Comparative analysis of silencing expression of myostatin (MSTN) and its two receptors (ACVR2A and ACVR2B) genes affecting growth traits in knock down chicken

Abstract

Myostatin (MSTN), a growth differentiation factor-8 regulates muscular development through its receptors, ACVR2A (Activin receptor type IIA) and ACVR2B (Activin receptor type IIB) by inhibiting cellular differentiation of developing somites during embryonic stage and diminishing myofibriller growth during post-embryonic period. The objective of this study was to compare the effect of knockdown of expression of myostatin, ACVR2A and ACVR2B genes on growth traits in chicken. The shRNAs for Myostatin, ACVR2A and ACVR2B genes were designed, synthesized and cloned in DEST vector. The recombinant molecules were transfected into the spermatozoa and transfected spermatozoa were inseminated artificially to the hens to obtain fertile eggs. The fertile eggs were collected, incubated in the incubator and hatched to chicks. Silencing of ACVR2B gene showed significantly higher body weight than other single, double and triple knock down of genes in transgenic birds. The carcass traits such as dressing%, leg muscle%, and breast muscle% were found with the highest magnitudes in birds with silencing of the ACVR2B gene as compared to the birds with that of other genes and control group. The lowest serum cholesterol and HDL content was found in ACVR2B silencing birds. The total RBC count was the highest in this group though the differential counts did not differ significantly among various silencing and control groups of birds. It is concluded that silencing of only one receptor of MSTN particularly, ACVR2B may augment the highest growth in chicken during juvenile stage. Our findings may be used as model for improving growth in other food animals and repairing muscular degenerative disorders in human and other animals.

Muscle Hyperplasia in Japanese Quail by Single Amino Acid Deletion in MSTN Propeptide

“…In C42del/C42del quail, increased muscle mass was caused by muscle fiber hyperplasia, rather than fiber hypertrophy. In addition, muscle fiber hyperplasia was also observed in MSTN knockdown chickens [22]. Although both C42del/C42del quail and MSTN knockdown chickens are not complete MSTN knockout avian models, the data suggests that disruption of the MSTN gene in the avian species will increase muscle mass with muscle fiber hyperplasia, rather than fiber hypertrophy or both.…”

“…The function of MSTN seems to be conserved in avian species as a recent study showed increased muscle mass in MSTN knockdown chickens [22]. To consolidate the anti-myogenic function of MSTN in avian species, an in vivo effect of MSTN mutation on avian muscle growth should be investigated.…”

DrosophilaActivin signaling promotes muscle growth through InR/dTORC1 dependent and independent processes

“…The Mstn/ Activin branch of the TGF- family is very ancient and is present in some pre-Bilateria groups including several cnidarian species (Watanabe, et al, 2014), but functional studies of its role in muscle size control in these animals are lacking. In fact, Mstn's role in muscle growth has only been studied in a few other non-mammalian vertebrates including chickens, turkeys and zebrafish where the results mirror the mammalian case (Bhattacharya, et al, 2019;Gao, et al, 2016;McFarland, et al, 2006). As for invertebrates, there are two published reports, one using the giant prawn Macrobrachium rosenbergii (Easwvaran, et al, 2019) and the other the penaeid shrimp Penaeus monodon (De Santis, et al, 2011).…”

“…Among these, Myostatin (Mstn), a member of the TGF- superfamily of growth and differentiation factors, has proven to be a prominent player. Loss-of-function mutations in mstn have been identified or induced in a large variety of mammals including mice, cattle, dogs, sheep and humans (Clop et al, 2006;Kambadur et al, 1997;Mosher et al, 2007;Schuelke et al, 2004) as well as birds and fish (Bhattacharya et al, 2019;Gao et al, 2016;McFarland et al, 2006). In all these species, loss of mstn results in larger skeletal muscles leading to the conclusion that Mstn is a negative regulator of muscle mass.…”

Development of Protocol for Production of Primary Antibody against Ovalbumin Protein in Chicken for Detection of the Protein through Western Blotting

“…The protein isolated from magnum tissues was run on SDS-PAGE following standard method (Bhattacharya et al, 2019). The proteins separated in SDS-PAGE were transferred into 0.45µm polyvinylidene fluoride (PVDF) membrane.…”