OBJECTIVETo analyze the activity of human sperm acrosin and semen parameters in male infertile patients and discuss the effect of sperm acrosin activity on semen quality.

METHODSActivity of human sperm acrosin, PMN-elastase, fructose, alpha-glucosidase, zinc, acid phosphatase levels and HOST of 214 male infertile patients were detected. Semen analysis was performed using WLJY-9000 WeiLi colorful semen quality analyzer. There were 111 cases of normal activity of human sperm acrosin (48.2 - 218.7 microIU/10(6) sperm), 103 cases abnormal (< 48.2 microIU/10(6) sperm). Using the group of normal activity of sperm acrosin to be the control, semen parameters was analyzed and compared with those of the group of abnormal activity of sperm acrosin.

RESULTSThere were significant difference (P < 0.001) between the 2 groups (normal and abnormal) in the areas of sperm density, motile sperm rate, percentage of grade (a + b) sperm and HOST. There were also significant difference in PMN-elastase, fructose and alpha-glucosidase (P < 0.05). There was no difference among sperm volume, zinc and acid phosphatase (P > 0.05).

CONCLUSIONThere was a strong correlation between the activity of human sperm sperm acrosin and semen quality. Activity of sperm acrosin is a reliable index of semen quality.

Acrosin ; metabolism ; Adult ; Humans ; Infertility, Male ; enzymology ; physiopathology ; Male ; Middle Aged ; Semen ; chemistry ; Sperm Count ; Sperm Motility ; Spermatozoa ; enzymology

OBJECTIVETo study the effect of Percoll selection technique on normal morphology and acrosin activity of human spermatoza.

METHODSThe sperm morphology and sperm acrosin activity were analyzed by automated sperm morphology analyzer(ASMA) and spectrocolorimetry.

RESULTSThe normal morphology sperm rate and acrosin activity were significantly increased after Percoll selection technique (P < 0.001).

CONCLUSIONPercoll selection technique could affect normal morphology sperm ratio and acrosin activity.

Acrosin ; metabolism ; Adult ; Centrifugation, Density Gradient ; Fertilization in Vitro ; Humans ; Male ; Spermatozoa ; cytology ; enzymology

Protein kinase C (PKC) is localized in the equatorial segment and the principal piece of the tail of spermatozoa. Activator of PKC results in increasing flagellar motility of sperm that is blocked by PKC inhibitors such as staurosporine. A good correlation (r = 0.9, P < 0.001) is found between the content of PKC in sperm and sperm motility. Zona pellucida (ZP) stimulates the spermatozoa binding the acrosome reaction resulting in the release of hydrolytic enzymes and in the exposure of new membrane domains. ZP binding to receptors in the plasma membrane can regulate adenyl cyclase (AC) leading to elevation of cAMP and protein kinase A (PKA) activation. The PKA activates a voltage-dependent Ca2+ channel in the outer acrosomal membrane which releases Ca2+ from the interior of the acrosome to the cytosol. Activation of the PLC resulted from the rise in Ca2+ hydrolyze phosphatidyl inositol bisphosphate. The product activate PCK to open a voltage-dependent Ca2+ channel (L) in the plasma membrane, leading to the second (II) Ca2+ higher increase which result in membrane fusion and acrosome reaction. It is proposed that PKC would be involved in the regulation of motility and acrosome reaction of sperm.
Acrosome Reaction ; physiology ; Humans ; Male ; Protein Kinase C ; metabolism ; Sperm Motility ; physiology ; Spermatozoa ; enzymology ; physiology

OBJECTIVESTo evaluate the application of Coomassie brilliant blue (CBB) G250 staining for the detection of human sperm deformity rate, rate of intact acrosome and acrosome reaction.

METHODSThe smear of spermatozoa before and after capacitation and induced acrosome reaction with progesterone (P) were stained with 0.05% CBB G250 and Wright-Giemisa solution respectively, and visualized with light microscopy. The deformity rate of spermatozoa, rate of intact acrosome and acrosome reaction were calculated.

RESULTSThere was no any difference in detection of deformity rate of spermatozoa and rate of intact acrosome with CBB G250 and Wright-Giemisa staining(P < 0.05). The sperm population of acrosome reaction with induced P was divided by CBB staining into two types: positive staining with dark violet blue on acrosome cap and pale or negative staining on the same area. The rate of the latter was increasing with increasing inductive time, maybe representative of the rate of acrosome reaction. The mean rate was(75.1 +/- 3.8)% after induced for 1 h.

CONCLUSIONSCBB G250 staining is a reliable method for assessment of the human sperm morphology and acrosome reaction.

