water permeability, the conventional view. We have discussed alternative possibilities for AQP3’s role in osmosensing or mechanosensing to regulate the subsequent RVD process (Acta Pharmacol Sin 2011).
Actually, in addition to our discovery, emerging evidence of other AQPs’ (such as AQP4, AQP5) having a role in cell volume regulation, can also not be fully explained by considering AQPs as inert pores simply for water permeability. We are now looking at the potential roles of AQPs as mechanosensors, especially in tissues with very low water permeability such as bladder, urethra etc.

AQP3's role as mechanosensor?