Acetyl-CoA carboxylase (
ACC), a rate-limiting enzyme was known to involve in
de novo fatty acid synthesis specifically in catalysing process of acetyl-CoA to malonyl-CoA
(Wakil 1958; Nugteren 1965; (
Abu-Elheiga et al., 2001). Two forms of
ACC genes were described such as
ACACA located in the cytosol and
ACACB associated/linked/connected with the mitochondrial membrane
(Munday 2002). The
ACACB gene was known to be associated with nephropathy, obesity, diabetes and end-stage renal diseases
(Tang et al., 2010; Riancho et al., 2011; Zain et al., 2017). This gene was first discovered in rat heart
(Thampy 1989). Mice lacking
ACACB gene are reported to be protected against obesity and diabetes
(Abu-Elheiga et al., 2003). The “Gallus Expression in Situ Hybridization Analysis” (GEISHA) database revealed non-availability of expression data for the gene. The human protein atlas (https://www.proteinatlas.org/ENSG000000 76555-
ACACB/tissue) showed that the gene expresses highly in skeletal muscle tissue compare to all other tissues. Hence, in chicken, skeletal muscle is targeted for its expression analysis.
Thus, we have chosen IWI line i.e. developed from white leghorn breed that was known for high egg production. These egg laying chickens provides good insight to understand various processes of lipid mobilization, transfer and utilization for yolk precursor synthesis, which is important in deposition of yolk and embryo development
(Saarela et al., 2008). Merkin et al. (2012) while studying evolutionary dynamics of gene regulation in mammalian and bird tissues, indicated that
ACACB gene expressed at low to medium level in colon, heart, kidney, lung, skeletal muscle tissue, spleen and testis while its expression is below cut off level in brain and liver. Similarly, a study by
Barborasa-Morais et al. (2012) also showed that
ACACB gene is low to medium in kidney, skeletal muscle and heart and below cut-off in brain and liver. There was no clear-cut idea how the gene is expressed over different developmental stages of chicken. There was no reports are available on expression of the
ACACB gene during the pre and post-hatch period in chicken. However, some other genes are studied and reported during pre and post-hatch period in chicken. In domestic chicken, the higher embryonic growth was reported during late embryonic stage
(Cogburn et al., 1989). In both control broiler and Aseel, the
ACTRIIB gene expressed at ED7, ED11 and ED16
(Vishnu et al., 2017). Similar results were also found in the post-hatch period in layer chicken line, where the highest and lowest expression level of the
SCD gene was found on the 2
nd (40.75) and 6th week (27.75), respectively
(Sagar et al., 2019). In the control layer, the expression of
ACACA gene was up-regulated during 2
nd, 4
th and 6
th weeks of age, when compared to the day one post-hatch period
(Prasad et al., 2018b). In two indigenous breeds
i.e. Ghagus and Aseel, the myoglobin (
Mb) gene expression was studied in different organs like muscle, bursa, heart, spleen and gizzard at day old and 4
th week, respectively
(Prasad et al., 2019). In both Aseel and control broiler, the
ACTRIIB gene expression was studied and showed up-regulatedon day one and downward upto 4
th week and gradually increased at 6
th week, respectively
(Vishnu et al., 2017). In Ghagus, the expression of
TNNC1 gene was studied in different organs such as muscle, bursa, heart, spleen and gizzard, respectively
(Prasad et al., 2018a). In this context, we explore the expression profile of
ACACB gene during pre and post-hatch period of IWI line as it no far reported.
Standard curve was plotted to optimize real time PCR, where a regression coefficient (slope) of -2.941 and -4.661 and coefficient of determination (R
2) of 0.9543 and 0.8374, respectively, was found for
ACACB and
GAPDH genes (Fig 2). A significant regression coefficient was found in the target and reference genes representing an optimum standardization of gene expression before carrying out the actual experiments. The amplification plot and dissociation curve of the respective target and reference genes were analyzed by qPCR (Fig 3; Fig 4). The common comparative threshold method of analysis
i.e. 2
-ΔΔCt was used to study the relative gene expression during pre and post-hatch period of IWI line. The ED5 and 18th week stage were used as a calibrator to determine the relative expression during pre and post-hatch periods, respectively. A significant (P<0.05) effect of
ACACB gene expression was observed in IWI line at different stages during different developmental periods (Fig 5; Fig 6). The expression of the
ACACB gene was down-regulated during embryonic period i.e. ED6 to ED18 (on ED6 by 0.40, ED7 by 0.57, ED8 by 0.58, ED9 by 0.38, ED10 by 0.57, ED11 by 0.35, ED12 by 0.26, ED13 by 0.24, ED14 by 0.35, ED15 by 0.62, ED16 by 0.93, ED17 by 0.80 and ED18 by 0.83 folds in this chicken line. During adult stage, the expression was also down-regulated by 0.32 fold on the 22
nd week and 0.40 fold on 40
th week of age. From this result, we may suggest that due to lower expression of the ACACB gene, the abdominal fat deposition is low in layer chicken. Earlier literature also revealed the same trend in mice where mice lacking
ACACB gene showed decreased fat storage in abdomen (
Abu-Elheiga et al., 2001). When metabolic fuel in the body is low and sufficient ATP is needed, ACACB pathway is turned off by phosphorylation. Consequently, the level of malonyl-CoA is reduced, which leads to generation of ATP through increased fatty acid oxidation and the decreased consumption of ATP for fatty acid synthesis. Thus,
ACACB gene expression is important only during
de novo fatty acid synthesis when lipids and triglycerides are synthesized in the body.