Acrosome ; metabolism ; Acrosome Reaction ; Humans ; Male ; Rosaniline Dyes ; chemistry ; metabolism ; Spermatozoa ; cytology ; enzymology ; Staining and Labeling

OBJECTIVETo detect succinate dehydrogenase (SDH) of sperm mitochondria by an improved method, and to evaluate the relationship between SDH and the motility and viability of sperm.

METHODSIn 46 fertile and infertile men (aged from 25 to 36), the motility, viability and mitochondrial SDH of sperm were detected by computer-assisted semen analysis system, dead/live sperm molecular fluorescent probe and an improved cytochemical method. Then, the correlation between the motility and viability of sperm and the positive percentage of sperm mitochondrial SDH were analyzed.

RESULTSIn the 46 fertile and infertile men, the motility and viability of sperm were (67.33 +/- 7.37)% and (79.78 +/- 7.65)%, and the positive percentage of sperm mitochondrial SDH was (74.74 +/- 8.29)%. The motility and viability of sperm and positive percentage of sperm mitochondrial SDH had significant correlation (r = 0.901, P < 0.01; r = 0.876, P < 0.01; r = 0.917, P < 0.01).

CONCLUSIONSDetection of sperm mitochondrial SDH has significance in evaluation of sperm mitochondrial function and may serve as an assisted marker of sperm viability.

Adult ; Case-Control Studies ; Cell Survival ; Humans ; Image Processing, Computer-Assisted ; Infertility, Male ; enzymology ; Male ; Mitochondria ; enzymology ; Sperm Motility ; Spermatozoa ; enzymology ; Succinate Dehydrogenase ; metabolism

OBJECTIVETo study the impacts of positive antisperm antibody (AsAb) in seminal plasma on acrosomal enzyme activity, nitric oxide synthase (NOS) and superoxide dismutase (SOD) levels of spermatozoa.

METHODSSwatch from 40 infertile patients with positive AsAb in seminal plasma as experimental group, and 40 fertile men as control group. Acrosomal enzyme activity was detected by the BAEE/ADH unitive method, NOS was detected by the redoxreaction assay, and SOD level was measured xanthine oxidase method.

RESULTSCompared with control group, acrosomal enzyme activity of spermatozoa of experimental group was significantly decreased (P <0.01), NOS activity was apparently increased (P < 0.01), and SOD level in seminal plasma was markedly decreased (P<0.01).

CONCLUSIONIt may be possible that the positive AsAb in seminal plasma beget infertility through the changes of acrosomal enzyme of spermatozoa, SOD and NOS activities in seminal plasma.

Acrosin ; metabolism ; Adult ; Autoantibodies ; analysis ; Case-Control Studies ; Humans ; Infertility, Male ; enzymology ; immunology ; Male ; Nitric Oxide Synthase ; metabolism ; Semen ; enzymology ; Spermatozoa ; enzymology ; immunology ; Superoxide Dismutase ; metabolism

The serine/threonine phosphatase (PP1) isoform PP1 gamma 2, predominantly expressed in the testis, is a key enzyme in spermatozoa. High PP1 gamma 2 catalytic activity holds motility in check in immature spermatozoa. Inhibition of PP1 gamma 2 causes motility initiation in immature spermatozoa and motility stimulation and changes in flagellar beat parameters in mature spermatozoa. The PP1 gamma 2 isoform is present in all mammalian spermatozoa studied: mouse, rat, hamster, bovine, non-human primate and man. We have now identified at least four of its regulatory proteins that regulate distinct pools of PP1 gamma 2 within spermatozoa. Our studies provide new insights into biochemical mechanisms underlying development and regulation of sperm motility. We hypothesize that changes in sperm PP1 gamma 2 activity as a result of phosphorylation and reversible binding of the regulatory proteins to the catalytic subunit are critical in the development and regulation of motility and the ability of sperm to fertilize eggs. Targeted disruption of the Ppp1cc gene, which encodes the PP1 gamma 1 or PP1 gamma 2 isoforms, causes male infertility in mice as a result of impaired spermiogenesis. Our observations suggest that, in addition to motility, the protein phosphatase PP1 gamma 2 might play an isoform-specific function in the development of specialized flagellar structures of mammalian spermatozoa.
Animals ; Cattle ; Cricetinae ; Epididymis ; enzymology ; physiology ; Homeostasis ; Humans ; Male ; Mice ; Phosphoprotein Phosphatases ; genetics ; metabolism ; Sperm Motility ; physiology ; Spermatozoa ; enzymology ; physiology ; Testis ; enzymology

