Molecular Investigations of the Effect of Thermal Manipulation During Embryogenesis on the Enzymatic Activity of Creatine Kinase and Lactate Dehydrogenase in Broiler Muscle

As Dalab,Km Alkhodair,Ta Althnaian,Sy Al-Ramadan,Am Ali

doi:10.1590/1806-9061-2020-1369

Abstract

Incubating temperature and timing or duration is critical to determine the optimum protocol of thermal manipulation (TM), which underlines muscle growth improvement. Therefore, the aim of the present study is to determine the optimum period of embryonic TM that may result in the improvement of pectoral and thigh muscle myogenesis. This is done by investigating the level of mRNA expression of creatine kinase (CK) and lactate dehydrogenase (LDH). An additional goal is measuring the blood levels of CK and LDH as a biomarker of muscle injury due to the experimental thermal challenge on post-hatch day 35. The study was conducted on 1,440 fertile eggs (Ross broilers) that were divided randomly and equally into a control group and four treatment groups (TM1, TM2, TM3, and TM4). The treatment groups were daily subjected to TM at 39 ºC for 18h with 65% relative humidity (RH) during embryonic days (EDs) 7-11, 11-15, 15-18, and 7-18, respectively. Among the thermally manipulated groups that were investigated, TM1 (ED 7-11) resulted in significant improvement of mRNA expression and enzymatic concentration of CK and LDH in muscle during embryogenesis, as compared to the control. Six hours of TC showed the highest significant CK and LDH expression and concentration levels in the control as compared to TM groups. Thus, the results of this study indicate that TM during ED 7-11 improves pectoral and thigh muscles’ response to heat stress without adversely affecting their performance. This finding could be used by commercial breeders to enhance local broiler production.

Highlights

During embryogenesis, creatine kinase (CK) is expressed at a high level in developing embryonic striated muscle
The 20μl reaction mix was prepared from 10μl of TM4 from embryonic days (EDs) 7 to ED 11 resulted in a higher significant master mix; 2μl forward primer pM/μl; 2μl reverse increase in mRNA expression of CK and lactate dehydrogenase (LDH) in both primer pm/μl (Table 1); 2μl cDNA of the sample; and 4μl pectoral and thigh muscles as compared to the control of nuclease-free water
This study shows that embryonic thermal manipulation has influenced the expression of CK for pectoral muscles more than for thigh muscles in TM1 and TM4 (ED 7–11)

Summary

Introduction

CK is expressed at a high level in developing embryonic striated muscle It has been used as an early marker for skeletal myogenesis in chickens by maintaining a high ATP/ADP ratio (Iyengar et al, 1983, Lyons et al, 1991). CK plays a role in cellular energy metabolism: it catalyzes the reversible transfer of high-energy phosphate from ATP to creatine, facilitating the storage of energy in the form of phosphocreatine (Hettling & van Beek 2011) Another proposed function of CK is the transportation of energy from ATPproducing to ATP-consuming sites via a shuttle mechanism involving a mitochondrial and a myofibrillar isoform of CK (Hettling & van Beek 2011). It is involved in the final steps of glycolysis eRBCA-2020-1369

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