1.Expression characteristics of the USP24 gene in the mouse testis during spermatogenesis.
Qi-Yi HU ; Qiong DENG ; Jian-Wen ZHANG ; Fan ZHI ; Rui SUN ; Hui LIANG
National Journal of Andrology 2017;23(11):963-968
Objective:
To investigate the expression characteristics of the USP24 gene in the mouse testis and its role in spermatogenesis.
METHODS:
We examined the expression characteristics of USP24 in the testis tissues of wild-type mice at different postnatal weeks (PNW) and androgen receptor (AR)-knockout (ARKO) adult mice using real-time quantitative PCR and immunofluorescence, and detected the transcriptional activity of the USP24 promoter by dual-luciferase reporter gene assay.
RESULTS:
The expression of the USP24 gene was low in the testis tissue of the wild-type mice at PNW 1, increased dramatically at PNW 3 and stayed at a similar level till PNW 8. The USP24 protein was located mainly in the cytoplasm of Sertoli and spermatogenic cells. Compared with the wild-type, the adult ARKO mice showed a decreased expression of USP24 localized in the posterior head and mid-piece of the mature sperm in the testis. Dual-luciferase reporter gene assay showed that the transcriptional activity of the USP24 promoter was increased after testosterone stimulation.
CONCLUSIONS
The increased expression of the USP24 gene was associated with the initiation of sexual development, and the USP24 protein was expressed in the mature sperm of the mice. USP24 is an AR-target gene, which may be involved in the regulation of spermatogenesis in mice.
Animals
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Male
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Mice
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Mice, Knockout
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Promoter Regions, Genetic
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Receptors, Androgen
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genetics
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Sertoli Cells
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Spermatogenesis
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genetics
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Spermatozoa
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metabolism
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Testis
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metabolism
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Testosterone
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administration & dosage
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Transcription, Genetic
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Ubiquitin Thiolesterase
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genetics
;
metabolism
3.Establishment of a CMTM2 transgenic mouse model and the alteration of its serum testosterone level.
Zhen-Hua LIU ; Yun-Bei XIAO ; Xiao-Wei ZHANG ; Jing XIE ; Tao XU ; Xiao-Feng WANG
National Journal of Andrology 2012;18(6):483-486
OBJECTIVETo establish a transgenic mouse model systemically expressing the CMTM2 gene and study the effect of the CMTM2 expression on the reproductive system of mice in vivo.
METHODSTransgenic mice were generated by microinjection of pRevTRE-CMTM2 and the genotype was detected by PCR. The expression of CMTM2 was determined by RT-PCR, Western blot and immunohistochemistry, and the serum testosterone level was measured by radioimmunoassay.
RESULTSThe CMTM2 gene was highly expressed in the testis of the transgenic mouse models and in their offspring as well. The level of serum testosterone was significantly increased in the transgenic models as compared with the wild-type mice ([46.04 +/- 3.72] vs [42.43 +/- 3.80] nmol/L, P < 0.05).
CONCLUSIONThe transgenic mouse model was established successfully, which could highly express the CMTM2 gene. It is indicated that CMTM2 may influence steroidogenesis and testosterone secretion in transgenic mice.
Animals ; Genotype ; MARVEL Domain-Containing Proteins ; genetics ; Mice ; Mice, Inbred C57BL ; Mice, Inbred ICR ; Mice, Transgenic ; Testosterone ; blood
4.Microdissection testicular sperm extraction for patients with non-mosaic Klinefelter's syndrome: An update.
Zhe YU ; Jun YANG ; Ji-Hong LIU
National Journal of Andrology 2017;23(9):842-847
Klinefelter's syndrome (KS) is a most frequent sex chromosomal disorder in males, which is characterized by hypogonadism and infertility. The development of assisted reproductive technology has made it possible for KS males to father children. Microdissection testicular sperm extraction (mTESE) is widely considered to be the best method for sperm retrieval in KS patients. This article presents an overview on mTESE for men with non-mosaic KS in the aspects of its predictors, sperm retrieval rate, operation procedure, preoperative hormonal therapy, and postoperative complications and testosterone reduction.
Adult
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Humans
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Klinefelter Syndrome
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genetics
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Male
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Microdissection
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adverse effects
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methods
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Postoperative Complications
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etiology
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Sperm Retrieval
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Spermatozoa
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Testis
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Testosterone
5.Association between polymorphism of CYP17 gene and serum hormone concentrations in aged men.
Jun-Qi WANG ; Xiang GU ; Jia-Cun CHEN ; Xiao-Qing SUN ; Hong-Tao MU ; Zeng-He WEI ; Ming LI ; Dian-Qi XIN ; Yan-Qun NA
National Journal of Andrology 2005;11(6):442-444
OBJECTIVETo investigate the association between polymorphism of CYP17 gene and serum hormone concentrations in aged men.
