Abstract
Serine proteases have been recognized as key factors in parasite physiology and disease development. We have identified a serine protease gene from Myxobolus cerebralis, MyxSP-1, the myxozoan parasite causing whirling disease in salmonid fishes. The amino acid sequence, as deduced from the cDNA sequence, included a catalytic residue arrangement similar to that of the chymotrypsin family of serine proteases. A real-time TaqMan polymerase chain reaction (PCR) analysis revealed differences in the transcription levels for the chymotrypsin-like protease as found in early, intermediate, and late developmental stages of the parasite in experimentally-infected rainbow trout Oncorhynchus mykiss. MyxSP-1 transcription differed between individual tissues at each sampling point and in the same tissues over time (p < 0.0001). A nonradioactive mRNA in situ hybridization (ISH) protocol was developed to detect MyxSP-1 transcripts. Using a mixture of 3 digoxigenin-labeled antisense mRNA probes, MyxSP-1 transcription was observed in developmental stages of the parasite during the acute and chronic phases of the disease over a 240 d time period in infected rainbow trout tissues. MyxSP-1 transcription observed by ISH in cartilage and as associated with cartilage destruction was consistent with our real-time TaqMan PCR findings that demonstrated high levels of MyxSP-1 transcription during lesion development. Identifying genes encoding these enzymes and characterization of their functions can lead to the development of new chemotherapeutic protocols and vaccine approaches to control parasitic diseases.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 235-248 |
| Number of pages | 14 |
| Journal | Diseases of Aquatic Organisms |
| Volume | 59 |
| Issue number | 3 |
| DOIs | |
| State | Published - Jun 11 2004 |
Fingerprint
Keywords
- mRNA in situ hybridization
- Myxobolus cerebralis
- Real-time Taq-Man polymerase chain reaction
- Salmonid whirling disease
- Serine protease
ASJC Scopus subject areas
- Ecology, Evolution, Behavior and Systematics
- Aquatic Science
Cite this
Identification of a serine protease gene expressed by Myxobolus cerebralis during development in rainbow trout Oncorhynchus mykiss. / Kelley, Garry O.; Zagmutt-Vergara, Francisco J.; Leutenegger, Christian M.; Adkison, Mark A.; Baxa, Dolores; Hedrick, Ronald.
In: Diseases of Aquatic Organisms, Vol. 59, No. 3, 11.06.2004, p. 235-248.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identification of a serine protease gene expressed by Myxobolus cerebralis during development in rainbow trout Oncorhynchus mykiss
AU - Kelley, Garry O.
AU - Zagmutt-Vergara, Francisco J.
AU - Leutenegger, Christian M.
AU - Adkison, Mark A.
AU - Baxa, Dolores
AU - Hedrick, Ronald
PY - 2004/6/11
Y1 - 2004/6/11
N2 - Serine proteases have been recognized as key factors in parasite physiology and disease development. We have identified a serine protease gene from Myxobolus cerebralis, MyxSP-1, the myxozoan parasite causing whirling disease in salmonid fishes. The amino acid sequence, as deduced from the cDNA sequence, included a catalytic residue arrangement similar to that of the chymotrypsin family of serine proteases. A real-time TaqMan polymerase chain reaction (PCR) analysis revealed differences in the transcription levels for the chymotrypsin-like protease as found in early, intermediate, and late developmental stages of the parasite in experimentally-infected rainbow trout Oncorhynchus mykiss. MyxSP-1 transcription differed between individual tissues at each sampling point and in the same tissues over time (p < 0.0001). A nonradioactive mRNA in situ hybridization (ISH) protocol was developed to detect MyxSP-1 transcripts. Using a mixture of 3 digoxigenin-labeled antisense mRNA probes, MyxSP-1 transcription was observed in developmental stages of the parasite during the acute and chronic phases of the disease over a 240 d time period in infected rainbow trout tissues. MyxSP-1 transcription observed by ISH in cartilage and as associated with cartilage destruction was consistent with our real-time TaqMan PCR findings that demonstrated high levels of MyxSP-1 transcription during lesion development. Identifying genes encoding these enzymes and characterization of their functions can lead to the development of new chemotherapeutic protocols and vaccine approaches to control parasitic diseases.
AB - Serine proteases have been recognized as key factors in parasite physiology and disease development. We have identified a serine protease gene from Myxobolus cerebralis, MyxSP-1, the myxozoan parasite causing whirling disease in salmonid fishes. The amino acid sequence, as deduced from the cDNA sequence, included a catalytic residue arrangement similar to that of the chymotrypsin family of serine proteases. A real-time TaqMan polymerase chain reaction (PCR) analysis revealed differences in the transcription levels for the chymotrypsin-like protease as found in early, intermediate, and late developmental stages of the parasite in experimentally-infected rainbow trout Oncorhynchus mykiss. MyxSP-1 transcription differed between individual tissues at each sampling point and in the same tissues over time (p < 0.0001). A nonradioactive mRNA in situ hybridization (ISH) protocol was developed to detect MyxSP-1 transcripts. Using a mixture of 3 digoxigenin-labeled antisense mRNA probes, MyxSP-1 transcription was observed in developmental stages of the parasite during the acute and chronic phases of the disease over a 240 d time period in infected rainbow trout tissues. MyxSP-1 transcription observed by ISH in cartilage and as associated with cartilage destruction was consistent with our real-time TaqMan PCR findings that demonstrated high levels of MyxSP-1 transcription during lesion development. Identifying genes encoding these enzymes and characterization of their functions can lead to the development of new chemotherapeutic protocols and vaccine approaches to control parasitic diseases.
KW - mRNA in situ hybridization
KW - Myxobolus cerebralis
KW - Real-time Taq-Man polymerase chain reaction
KW - Salmonid whirling disease
KW - Serine protease
UR - http://www.scopus.com/inward/record.url?scp=4344565771&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=4344565771&partnerID=8YFLogxK
U2 - 10.3354/dao059235
DO - 10.3354/dao059235
M3 - Article
C2 - 15264720
AN - SCOPUS:4344565771
VL - 59
SP - 235
EP - 248
JO - Diseases of Aquatic Organisms
JF - Diseases of Aquatic Organisms
SN - 0177-5103
IS - 3
ER -