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Characterization of larval neurons in Nematostella vectensis

Authors: Brandic, Ana;

Characterization of larval neurons in Nematostella vectensis

Abstract

Der zu den Cnidaria gehörenden Modellorganismus Nematostella vectensis, eine Seeanemone, durchläuft eine leichte Form von Metamorphose von der frei-schwimmenden Planula-Larve bis hin zum sessilen Polypen, wobei die Fähigkeit zu schwimmen verloren geht. Das Nervensystem von Nematostella besteht aus einem diffusen Nervennetz. Cole et al., 2024 veröffentlichte Single-Cell RNA Sequencing Daten, die prognostizieren, dass in Nematostella larvenspezifische Neuronen existieren, die nach dem Übergang ins Polypenstadium nicht mehr vorhanden sind. Es wurden drei larvenspezifische neuronale Zustände (N1.L1, N1.L2 und N1.L3) und ein larvenspezifischer sekretorischer Zustand (S1.L1) beschrieben, von denen nur einige Zellen des larvalen neuronalen Zustands N1.L3 im adulten Polypen bestehen bleiben. In dieser Arbeit setzen wir die ersten Schritte zur Charakterisierung dieser larvalen Neuronen. Dazu wurde die grundlegende Arbeit zur Erzeugung von sieben verschiedenen Larven-spezifischen neuronalen transgenen Reporterlinien durchgeführt. Anhand eines repräsentativen Gens pro Larvenstadium zeigen wir, dass diese Neuronen tatsächlich nur in Nematostella-Larven exprimiert werden. Dies wurde quantitativ durch Fluorescence-Live-Cell-Imaging verifiziert und zusätzlich durch Immunostaining-Experimente bestätigt. Mittels In-situ-Hybridisierungen verschiedener Marker-Gene der drei larvalen neuronalen Zustände bestätigen wir die Expression im Larvenstadium, zeigen aber, dass Gene der larvalen neuronalen Zustände N1.L1 und N1.L2 nicht in primären Polypen exprimiert werden. Darüber hinaus zeigen wir, dass es zusätzlich zur Genexpression im Larvenstadium ein für den larvalen neuronalen Zustand N1.L3 spezifisches Marker-Gen gibt, das auch im primären Polypen exprimiert wird.

The cnidarian model organism Nematostella vectensis, the starlet sea anemone, undergoes a subtle type of metamorphosis from the free-swimming planula stage to the sessile polyp stage where the ability to swim is lost. The nervous system of Nematostella consists of a diffuse nerve net. Single cell RNA sequencing data published by Cole et al., 2024 proposed that in Nematostella larva-specific neurons exist which disappear after transition into the polyp stage. There were three larval-specific neural states (N1.L1, N1.L2 and N1.L3) and one larval-specific secretory state (S1.L1) described of which only some cells of the larval neural state N1.L3 persist into the adult polyp. Here we aim to initiate the process of characterizing these larval neurons. Therefore, the fundamental work to generate seven different larval neural specific transgenic reporter lines was performed. Focusing on one representative gene candidate per larval state we show that these neurons are truly only expressed in Nematostella larvae. This was quantitatively verified by fluorescence live-cell imaging and additionally confirmed through immunostaining assays. Using in situ hybridizations of various marker genes of the three larval neural states we confirm expression in larvae but show that there is no expression of the larval neural states N1.L1 and N1.L2 in primary polyps. Moreover, we show that in addition to gene expression at the larval stage, there is one marker gene specific for the larval neural state N1.L3 that is also expressed in the primary polyp.

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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