N-carbamylglutamate restores nitric oxide synthesis and attenuates high altitudeinduced pulmonary hypertension in Holstein heifers ascended to high altitude

Background: High-altitude pulmonary hypertension(HAPH) is a life-threating condition for animals in high altitude,and disturbance of endothelial nitric oxide(NO) synthesis contributes to its pathogenesis. N-carbamylglutamate(NCG), which enhances arginine synthesis, promotes endogenous synthesis of NO. In this study, we determined the effects of NCG on alleviating HAPH in Holstein heifers that ascended to Tibet(Lhasa, 3,658 m).Methods: Exp. 1, 2,000 Holstein heifers were transported from low elevation(1,027 m) to Lhasa. After being exposed to hypoxia for 1 yr, Holstein heifers were assigned to a healthy group(Control, n = 6) with mean pulmonary hypertension(mPAP) < 41 mmHg, and an HAPH affected group(HAPH, n = 6) with mPAP > 49 mmHg.Lung tissues were collected to evaluate histopathological changes and the expression of endothelial nitric oxide synthase(eNOS). Exp. 2, ten healthy heifers and 10 HAPH affected heifers were supplemented with NCG(20 g/d per heifer) for 4 wk. Physiological parameters were determined and blood samples were collected on d-1 and d 28 of the feeding trial.Results: Expression of e NOS in small pulmonary arteriole intima was higher in the healthy than HAPH group(P = 0.006), whereas HAPH group had significantly thicker media and adventitia than healthy group(all P < 0.05). The mRNA of eNOS and protein level of eNOS were higher in the lungs of heifers in the healthy group than in the HAPH group(both P < 0.001), whereas endothelin-1 protein levels were higher in HAPH group than in the healthy group(P = 0.025). NCG supplementation decreased mPAP and ammonia(both P = 0.001), whereas it increased the expression of eNOS, arginine, and plasma NO(all P < 0.05).Conclusions: The expression of eNOS was decreased in Holstein heifers with HAPH. NCG supplementation decreased m PAP through the restoration of eNOS and endogenous NO synthesis.

Association of a missense mutation in the luteinizing hormone/choriogonadotropin receptor gene (LHCGR) with superovulation traits in Chinese Holstein heifers

Background： Upon binding luteinizing hormone in the ovary, the luteinizing hormone/choriogonadotropin receptor （LHCGR） is necessary for follicular maturation and ovulation, as well as luteal function. We detected mutations in the LHCGR gene and evaluated their association with superovulation. Methods： Using polymerase chain reaction-single strand sequencing, we examined polymorphisms in LHCGR and Chinese Holstein heifers. conformation polymorphism （PCR-SSCP） and DNA the genotypes associated with superovulation traits in 127 Results： A G/T polymorphism （ss52050737） in exon 11 was significantly associated with the total number of ova and the number of transferable embryos. Conclusions： LHCGR may be a new predictor for superovulation in Chinese Holstein heifers.

The crucial role of lysine in the hepatic metabolism of growing Holstein dairy heifers as revealed by LC-MS-based untargeted metabolomics

The objective of this experiment was to evaluate the effect of supplementing rumen-protected Lys based on a Lys-deficient diet on liver metabolism in growing Holstein heifers.The experiment was conducted for 3 months with 36 Holstein heifers(initial body weight:200±9.0 kg;7-month-old).Heifers were randomly assigned to 2 diets based on corn,soybean meal,alfalfa hay,and wheat bran:control,Lysdeficient diet(LD;0.66%Lys in diet),and Lys-adequate diet(LA;1.00%Lys in diet).The results showed no difference in growth performance between the 2 groups(P>0.05).However,there was a clear trend of increasing feed conversion rate with Lys supplementation(0.05<P<0.01).The serum urea nitrogen concentration was significantly decreased,and the aspartate aminotransferase-to-alanine aminotransferase ratio was significantly decreased by Lys supplementation(P<0.05).Moreover,growing heifers fed a Lys-adequate diet had lower levels of urine nitrogen excretion and higher levels of the biological value of nitrogen(P<0.05).Metabolomic analysis revealed that 5 types of phosphatidylcholine and 3 types of ceramide were significantly increased and enriched in sphingolipid metabolism and glycero phospholipid metabolism(P<0.05).His,Leu,and Asp levels were significantly decreased in the liver following Lys supplementation(P<0.05).In conclusion,Lys supplementation may promote the synthesis of body tissue proteins,as evidenced by significantly decreased amino acids in the liver and urine N excretion,it also improves hepatic lipid metabolism by providing lipoprotein precursors.