Saxitoxin Modulates Immunological Parameters and Gene Transcription in Mytilus chilensis Hemocytes
Abstract
:1. Introduction
2. Results
2.1. Primary Culture of Hemocytes from M. chilensis
2.2. Effects of Saxitoxin (STX) on Cellular Parameters
2.3. Effects of STX on the Gene Expression of Antioxidant Enzymes, Oxidative Phosphorylation Transcripts, Ion Channels and Pattern Recognition Receptors
3. Discussion
3.1. Characterization of Hemocyte Primary Cell Cultures
3.2. Cellular Response of Hemocytes to STX Exposure
3.3. Effects of STX on Gene Transcription in M. chilensis Hemocytes
3.4. Use of Primary Hemocyte Cultures in Toxicological Studies
4. Experimental Section
4.1. Primary Culture of Hemocytes
4.2. Experimental Design
4.3. Viability and Phagocytosis Assays Using Light and Fluorescent Microscopy
4.4. Hemocyte Phagocytosis Using Flow Cytometry
4.5. Intracellular ROS Production Using Fluorescent Microscopy
4.6. Transcription Expression of Candidate Genes
Gene | Primers (5ʹ–3ʹ) | Reference |
---|---|---|
Catalase | CCAGGTGTCCTTCCTGTTTTCT | Núñez-Acuña et al., 2012 [60] |
TGTCCATCCTTGTTGACTGTCTTAA | ||
Ec-SOD Cu/Zn | TCGCTTTCAGTCAACAGAATGG | Núñez-Acuña et al., 2012 [60] |
CCAAACTCGTGAACGTGGAA | ||
Ferritin | CAAGTGAACGCCATCAAGGA | Núñez-Acuña et al., 2012 [60] |
CGTTGATGCTCTCTTTGTCATACA | ||
Metallothionein | CTGTGGTGACGCCTGCAAGT | Núñez-Acuña et al., 2013 [20] |
CTGTGGTGACGCCTGCAAGT | ||
NADH dehydrogenase I | CTTACTAGGTGCAGTCCGTG | Primers designed in this study |
ACATAAACACGCCTGAAGATAGT | ||
ATP6 | TGGGCATACCGCTGTGACTTACT | Primers designed in this study |
GTGCAGGGGCACCCATAGGA | ||
Cytochrome oxidase subunit I | TCAGCTGCTGTAGAGAGTGGGG | Primers designed in this study |
TCACGCCGGTGGTTCAGTTG | ||
Cytochrome oxidase subunit III | TATGTACCAGGCCCAAGTCC | Primers designed in this study |
AAAATCTGGGGGTTCGATGC | ||
Cytochrome B | GGAAGATGCCCGTTGGAGGC | Primers designed in this study |
CCCTGCGCTCCAGAAATTATGGG | ||
Voltage-dependent anion channel | CAGACAGGAAAGAAGCAGGG | Primers designed in this study |
CTAAGGCACCATTCACAGCT | ||
Calcium channel | TGTAAGCAAGTCGTGTGGGGCAC | Primers designed in this study |
ACGAGTTTGGGCAGACTATGATCCTG | ||
Potassium channel | CGGCAATTTGGACCGGGCTTGTG | Primers designed in this study |
GCGCCCTCTGTCCATTACTCGGC | ||
C-type lectin | GATATCGCTGTGCAGAACCA | Primers designed in this study |
AAACACAAGGCCAGTGGAAA | ||
TLR | CTTTCTTAGCCACGCCAGAC | Primers designed in this study |
TGTCATTGTCATTGTCAGTTTCGTCA | ||
α-Tubulin | GAGCCGTCTGCATGTTGAGC | Núñez-Acuña et al., 2012 [60] |
TGGACGAAAGCACGTTTGGC |
4.7. Statistical Analyses
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Astuya, A.; Carrera, C.; Ulloa, V.; Aballay, A.; Núñez-Acuña, G.; Hégaret, H.; Gallardo-Escárate, C. Saxitoxin Modulates Immunological Parameters and Gene Transcription in Mytilus chilensis Hemocytes. Int. J. Mol. Sci. 2015, 16, 15235-15250. https://doi.org/10.3390/ijms160715235
Astuya A, Carrera C, Ulloa V, Aballay A, Núñez-Acuña G, Hégaret H, Gallardo-Escárate C. Saxitoxin Modulates Immunological Parameters and Gene Transcription in Mytilus chilensis Hemocytes. International Journal of Molecular Sciences. 2015; 16(7):15235-15250. https://doi.org/10.3390/ijms160715235
Chicago/Turabian StyleAstuya, Allisson, Crisleri Carrera, Viviana Ulloa, Ambbar Aballay, Gustavo Núñez-Acuña, Hélène Hégaret, and Cristian Gallardo-Escárate. 2015. "Saxitoxin Modulates Immunological Parameters and Gene Transcription in Mytilus chilensis Hemocytes" International Journal of Molecular Sciences 16, no. 7: 15235-15250. https://doi.org/10.3390/ijms160715235