Pregnancy Determination And Sex Identification With Urinary Glycopatterns Of Two Mammal Species

The purpose of feeding animal is satisfying the supply and demand for human beings,or increasing the number of wild animals.From the reproductive theory to the practical application,many efforts have been made to know and improve the reproductive capacity of captive and wild animals,especially endangered species.The methods of pregnancy diagnosis and sex identification are of wide application value,but the results up to now are still far from the desired.Here,we take GSMs and rats as examples using the lectin microarray which is simple,rapid and high throughput technique belonging to the glycomics,to assess glycopatterns in these two mammals' urine samples.The samples are divided into different groups based on the specificity of lectins.Meanwhile,the urinary biomarkers that could be used for pregnancy diagnosis and sex identification are identified in these two species.We utilized the microarrays manufactured by 37 kinds of lectins to analyze and compare the urinary glycopatterns from four adult pregnant females,three adult non-pregnant females,five adult male GSMs.Our results indicate:(1)the groups of pregnant and non-pregnant females,normal females and males can be clustered by the detection model consisting of 17 lectins(such as MAL-I?BPL?DBA?PNA?VVA,etc.),according to the differences among them.(2)Four lectins show the significant differences between pregnant and normal females including BS-I and PSA expressing higher in pregnant females,PHA-E+L and DBA expressing higher in non-pregnant females.(3)The gender significant differences in the glycopatterns are found in six lectins including MAL-I,BPL and DBA expressing higher in normal females,PNA,VVA,LCA expressing higher in male GSMs.These lectins' fold changes of normalized fluorescent intensities are more than two times between pregnant and non-pregnant females,formal females and males,respectively.The glycans bound with these lectins can be used as biomarkers for pregnancy diagnosis and sex identification of Sichuan golden snub-nosed monkeys.We also used rats as animal models to test the feasibility of this method.Five adult male rats and five female rats were detected by the lectin microarray manufactured by the same 37 kinds of lectins.The samples of these five female rats were collected before pregnancy,and at the stage of early,medium and late pregnant periods,respectively.The results indicate:(1)the groups of pregnant and non-pregnant females can be clustered bythe detection model consisting of 16 lectins(such as PHA-E,WGA,Jacalin,GNA,VVA etc.)according to the differences between them.(2)Females and males can be clustered by15 lectins(such as EEL,WGA,DBA,ECA,MAL-I etc.).(3)Three lectins show the significant differences between pregnant and normal females including Jacalin,PWM,HHL.(4)The gender significant differences in the glycopatterns are found in five lectins including EEL,PTL-II,Jacalin,AAL,DSA.These lectins' fold changes of normalized fluorescent intensities are more than two times between pregnant and non-pregnant females,formal females and males,respectively.The glycans bound with these lectins can be used as biomarkers for pregnancy diagnosis and sex identification of rats.For the first time,the golden snub-nosed monkey urine was used for glycomics gestation research,which proves that the lectin microarray technique can be used to detect differences between pregnant and non-pregnant,female and male Sichuan golden snub-nosed monkeys,respectively.The study of rats also demonstrates the differences of the urinary glycopatterns between different pregnant stages.This method and results represent a new rapid method of diagnosing pregnancy and identifying gender,which can also be used as a reference for the study and application of other mammals.