| Objective: To study the reasons, related indicators of changes, the origin ofabnormal karyotypes and the influence on offspring of chromosome abnormalities ofmale patients with testicular spermatogenesis impairment, and to provide moreaccurate genetic counseling for different types of chromosomal abnormalities ofspermatogenesis dysfunctions. In addition, to find related spermatogenesis failuregenes caused by copy number variations in order to provide theoretical basis forfurther research.Methods: From September2009to December2013, infertility males came fromFirst Hospital, Jilin University, who were diagnosed as azoospermia,cryptozoospermia or oligozoospermia and performed chromosome karyotypeanalyses. Patients with obstructive azoospermia were excluded. Of all the2411patients with abnormal semen quality,365cases were detected with chromosomalabnormalities (age from20to48). Basic information was collected by questionnaireincluding ages, height, weight, family histories, medical histories, parents’reproductive ages and others. Clinical information was recorded by doctors’examinations containing testicular volumes, texture, secondary development, etc.Patients’ family members were recalled for chromosome karyotype analyses andsurveillance of their detailed reproductive histories. Sperm concentration wasdetected by WLJY-9000color image analysis system. Semen cytology examinationswere carried out though Diff-Quick staining. Reproductive hormone levels weretested by Roche Electrochemical Luminescence Meter2010. The frequency ofsperm aneuploidy was detected by fluorescence in situ hybridization technology(FISH). Y chromosome microdeletions and SRY gene were examinationed by PCR.Data collection by means of telephone interview focused on the reproductiveoutcomes of couples after chromosome analyses.50cases with normal chromosome karyotypes and semen quality whose infertility reasons from their wives were ascontrol group (age from22to42), and they had fathered normal offspring throughartificial insemination with husband’s semen (AIH) or in vitro fertilization-embryotransfer (IVF-ET). Statistical analysis was performed with SPSS version17.0statistical package. In addition,13cases of patients with abnormal karyotypes foundby G banding were preformed high throughput genome-wide sequencing to findCNVs, and the results were analyzed by retrieving DGV database, OMIM databaseand PubMed.Results:1.365cases with chromosomal abnormalities were detected in2411patients withabnormal semen quality (15.14%). The main types of chromosome abnormalitieswere201cases of Klinefelter syndrome,62cases of chromosomepolymorphisms,41cases of chromosome translocations,23cases ofchromosomal inversions,12cases of XYY syndrome,9cases of chromosomemosaicisms and7cases of male sex reversal syndrome.2. Klinefelter syndrome included184cases of azoospermia, six cases ofcryptozoospermia and seven cases of oligozoospermia.13.05%azoospermicpatients showed mature sperm through semen cytology examinations. There wassignificantly different in height, testicular volumes, FSH, LH, T, and T/LHbetween patients and control group (P<0.05). Only testicular volumes weredifferent in varying degrees of spermatogenesis impairment groups (P<0.05).The oligozoospermic patient with the karyotype of47,XXY[10]/46,XY[90] hadfathered a healthy offspring naturally. The karyotypes of the probands’ parentswere normal. Compared with control group, parents’ reproductive ages were notstatistically different (P>0.05).3. XYY syndrome included four cases of azoospermia and eight cases ofoligozoospermia. The patient had mature sperm by semen cytology examination.The height and testicular volumes were significantly different between patientsand control group (P<0.05). There was not significantly different in age,testicular volumes and reproductive hormone levels in varying degrees ofspermatogenesis impairment groups (P>0.05). The frequency of sperm aneuploidy was not more significantly increased than control (P>0.05). Thekaryotypes of the probands’ parents were normal. Compared with control group,parents’ reproductive ages were not statistically different (P>0.05).4. Chromosome translocations included13cases of Robertsonian translocations (4azoospermia,9oligozoospermia) and28cases of reciprocal translocations (7azoospermia,21oligozoospermia). Semen cytology examinations showeddifferent translocations with different stages of maturation arrest. Thetranslocational breaking point involving1q21in three patients showedazoospermia. There was not significantly