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The Rise and Fall of TRP-N, an Ancient Family of Mechanogated Ion Channels, in Metazoa.

Schüler A, Schmitz G, Reft A, Özbek S, Thurm U, Bornberg-Bauer E - Genome Biol Evol (2015)

Bottom Line: In flies, the transient-receptor-potential N protein (TRP-N) was found to be a cilia-associated mechanoreceptor.We propose that these new candidate proteins help explain the sensory complexity of Cnidaria which has been previously observed but so far has lacked a molecular underpinning.Also, the ancient appearance of TRP-N supports a common origin of important components of the nervous systems in Ctenophores, Cnidaria, and Bilateria.

View Article: PubMed Central - PubMed

Affiliation: Institute for Evolution and Biodiversity, University of Muenster, Germany.

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Expression and localization of individual TRP-N molecules in Hydra magnipapillata. (A–F) In situ hybridizations. (A) In situ hybridization for TRP-N1 showing expression of transcripts in the nests of developing nematocytes. Scale bar = 100 µm. Inset shows a close up of TRP-N1 positive individual nematocyte nests in the body column of the same animal as in (A). Scale bar = 50 µm. (B) In situ hybridization for TRP-N2 showing expression of transcripts in developing nematocytes, Scale bar = 100 µm. (C–F) Immunostaining with TRP-N4 antibodies. (C) Overview of localization of TRP-N4 protein in tentacles and body column by antibody staining. Scale bar = 100 µm. (D) Localization of TRP-N4 in the nests of developing nematocysts in the body column. Scale bar = 20 µm. (E) Localization of TRP-N4 in tentacles where it surrounds individual mature nematocyst capsules. Scale bar = 10 µm. (F) Transmitted light view on nematocysts in (E) showing localization around various nematocysts. Scale bar = 10 µm. (G–M) Immunostaining of cnidocils with pan-TRP-N antibody. (G) A single nematocyte with desmoneme and stained cnidocil. Scale bar = 5 µm. (H) Staining of cnidocils in tentacle. Scale Bar = 10 µm. (I) Surface view of tentacle indicating spot-like staining in the basal part of cnidocils. Scale Bar = 10 µm. (J) Staining of isolated cnidocil showing a gradient toward the base of the cilium. Scale bar = 10 µm. (K) Costaining of TRP-N in the cnidocil (red) and of phalloidin in the stereocilia (green). Scale bar = 5 µm. (L) Same nematocyte as in (E), showing phalloidin staining of the stereocilia only. Scale bar = 5 µm. (M) Scanning electron microscope of tentacle surface with cnidocil. Scale bar = 2 µm.
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evv091-F3: Expression and localization of individual TRP-N molecules in Hydra magnipapillata. (A–F) In situ hybridizations. (A) In situ hybridization for TRP-N1 showing expression of transcripts in the nests of developing nematocytes. Scale bar = 100 µm. Inset shows a close up of TRP-N1 positive individual nematocyte nests in the body column of the same animal as in (A). Scale bar = 50 µm. (B) In situ hybridization for TRP-N2 showing expression of transcripts in developing nematocytes, Scale bar = 100 µm. (C–F) Immunostaining with TRP-N4 antibodies. (C) Overview of localization of TRP-N4 protein in tentacles and body column by antibody staining. Scale bar = 100 µm. (D) Localization of TRP-N4 in the nests of developing nematocysts in the body column. Scale bar = 20 µm. (E) Localization of TRP-N4 in tentacles where it surrounds individual mature nematocyst capsules. Scale bar = 10 µm. (F) Transmitted light view on nematocysts in (E) showing localization around various nematocysts. Scale bar = 10 µm. (G–M) Immunostaining of cnidocils with pan-TRP-N antibody. (G) A single nematocyte with desmoneme and stained cnidocil. Scale bar = 5 µm. (H) Staining of cnidocils in tentacle. Scale Bar = 10 µm. (I) Surface view of tentacle indicating spot-like staining in the basal part of cnidocils. Scale Bar = 10 µm. (J) Staining of isolated cnidocil showing a gradient toward the base of the cilium. Scale bar = 10 µm. (K) Costaining of TRP-N in the cnidocil (red) and of phalloidin in the stereocilia (green). Scale bar = 5 µm. (L) Same nematocyte as in (E), showing phalloidin staining of the stereocilia only. Scale bar = 5 µm. (M) Scanning electron microscope of tentacle surface with cnidocil. Scale bar = 2 µm.

Mentions: We next aimed to localize expression of TRP-Ns and distinguish possible spatial differences in their expression. In situ hybridization (see Materials and Methods for details) was used to localize TRP-N1 and TRP-N2 mRNAs. Due to a lack of unique sequences, specific probes for the other TRP-N mRNAs were not feasible. Expression patterns of both hydra-TRP-N1 and hydra-TRP-N2 reveal a clear restriction to developing nematocytes in the body column of hydra, with hydra-TRP-N1 showing a stronger signal than Hydra-TRP-N2 (see fig. 3A and B). This strong hydra-TRP-N1 signal (see fig. 3A) refers to cell clusters of nascent, premature nematocytes (Fawcett et al. 1959). These clusters are known to break up upon maturation and the isolated nematocytes subsequently migrate toward the tentacles (Campbell and Marcum 1980). This expression pattern of hydra-TRP-N1 and hydra-TRP-N2 resembles the ones of most nematocyst-associated genes, such as minicollagens, which are downregulated in the head region (Beckmann and Özbek 2012).Fig. 3.—


The Rise and Fall of TRP-N, an Ancient Family of Mechanogated Ion Channels, in Metazoa.

