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Dynamically-expressed prion-like proteins form a cuticle in the pharynx of Caenorhabditis elegans.

George-Raizen JB, Shockley KR, Trojanowski NF, Lamb AL, Raizen DM - Biol Open (2014)

Bottom Line: We found strong transcriptional induction during the molt of 12 of 15 previously identified abu genes encoding Prion-like (P) glutamine (Q) and asparagine (N) rich PQN proteins, as well as 15 additional genes encoding closely related PQN proteins. abu/pqn genes, which we name the abu/pqn paralog group (APPG) genes, were expressed in pharyngeal cells and the proteins encoded by two APPG genes we tested localized to the pharyngeal cuticle.Deleting the APPG gene abu-14 caused abnormal pharyngeal cuticular structures and knocking down other APPG genes resulted in abnormal cuticular function.The strong developmental regulation of the APPG genes raises the possibility that such genes would be identified in transcriptional profiling experiments in which the animals' developmental stage is not precisely staged.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.

No MeSH data available.


Related in: MedlinePlus

abu/pqn genes are expressed in the pharyngeal cuticle.(A) Expression of abu-5 (red) and abu-11 (green) in pharyngeal cells. Scale bar is 10 µm. (B) Differential interference contrast (DIC) and fluorescence image of the anterior pharynx of an adult transgenic animal containing the transgene abu-14>abu-14::sfGFP. The fluorescence localizes to the pharyngeal corpus and buccal cavity cuticle (arrows). A non-transgenic sister (marked with “non-Tg”) shows no fluorescence. Anterior is to the left. Scale bar is 10 µm. (C) DIC and fluorescence image of the pharynx of a fourth larval stage molting animal containing the transgene abu-14>abu-14::sfGFP. Green fluorescence is seen in both the L4 and adult pharyngeal grinders, in the pharyngeal cuticle, and in the buccal cap. Anterior is to the left. Scale bar is 10 µm. (D) DIC (left) and fluorescence (right) images of a larval animal containing the transgene abu-7>abu-7::GFP. The pharyngeal grinder is brightly fluorescent. Scale bar is 10 µm.
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f02: abu/pqn genes are expressed in the pharyngeal cuticle.(A) Expression of abu-5 (red) and abu-11 (green) in pharyngeal cells. Scale bar is 10 µm. (B) Differential interference contrast (DIC) and fluorescence image of the anterior pharynx of an adult transgenic animal containing the transgene abu-14>abu-14::sfGFP. The fluorescence localizes to the pharyngeal corpus and buccal cavity cuticle (arrows). A non-transgenic sister (marked with “non-Tg”) shows no fluorescence. Anterior is to the left. Scale bar is 10 µm. (C) DIC and fluorescence image of the pharynx of a fourth larval stage molting animal containing the transgene abu-14>abu-14::sfGFP. Green fluorescence is seen in both the L4 and adult pharyngeal grinders, in the pharyngeal cuticle, and in the buccal cap. Anterior is to the left. Scale bar is 10 µm. (D) DIC (left) and fluorescence (right) images of a larval animal containing the transgene abu-7>abu-7::GFP. The pharyngeal grinder is brightly fluorescent. Scale bar is 10 µm.

Mentions: Fluorescent transcriptional reporters of all APPG genes we tested (abu-5, abu-6, abu-11, abu-14, abu-15, pqn-57) were expressed in pharyngeal muscle (Fig. 2; supplementary material Fig. S3), suggesting that most or all of the APPG genes are expressed in pharyngeal muscle. This suggestion is supported by prior reporter transgene expression analysis of abu-1 (Urano et al., 2002) and abu-14 (Ao et al., 2004) and by RNA in situ hybridization analysis of abu-8, abu-14, and pqn-13 expression (http://nematode.lab.nig.ac.jp), all of which have been demonstrated to be expressed in pharyngeal muscle. In rare cases, expression was seen outside the pharynx: the pqn-57 reporter was also expressed near the rectum (supplementary material Fig. S3) and RNA in situ analysis showed rare abu-14 staining in the intestine (http://nematode.lab.nig.ac.jp). Consistent with the microarray data, indicating dynamic expression of APPG genes and a role during the molt, we observed dynamic expression of a fluorescent transcriptional reporter for abu-11 (supplementary material Fig. S4). Consistent with a larval-specific function of APPG genes, transcriptional reporters for abu-5, abu-11, abu-14, and abu-15 showed little to no adult fluorescence (supplementary material Fig. S5). In contrast, expression of a pqn-57 reporter persisted in adult pharynxes (supplementary material Fig. S5).


