OBJECTIVETo identify proteins in the seminal plasma of healthy fertile men.

METHODSThree seminal plasma samples were collected from healthy fertile volunteers by Percoll isolation, and then the balanced mixture of the seminal plasma was separated by SDS-PAGE. The proteins in the gel band underwent enzymoloysis, and was extracted and identified by shotgun proteomic strategy.

RESULTSA total of 331 proteins were identified, with the molecular weight (MW) ranging from 8 000 to 572 068 and the isoelectric point (pI) from 4.36 to 11.05. Based on the molecular function and biological process of the proteins, 51 (15.4%) were classified as transport proteins, 11 (3.32%) as cell movement proteins, 63 (19.03%) as signal transduction proteins, 147 (44.4%) as proteases, 38 (11.5%) as enzyme regulator proteins, 21 (6.3%) as programmed cell death proteins, 12 (3.62%) as structural proteins and 59 (17.8%) as proteins with unknown molecular function.

CONCLUSIONShotgun proteomic strategy is a good method for protein identification. Annexin A, Annexin-associated proteins and the Ras-related protein Rab were the major members of the signal transducer proteins identified. Ca2+ and G protein signal pathways may play a most important role in the extracellular signal transduction into cells, but the interactions between these proteins remain unknown. The great quantity of enzymes and enzyme regulator proteins identified in the seminal plasma may be closely related with the maintenance of sperm motility and metabolism.

Adult ; Fertility ; Humans ; Male ; Proteomics ; methods ; Semen ; chemistry ; Seminal Plasma Proteins ; isolation & purification ; Sperm Motility

Microdeletion of the azoospermia factor in the Yq of the Y chromosome is one of the important causes of male infertility. Complete deletion of the AZFc region (b2/b4 deletion) is the most common type of AZF deletion. Recent studies have shown a variety of deletions of the AZFc region, including partial deletions, such as gr/gr deletion, b1/b3 deletion and b2/b3 deletion, which may also be associated with male infertility.
Chromosome Deletion ; Chromosomes, Human, Y ; Genetic Loci ; Humans ; Infertility, Male ; genetics ; Male ; Seminal Plasma Proteins ; genetics

Azoospermia factor (AZF) microdeletions of the Y chromosome, which occur in 1 - 55% of infertile men, are closely associated with severe spermatogenic failure and represent the most frequent molecular genetic causes of azoospermia and severe oligozoospermia. Researches on AZF and its related genes, approaching the mechanisms of spermatogenic failure at the molecular level, are of great significance for the diagnosis, treatment and prognosis of male infertility. The detection of AZF microdeletions can provide scientific basis for correct diagnosis and reasonable therapy. This article outlines the structure and functional characteristics of AZF, as well as its relationship with male infertility, cryptorchidism, varicocele, Klinefelter syndrome, seminoma, and recurrent abortion.
Genetic Loci ; Humans ; Infertility, Male ; Male ; Seminal Plasma Proteins ; Sequence Deletion

OBJECTIVETo investigate the effects of recombinant human testis sperm binding protein (TSBP) on human sperm motility parameters in vitro.

METHODSSperm specimens obtained from 22 healthy fertile men were prepared by the Percoll gradient-centrifugation technique. The sperm suspension was incubated with recombinant His6-TSBP at the concentration of 0.01 mg/ml or 0.1 mg/ml at 37 degrees C for 1 or 3 hours in vitro. The combination of the recombinant protein and sperm membrane was determined by Western blot, and the sperm motility parameters were analyzed by computer-aided sperm analysis (CASA). The same procedure was performed for 12 asthenospermia patients.

RESULTSIn the 22 healthy volunteers, the percentage of forward motile sperm was increased after incubated with 0.1 mg/ml recombinant protein for 1 h (P < 0.05), both forward motile sperm percentage and motility were increased after incubated with recombinant protein at the same concentration for 3 h (P < 0.05), but no effect was observed after incubation with 0.01 mg/ml recombinant protein. In the 12 asthenospermia patients, the forward motile sperm percentage was increased after incubated with 0.1 mg/ml recombinant protein for 3 h (P < 0.05), but no statistically significant difference was observed in sperm motility.

CONCLUSIONRecombinant His6-TSBP at the concentration of 0.1 mg/ml can increase sperm motility in healthy fertile men and the forward motile sperm percentage in both healthy fertile men and asthenospermia patients in vitro.

Adult ; Humans ; Infertility, Male ; metabolism ; Male ; Recombinant Proteins ; pharmacology ; Seminal Plasma Proteins ; pharmacology ; Sperm Motility ; drug effects

OBJECTIVETo study the morphology of Y chromosome and microdeletion of the correlated specific azoospermia factor(AZF) region on Y chromosome in cases of azoospermia and to identify the genetic diagnosis made for male infertility patients.

METHODSPeripheral blood samples were taken from two patients with azoospermia, and then were examined by use of G banding, C banding cytogenetic analysis and multiplex polymerase chain reaction (PCR) microdeletion analysis.

RESULTSThe karyotypes of the two cases were 45, X, -Y, -22, +der(Y)t(Y;22)(q11.2;q11.2) and 46, XY, del(Y)(q11.2) respectively. In 12 sequence-tagged sites(STS) of AZFa, AZFb, AZFd, AZFc, only one was detected in the first case and two were detected in the other case.

CONCLUSIONThe cytogenetic analysis and the detection of AZF microdeletion on Y chromosome are essential to the final genetic diagnosis to be made for male infertility patients.

Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Genetic Loci ; Humans ; Infertility, Male ; genetics ; Male ; Seminal Plasma Proteins ; genetics

OBJECTIVETo analyse the variability of proteins in the seminal plasma of severe oligospermic and healthy fertile men.

METHODSSpermatic fluid samples were collected from 11 healthy fertile men and 6 severe oligospermic male volunteers and tested by SELDI-TOF-MS with the CM10 protein chip to get the protein spectra maps, which were automatically treated with the special softwares of Ciphergen Inc.

RESULTSThe mean peak heights of 2 lower-abundance proteins expressed in the seminal plasma of the severe oligospermic men were statistically different from the healthy fertile males (P<0.05). Fifteen different proteins existed between the nonobstructive azoospermic and the severe oligospermic group, 7 of which, with m/z of 7,196.058, 7,547.610, 5,780.493, 7,059.844, 7,409.589, 5,379.173 and 10,778.810, also between the non-obstructive azoospermic and the healthy fertile males (P<0.05). Except the latter two, the contents of the other 5 proteins were decreased in the non-obstructive azoospermic men (P<0.05).

CONCLUSIONThe finger prints of the seminal plasma proteins of the severe oligospermic group were similar to those of the healthy fertile males, both significantly different from the non-obstructive azoospermic men. It is suggested that pathogenesis mechanisms differ exist between non-obstructive azoospermia and severe oligospermia but are not the simple accumulation of genetic factors.

Adult ; Humans ; Male ; Oligospermia ; metabolism ; Semen ; metabolism ; Seminal Plasma Proteins ; analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Recently, intracytoplasmic sperm injection (ICSI) has been extremely successful in the treatment of male infertility. However, the consequent transmission of sperm cytogenetic defects and genetic defects to the offspring has aroused considerable concern. Among infertile men, those with severe spermatogenic defects, including oligozoospermia and azoospermia, are mostly the subjects for ICSI. Therefore it is very important to obtain cytogenetic and chromosomal information on these infertile patients and prevent the inheritance of these genetic defects. This review offers an analysis on the genetic defects among infertile men.
Chromosomes, Human, Y ; genetics ; Genetic Loci ; Humans ; Infertility, Male ; genetics ; therapy ; Male ; Seminal Plasma Proteins ; genetics ; Sperm Injections, Intracytoplasmic

OBJECTIVESTo evaluate the application of two-dimensional electrophoresis in the research of differentially expressed proteins in human spermatozoa.

METHODSTwo-dimensional gel electrophoresis was performed on 4 sperm samples from normal healthy men and another 4 from asthenospermia patients. After silver staining, the differential expression proteins were analyzed by PDQuest 2D analysis software.

RESULTSSeven differential protein spots were identified, 2 expressed highly in the asthenospermia sperm but lowly in the normal spermatozoa, while the other 5 expressed just the opposite way.

CONCLUSIONThe protein profiles of differential expression between the normal spermatozoa and idiopathic asthenospermia were established and some differential proteins were found. The data obtained in this study may help prepare the ground for further studies on the isolation and identification of differentially expressed proteins in human asthenospermia sperm.

Case-Control Studies ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Male ; Oligospermia ; metabolism ; Seminal Plasma Proteins ; analysis ; biosynthesis ; Spermatozoa ; chemistry

OBJECTIVETo explore the effect of seminal plasma lipoprotein (a) in abnormal semen liquefaction and its clinical significance.

METHODSAccording to The WHO Laboratory Manual for the Examination and Processing of Human Semen, we conducted semen routine analyses of 101 patients with abnormal semen liquefaction and 26 normal healthy controls. We added chymotrypsin to the semen for 30 minutes of incubation at 37 degrees C. When there were filaments, we centrifuged the semen and obtained the upper seminal plasma to determine the level of lipoprotein (a).

RESULTSThe level of lipoprotein (a) was significantly higher in the 101 patients ([526.2 +/- 243.5] mg/L) than in the 26 normal controls ([296.9 +/- 105.2] mg/L) (P < 0.01) .

CONCLUSIONLipoprotein (a) can inhibit fibrin dissolution, and delayed fibrin dissolution in semen liquefaction may be related to the increased level of seminal plasma lipoprotein (a). The seminal plasma lipoprotein (a) level should be taken into account in the clinical diagnosis of male infertility caused by abnormal semen liquefaction.

Adult ; Case-Control Studies ; Humans ; Infertility, Male ; metabolism ; physiopathology ; Lipoprotein(a) ; analysis ; Male ; Semen ; metabolism ; Seminal Plasma Proteins ; analysis ; Young Adult

OBJECTIVETo identify the proteins that could improve the resistance of human sperm to cryopreservation using comparative proteomics.

METHODSA total of 31 semen samples from 10 donors were divided into a high recovery and a low recovery group. Differentially expressed proteins in sperm and seminal plasma were detected and compared between the two groups by two-dimensional differential gel electrophoresis and mass spectrometry.

RESULTSTotally, 22 differentially expressed proteins were found in the two groups, 12 seminal plasma proteins, 9 sperm proteins, and 1 belonging to both. These identified proteins were involved in the maturation, movement, energy metabolism, DNA repair and other activities of spermatozoa.

CONCLUSIONMany proteins were identified in sperm and seminal plasma that might influence the resistance of human sperm to cryopreservation.

Adult ; Cryopreservation ; Humans ; Male ; Proteomics ; Semen ; metabolism ; Seminal Plasma Proteins ; metabolism ; Sperm Motility ; Spermatozoa ; metabolism ; Young Adult