Hydrozoan sperm-specific SPKK motif-containing histone H2B variants stabilise chromatin with limited compaction.

Many animals achieve sperm chromatin compaction and stabilisation by replacing canonical histones with sperm nuclear basic proteins (SNBPs) such as protamines during spermatogenesis. Hydrozoan cnidarians and echinoid sea urchins lack protamines and have evolved a distinctive family of sperm-specific histone H2Bs (spH2Bs) with extended N termini rich in SPK(K/R) motifs. Echinoid sperm packaging is regulated by spH2Bs. Their sperm is negatively buoyant and fertilises on the sea floor. Hydroid cnidarians undertake broadcast spawning but their sperm properties are poorly characterised. We show that Hydractinia echinata and H. symbiolongicarpus sperm chromatin possesses higher stability than somatic chromatin, with reduced accessibility to transposase Tn5 integration and to endonucleases in vitro. In contrast, nuclear dimensions are only moderately reduced in mature Hydractinia sperm. Ectopic expression of spH2B in the background of H2B.1 knockdown results in downregulation of global transcription and cell cycle arrest in embryos, without altering their nuclear density. Taken together, SPKK-containing spH2B variants act to stabilise chromatin and silence transcription in Hydractinia sperm with only limited chromatin compaction. We suggest that spH2Bs could contribute to sperm buoyancy as a reproductive adaptation.

A genetically encoded sensor for visualizing leukotriene B4 gradients in vivo.

Leukotriene B4 (LTB4) is a potent lipid chemoattractant driving inflammatory responses during host defense, allergy, autoimmune and metabolic diseases. Gradients of LTB4 orchestrate leukocyte recruitment and swarming to sites of tissue damage and infection. How LTB4 gradients form and spread in live tissues to regulate these processes remains largely elusive due to the lack of suitable tools for monitoring LTB4 levels in vivo. Here, we develop GEM-LTB4, a genetically encoded green fluorescent LTB4 biosensor based on the human G-protein-coupled receptor BLT1. GEM-LTB4 shows high sensitivity, specificity and a robust fluorescence increase in response to LTB4 without affecting downstream signaling pathways. We use GEM-LTB4 to measure ex vivo LTB4 production of murine neutrophils. Transgenic expression of GEM-LTB4 in zebrafish allows the real-time visualization of both exogenously applied and endogenously produced LTB4 gradients. GEM-LTB4 thus serves as a broadly applicable tool for analyzing LTB4 dynamics in various experimental systems and model organisms.

Sperm Nuclear Basic Proteins of Marine Invertebrates.

In this chapter, a short evolutionary history and comparative analysis of sperm nuclear basic proteins (SNBPs) in marine invertebrates are presented based on some of the most recent publications in the field and building upon previously published reviews on the topic. Putative functions of SNBPs in sperm chromatin beyond DNA packaging will also be discussed with a primary focus on outstanding research questions.In somatic cells of all metazoans, DNA is packaged into tightly folded and dynamically accessible chromatin by canonical histones H2A, H2B, H3 and H4. Sperm chromatin of many animals, on the other hand, is organised by small yet structurally highly heterogeneous proteins called SNBPs, which can package sperm DNA on their own or in combination with each other. In extreme cases, sperm chromatin is condensed into a volume 6-10 times smaller than that of a somatic nucleus. SNBPs are classified into three major groups: H1 histone-type proteins (H-type SNBPs), protamines (P-type SNBPs) and protamine-like proteins (PL-type SNBPs). P-type SNBPs are mostly found in vertebrates, while PL-type SNBPs are ubiquitous in many invertebrate phyla. PL-type and P-type SNBPs evolved from histone H-type SNBP precursors through vertical evolution. Porifera, Ctenophora and Crustacea, Echinoidea (phylum Echinodermata) and Hydrozoa (phylum Hydrozoa) lack SNBPs. Echinoidea and Hydrozoa, however, evolved novel nucleosomal histone variants with specific roles during spermatogenesis. Seemingly, chromatin condensation plays a critical role in the silencing and tight packing of the genome within the sperm nucleus of most animals. However, the question of what necessitates the compaction of some sperm DNA beyond classical nucleosomal packaging while other sperm function using 'normal' histones remains unanswered to date.

The cnidarian Hydractinia echinata employs canonical and highly adapted histones to pack its DNA.

BACKGROUND: Cnidarians are a group of early branching animals including corals, jellyfish and hydroids that are renowned for their high regenerative ability, growth plasticity and longevity. Because cnidarian genomes are conventional in terms of protein-coding genes, their remarkable features are likely a consequence of epigenetic regulation. To facilitate epigenetics research in cnidarians, we analysed the histone complement of the cnidarian model organism Hydractinia echinata using phylogenomics, proteomics, transcriptomics and mRNA in situ hybridisations. RESULTS: We find that the Hydractinia genome encodes 19 histones and analyse their spatial expression patterns, genomic loci and replication-dependency. Alongside core and other replication-independent histone variants, we find several histone replication-dependent variants, including a rare replication-dependent H3.3, a female germ cell-specific H2A.X and an unusual set of five H2B variants, four of which are male germ cell-specific. We further confirm the absence of protamines in Hydractinia. CONCLUSIONS: Since no protamines are found in hydroids, we suggest that the novel H2B variants are pivotal for sperm DNA packaging in this class of Cnidaria. This study adds to the limited number of full histone gene complements available in animals and sets a comprehensive framework for future studies on the role of histones and their post-translational modifications in cnidarian epigenetics. Finally, it provides insight into the evolution of spermatogenesis.