Identification of a serine protease gene expressed by Myxobolus cerebralis during development in rainbow trout Oncorhynchus mykiss

Garry O. Kelley, Francisco J. Zagmutt-Vergara, Christian M. Leutenegger, Mark A. Adkison, Dolores Baxa, Ronald Hedrick

Research output: Contribution to journalArticle

21 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)235-248
Number of pages14
JournalDiseases of Aquatic Organisms
Volume59
Issue number3
DOIs
StatePublished - Jun 11 2004

Fingerprint

Myxobolus cerebralis
serine proteinases
rainbow
Oncorhynchus mykiss
parasite
transcription (genetics)
gene
parasites
developmental stage
polymerase chain reaction
chymotrypsin
genes
cartilage
in situ hybridization
parasitic disease
quantitative polymerase chain reaction
whirling disease
developmental stages
vaccine
salmonid

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 journalArticle

Kelley, Garry O. ; Zagmutt-Vergara, Francisco J. ; Leutenegger, Christian M. ; Adkison, Mark A. ; Baxa, Dolores ; Hedrick, Ronald. / Identification of a serine protease gene expressed by Myxobolus cerebralis during development in rainbow trout Oncorhynchus mykiss. In: Diseases of Aquatic Organisms. 2004 ; Vol. 59, No. 3. pp. 235-248.
@article{20680ca6e93a49e2beeb8fdafb2083e6,
title = "Identification of a serine protease gene expressed by Myxobolus cerebralis during development in rainbow trout Oncorhynchus mykiss",
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.",
keywords = "mRNA in situ hybridization, Myxobolus cerebralis, Real-time Taq-Man polymerase chain reaction, Salmonid whirling disease, Serine protease",
author = "Kelley, {Garry O.} and Zagmutt-Vergara, {Francisco J.} and Leutenegger, {Christian M.} and Adkison, {Mark A.} and Dolores Baxa and Ronald Hedrick",
year = "2004",
month = "6",
day = "11",
doi = "10.3354/dao059235",
language = "English (US)",
volume = "59",
pages = "235--248",
journal = "Diseases of Aquatic Organisms",
issn = "0177-5103",
publisher = "Inter-Research",
number = "3",

}

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 -