OBJECTIVESTo investigate possible abnormalities of the sperm neck in eight infertile painters observed by transmission electron microscope.

METHODSSpermatozoa from 8 infertile painters (21-32 years old) were observed by transmission electron microscope.

RESULTSThe abnormalities of the connecting piece could be divided into 6 types, among which the anomaly of the connecting piece concomitant with a true nuclemalamic condition was the most serious.

CONCLUSIONSThe abnormalities may be related to expression obstacles of nuclear genome and mitochondrial genome, and to anomalous cellular signal transduction during spermiogenesis as well.

Adult ; Humans ; Infertility, Male ; pathology ; Male ; Microscopy, Electron, Transmission ; Occupational Exposure ; Paint ; adverse effects ; Spermatozoa ; ultrastructure

The ultrastructural abnormalities of human sperm flagella can cause sperm movement disorders. Dysplasia of the fibrous sheath (DFS) is an autosomal recessive genetic disease. The affected sperm in 95-100% of the patients display short, thick and irregular tails. Transmission electron microscopy can be used to confirm the diagnosis, which reveals gross abnormal flagella, with hypertrophy and hyperplasia of the fibrous sheath, without orderly disposition in longitudinal columns and transversal ribs. The axoneme shows variable distortion or almost complete obliteration. Microtubular doublets may exhibit partial or total lack of dynein arms. The genetic etiology of DFS is not yet clear. DFS does not affect the rates of fertilization and clinical pregnancy in ICSI, but due attention should be paid to the genetic risks in the offspring of the patient.
Humans ; Hyperplasia ; complications ; pathology ; Hypertrophy ; complications ; pathology ; Infertility, Male ; Male ; Microscopy, Electron ; Sperm Motility ; physiology ; Sperm Tail ; pathology ; ultrastructure ; Spermatozoa

OBJECTIVETo examine and analyze semen quality and sperm ultrastructural characteristics of infertile patients with varicocele.

METHODSThis study included 118 infertile patients with varicocele (the VC group) and 76 normal semen donors (the control group). We obtained routine semen parameters, seminal plasma biochemical markers and the levels of reproductive hormones in the subjects, and observed the changes in sperm structure under the scanning electron microscope and transmission electron microscope.

RESULTSCompared with the normal control, the VC patients showed significantly decreased sperm concentration, sperm progressive motility, sperm viability (P < 0.05), but no remarkable difference in semen volume and non-progressive motility (P > 0.05). The concentrations of zinc and alpha-glycoside enzyme in the seminal plasma were markedly reduced in the VC group in comparison with the controls (P < 0.05), but there was no significant difference in the level of fructose (P > 0.05), nor in such seminal plasma biochemical markers as FSH, LH, T and E2 between the two groups (P > 0.05). The percentage of morphologically normal sperm was dramatically lower in the VC than in the control group ([56.76 +/- 15.32]% vs [12.34 +/- 6.58]%, P < 0.05), and the sperm deformities were mostly in the head and neck, mainly tapering pin head accompanied by complex abnormal differentiation.

CONCLUSIONThis study demonstrated that VC may lead to oligo-astheno-terato zoospermia, and hence male infertility, which may be attributed to the changes of seminal plasma microenvironment and sperm ultrastructure.

Adult ; Case-Control Studies ; Humans ; Infertility, Male ; etiology ; pathology ; Male ; Semen Analysis ; Sperm Motility ; Spermatozoa ; ultrastructure ; Varicocele ; complications ; pathology

OBJECTIVEGlobozoospermia is mostly associated with homozygous deletion of the DPY19L2 gene. This study aimed to investigate the DPY19L2 gene mutation in a globozoospermia patient.

METHODSWe observed the sperm histomorphology of a patient with globozoospermia using Wright-Giemsa's staining and transmission electron microscopy, detected the mutation of the DPY19L2 gene by PCR amplification and DNA sequencing, and compared the findings with the sequences issued in the Genbank.

RESULTSWright-Giemsa's staining showed that all the spermatozoa were round-headed and lacked the acrosome, with the head nucleus darkly, fully and densely stained. Transmission electron microscopy revealed larger round sperm heads, with an even layer of unit membrane surrounding the nuclei and dispersed cytoplasmic vacuoles but no acrosomal structure. No DPY19L2 gene mutation was found by PCR amplification and DNA sequencing.

CONCLUSIONNo homozygous mutation of the DPY19L2 gene was found in the globozoospermia patient, and therefore some other disease-causing genes might be involved.

Acrosome ; pathology ; ultrastructure ; DNA Mutational Analysis ; Gene Deletion ; Humans ; Infertility, Male ; genetics ; Male ; Membrane Proteins ; genetics ; Microscopy, Electron, Transmission ; Spermatozoa ; pathology ; ultrastructure

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.
Acrosome/*pathology/ultrastructure ; Animals ; Cattle/*physiology ; Cell Membrane/*pathology/ultrastructure ; Cell Separation/veterinary ; Centrifugation, Density Gradient/veterinary ; Cryopreservation/veterinary ; Male ; Microscopy, Electron, Transmission/veterinary ; Povidone/*adverse effects ; Silicon Dioxide/*adverse effects ; Spermatozoa/pathology/ultrastructure

AIMTo investigate the changes of the spermatozoa ultrastructures before and after renal transplantation in uremic patients.

