Ontogeny and tissue-specific expression of immune-relevant genes in Catla catla (Hamilton)
Introduction
India holds the second rank globally in total fish production with an annual fish production of about 12.6 million tonnes in 2017–18 (NFDB, 2018). Freshwater aquaculture contributes over 89% of the total aquaculture production in the country and three Indian major carps (IMC), namely rohu Labeo rohita, catla Catla catla and mrigal Cirrhinus mrigala contribute to the extent of 74% of the total freshwater fish production. Among IMCs, catla is the fastest growing species (Jena, 2006). However, the survival rate of catla during larval rearing is normally lower than that of rohu and mrigal (Jena, 2006). The immune competence that the fish acquire during their early life stages, could be a significant factor for better survival in later stages or when they are reared in aquaculture facilities. Vaccination of fish in the early stages of culture can help in increasing survival and decreasing the use of anti-bacterial drugs (Zapata et al., 1997). However, if fish are vaccinated prior to becoming immunologically competent, a phenomenon known as immunological tolerance can occur, leading to a decrease in the subsequent secondary immune response and efficacy of vaccination (Magnadóttir et al., 2005). Therefore, it is important that vaccination of cultured fish is done following development of functional response. Understanding the development of immunity in fish larvae and juveniles may help in developing effective immune intervention practices (Mulero et al., 2007).
In recent decades, the importance of cytokine genes has been widely accepted as effective parameters for the evaluation of immune response (Cecchini et al., 2013). Interleukin-1β (IL-1β) is an important early response pro-inflammatory cytokine that evades pathogen evasion and induces an inflammatory cascade with the help of other defensive responses (Mohanty and Sahoo, 2007). Interleukin-10 (IL-10) is known as a potent anti-inflammatory as well as a regulatory cytokine which plays a pivotal role in preventing inflammatory and autoimmune pathologies (Ouyang et al., 2011). The inducible Nitric Oxide Synthase (iNOS) gene helps in inhibiting the invading pathogens through the production of reactive nitrogen intermediates (RNI) and is considered as a crucial component of host immune response (Nathan, 1992). Complement component 3 (C3), a major complement component present in the plasma, facilitates inflammatory reactions and lysis of pathogens (Pushpa et al., 2014). In addition, expression of adaptive immune genes is of significance for indicating the state of lymphopoiesis and maturation of adaptive immune system during the development of an organism (Saravanan et al., 2017). The recombination activating gene (RAG-1) is a useful marker for the developmental study of the immune system (Mao et al., 2012). Ikaros gene, a principal component of the adaptive immune system is used as an early lymphoid marker (Willett et al., 2001). The presence of a functional immunoglobulin (Ig) molecule indicates the existence of an adaptive immune response, and thus provides insight into the earliest possible time at which fish can be vaccinated. IgM has been considered as an important marker in the study of the physiological maturity of the immune system (Mao et al., 2012). In addition, the early expression of IgZ in different fish indicates their protective role during larval development (Dash and Sahoo, 2015).
Therefore, the aim of the present study was to analyze the expression of some of the important innate and adaptive immune genes during the ontogenic development of catla and, further evaluating their tissue-specific expression in adult specimens. The information generated will improve knowledge of the maturation of the immune system during the larval development of catla, one of the important aquaculture species of the Indian sub-continent.
Section snippets
Results and discussion
Expression levels of innate and adaptive immune-relevant genes in catla were determined by qRT-PCR, in different developmental stages starting from unfertilized eggs and fertilized eggs up to 54 dph larvae using EF-1α as reference gene. Transcripts of all the studied genes were found to be expressed from early fertilized egg stage except for IL-1β, C3, IgM and IgZ which were detected even in unfertilized eggs. Expression profiles of these genes were similar as reported earlier in rohu, (Dash et
Sample collection
For the ontogenetic study, the eggs/larvae were collected from Indian major carp hatchery of ICAR-National Bureau of Fish Genetic Resources, Lucknow. Briefly, catla broodfish were induced bred using Ovaprim, in the Chinese circular hatchery system. After 8–12 h of breeding, the fertilized eggs were collected and transferred to the hatching tank. The larvae hatched within 20–26 h of fertilization at water temperature of 27–29 °C. Forty eight hours after hatching, the larvae were stocked at a
Acknowledgements
The authors are thankful to the Director, ICAR-National Bureau of Fish Genetic Resources, Lucknow for providing necessary facilities to carry out this work. The first author is thankful to Integral University, Lucknow for supporting this work under manuscript number IU/R&D/2018-MCN000405 and also thankful to ICAR-Consortium Research Platform on Vaccines and Diagnostics for providing the fellowship.
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2021, Fish and Shellfish ImmunologyCitation Excerpt :The C3 expression level increased at advanced fry stage which is in agreement with [32] in Catla catla and in other teleosts [34,35]. The enhanced expression of C3 during the advanced developmental stages may be attributed to its involvement in adaptive immune gene's complement lysis [32]. The expression of complement C3 at different levels during ontogeny of golden mahseer also indicates its probable association with non-immune-related events during ontogeny [36].