Dynamically-expressed prion-like proteins form a cuticle in the pharynx of Caenorhabditis elegans.

George-Raizen JB, Shockley KR, Trojanowski NF, Lamb AL, Raizen DM - Biol Open (2014)

abu/pqn genes are expressed in the pharyngeal cuticle.(A) Expression of abu-5 (red) and abu-11 (green) in pharyngeal cells. Scale bar is 10 µm. (B) Differential interference contrast (DIC) and fluorescence image of the anterior pharynx of an adult transgenic animal containing the transgene abu-14>abu-14::sfGFP. The fluorescence localizes to the pharyngeal corpus and buccal cavity cuticle (arrows). A non-transgenic sister (marked with “non-Tg”) shows no fluorescence. Anterior is to the left. Scale bar is 10 µm. (C) DIC and fluorescence image of the pharynx of a fourth larval stage molting animal containing the transgene abu-14>abu-14::sfGFP. Green fluorescence is seen in both the L4 and adult pharyngeal grinders, in the pharyngeal cuticle, and in the buccal cap. Anterior is to the left. Scale bar is 10 µm. (D) DIC (left) and fluorescence (right) images of a larval animal containing the transgene abu-7>abu-7::GFP. The pharyngeal grinder is brightly fluorescent. Scale bar is 10 µm.
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Related In: Results  -  Collection

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f02: abu/pqn genes are expressed in the pharyngeal cuticle.(A) Expression of abu-5 (red) and abu-11 (green) in pharyngeal cells. Scale bar is 10 µm. (B) Differential interference contrast (DIC) and fluorescence image of the anterior pharynx of an adult transgenic animal containing the transgene abu-14>abu-14::sfGFP. The fluorescence localizes to the pharyngeal corpus and buccal cavity cuticle (arrows). A non-transgenic sister (marked with “non-Tg”) shows no fluorescence. Anterior is to the left. Scale bar is 10 µm. (C) DIC and fluorescence image of the pharynx of a fourth larval stage molting animal containing the transgene abu-14>abu-14::sfGFP. Green fluorescence is seen in both the L4 and adult pharyngeal grinders, in the pharyngeal cuticle, and in the buccal cap. Anterior is to the left. Scale bar is 10 µm. (D) DIC (left) and fluorescence (right) images of a larval animal containing the transgene abu-7>abu-7::GFP. The pharyngeal grinder is brightly fluorescent. Scale bar is 10 µm.
Mentions: Fluorescent transcriptional reporters of all APPG genes we tested (abu-5, abu-6, abu-11, abu-14, abu-15, pqn-57) were expressed in pharyngeal muscle (Fig. 2; supplementary material Fig. S3), suggesting that most or all of the APPG genes are expressed in pharyngeal muscle. This suggestion is supported by prior reporter transgene expression analysis of abu-1 (Urano et al., 2002) and abu-14 (Ao et al., 2004) and by RNA in situ hybridization analysis of abu-8, abu-14, and pqn-13 expression (http://nematode.lab.nig.ac.jp), all of which have been demonstrated to be expressed in pharyngeal muscle. In rare cases, expression was seen outside the pharynx: the pqn-57 reporter was also expressed near the rectum (supplementary material Fig. S3) and RNA in situ analysis showed rare abu-14 staining in the intestine (http://nematode.lab.nig.ac.jp). Consistent with the microarray data, indicating dynamic expression of APPG genes and a role during the molt, we observed dynamic expression of a fluorescent transcriptional reporter for abu-11 (supplementary material Fig. S4). Consistent with a larval-specific function of APPG genes, transcriptional reporters for abu-5, abu-11, abu-14, and abu-15 showed little to no adult fluorescence (supplementary material Fig. S5). In contrast, expression of a pqn-57 reporter persisted in adult pharynxes (supplementary material Fig. S5).

Bottom Line: We found strong transcriptional induction during the molt of 12 of 15 previously identified abu genes encoding Prion-like (P) glutamine (Q) and asparagine (N) rich PQN proteins, as well as 15 additional genes encoding closely related PQN proteins. abu/pqn genes, which we name the abu/pqn paralog group (APPG) genes, were expressed in pharyngeal cells and the proteins encoded by two APPG genes we tested localized to the pharyngeal cuticle.Deleting the APPG gene abu-14 caused abnormal pharyngeal cuticular structures and knocking down other APPG genes resulted in abnormal cuticular function.The strong developmental regulation of the APPG genes raises the possibility that such genes would be identified in transcriptional profiling experiments in which the animals' developmental stage is not precisely staged.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.

No MeSH data available.


Related in: MedlinePlus