Characterization of longitudinal canal tissue in the acorn barnacle Amphibalanus amphitrite

Abstract

The morphology and composition of tissue located within parietal shell canals of the barnacle Amphibalanus amphitrite are described. Longitudinal canal tissue nearly spans the length of side shell plates, terminating near the leading edge of the specimen basis in proximity to female reproductive tissue located throughout the peripheral sub-mantle region, i.e. mantle parenchyma. Microscopic examination of stained longitudinal canal sections reveal the presence of cell nuclei as well as an abundance of micron-sized spheroids staining positive for basic residues and lipids. Spheroids with the same staining profile are present extensively in ovarioles, particularly within oocytes which are readily identifiable at various developmental stages. Mass spectrometry analysis of longitudinal canal tissue compared to tissue collected from the mantle parenchyma reveals a nearly 50% overlap of the protein profile with the greatest number of sequence matches to vitellogenin, a glycolipoprotein playing a key role in vitellogenesis–yolk formation in developing oocytes. The morphological similarity and proximity to female reproductive tissue, combined with mass spectrometry of the two tissues, provides compelling evidence that one of several possible functions of longitudinal canal tissue is supporting the female reproductive system of A. amphitrite, thus expanding the understanding of the growth and development of this sessile marine organism.

Proteomics Reveals Distinct Changes Associated with Increased Gamma Radiation Resistance in the Black Yeast Exophiala dermatitidis

“…PCT facilitates protein unfolding and enzymatic access to cleavage sites, reducing the time needed for digestion [ 32 , 33 , 34 , 35 , 36 ]. Lysate containing ten micrograms of protein was added to a tube designed for use in the Barocycler (Pressure Biosciences Inc., Easton, MA, USA) with a 1:50 ratio of sequencing grade modified trypsin (Promega, Madison, WI, USA) as previously described [ 32 , 37 , 38 ]. The volume was adjusted to 150 µL 100 mM ammonium bicarbonate + 10% n-propanol.…”

Distribution of Select Cement Proteins in the Acorn Barnacle Amphibalanus amphitrite

“…Formalin fixed paraffin embedded tissue sections were prepared for proteome profile analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS) with analytical-grade reagents (Fisher Optima grade, etc.). A method for protein sample recovery from FFPE sections (Föll et al, 2018) was adapted for use with pressure cycling technology (PCT) (Tao et al, 2007;López-Ferrer et al, 2008) for barnacle protein extracts as previously described (Fears et al, 2018;Wang et al, 2018;Schultzhaus et al, 2019). Tissue area was measured and sections were deparaffinized and rehydrated (3 × 5 min in xylene; 2 × 5 min in 99% EtOH; 20 s each in 99, 96, 70, and 50% EtOH) and scraped off glass slides into 1.5-mL protein low-bind Eppendorf tubes.…”

The complex barnacle perfume: identification of waterborne pheromone homologues inBalanus improvisusand their differential expression during settlement

“…Originally, Endo et al (2009) reported the results of a preliminary study that detected several proteins with weights around 32 kDa in B. amphitrite-conditioned seawater that exhibited settlement-inducing activity. In addition, several WSP sequences were found in the water-soluble fraction of proteins rinsed from the cement (So et al 2017) and among the proteins from longitudinal canal and submantle tissues of B. amphitrite (Wang et al 2018). In adults of T. j. formosana, Lin et al (2014) reported the presence of two different WSP sequences displaying different expression in prosoma and basis, suggesting the existence of more than one WSP also in this species.…”

“…Since the initial publication of the discovery of WSP in 2009 (Endo et al 2009), there have been several studies indicating that more than one WSP gene is present in some barnacle species (Lin et al 2014;So et al 2017;Wang et al 2018). Originally, Endo et al (2009) reported the results of a preliminary study that detected several proteins with weights around 32 kDa in B. amphitrite-conditioned seawater that exhibited settlement-inducing activity.…”

“…Since the original publication of the discovery of WSP in 2009 (Endo et al 2009), there have been no follow-up studies to further elucidate the structure and function of the WSP. However, it was recently indicated that more than one WSP gene is present in Tetraclita japonica formosana (Lin et al 2014) and B. amphitrite (So et al 2017;Wang et al 2018), but the available information is extremely scarce and only one of the sequences has been published (Lin et al 2014) and none are deposited in public databases. Moreover, the study by Endo et al (2009) mentioned preliminary unpublished data showing that several proteins with a molecular mass of around 32 kDa detected in barnacle-conditioned seawater have settlement-inducing activity.…”