| For cherishing endangered wild animals,improving the success rate of reproduction is particularly important for their protection and management.Wild animal pregnancy diagnosis and sex identification help to accurately calculate female reproductive rate and sex ratio,which is essential for establishing population growth models and conducting viability analysis.However,there is no simple,fast,and accurate pregnancy diagnosis and sex identification technology applied to non-human primates.In this study,we used lectin microarray and MS to identify the urinary glycopattern differences of BLMs pregnancy diagnosis and gender identification,and use GSM to further verify whether this method is applicable to other nonhuman primates.To assess the feasibility of urinary glycopattern as potential biomarkers for pregnancy diagnosis and gender identification.Using lectin microarray to screen the differences in urinary protein glycosylation of BLMs pregnant(5)and non-pregnant(including non-pregnant females(10)and postpartum females(4),females and males(10).Secondly,for females of pregnant and non-pregnant,females and males of BLM,we screened STL and SBA respectively,and used MALDI-TOF-MS to identify the changes in the complete N-glycan.Finally,combined with the results of Zhang Simeng's 2019 GSMs glycopattern study,screened out BS-I and MAL-I,and used the same method to identify pregnant females and non-pregnant females,and females and males related N-glycan.(1)The results showed that the STL expressing higher in pregnant females,the PTL-I,PNA,SNA expressing higher in non-pregnant females;and the Jacalin,BS-I,SBA expressing higher in females.(2)Pregnancy and non-pregnancy identification results show that the glycan structures and sialylated N-glycan in pregnant females are more than non-pregnant females.In BLM,16 kinds of N-glycan(m/z 1079.375,1316.630,1589.545,etc.)only appeared in pregnant females,while 3 kinds of N-glycan(m/z 1095.370,1479.547,1641.599)only appeared in non-pregnant females.The relative intensity of N-glycan annotated by m/z 1584.578,1606.560 was higher in pregnant females than in non-pregnant females,ratio value >1.5.In GSM,the 19 kinds of Nglycan(m/z 1707.658,1729.616,1751.597,etc.)only appeared in pregnant females,and 7 Nglycan(m/z 1501.529,1542.555,1549.538,etc.)only appeared in non-pregnant females.The relative intensity of 11 kinds of N-glycan(m/z 1515.508,1622.555,1762.583,etc.)in pregnant females is higher than that in non-pregnant females,ratio value> 1.5.The relatively intensity of 3 kinds N-glycan(m/z ? 1606.560,1730.636,2412.890)in pregnant females is opponent,ratio value <0.67.(3)Female and male identification results show that in BLM,more N-glycan in females than in males.11 kinds of N-glycan(m/z 1460.502,1751.597,1770.642,etc.)only appeared in females,and 6 kinds of N-glycan(m/z 1704.608,1793.644,1812.653,etc.)only appeared in males.The relative intensity of 5 kinds of N-glycan(m/z 1606.560,1647.587,2021.731,etc.)in females is higher than in males,ratio value> 1.5,the relative intensity of m/z 1743.538 in females is lower than in males,ratio value <0.67.In GSM,the m/z 1438.520,1752.618 only appeared in the males,and the relative intensity of m/z 1905.548 in females is higher than males,ratio value >1.5.Pregnancy and gender identification In of BLMs,the N-glycan exist differences in the number of glycan,glyan types,glycan structure,relative abundance,and the degree of sialylation and fucosylation,and the same changes in the GSMs.The corresponding differences indicate that glycomics can be used to characterize pregnancy and gender,which will provide references for the pregnancy diagnosis and gender identification of non-human primates,indicating that urine glycopattern has the feasibility as a potential biomarker for pregnancy diagnosis and gender identification. |
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