METHODSEighty-three healthy men at the average age of 66.7 were divided into a < 66.7 group (n = 36) and a > 66.7 group (n = 47), and the polymorphism of CYP17 gene in the 5' promoter region was investigated by PCR using DNA from the men's peripheral blood lymphocytes. A new recognition site was created for the restriction enzyme MspA1 I by transition (T --> C) in the risk allele (A2). Three genotypes A1/A1, A1/A2, A2/A2 were established, serum sex-hormone levels measured, and mean hormone concentration evaluated in each genotype and age group.
RESULTSNo evidence was found that the testosterone (T) level, estrogen (E2) level and T/E2 ratio were associated with the genotype of CYP17 gene. There was no significant difference in T and E2 levels between the two groups, but there was a significant increase in the T/E2 ratio (P < 0.05).
CONCLUSIONA2 allele does not increase sex hormone levels in aged men, but the T/E, ratio was higher in the > 66.7 group than in the < 66.7 group. This may be closely associated with the mechanism of benign prostate hyperplasia and prostate cancer in aged men.
Adult ; Aged ; Aged, 80 and over ; Estradiol ; blood ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Steroid 17-alpha-Hydroxylase ; genetics ; Testosterone ; blood
7.An assessment of androgenic/anti-androgenic effects of GH transgenic carp by Hershberger assay.
YuMei LIU ; WenZhong ZHANG ; Ling YONG ; XiaoHong ZHAO ; XuDong JIA ; Ning LI
Biomedical and Environmental Sciences 2011;24(4):445-449
OBJECTIVETo evaluate the androgenic and anti-androgenic effects of GH (growth hormone) transgenic carp in male rats.
METHODSHershberger assay was carried out in castrated male SD rats aged 4-5 weeks. Testosterone propionate (TP) (0.4 mg/kg BW) was administrated for a positive control, GH transgenic carp (3.0 g/kg BW)+TP (0.4 mg/kg BW), parental carp (3.0 g/kg BW) + TP (0.4 mg/kg BW), and flutamide (Flu) (3.0 g/kg BW) were used for negative controls, and vehicle was administered orally for a blank control. All groups were administrated for 10 consecutive days. At the end of the test, animals were anesthetized, then weights of accessory sex organ were measured. Serum testosterone (T), luteinizing hormone (LH), and Follicle-Stimulating Hormone (FSH) levels were detected.
RESULTSThe weights ratios of the accessory sex organs and body weights showed no significant differences between the solvent control and the GH transgenic carp-treated groups. Serum concentrations of FSH, LH, and T of the rats treated with GH transgenic carp + TP showed no significant changes, compared with those treated with TP only.
CONCLUSIONGH transgenic carp does not have any androgenic agonist or antagonist properties in vivo screening tests.
Animals ; Animals, Genetically Modified ; Carps ; genetics ; Follicle Stimulating Hormone ; blood ; Genitalia, Male ; drug effects ; Growth Hormone ; genetics ; metabolism ; pharmacology ; Luteinizing Hormone ; blood ; Male ; Rats ; Testosterone ; blood
8.Microdeletions in the Y chromosome of patients with idiopathic azoospermia.
Akiyuki SHIMIZU ; Tomohiko ICHIKAWA ; Noriyuki SUZUKI ; Takako YAMAZAKI ; Takashi IMAMOTO ; Satoko KOJIMA ; Yukio NAYA ; Akira KOMIYA ; Hiroyoshi SUZUKI ; Koichi NAGAO ; Kazukiyo MIURA ; Haruo ITO
Asian Journal of Andrology 2002;4(2):111-115
AIMTo evaluate the occurrence and prevalence of microdeletions in the gamma chromosome of patients with azoospermia.
METHODSDNA from 29 men with idiopathic azoospermia was screened by polymerase chain reaction (PCR) analysis with a set of gamma chromosome specific sequence-tagged sites (STSs) to determine microdeletions in the gamma chromosome.
RESULTSDeletions in the DAZ (deleted in azoospermia) loci sgamma254 and sgamma255 were found in three patients with idiopathic azoospermia, resulting in an estimated frequency of deletions of 10.7% in idiopathic azoospermia men.
CONCLUSIONWe conclude that PCR analysis is useful for the diagnosis of microdeletions in the Y chromosome, which is important when deciding the suitability of a patient for assisted reproductive technology such as testicular sperm extracion-intracytoplasmic sperm injection (TESE-ICSI).
