Simultaneous expression of growth hormone releasing hormone (GHRH) and hepatitis B surface antigen/somatostatin (HBsAg/SS) fusion genes in a construct in the skeletal muscle enhances rabbit weight gain.
Somatostatin (SS) and growth hormone-releasing hormone (GHRH) are synthesized and secreted by the hypothalamus, which can control the synthesis and secretion of the growth hormone (GH) from the hypophysis as well as regulate the GH concentrations in animals and humans. In this article, we describe the regulation of animal growth using plasmid DNA encoding both the GHRH gene and the SS gene fused with the hepatitis B surface antigen (HBsAg) gene. We constructed a series of expression plasmids to express the GHRH and HBsAg-SS fusion genes individually as well as collectively. The fusion gene and GHRH were successfully expressed in Chinese hamster ovary (CHO) cells, as proven by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting tests. Poly D, L-lactide-co-glycolic acid (PLGA) plasmid-encapsulating microspheres were prepared and injected intramuscularly into the leg skeletal muscles of rabbits. Weight gain/day and the levels of insulinlike growth factor-I (IGF-I), SS, and hepatitis B surface antibody (HBsAb) were monitored. During days 30 postinjection, increase in weight gain/day and IGF- I concentration and decrease in SS were observed in treatment groups. From days 15 to 30 postinjection, the weight gain/day significantly increased (P < 0.05) by 129.13%, 106.8%, and 72.82% relative to the control group in the co-expression GHRH and fusion gene (named P-G-HS), fusion gene (named P-HS), and GHRH (named P-G) groups, respectively. And most importantly, the P-G-HS group showed significant weight gain/day (P < 0.05) relative to the P-G and P-HS groups. A significant increase in the IGF-I concentration and decrease in the SS level relative to the control group were also observed. The results indicated that the combination of plasmid-mediated GHRH supplementation and positive immunization against SS led to more robust weight gain/day in rabbits.
Dai
Jian-wei
JW
Department of Biochemistry, Veterinary College, Jinlin University, Changchun, People's Republic of China.
Liu
Song-cai
SC
Hao
Lin-lin
LL
Zhang
Yong-liang
YL
Zhang
Qianqian
Q
Ren
Xiao-hui
XH
Jiang
Qing-yan
QY
D000818:Animals; D001835:Body Weight / Q000187:drug effects / Q000502:physiology; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry / Q000235:genetics; D050939:Gene Fusion; D007987:Gonadotropin-Releasing Hormone / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D019344:Lactic Acid / Q000008:administration & dosage; D008297:Male; D008863:Microspheres; D018482:Muscle, Skeletal / Q000502:physiology*; D010957:Plasmids / Q000235:genetics; D011100:Polyglycolic Acid / Q000008:administration & dosage; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D016133:Polymerase Chain Reaction; D011817:Rabbits / Q000254:growth & development* / Q000502:physiology; D013004:Somatostatin / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D014162:Transfection / Q000662:veterinary
D006514:Hepatitis B Surface Antigens; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D007987:Gonadotropin-Releasing Hormone; D019344:Lactic Acid; D013004:Somatostatin; D007334:Insulin-Like Growth Factor I; D004247:DNA