Single-Molecule Fluorescent In Situ Hybridization (smFISH) on Whole-Mount Planarians.

Since the establishment of planarian species as laboratory models, investigation of molecular pathways has relied heavily on visualization of transcripts using in situ hybridization (ISH). ISH has revealed various aspects ranging from anatomical details of different organs to distribution of planarian stem cell populations and signaling pathways involved in their unique regenerative response. High-throughput sequencing techniques including single-cell approaches have allowed us to investigate gene expression and cell lineages in more detail.

Isolation and Processing of Bovine Oocytes for Small RNA Sequencing.

In modern biomedical research, mice have been the mammalian model system of choice to investigate molecular pathways for potential future medical applications. Over the last years, it has become clear that female mice employ an exceptional piRNA pathway-independent mechanism to neutralize transposon activity in the ovary. In other model organisms studied to date, the piRNA pathway is indispensable for efficient targeting of transposable elements and fertility in both males and females.

Ischemic tolerance and cardiac repair in the spiny mouse (Acomys).

Ischemic heart disease and by extension myocardial infarction is the primary cause of death worldwide, warranting regenerative therapies to restore heart function. Current models of natural heart regeneration are restricted in that they are not of adult mammalian origin, precluding the study of class-specific traits that have emerged throughout evolution, and reducing translatability of research findings to humans. Here, we present the spiny mouse (Acomys spp.

PIWIL3 Forms a Complex with TDRKH in Mammalian Oocytes.

P-element induced wimpy testis (PIWIs) are crucial guardians of genome integrity, particularly in germ cells. While mammalian PIWIs have been primarily studied in mouse and rat, a homologue for the human gene is absent in the Muridae family, and hence the unique function of PIWIL3 in germ cells cannot be effectively modeled by mouse knockouts. Herein, we investigated the expression, distribution, and interaction of PIWIL3 in bovine oocytes.

Tdrd6a Regulates the Aggregation of Buc into Functional Subcellular Compartments that Drive Germ Cell Specification.

Phase separation represents an important form of subcellular compartmentalization. However, relatively little is known about how the formation or disassembly of such compartments is regulated. In zebrafish, the Balbiani body (Bb) and the germ plasm (Gp) are intimately linked phase-separated structures essential for germ cell specification and home to many germ cell-specific mRNAs and proteins.

Live Cell Imaging Reveals the Relocation of dsRNA Binding Proteins Upon Viral Infection.

Viral infection triggers a range of plant responses such as the activation of the RNA interference (RNAi) pathway. The double-stranded RNA binding (DRB) proteins DRB3 and DRB4 are part of this pathway and aid in defending against DNA and RNA viruses, respectively. Using live cell imaging, we show that DRB2, DRB3, and DRB5 relocate from their uniform cytoplasmic distribution to concentrated accumulation in nascent viral replication complexes (VRC) that develop following cell invasion by viral RNA.

Piwi proteins and piRNAs in mammalian oocytes and early embryos: From sample to sequence.

The role of the Piwi/piRNA pathway during mammalian oogenesis has remained enigmatic thus far, especially since experiments with Piwi knockout mice did not reveal any phenotypic defects in female individuals. This is in striking contrast with results obtained from other species including flies and zebrafish. In mouse oocytes, however, only low levels of piRNAs are found and they are not required for their function.

Piwi proteins and piRNAs in mammalian oocytes and early embryos.

Germ cells of most animals critically depend on piRNAs and Piwi proteins. Surprisingly, piRNAs in mouse oocytes are relatively rare and dispensable. We present compelling evidence for strong Piwi and piRNA expression in oocytes of other mammals.

MUT-14 and SMUT-1 DEAD box RNA helicases have overlapping roles in germline RNAi and endogenous siRNA formation.

More than 2,000 C. elegans genes are targeted for RNA silencing by the mutator complex, a specialized small interfering RNA (siRNA) amplification module which is nucleated by the Q/N-rich protein MUT-16. The mutator complex localizes to Mutator foci adjacent to P granules at the nuclear periphery in germ cells.

Extremely stable Piwi-induced gene silencing in Caenorhabditis elegans.

In recent years, the Piwi pathway has been shown to regulate the silencing of mobile genetic elements. However, we know little about how Piwi pathways impose silencing and even less about trans-generational stability of Piwi-induced silencing. We demonstrate that the Caenorhabditis elegans Piwi protein PRG-1 can initiate an extremely stable form of gene silencing on a transgenic, single-copy target.