Adult ; Base Sequence ; Chromosomes, Human, Y ; DNA Primers ; Euchromatin ; genetics ; Follicle Stimulating Hormone ; blood ; Heterochromatin ; genetics ; Humans ; Luteinizing Hormone ; blood ; Male ; Oligospermia ; blood ; etiology ; genetics ; Polymerase Chain Reaction ; Prolactin ; blood ; Sequence Deletion ; genetics ; Sequence Tagged Sites ; Testosterone ; blood
9.Androgen insensitivity syndrome: do trinucleotide repeats in androgen receptor gene have any role?
Singh RAJENDER ; Nalini J GUPTA ; Baidyanath CHAKRAVARTY ; Lalji SINGH ; Kumarasamy THANGARAJ
Asian Journal of Andrology 2008;10(4):616-624
AIMTo investigate the role of CAG and GGN repeats as genetic background affecting androgen insensitivity syndrome (AIS) phenotype.
METHODSWe analyzed lengths of androgen receptor (AR)-CAG and GGN repeats in 69 AIS cases, along with 136 unrelated normal male individuals. The lengths of repeats were analyzed using polymerase chain reaction (PCR) amplification followed by allelic genotyping to determine allele length.
RESULTSOur study revealed significantly shorter mean lengths of CAG repeats in patients (mean 18.25 repeats, range 14-26 repeats) in comparison to the controls (mean 22.57 repeats, range 12-39 repeats) (two-tailed P < 0.0001). GGN repeats, however, did not differ significantly between patients (mean 21.48 repeats) and controls (mean 21.21 repeats) (two-tailed P = 0.474). Among patients' groups, the mean number of CAG repeats in partial androgen insensitivity cases (mean 15.83 repeats) was significantly less than in complete androgen insensitivity cases (mean 19.46 repeats) (two-tailed P < 0.0001).
CONCLUSIONThe findings suggest that shorter lengths of repeats in the AR gene might act as low penetrance genetic background in varying manifestation of androgen insensitivity.
Androgen-Insensitivity Syndrome ; genetics ; DNA ; genetics ; Follicle Stimulating Hormone ; blood ; Genetic Predisposition to Disease ; Humans ; Luteinizing Hormone ; blood ; Male ; Phenotype ; Receptors, Androgen ; genetics ; Regression Analysis ; Testosterone ; blood ; Trinucleotide Repeats ; genetics
10.Routine screening for classical azoospermia factor deletions of the Y chromosome in azoospermic patients with Klinefelter syndrome.
Jin Ho CHOE ; Jong Woo KIM ; Joong Shik LEE ; Ju Tae SEO
Asian Journal of Andrology 2007;9(6):815-820
AIMTo evaluate the occurrence of classical azoospermia factor (AZF) deletions of the Y chromosome as a routine examination in azoospermic subjects with Klinefelter syndrome (KS).
METHODSBlood samples were collected from 95 azoospermic subjects with KS (91 subjects had a 47,XXY karyotype and four subjects had a mosaic 47,XXY/46,XY karyotype) and a control group of 93 fertile men. The values of testosterone, follicle stimulating hormone (FSH) and luteinizing hormone (LH) were measured. To determine the presence of Y chromosome microdeletions, polymerase chain reaction (PCR) of five sequence-tagged site primers (sY84, sY129, sY134, sY254, sY255) spanning the AZF region, was performed on isolated genomic DNA.
RESULTSY chromosome microdeletions were not found in any of the 95 azoospermic subjects with KS. In addition, using similar conditions of PCR, no microdeletions were observed in the 93 fertile men evaluated. The level of FSH in KS subjects was higher than that in fertile men (38.2 +/- 10.3 mIU/mL vs. 5.4 +/- 2.9 mIU/mL, P < 0.001) and the testosterone level was lower than that in the control group (1.7 +/- 0.3 ng/mL vs. 4.3 +/- 1.3 ng/mL, P < 0.001).
CONCLUSIONOur data and review of the published literature suggest that classical AZF deletions might not play a role in predisposing genetic background for the phenotype of azoospermic KS subjects with a 47,XXY karyotype. In addition, routine screening for the classical AZF deletions might not be required for these subjects. Further studies including partial AZFc deletions (e.g. gr/gr or b2/b3) are necessary to establish other mechanism underlying severe spermatogenesis impairment in KS.
Adult ; Azoospermia ; blood ; etiology ; genetics ; Case-Control Studies ; Chromosomes, Human, Y ; genetics ; Gene Deletion ; Genetic Loci ; Genetic Testing ; methods ; Humans ; Karyotyping ; Klinefelter Syndrome ; blood ; complications ; genetics ; Male ; Seminal Plasma Proteins ; genetics ; metabolism ; Testis ; metabolism ; Testosterone ; blood