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Cathepsin Gene Family Reveals Transcriptome Patterns Related to the Infective Stages of the Salmon Louse Caligus rogercresseyi.

Maldonado-Aguayo W, Chávez-Mardones J, Gonçalves AT, Gallardo-Escárate C - PLoS ONE (2015)

Bottom Line: Using high-throughput transcriptome sequencing, 56 cathepsin-like sequences were found distributed in five cysteine protease groups (B, F, L, Z, and S) as well as in an aspartic protease group (D).Putative functional roles of cathepsins were conjectured based on the differential expressions found and on roles previously described in other phylogenetically related species.The identification of cathepsins together with functional validations represents a valuable strategy for pinpointing target molecules that could be used in the development of new delousing drugs or vaccines against C. rogercresseyi.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Sustainable Aquaculture Research (INCAR), University of Concepción, Concepción, Chile.

ABSTRACT
Cathepsins are proteases involved in the ability of parasites to overcome and/or modulate host defenses so as to complete their own lifecycle. However, the mechanisms underlying this ability of cathepsins are still poorly understood. One excellent model for identifying and exploring the molecular functions of cathepsins is the marine ectoparasitic copepod Caligus rogercresseyi that currently affects the Chilean salmon industry. Using high-throughput transcriptome sequencing, 56 cathepsin-like sequences were found distributed in five cysteine protease groups (B, F, L, Z, and S) as well as in an aspartic protease group (D). Ontogenic transcriptome analysis evidenced that L cathepsins were the most abundant during the lifecycle, while cathepsins B and K were mostly expressed in the larval stages and adult females, thus suggesting participation in the molting processes and embryonic development, respectively. Interestingly, a variety of cathepsins from groups Z, L, D, B, K, and S were upregulated in the infective stage of copepodid, corroborating the complexity of the processes involved in the parasitic success of this copepod. Putative functional roles of cathepsins were conjectured based on the differential expressions found and on roles previously described in other phylogenetically related species. Moreover, 140 single nucleotide polymorphisms (SNP) were identified in transcripts annotated for cysteine and aspartic proteases located into untranslated regions, or the coding region. This study reports for the first time the presence of cathepsin-like genes and differential expressions throughout a copepod lifecycle. The identification of cathepsins together with functional validations represents a valuable strategy for pinpointing target molecules that could be used in the development of new delousing drugs or vaccines against C. rogercresseyi.

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Heat maps of cathepsin-like transcripts from the six developmental stages of Caligus rogercresseyi representing all cathepsins found (A), and a magnification of the cathepsins found in cluster A and B (B).Transcript abundance is represented as RPKM values and color scales show relative transcript expression.
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pone.0123954.g004: Heat maps of cathepsin-like transcripts from the six developmental stages of Caligus rogercresseyi representing all cathepsins found (A), and a magnification of the cathepsins found in cluster A and B (B).Transcript abundance is represented as RPKM values and color scales show relative transcript expression.

Mentions: The 56 cathepsin-related transcripts found in C. rogercresseyi were used for RNA-seq analysis, including cathepsin-like transcripts from groups L, D, F, B, Z, H, K, and S (see S1A Table for transcript details). Analysis revealed that the transcripts were differentially expressed between six developmental stages of C. rogercresseyi (Fig 4A). A sub-experiment with the 42 sequences included in clusters A and B revealed underlying differential expression patterns mainly associated with the copepodid stage, where several cathepsin B, D, F, K, L, S, and Z-like genes were upregulated (Fig 4B). A total of 26 cathepsin-like genes were differentially expressed throughout the different stages. Between the nauplius and copepodid stages, five genes were found to be up-regulated while another five were down-regulated (Table 2), with Cr-CatL2 being highly downregulated. On the other hand, between the copepodid and chalimus stages, five genes were upregulated while 13 were downregulated (Table 3). Of these, Cr-CatL2 was the most significantly expressed with an up-regulation of 198-fold. The analysis between adult females and males evidenced that 19 genes were upregulated while five cathepsin-like sequences were downregulated (Table 4), and among these, Cr-CatL26 was the most downregulated by more than 60-fold. The majority of the genes that were significantly regulated were those associated with the adult stage, while for the nauplius and copepodid stages, only ten cathepsin-like transcripts were differentially expressed. Finally, the seven cathepsin-like selected genes mRNA expression (Fig 5) validated the in silico differential expression observed in the study.


