| ObjectiveIn this project,through the study of the effects of feeding different levels of methionine on hybrid chicken serum on the expression of genes related to the intake of chicken hypothalamic neurons,it was clarified whether the dietary methionine levels regulate the expression of genes related to hypothalamic feeding through muscle secretion of cytokines.Methods1、126 3-week-old hybrid chickens(♀Big bone chicken×♂Recessive white feather broiler)half cock and hen,were randomly divided into three groups according to 2 × 3 factor,each treatment was divided into 3 groups.Three levels of methionine were fed to each treatment group.The weigh and feed intake of chickens in each repeat group were accurately recorded.At 91 days of age,3 roosters and 3 hens were randomly selected from each repeat group,and the serum was separated from the whole blood of the neck.2 、 Determination of Some Biochemical Indicators of Rooster and Hen Serum by Automatic Biochemical Analyzer : total protein,albumin,uric acid,blood sugar,urea,calcium,white ball ratio,amylase,total bilirubin,carbon dioxide.3、The hypothalamus tissue of 15-17 embryo-old chickens was extracted and the neurons were isolated.In vitro culture and identification of neuronal cells.The hypothalamic neuron cells of 48 h were cultured with the serum of rooster and hen,and continue to cultivate 24 h.Real-time PCR and WesternBlot methods were used to detect hypothalamic intake-related gene(NPY、Ag RP、GABAA-ɑ1、MC4R and POMC)and protein expression.Results1、Different dietary methionine levels have a significant effect on ADFI of chickens aged 29-91 days.The ADFI of cock and hen L group was significantly higher than that of M group and L group(P <0.05).2、There was no significant difference in total serum protein,albumin,uric acid,blood sugar,urea,calcium,white ball ratio,amylase,total bilirubin and carbon dioxide of 91-day-old chickens in each treatment group(P>0.05).3 、 Different dietary methionine levels of chicken serum significantly affected NPY m RNA、Ag RP m RNA、GABAA-α1 m RNA expression(P<0.01).L group was significantly higher than H group and M group(P<0.01),there was no significant difference between H and M groups(P>0.05).Chicken serum treated with dietary methionine level had no significant effect on MC4 R m RNA、POMC m RNA(P>0.05).NPY m RNA、Ag RP m RNA、GABAA-α1 m RNA of hen serum treatment was significantly higher than that of cock serum treatment(P<0.05).Effects of serum and sex on NPY m RNA、Ag RP m RNA、GABAA-α1 m RNA of chickens treated with different dietary methionine levels(P<0.05).There was significant interaction between serum and sex of chickens treated with different dietary methionine levels(P<0.05).4 、 Chicken serum treated with dietary methionine level significantly affected NPY、Ag RP、GABAA-α1 protein expression(P<0.05).The serum NPY、Ag RP、GABAA-α1 proteins in L groups were significantly or significantly higher than those in H and M groups(P<0.01),there was no significant difference in MC4 R and POMC proteins between treatment groups(P>0.05).The serum of chickens treated with dietary methionine significantly affected NPY 、 Ag RP protein(P<0.01)and GABAA-α1protein(P<0.05).No significant effect on MC4 R and POMC proteins(P>0.05).Sex had no significant effect on NPY、Ag RP、GABAA-α1、MC4R and POMC proteins(P>0.05).No interaction between dietary methionine level and sex(P>0.05).Conclusions1、The dietary methionine level significantly affected the feed intake of chickens.A low level of dietary methionine is more beneficial to increase the feed intake of 29~91 day old chickens.2 、 The dietary methionine level had no significant effect on serum biochemical indexes of 91 day old chickens.3、The gene(NPY、Ag RP、GABAA-ɑ1)expression of hypothalamic neuron cells was significantly affected by serum of chickens fed with different methionine levels.No significant effect on gene(MC4R and POMC)expression.The expression level of NPY、Ag RP、GABAA-ɑ1 gene m RNA was significantly higher than that of rooster serum.There was a significant interaction between serum and sex of chickens treated with different dietary methionine levels.4 、 The protein(NPY 、 Ag RP 、 GABAA-ɑ1)expression of hypothalamic neuron cells was significantly affected by serum of chickens fed with different methionine levels.No significant effect on protein(MC4R and POMC)expression.There was no significant difference between treatment of cock serum and hen serum.Sex had no significant effect on NPY、Ag RP、GABAA-α1、MC4R and POMC protein,and there was no interaction between dietary methionine level and sex. |
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