Laser capture microdissection in combination with mass
spectrometry: Approach to characterization of tissue-specific
proteomes of Eudiplozoon nipponicum (Monogenea,
Polyopisthocotylea)

J 2020

Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes of Eudiplozoon nipponicum (Monogenea, Polyopisthocotylea)

ROUDNICKÝ, Pavel, David POTĚŠIL, Zbyněk ZDRÁHAL, Milan GELNAR, Martin KAŠNÝ et. al.

Basic information

Original name

Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes of Eudiplozoon nipponicum (Monogenea, Polyopisthocotylea)

Authors

ROUDNICKÝ, Pavel (203 Czech Republic, guarantor, belonging to the institution), David POTĚŠIL (203 Czech Republic, belonging to the institution), Zbyněk ZDRÁHAL (203 Czech Republic, belonging to the institution), Milan GELNAR (203 Czech Republic, belonging to the institution) and Martin KAŠNÝ (203 Czech Republic, belonging to the institution)

Edition

PLOS ONE, San Francisco, Public Library of Science, 2020, 1932-6203

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10613 Zoology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 3.240

RIV identification code

RIV/00216224:14310/20:00114261

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1371/journal.pone.0231681

UT WoS

000542759600012

Keywords in English

helminths; parasite; peptidase; inhibitor; laser microdissection; mass spectrometry

Abstract

V originále

Eudiplozoon nipponicum (Goto, 1891) is a hematophagous monogenean ectoparasite which inhabits the gills of the common carp (Cyprinus carpio). Heavy infestation can lead to anemia and in conjunction with secondary bacterial infections cause poor health and eventual death of the host. This study is based on an innovative approach to protein localization which has never been used in parasitology before. Using laser capture microdissection, we dissected particular areas of the parasite body without contaminating the samples by surrounding tissue and in combination with analysis by mass spectrometry obtained tissue-specific proteomes of tegument, intestine, and parenchyma of our model organism, E. nipponicum. We successfully verified the presence of certain functional proteins (e.g. cathepsin L) in tissues where their presence was expected (intestine) and confirmed that there were no traces of these proteins in other tissues (tegument and parenchyma). Additionally, we identified a total of 2,059 proteins, including 72 peptidases and 33 peptidase inhibitors. As expected, the greatest variety was found in the intestine and the lowest variety in the parenchyma. Our results are significant on two levels. Firstly, we demonstrated that one can localize all proteins in one analysis and without using laboratory animals (antibodies for immunolocalization of single proteins). Secondly, this study offers the first complex proteomic data on not only the E. nipponicum but within the whole class of Monogenea, which was from this point of view until recently neglected.

Links

GAP506/12/1258, research and development project

Name: Interakce hostitel-parazit u krevsajících diplozoidních monogeneí: Výzkum vysoce specializovaných adaptací k parazitismu

Investor: Czech Science Foundation

GBP505/12/G112, research and development project

Name: ECIP - Evropské centrum ichtyoparazitologie

Investor: Czech Science Foundation

LM2018140, research and development project

Name: e-Infrastruktura CZ (Acronym: e-INFRA CZ)

Investor: Ministry of Education, Youth and Sports of the CR

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

MUNI/A/0918/2018, interní kód MU

Name: Studium evolučních a ekologických vztahů a procesů u akvatických bezobratlých živočichů a v hostitelsko-parazitických systémech (Acronym: EvolEcolParaHydro)

Investor: Masaryk University, Category A

90127, large research infrastructures

Name: CIISB II

90132, large research infrastructures

Name: NCMG II

Citovat

ROUDNICKÝ, Pavel, David POTĚŠIL, Zbyněk ZDRÁHAL, Milan GELNAR and Martin KAŠNÝ. Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes of Eudiplozoon nipponicum (Monogenea, Polyopisthocotylea). PLOS ONE. San Francisco: Public Library of Science, 2020, vol. 15, No 6, p. 1-15. ISSN 1932-6203. Available from: https://dx.doi.org/10.1371/journal.pone.0231681.

@article{1673338,
   author = {Roudnický, Pavel and Potěšil, David and Zdráhal, Zbyněk and Gelnar, Milan and Kašný, Martin},
   article_location = {San Francisco},
   article_number = {6},
   doi = {http://dx.doi.org/10.1371/journal.pone.0231681},
   keywords = {helminths; parasite; peptidase; inhibitor; laser microdissection; mass spectrometry},
   language = {eng},
   issn = {1932-6203},
   journal = {PLOS ONE},
   title = {Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes of Eudiplozoon nipponicum (Monogenea, Polyopisthocotylea)},
   url = {https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0231681},
   volume = {15},
   year = {2020}
}

TY  - JOUR
ID  - 1673338
AU  - Roudnický, Pavel - Potěšil, David - Zdráhal, Zbyněk - Gelnar, Milan - Kašný, Martin
PY  - 2020
TI  - Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes of Eudiplozoon nipponicum (Monogenea, Polyopisthocotylea)
JF  - PLOS ONE
VL  - 15
IS  - 6
SP  - 1-15
EP  - 1-15
PB  - Public Library of Science
SN  - 19326203
KW  - helminths
KW  - parasite
KW  - peptidase
KW  - inhibitor
KW  - laser microdissection
KW  - mass spectrometry
UR  - https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0231681
L2  - https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0231681
N2  - Eudiplozoon nipponicum (Goto, 1891) is a hematophagous monogenean ectoparasite which inhabits the gills of the common carp (Cyprinus carpio). Heavy infestation can lead to anemia and in conjunction with secondary bacterial infections cause poor health and eventual death of the host. This study is based on an innovative approach to protein localization which has never been used in parasitology before. Using laser capture microdissection, we dissected particular areas of the parasite body without contaminating the samples by surrounding tissue and in combination with analysis by mass spectrometry obtained tissue-specific proteomes of tegument, intestine, and parenchyma of our model organism, E. nipponicum. We successfully verified the presence of certain functional proteins (e.g. cathepsin L) in tissues where their presence was expected (intestine) and confirmed that there were no traces of these proteins in other tissues (tegument and parenchyma). Additionally, we identified a total of 2,059 proteins, including 72 peptidases and 33 peptidase inhibitors. As expected, the greatest variety was found in the intestine and the lowest variety in the parenchyma. Our results are significant on two levels. Firstly, we demonstrated that one can localize all proteins in one analysis and without using laboratory animals (antibodies for immunolocalization of single proteins). Secondly, this study offers the first complex proteomic data on not only the E. nipponicum but within the whole class of Monogenea, which was from this point of view until recently neglected.
ER  -

ROUDNICKÝ, Pavel, David POTĚŠIL, Zbyněk ZDRÁHAL, Milan GELNAR and Martin KAŠNÝ. Laser capture microdissection in combination with mass spectrometry: Approach to characterization of tissue-specific proteomes of Eudiplozoon nipponicum (Monogenea, Polyopisthocotylea). \textit{PLOS ONE}. San Francisco: Public Library of Science, 2020, vol.~15, No~6, p.~1-15. ISSN~1932-6203. Available from: https://dx.doi.org/10.1371/journal.pone.0231681.

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