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Bacteria-Induced Dscam Isoforms of the Crustacean, Pacifastacus leniusculus.

Watthanasurorot A, Jiravanichpaisal P, Liu H, Söderhäll I, Söderhäll K - PLoS Pathog. (2011)

Bottom Line: The cytoplasmic tail can generate multiple isoforms.PlDscam can generate more than 22,000 different unique isoforms.Furthermore, PlDscam silencing did not have any effect on the replication of the WSSV.

View Article: PubMed Central - PubMed

Affiliation: Department of Comparative Physiology, Uppsala University, Uppsala, Sweden.

ABSTRACT
The Down syndrome cell adhesion molecule, also known as Dscam, is a member of the immunoglobulin super family. Dscam plays an essential function in neuronal wiring and appears to be involved in innate immune reactions in insects. The deduced amino acid sequence of Dscam in the crustacean Pacifastacus leniusculus (PlDscam), encodes 9(Ig)-4(FNIII)-(Ig)-2(FNIII)-TM and it has variable regions in the N-terminal half of Ig2 and Ig3 and the complete Ig7 and in the transmembrane domain. The cytoplasmic tail can generate multiple isoforms. PlDscam can generate more than 22,000 different unique isoforms. Bacteria and LPS injection enhanced the expression of PlDscam, but no response in expression occurred after a white spot syndrome virus (WSSV) infection or injection with peptidoglycans. Furthermore, PlDscam silencing did not have any effect on the replication of the WSSV. Bacterial specific isoforms of PlDscam were shown to have a specific binding property to each tested bacteria, E. coli or S. aureus. The bacteria specific isoforms of PlDscam were shown to be associated with bacterial clearance and phagocytosis in crayfish.

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Related in: MedlinePlus

Effect of PlDscam silencing on viral replication of WSSV in vitro.A) Silencing of PlDscam using dsRNA in cultured HPT cells was confirmed by RT-PCR before WSSV inoculation. B) WSSV VP28 expression in PlDscam silenced animals (lane 7–9) and the control (lane 4–6). UV-killed WSSV was served as control (lane 1–3), analyzed by semi-quantitative RT-PCR. C) Statistical analysis after comparing different band intensity of the results shown in (B). Significant differences are indicated by asterisks (P<0.05).
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ppat-1002062-g007: Effect of PlDscam silencing on viral replication of WSSV in vitro.A) Silencing of PlDscam using dsRNA in cultured HPT cells was confirmed by RT-PCR before WSSV inoculation. B) WSSV VP28 expression in PlDscam silenced animals (lane 7–9) and the control (lane 4–6). UV-killed WSSV was served as control (lane 1–3), analyzed by semi-quantitative RT-PCR. C) Statistical analysis after comparing different band intensity of the results shown in (B). Significant differences are indicated by asterisks (P<0.05).

Mentions: The role of PlDscam during a WSSV infection was investigated using PlDscam RNAi to suppress the PlDscam expression. The PlDscam gene was completely knocked down in an HPTcell culture (Figure 7A) whereas the 40S ribosomal gene was unaffected. However, PlDscam silencing did not have any effect on WSSV replication as shown with no changes in transcription level of WSSV structural protein transcript VP28 between control and PlDscam silenced groups (Figure 7B and 7C). Moreover, this result also indicates that the expression of PlDscam was not affected by WSSV infection. This agrees with our previous experiment where we injected WSSV to live crayfish and the transcript level of Dscams was not affected (Figure 4E).


Bacteria-Induced Dscam Isoforms of the Crustacean, Pacifastacus leniusculus.

Watthanasurorot A, Jiravanichpaisal P, Liu H, Söderhäll I, Söderhäll K - PLoS Pathog. (2011)

Effect of PlDscam silencing on viral replication of WSSV in vitro.A) Silencing of PlDscam using dsRNA in cultured HPT cells was confirmed by RT-PCR before WSSV inoculation. B) WSSV VP28 expression in PlDscam silenced animals (lane 7–9) and the control (lane 4–6). UV-killed WSSV was served as control (lane 1–3), analyzed by semi-quantitative RT-PCR. C) Statistical analysis after comparing different band intensity of the results shown in (B). Significant differences are indicated by asterisks (P<0.05).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3111544&req=5

ppat-1002062-g007: Effect of PlDscam silencing on viral replication of WSSV in vitro.A) Silencing of PlDscam using dsRNA in cultured HPT cells was confirmed by RT-PCR before WSSV inoculation. B) WSSV VP28 expression in PlDscam silenced animals (lane 7–9) and the control (lane 4–6). UV-killed WSSV was served as control (lane 1–3), analyzed by semi-quantitative RT-PCR. C) Statistical analysis after comparing different band intensity of the results shown in (B). Significant differences are indicated by asterisks (P<0.05).
Mentions: The role of PlDscam during a WSSV infection was investigated using PlDscam RNAi to suppress the PlDscam expression. The PlDscam gene was completely knocked down in an HPTcell culture (Figure 7A) whereas the 40S ribosomal gene was unaffected. However, PlDscam silencing did not have any effect on WSSV replication as shown with no changes in transcription level of WSSV structural protein transcript VP28 between control and PlDscam silenced groups (Figure 7B and 7C). Moreover, this result also indicates that the expression of PlDscam was not affected by WSSV infection. This agrees with our previous experiment where we injected WSSV to live crayfish and the transcript level of Dscams was not affected (Figure 4E).

Bottom Line: The cytoplasmic tail can generate multiple isoforms.PlDscam can generate more than 22,000 different unique isoforms.Furthermore, PlDscam silencing did not have any effect on the replication of the WSSV.

View Article: PubMed Central - PubMed

Affiliation: Department of Comparative Physiology, Uppsala University, Uppsala, Sweden.

ABSTRACT
The Down syndrome cell adhesion molecule, also known as Dscam, is a member of the immunoglobulin super family. Dscam plays an essential function in neuronal wiring and appears to be involved in innate immune reactions in insects. The deduced amino acid sequence of Dscam in the crustacean Pacifastacus leniusculus (PlDscam), encodes 9(Ig)-4(FNIII)-(Ig)-2(FNIII)-TM and it has variable regions in the N-terminal half of Ig2 and Ig3 and the complete Ig7 and in the transmembrane domain. The cytoplasmic tail can generate multiple isoforms. PlDscam can generate more than 22,000 different unique isoforms. Bacteria and LPS injection enhanced the expression of PlDscam, but no response in expression occurred after a white spot syndrome virus (WSSV) infection or injection with peptidoglycans. Furthermore, PlDscam silencing did not have any effect on the replication of the WSSV. Bacterial specific isoforms of PlDscam were shown to have a specific binding property to each tested bacteria, E. coli or S. aureus. The bacteria specific isoforms of PlDscam were shown to be associated with bacterial clearance and phagocytosis in crayfish.

Show MeSH
Related in: MedlinePlus