Cathepsin Gene Family Reveals Transcriptome Patterns Related to the Infective Stages of the Salmon Louse Caligus rogercresseyi.

Maldonado-Aguayo W, Chávez-Mardones J, Gonçalves AT, Gallardo-Escárate C - PLoS ONE (2015)

Heat maps of cathepsin-like transcripts from the six developmental stages of Caligus rogercresseyi representing all cathepsins found (A), and a magnification of the cathepsins found in cluster A and B (B).Transcript abundance is represented as RPKM values and color scales show relative transcript expression.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414500&req=5

pone.0123954.g004: Heat maps of cathepsin-like transcripts from the six developmental stages of Caligus rogercresseyi representing all cathepsins found (A), and a magnification of the cathepsins found in cluster A and B (B).Transcript abundance is represented as RPKM values and color scales show relative transcript expression.
Mentions: The 56 cathepsin-related transcripts found in C. rogercresseyi were used for RNA-seq analysis, including cathepsin-like transcripts from groups L, D, F, B, Z, H, K, and S (see S1A Table for transcript details). Analysis revealed that the transcripts were differentially expressed between six developmental stages of C. rogercresseyi (Fig 4A). A sub-experiment with the 42 sequences included in clusters A and B revealed underlying differential expression patterns mainly associated with the copepodid stage, where several cathepsin B, D, F, K, L, S, and Z-like genes were upregulated (Fig 4B). A total of 26 cathepsin-like genes were differentially expressed throughout the different stages. Between the nauplius and copepodid stages, five genes were found to be up-regulated while another five were down-regulated (Table 2), with Cr-CatL2 being highly downregulated. On the other hand, between the copepodid and chalimus stages, five genes were upregulated while 13 were downregulated (Table 3). Of these, Cr-CatL2 was the most significantly expressed with an up-regulation of 198-fold. The analysis between adult females and males evidenced that 19 genes were upregulated while five cathepsin-like sequences were downregulated (Table 4), and among these, Cr-CatL26 was the most downregulated by more than 60-fold. The majority of the genes that were significantly regulated were those associated with the adult stage, while for the nauplius and copepodid stages, only ten cathepsin-like transcripts were differentially expressed. Finally, the seven cathepsin-like selected genes mRNA expression (Fig 5) validated the in silico differential expression observed in the study.

Bottom Line: Using high-throughput transcriptome sequencing, 56 cathepsin-like sequences were found distributed in five cysteine protease groups (B, F, L, Z, and S) as well as in an aspartic protease group (D).Putative functional roles of cathepsins were conjectured based on the differential expressions found and on roles previously described in other phylogenetically related species.The identification of cathepsins together with functional validations represents a valuable strategy for pinpointing target molecules that could be used in the development of new delousing drugs or vaccines against C. rogercresseyi.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Sustainable Aquaculture Research (INCAR), University of Concepción, Concepción, Chile.

ABSTRACT
Cathepsins are proteases involved in the ability of parasites to overcome and/or modulate host defenses so as to complete their own lifecycle. However, the mechanisms underlying this ability of cathepsins are still poorly understood. One excellent model for identifying and exploring the molecular functions of cathepsins is the marine ectoparasitic copepod Caligus rogercresseyi that currently affects the Chilean salmon industry. Using high-throughput transcriptome sequencing, 56 cathepsin-like sequences were found distributed in five cysteine protease groups (B, F, L, Z, and S) as well as in an aspartic protease group (D). Ontogenic transcriptome analysis evidenced that L cathepsins were the most abundant during the lifecycle, while cathepsins B and K were mostly expressed in the larval stages and adult females, thus suggesting participation in the molting processes and embryonic development, respectively. Interestingly, a variety of cathepsins from groups Z, L, D, B, K, and S were upregulated in the infective stage of copepodid, corroborating the complexity of the processes involved in the parasitic success of this copepod. Putative functional roles of cathepsins were conjectured based on the differential expressions found and on roles previously described in other phylogenetically related species. Moreover, 140 single nucleotide polymorphisms (SNP) were identified in transcripts annotated for cysteine and aspartic proteases located into untranslated regions, or the coding region. This study reports for the first time the presence of cathepsin-like genes and differential expressions throughout a copepod lifecycle. The identification of cathepsins together with functional validations represents a valuable strategy for pinpointing target molecules that could be used in the development of new delousing drugs or vaccines against C. rogercresseyi.

Show MeSH
Related in: MedlinePlus