Schüler A, Schmitz G, Reft A, Özbek S, Thurm U, Bornberg-Bauer E - Genome Biol Evol (2015)

Expression and localization of individual TRP-N molecules in Hydra magnipapillata. (A–F) In situ hybridizations. (A) In situ hybridization for TRP-N1 showing expression of transcripts in the nests of developing nematocytes. Scale bar = 100 µm. Inset shows a close up of TRP-N1 positive individual nematocyte nests in the body column of the same animal as in (A). Scale bar = 50 µm. (B) In situ hybridization for TRP-N2 showing expression of transcripts in developing nematocytes, Scale bar = 100 µm. (C–F) Immunostaining with TRP-N4 antibodies. (C) Overview of localization of TRP-N4 protein in tentacles and body column by antibody staining. Scale bar = 100 µm. (D) Localization of TRP-N4 in the nests of developing nematocysts in the body column. Scale bar = 20 µm. (E) Localization of TRP-N4 in tentacles where it surrounds individual mature nematocyst capsules. Scale bar = 10 µm. (F) Transmitted light view on nematocysts in (E) showing localization around various nematocysts. Scale bar = 10 µm. (G–M) Immunostaining of cnidocils with pan-TRP-N antibody. (G) A single nematocyte with desmoneme and stained cnidocil. Scale bar = 5 µm. (H) Staining of cnidocils in tentacle. Scale Bar = 10 µm. (I) Surface view of tentacle indicating spot-like staining in the basal part of cnidocils. Scale Bar = 10 µm. (J) Staining of isolated cnidocil showing a gradient toward the base of the cilium. Scale bar = 10 µm. (K) Costaining of TRP-N in the cnidocil (red) and of phalloidin in the stereocilia (green). Scale bar = 5 µm. (L) Same nematocyte as in (E), showing phalloidin staining of the stereocilia only. Scale bar = 5 µm. (M) Scanning electron microscope of tentacle surface with cnidocil. Scale bar = 2 µm.
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evv091-F3: Expression and localization of individual TRP-N molecules in Hydra magnipapillata. (A–F) In situ hybridizations. (A) In situ hybridization for TRP-N1 showing expression of transcripts in the nests of developing nematocytes. Scale bar = 100 µm. Inset shows a close up of TRP-N1 positive individual nematocyte nests in the body column of the same animal as in (A). Scale bar = 50 µm. (B) In situ hybridization for TRP-N2 showing expression of transcripts in developing nematocytes, Scale bar = 100 µm. (C–F) Immunostaining with TRP-N4 antibodies. (C) Overview of localization of TRP-N4 protein in tentacles and body column by antibody staining. Scale bar = 100 µm. (D) Localization of TRP-N4 in the nests of developing nematocysts in the body column. Scale bar = 20 µm. (E) Localization of TRP-N4 in tentacles where it surrounds individual mature nematocyst capsules. Scale bar = 10 µm. (F) Transmitted light view on nematocysts in (E) showing localization around various nematocysts. Scale bar = 10 µm. (G–M) Immunostaining of cnidocils with pan-TRP-N antibody. (G) A single nematocyte with desmoneme and stained cnidocil. Scale bar = 5 µm. (H) Staining of cnidocils in tentacle. Scale Bar = 10 µm. (I) Surface view of tentacle indicating spot-like staining in the basal part of cnidocils. Scale Bar = 10 µm. (J) Staining of isolated cnidocil showing a gradient toward the base of the cilium. Scale bar = 10 µm. (K) Costaining of TRP-N in the cnidocil (red) and of phalloidin in the stereocilia (green). Scale bar = 5 µm. (L) Same nematocyte as in (E), showing phalloidin staining of the stereocilia only. Scale bar = 5 µm. (M) Scanning electron microscope of tentacle surface with cnidocil. Scale bar = 2 µm.
Mentions: We next aimed to localize expression of TRP-Ns and distinguish possible spatial differences in their expression. In situ hybridization (see Materials and Methods for details) was used to localize TRP-N1 and TRP-N2 mRNAs. Due to a lack of unique sequences, specific probes for the other TRP-N mRNAs were not feasible. Expression patterns of both hydra-TRP-N1 and hydra-TRP-N2 reveal a clear restriction to developing nematocytes in the body column of hydra, with hydra-TRP-N1 showing a stronger signal than Hydra-TRP-N2 (see fig. 3A and B). This strong hydra-TRP-N1 signal (see fig. 3A) refers to cell clusters of nascent, premature nematocytes (Fawcett et al. 1959). These clusters are known to break up upon maturation and the isolated nematocytes subsequently migrate toward the tentacles (Campbell and Marcum 1980). This expression pattern of hydra-TRP-N1 and hydra-TRP-N2 resembles the ones of most nematocyst-associated genes, such as minicollagens, which are downregulated in the head region (Beckmann and Özbek 2012).Fig. 3.—

Bottom Line: In flies, the transient-receptor-potential N protein (TRP-N) was found to be a cilia-associated mechanoreceptor.We propose that these new candidate proteins help explain the sensory complexity of Cnidaria which has been previously observed but so far has lacked a molecular underpinning.Also, the ancient appearance of TRP-N supports a common origin of important components of the nervous systems in Ctenophores, Cnidaria, and Bilateria.

View Article: PubMed Central - PubMed

Affiliation: Institute for Evolution and Biodiversity, University of Muenster, Germany.

Show MeSH
Related in: MedlinePlus