METHODSThe sperm of five uremic patients before and after transplantation and four healthy volunteers were collected and examined by scanning electron microscopy.

RESULTSAbnormal spermatozoa were found in patients pre-transplantation; abnormalities included deletion of the acrosome, absence of the postacrosomal and postnuclear ring, dumbbell-like changes of the head, tail curling, and absence of the mitochondrial sheath in the mid-segment. After renal transplantation, most of the spermatozoa became normal.

CONCLUSIONThere are many abnormalities with regard to the appearance and structure of the head, acrosome, mitochondria and tail of the spermatozoa in uremic patients. The majority of the spermatozoa returned to normal after renal transplantation, but a few still presented some abnormalities possibly relating to the administration of immunosuppressants.

Acrosome ; pathology ; Adult ; Case-Control Studies ; Humans ; Kidney Failure, Chronic ; complications ; Kidney Transplantation ; Male ; Microscopy, Electron ; Renal Dialysis ; Sperm Head ; pathology ; Sperm Tail ; pathology ; Spermatozoa ; pathology ; ultrastructure

OBJECTIVETo investigate the effects of Jujingwan on the spermatozoal ultrastructure and apoptosis of germ cells in oligospermia patients.

METHODSWe treated 50 oligospermia patients with Jujingwan and observed the spermatozoal ultrastructure, the apoptosis of germ cells and the changes in the DNA ploidy proportion of spermatogenic cells by electron microscopy and FCM before the treatment and 3, 6, 9 and 12 months after it.

RESULTSJujingwan increased sperm acrosome base density 6 months after the treatment and remarkably improved the integrity of acrosome membrane 12 months after it, with no obvious pathological changes in the nuclei and tails. Three months after the treatment, cell debris and apoptotic cells decreased significantly as compared with pre-treatment (P < 0. 05) , and very significantly 12 months after the treatment (P <0. 01). The proportion of haploid spermatozoa increased very significantly (P <0.01) , and the lost primary spermatocytes decreased significantly (P <0. 05) compared with pre-treatment.

CONCLUSIONJujingwan can increase the density of sperm acrosome base and improve the pathological changes of acrosome membrane in oligospermia patients; it can improve the activity of acrosome enzyme and the integrity of acrosome membrane, decrease the apoptosis rate of germ cells and sperm and increase the percentage of haploid spermatozoa; it can also reduce the percentage of apoptotic bodies and diploid sperm cells. It is indicated that Jujingwan can inhibit the apoptosis of germ cells and sperm and improve spermatogenesis in oligospermia patients.

Acrosome ; pathology ; Adult ; Apoptosis ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Infertility, Male ; drug therapy ; pathology ; Male ; Oligospermia ; drug therapy ; pathology ; Phytotherapy ; Sperm Count ; Spermatocytes ; cytology ; Spermatozoa ; ultrastructure

OBJECTIVETo study the effects of Wuzi Yanzong Pills (WYP) on sperm mitochondrial membrane potential (MMP) and its ultrastructure in oligo-asthenozoospermia model rats.

METHODSOligo-asthenozoospermia models were made in 50 male rats weighing 200 - 220 g by intragastric administration of Tripterygium Glucosides at 30 mg per kg per d for 8 weeks, and then equally allocated to a model control, a Huangjing Zanyu Capsule (HZC) control, a low-dose WYP, a medium-dose WYP, and a high-dose WYP group. Another 10 age-matched normal male rats were included as normal controls. The rats in the model and normal control groups were given intragastrically distilled water at 10 ml/kg, those in the HZC group administered HZC at 3.01 g/kg, and those in the low-, medium- and high-dose WYP groups medicated with WYP at 2.30, 4.60 and 9.20 g/kg, respectively, once daily for 30 days. At 30 minutes after the last administration, we detected the sperm MMP by JC-1 fluorescent staining and flow cytometry, and examined the sperm ultrastructure under the JEM-1230 transmission electron microscope.

RESULTSJC-1 + % and its fluorescence intensity were (33.77 +/- 6.19)% and 1 468 +/- 496 in the model control, (56.34 +/- 10.35)% and 3 277 +/- 895 in the HZC control, (40.80 +/- 10.40)% and 2 016 +/- 767 in the low-dose WYP, (59.40 +/- 6.51)% and 3 897 +/- 643 in the medium-dose WYP, and (60.71 +/- 7.81)% and 3 371 +/- 647 in the high-dose WYP group, significantly reduced in comparison with (70.80 +/- 4.92)% and 4 360 +/- 945 in the normal control group (P < 0.05), but remarkably higher in the medium- and high-dose WYP groups than in the model controls (P < 0. 05). After modeling, the sperm membrane was loose and degenerated, the mitochondria swelling, variously sized and with incomplete membrane, and the axonemal structure unclear or ruptured. After 30 days of WYP administration, compared with the model control group, the rats exhibited integrated sperm membrane and mitochondrial membrane, reduced mitochondrial swelling and basically normal axonemal and microtubular structures.