different in age, testicular volumes andreproductive hormone levels in varying degrees of spermatogenesis impairmentgroups (P>0.05). The frequency of sperm aneuploidy was not more significantlyincreased than control (P>0.05).36.4%patients with Robertsonian translocationand11.1%patients with reciprocal translocation had fathered their own offspringnaturally or through intracytoplasmic sperm injection (ICSI). One of thekaryotypes of the probands’s fathers or mothers were as the same as theprobands’. Compared with control group, parents’ reproductive ages were notstatistically different (P>0.05).5. Chromosome inversions included10cases of azoospermia and13cases ofoligozoospermia. Inv(9) was the most common types of inversions. All thespermatogenic cells and sperm were not found by semen cytology examinations.The testicular volumes and T/LH were significantly different between patientsand control group (P<0.05). There was not significantly different in age,testicular volumes and reproductive hormone levels in varying degrees ofspermatogenesis impairment groups (P>0.05).23.1%inv(9) patients had fatheredtheir own offspring naturally or through ICSI. One of the karyotypes of theprobands’s fathers or mothers were as the same as the probands’. Compared withcontrol group, parents’ reproductive ages were not statistically different (P>0.05).6. Chromosome polymorphisms included34cases of azoospermia and28cases ofoligozoospermia. The azoospermic patient showed mature sperm by semencytology examination. Only testicular volumes were different in varying degreesof spermatogenesis impairment groups (P<0.05). The frequency of spermaneuploidy was not more significantly increased than control (P>0.05). Patients with Yqh-had higher rate of Y chromosome microdeletions (54.55%).25.0%chromosome polymorphism patients had fathered their own offspring naturally orthrough ICSI. One of the karyotypes of the probands’s fathers or mothers were asthe same as the probands’. Compared with control group, parents’ reproductiveages were not statistically different (P>0.05).7. Seven cases of male sex reversal syndrome were azoospermia and no maturesperm were detected by semen cytology examinations. Y chromosomemicrodeletions were existed in all the patients. SRY gene examinations werepositive in six cases and negative in one case. There was not significantlydifferent in height, testicular volumes, FSH, LH, T, and T/LH between patientsand control group (P<0.05). Patients can not obtain their own offspring naturallyor through ART, but only through AID. The karyotypes of the probands’ parentswere normal. Parents’ reproductive ages were significantly higher than controlgroup (P<0.05).8. Eleven patients with chromosome abnormalities were detected CNVs by usinghigh throughput sequencing. Some CNVs associated with male spermatogenesisimpairment involving the changes of genetic structure or function, such as ALFon2p16.3, NPHP1on2q13, HLA-DRB1on6p21.32, BCL8on15q11.2, COX10,MAK, DNEL1on17p12and DNMT3L on21q22.3.Conclusions:1. Testicular volumes were associated with varying degrees of spermatogenesisimpairment in Klinefelter syndrome and chromosome polymorphisms. However,ages, testicular volumes and reproductive hormone levels were no related withvarying degrees of spermatogenesis impairment in XYY syndrome, chromosometranslocations and inversions.2. The incidence of sperm aneuploidy in patients with XYY syndrome, chromosometranslocations and polymorphisms was not significantly increased than control. Itwas considered as no interchromosomal effect in these three types ofchromosome abnormalities.3. Translocational breaking point occurred on1q21could lead to azoospermia.4. Some patients with mosaic Klinefelter syndrome, chromosome translocations, inv(9), and polymorphisms had the chance of having their own offspringnaturally or through ICSI.5. The abnormal karyotypes of Klinefelter syndrome, XYY syndrome, and male sexreversal syndrome were usually de novo. Chromosome translocations, inversionsand polymorphisms were usually inherited by one of their parents. Theoccurrence of male sex reversal syndrome may be associated with elderly parents,but other types of abnormalities had no concern with older parents.6. Some CNVs detected by High throughput sequencing may involve certainchanges of genetic structures or functions, which may affect gene expression intesticular tissues such as ALF, NPHP1, HLA-DRB1, BCL8, COX10, MAK,DNEL1, DNMT3L and lead to spermatogenesis impairment. |
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