To study the expression of mTERT gene in the testis of SD rats and its significance, in situ hybridization (ISH) techniques were used to detect the expression of telomerase gene mTERT mRNA in the testis of SD rats. The expression of mTERT was detectable in different-age male SD rats testis. There was a positive correlation between the expression of mTERT and the location of germ cells (spermatogonia, spermatocyte, spermatid). In Sertoli cells, leydig cell and spermatozoa, telomerase mTERT was not detected. Type A spermatogonia expressed the highest level of telomerase mTERT mRNA. Our results suggest that the expression of mTERT gene in the testis of SD rats is of lifetime and coincide with the telomerase activity.
Animals ; Catalysis ; Cell Differentiation ; DNA-Binding Proteins ; Gene Expression Regulation, Developmental ; Male ; RNA ; Rats ; Rats, Sprague-Dawley ; Spermatids ; enzymology ; ultrastructure ; Spermatogenesis ; genetics ; Spermatogonia ; enzymology ; ultrastructure ; Spermatozoa ; enzymology ; ultrastructure ; Telomerase ; biosynthesis ; genetics ; metabolism ; Testis ; enzymology ; growth & development

AIMTo estimate the dissipation of mitochondrial transmembrane potential (mTMP, DeltaPsim) and activation of sperm caspases (aCP) as signs of apoptosis in human spermatozoa during cryopreservation and to evaluate the efficiency of immunomagnetic cell separation (MACS) of these spermatozoa via annexin V-binding.

METHODSThe mTMP and aCP in fresh and cryopreserved spermatozoa were detected by fluorescence microscopy and by Western blots. The sperm suspensions were divided into two sperm fractions (with intact and deteriorated membranes) by magnetic cell separation (MiniMACS, Miltenyi Biotec, Bergisch Gladbach, Germany) in dependence on their binding to superparamagnetic annexin V-microbeads (AN-MB).

RESULTSThe cryopreservation decreased the portion of spermatozoa with intact mTMP from 80.1 % +/- 7.2 % to 53.5 % +/- 13.1 % and increased the spermatozoa with activated pan-caspases (aCP) from 21.8 % +/- 2.6 % to 47.7 % +/- 5.8 % (n=10; mean +/- SEM; P <0.01). The activation of caspases 1, 3, 8, and 9 in the cryopreserved spermatozoa was confirmed by Western blots (n=22). MACS reduced significantly the percentage of cryopreserved spermatozoa with dissipated mTMP to 8.1 +/- 3.9 (P <0.01) and also those with aCP to 9.3 % +/- 2.2 %. Western blot analyses confirmed the increase of the activated caspase3, 9, and 8 in the AN-MB-positive fraction (P <0.05) compared with the AN-MB-negative fraction. The MACS separation effect was confirmed by anti-annexin V-antibodies. There was no significant influence of the separation column and the magnetic field on the sperm functions.

CONCLUSIONThe cryopreservation impaired the mTMP and enhanced the activation status of caspases in human spermatozoa. The immunomagnetic sperm separation via binding of AN-MB could deplete low quality spermatozoa from cryopreserved semen samples.

Apoptosis ; Blotting, Western ; Caspases ; metabolism ; Cold Temperature ; adverse effects ; Cryopreservation ; Humans ; Immunomagnetic Separation ; Intracellular Membranes ; enzymology ; physiology ; Male ; Microscopy, Fluorescence ; Spermatozoa ; enzymology ; physiology

OBJECTIVETo observe the clinical efficacy of Shugan Yiyang Capsules in the treatment of asthenospermia and its action mechanisms.

METHODSWe randomly assigned 135 asthenospermia patients to groups A (n = 47), B (n = 45), and C (n = 43) to be treated with Shugan Yiyang Capsules, oral levocarnitine, or combination of the two. We observed sperm quality and the level of α-glucosidase in the seminal plasma before and after medication.

RESULTSThe total effectiveness rate was 70.21% in group A (markedly effective in 16 cases and effective in 17), 68.89% in group B (markedly effective in 15 cases and effective in 16), and 83.72% in group C (markedly effective in 16 cases and effective in 20), significantly higher in C than in A and B (P < 0.05). Both sperm quality and the level of α-glucosidase in the seminal plasma were improved in the three groups of patients, most obviously in group C.

CONCLUSIONShugan Yiyang Capsules can be used for the treatment of asthenospermia, and its effect can be enhanced in combination with oral levocarnitine.

Asthenozoospermia ; drug therapy ; enzymology ; Biomedical Research ; Capsules ; Carnitine ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Semen ; enzymology ; Spermatozoa ; alpha-Glucosidases ; analysis