The Effects Of Autism Associated CNTNAP2 Variations On Neurons Development And Analysis Of Autism Children’ Symptoms And Their Parents’ Personality Characteristics

Part1 Effects of autism associated CNTNAP2 variations on neurons developmentObjectivesContactin-associated protein-like2(CNTNAP2)is one of the synaptic related proteins,which has been found that disfunction of the CNTNAP2 gene could affect the development of the nervous system.In the previous linkage analysis and genome wide association study,CNTNAP2 gene was associated with clinical manifestaitons of autism.In our previous research,we have been screened 180 cases of autism children in the Chinese Han population via whole exons sequencing,and found 7 CNTNAP2 nonsynonymous mutations in 8 autism children.This study aims to explore the genetic way of these mutations through compare the DNA sequence of children with their parents and discuss the effect of variations to neural development in cellular level by method of neurobiology.Methods1.The Sanger sequencing was used to verify the presence of CNTNAP2 mutations in eight autism children which we screened from the WES by the previous study,and DNA samples of their biological parents was also sequenced by Sanger sequencing to determine the source of the CNTNAP2 mutations.2.We build anthropogenic wild type and mutant forms CNTNAP2 plasmid and transfect the 293 T cells with CNTNAP2 plasmid by Lipofectamine.Using Western blot technique to detect the exogenous CNTNAP2 protein expression in 293 T cells.3.Mouse cortical neurons were transfected with CNTNAP2 wild type or mutant forms plasmid by electroporation and calcium phosphate transfection methods.After fixed by immunohistochemistry,neurons was observed through fluorescence microscope.Results1.Eight autism patients were found to carry seven CNTNAP2 nonsynonymous mutations by whole exons sequencing(c.G209 C,p.G70A;c.T515 C,p.I172T;c.C2328 A,p.D776E;c.A2353 G,p.S785G;c.C2492 T,p.T831L;c.C3517 A,p.P1173T;c.G3709 A,p.D1237 N,there were two children carried the same mutation).All the CNTNAP2 mutations was heterozygous and derived from one of their parent.2.Exogenous wild-type and seven mutants forms of CNTNAP2 plasmids can normally expressed in the 293 T cell and the expression of wild type and mutants plasmids have no statistical difference.3.We found mutant forms CNTNAP2 plasmids could affect neuronal development in vitro culture of mouse cortical neurons.Compared with wild-type group,D776 E,T831L,D1237 N mutation groups had a significantly shorter axons,p<0.05,T831 L and D1237 N mutation groups had a significantly shorter length of neurons and decreased of spine density,p<0.05.ConclusionsThree Non-synonymous mutations(D776E,T831 L,D1237N)found in Chinese han population autism children could affect the neurites outgrouth of neuron.Part 2 Analysis of autism children’ symptoms and their parents’ personality characteristicsObjectivesAutism is a neurodevelopmental disorder which was caused by the environmental and genetic factors.Parental personality is an important part of family environment factors,which could affect the occurrence and the clinical symptoms of autism.The purpose of this study is to investigate the relationship between the autism children’ symptoms and their parents’ personality characteristicsMethods250 cases of children with autism were selected in our cross-sectional study.Generally demographic data of the children and their parents was collected.Autisms patients completed Social Communication Questionnaire(SCQ),parents of autisms patients completed Eysenk Personality Questionnaire(EPQ).The SCQ and EPQ score were compared between different age and gender in autism children.According to whether children SCQ score was higher than 22 points,the patients of autistic children were divided into two groups,then the various dimension of the parents’ EPQ scores were analyzed between two groups.Finally,we analysis the correlation between the symptoms of autism children and their parents’ personality.Results1.Compared to male children,the female children was more serious with repetitive behaviors,the difference was statistically significant(p<0.05).Among them,the mother’s mood of female children was less stable than that who has male children,the difference was statistically significant(p<0.05).2.The patients of autistic children were divided into two groups according to whether their children’s SCQ score was higher than 22 points,the results showed that there was no significant difference between the two groups in four dimensions,which including the emotional stability,the introversion and extroversion,psychoticism and deception.3.Through the analysis of the total score of SCQ,the social communication ability,the language ability,the repetitive and stereotyped behavior of autistic children with the four dimensions of their parents,We found there’s a correlation between the autistic children and their mothers,which contained the social communication ability with the psychoticism;the repetitive and stereotyped behavior with the emotional stability;the repetitive and stereotyped behavior with the psychoticism(p<0.05),however,the correlation coefficient is low(R=0.138,R=0.185,R=0.132),so we had no sufficient proof to indicate there’s a correlation between them.There’s no correlation between the other groupsConclusionsThe female children was more serious with repetitive behaviors than the male one,and the mother’s mood of female children was less stable than that who has male children.There was no correlation between the symptoms of autistic children and their parents’ personality characteristics.