CONCLUSIONTripterygium Glucosides could decrease the sperm mitochondrial membrane potential and damage the mitochondrial structure, while WYP could significantly increase the sperm mitochondrial membrane potential and reduce the sperm mitochondrial structure damage. The protection of the integrity of sperm mitochondrial structure and function is one of the mechanisms of WYP acting on oligo-asthenozoospermia.

Animals ; Asthenozoospermia ; pathology ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Oligospermia ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Spermatozoa ; drug effects ; physiology ; ultrastructure

In this study, the effects of mono(2-ethylhexyl) phthalate (MEHP), one of metabolites of di(2-ethylhexyl) phthalate, on immature Shiba goat testes in vitro were examined. The testes of 2-month-old Shiba goats were cut into smaller pieces, and seeded in medium. At 1, 3, 6 and 9 hr after administration of MEHP at various concentrations (0, 100 nmol ml-1, 1 nmol ml-1, and 1 x 10-3 nmol ml-1, respectively), the specimens were obtained for light and transmission electron microscopic observations. As a result, at 1 hr after exposure to MEHP, the vacuolization and nuclear membrane rupture appeared in Sertoli cells. Such alterations tended to gradually increase in number in timeand dose-dependent manners. Moreover, by MEHP treatment, apoptotic spermatogenic cells (characterized with chromatin condensation, cytoplasm shrinkage without membrane rupture, still functioning cell organelles, and packed cell contents in membrane-bounded bodies), apoptotic Sertoli cells (characterized with nuclear membrane lysis, nuclear condensation), necrotic spermatogenic cells (characterized with swollen and ruptured mitochondria, plasma membrane lysis, spilt cell contents, and chromatin clumps), and necrotic Sertoli cells (characterized with marginated chromatins along the nuclear membrane, ruptured vesicles within the MNB, some swollen and ruptured cell organelles, e.g. mitochondria) could be identified. Conclusively, ultrastructurally the treatment with MEHP at low concentration tends to lead spermatogenic and Sertoli cells to apoptosis, whereas that at high concentration tends to lead spermatogenic and Sertoli cells to necrosis. Thus, the testicular tissue culture is advantageous for screening testicular toxicity of chemicals.
Animals ; Apoptosis/drug effects/physiology ; Diethylhexyl Phthalate/*analogs&derivatives/*toxicity ; Goat Diseases/*chemically induced/pathology ; Goats ; Male ; Microscopy, Electron, Transmission/veterinary ; Necrosis ; Sertoli Cells/ultrastructure ; Spermatozoa/drug effects/pathology/ultrastructure ; Testicular Diseases/*chemically induced/pathology ; Testis/*drug effects/metabolism/pathology/ultrastructure ; Vacuoles/physiology/ultrastructure

Peroxynitrite is a highly reactive nitrogen species and a potent inducer of apoptosis and necrosis in somatic cells. Peroxynitrite-induced nitrosative stress has emerged as a major cause of impaired sperm function; however, its ability to trigger cell death has not been described in human spermatozoa. The objective here was to characterize biochemical and morphological features of cell death induced by peroxynitrite-mediated nitrosative stress in human spermatozoa. For this, spermatozoa were incubated with and without (untreated control) 3-morpholinosydnonimine (SIN-1), in order to generate peroxynitrite. Sperm viability, mitochondrial permeability transition (MPT), externalization of phosphatidylserine, DNA oxidation and fragmentation, caspase activation, tyrosine nitration, and sperm ultrastructure were analyzed. The results showed that at 24 h of incubation with SIN-1, the sperm viability was significantly reduced compared to untreated control (P < 0.001). Furthermore, the MPT was induced (P < 0.01) and increment in DNA oxidation (P < 0.01), DNA fragmentation (P < 0.01), tyrosine nitration (P < 0.0001) and ultrastructural damage were observed when compared to untreated control. Caspase activation was not evidenced, and although phosphatidylserine externalization increased compared to untreated control (P < 0.001), this process was observed in <10% of the cells and the gradual loss of viability was not characterized by an important increase in this parameter. In conclusion, peroxynitrite-mediated nitrosative stress induces the regulated variant of cell death known as MPT-driven necrosis in human spermatozoa. This study provides a new insight into the pathophysiology of nitrosative stress in human spermatozoa and opens up a new focus for developing specific therapeutic strategies to better preserve sperm viability or to avoid cell death.
Adult ; Caspases/metabolism* ; Cell Death ; Enzyme Activation ; Humans ; Male ; Mitochondria/pathology* ; Necrosis ; Nitrosative Stress/physiology* ; Permeability ; Peroxynitrous Acid/pharmacology* ; Phosphatidylserines/metabolism* ; Spermatozoa/ultrastructure*