Gene Diagnosis And Assisted Reproductive Therapy Of Globozoospermic Patients

Globozoospermia is a kind of teratozoospermia which causes male infertility,And its cases occur in less than 0.1% among male infertility.The feature of semen is unique,which acrosome of the sperm is completely absent.However,identification of causative genes for globozoospermia is difficult.With the development of global medical technology and genome sequencing technology,the reports from both at home and abroad identified a few genes responsible for globozoospermia,also with the animal model studies,gene diagnosis and treatment for globozoospermic patients become available.At the same time,we can provide reproductive counseling and prevent children of these patients from male infertility.The patients were diagnosed as globozoospermia in the outpatient department of our reproductive center from 2013 to 2018.The criteria of selection are followed: male infertile patients were diagnosed as male infertility for had been not pregnant for more than 1 year in the male infertility clinic of our reproductive center;semen analysis and papanicolaou staining morphological examination showed type Ⅰ globozoospermia or miniacrosome sperm defect which were all round-headed sperm or miniacrosome sperm in semen;the patients were all underwent intracytoplasmic injection of single sperm in our reproductive center.Patients in our experiment are type Ⅰ globozoospermia or miniacrosome sperm defect.We extracted DNA in peripheral blood,amplified all exons of DPY19L2 using PCR by specific primers of DPY19L2 and analysed products by Sanger sequencing to detect DPY19L2 deletions or mutations in these patients.Semen samples of these patients were stained with Papanicolaou,which were then observed under the microscope.It was found that sperm of these patients is round-headed or miniacrosome.PCR of LCR₁ a,LCR₁b,LCR₁ c,LCR₂d,LCR₂ e and exon 1,exon 11,exon 22 in control and 6 globozoospermia and 2 miniacrosome sperm defect patients showed whether there were deletion of DPY19L2 gene or not.We found deletion of DPY19L2 in 3 patients,then PCR of the remaining 5 patients for all 22 exons were sequenced.The results showed that patient with globozoospermia（P7）was found mutation in exon 3 of DPY19L2 c.368A>G;p.His123 Arg and patient with miniacrosome sperm defect for mutation in exon 16 of DPY19L2 gene c.1541 G >A;p.Arg514 His.Complete deletions of DPY19L2 gene were found in P3,P5 and P8.Conclusion: DPY19L2 gene deletion or mutation is the main cause of round head sperm disease.Three patients with globozoospermia had complete deletion of DPY19L2 gene,and one patient with globozoospermia had one homozygous missense mutation of DPY19L2,one miniacrsome sperm defect patient had one homozygous missense mutation of DPY19L2;ICSI combined with AOA is an effective assisted reproductive treatment strategy for patients with globozoospermia and miniacrosome sperm defect.