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Journal of Andrology 1984This study compared the cryoprotective effect of glycerol with that of a zwitter ion buffer system (TESTCY). Spermatozoa that are cryopreserved in the presence of... (Comparative Study)
Comparative Study
Comparison of glycerol and a zwitter ion buffer system as cryoprotective media for human spermatozoa. Effect on motility, penetration of zona-free hamster oocytes, and acrosin/proacrosin.
This study compared the cryoprotective effect of glycerol with that of a zwitter ion buffer system (TESTCY). Spermatozoa that are cryopreserved in the presence of glycerol possess a somewhat higher progressive motility immediately after thawing than those preserved in the presence of TESTCY. However, after a 1-hour incubation in glycerol-free medium, the progressive motilities of the glycerol- and TESTCY-treated spermatozoa become essentially identical. After 2 hours in culture medium, TESTCY-treated spermatozoa possess a higher motility than glycerol-treated spermatozoa, indicating that TESTCY is a better preservative than glycerol for the long-term motility of human spermatozoa. The fertilizing potential of the cryopreserved spermatozoa was assessed by their ability to penetrate zona-free hamster oocytes in vitro. Spermatozoa that are cryopreserved in the presence of TESTCY produce three- to four-fold higher penetration rates than glycerol-treated, cryopreserved spermatozoa. Cryopreservation in the presence of TESTCY also results in a higher stability of the acrosin/proacrosin system than when the spermatozoa are preserved in glycerol, since about two- to three-fold higher levels of proacrosin are retained. These results indicate that TESTCY is a better cryopreservative for human spermatozoa than glycerol.
Topics: Acrosin; Animals; Buffers; Cricetinae; Cryoprotective Agents; Culture Media; Drug Combinations; Enzyme Precursors; Fertilization in Vitro; Freezing; Glycerol; Humans; In Vitro Techniques; Male; Sperm Motility; Spermatozoa; Time Factors; Tromethamine
PubMed: 6423595
DOI: 10.1002/j.1939-4640.1984.tb00770.x -
Asian Journal of Andrology 2022Chlamydia trachomatis (CT) infection is the most prevalent sexually transmitted bacterial disease worldwide. However, unlike that in female infertility, the role of CT...
Chlamydia trachomatis (CT) infection is the most prevalent sexually transmitted bacterial disease worldwide. However, unlike that in female infertility, the role of CT infection in male infertility remains controversial. The objective of this retrospective study was to explore the impacts of CT infection in the genital tract on sperm quality, sperm acrosin activity, antisperm antibody levels, and inflammation in a large cohort of infertile males in China. A total of 7154 semen samples were collected from infertile male subjects, 416 of whom were CT positive (CT+ group) and 6738 of whom were CT negative (CT- group), in our hospital between January 2016 and December 2018. Routine semen parameters (semen volume, pH, sperm concentration, viability, motility, morphology, etc.), granulocyte elastase levels, antisperm antibody levels, and sperm acrosin activity were compared between the CT+ and CT- groups. Our results showed that CT infection was significantly correlated with an abnormally low semen volume, as well as an increased white blood cell count and granulocyte elastase level (all P < 0.05) in the semen of infertile males; other routine semen parameters were not negatively impacted. The antisperm antibody level and sperm acrosin activity were not affected by CT infection. These findings suggested that CT infection might contribute to inflammation and hypospermia but does not impair sperm viability, motility morphology, and acrosin activity or generate antisperm antibodies in the infertile males of China.
Topics: Chlamydia trachomatis; Female; Genitalia; Humans; Infertility, Male; Inflammation; Male; Retrospective Studies; Semen; Spermatozoa
PubMed: 34145079
DOI: 10.4103/aja.aja_54_21 -
The International Journal of... 2008Mammalian sperm must have properly formed acrosomes to be fully functional in the process of binding and penetrating the zona pellucida (ZP), the extracellular matrix... (Review)
Review
Mammalian sperm must have properly formed acrosomes to be fully functional in the process of binding and penetrating the zona pellucida (ZP), the extracellular matrix surrounding the egg. There is much evidence to raise doubts about the old "bag of enzymes" paradigm of acrosomal function, although this is the model that seems to prevail. We concur with other scientists that acrosomal exocytosis is not an all or none event where the acrosome is either "intact" or "reacted". As determined by transmission electron microscopy of human sperm undergoing acrosomal exocytosis, six stages can be identified, with the intermediate ones involving loss of acrosomal matrix material. In the mouse, there is a temporal relationship among four stages of acrosomal exocytosis. Numerous evidences suggest a more complex role for the acrosome in fertilization in which the acrosomal matrix is a scaffold for sperm-ZP interactions that self-regulates by a controlled disassembly mechanism.
Topics: Acrosin; Acrosome; Animals; Enzyme Precursors; Exocytosis; Fertilization; Guinea Pigs; Male; Membrane Proteins; Mice; Mice, Transgenic; Microscopy, Electron, Scanning; Models, Biological; Receptors, Cell Surface; Spermatozoa; Zona Pellucida
PubMed: 18649264
DOI: 10.1387/ijdb.072532mb -
Journal of Andrology 1987Previous results, simultaneously confirmed by others, suggest that a relationship exists between sperm acrosin levels and fertility in man. The assessment of sperm...
Previous results, simultaneously confirmed by others, suggest that a relationship exists between sperm acrosin levels and fertility in man. The assessment of sperm acrosin may therefore be a useful addition to the semen analysis, but only if the more standard semen parameter measurements cannot predict acrosin levels. Ejaculates from 102 men were analyzed and the relationship of the sperm acrosin system (acrosin, proacrosin, and acrosin inhibitor) to other seminal characteristics was determined. Very little correlation was observed between enzymatic and nonenzymatic parameters. Only five non-enzymatic parameters, all of which were morphologic, showed correlation coefficients of greater than or equal to 0.35 with acrosin and proacrosin, but none had an r-value above 0.48. The total acrosin and proacrosin levels were highly correlated to each other (r = 0.93). It is concluded that sperm acrosin/proacrosin levels cannot be predicted by other seminal parameters; thus, measurement of sperm acrosin/proacrosin may be clinically useful as a diagnostic parameters.
Topics: Acrosin; Endopeptidases; Enzyme Precursors; Humans; Infertility, Male; Male; Semen; Spermatozoa
PubMed: 3114204
DOI: 10.1002/j.1939-4640.1987.tb03320.x -
PloS One 2023Mouse spermatogenesis, from spermatogonial stem cell proliferation to sperm formation, can be reproduced in vitro by culturing testis tissue masses of neonatal mice....
Mouse spermatogenesis, from spermatogonial stem cell proliferation to sperm formation, can be reproduced in vitro by culturing testis tissue masses of neonatal mice. However, it remains to be determined whether this method is also applicable when testis tissues are further divided into tiny fragments, such as segments of the seminiferous tubule (ST), a minimal anatomical unit for spermatogenesis. In this study, we investigated this issue using the testis of an Acrosin-GFP/Histone H3.3-mCherry (Acr/H3) double-transgenic mouse and monitored the expression of GFP and mCherry as indicators of spermatogenic progression. Initially, we noticed that the cut and isolated stretches of ST shrunk rapidly and conglomerated. We therefore maintained the isolation of STs in two ways: segmental isolation without truncation or embedding in soft agarose. In both cases, GFP expression was observed by fluorescence microscopy. By whole-mount immunochemical staining, meiotic spermatocytes and round and elongating spermatids were identified as Sycp3-, crescent-form GFP-, and mCherry-positive cells, respectively. Although the efficiency was significantly lower than that with tissue mass culture, we clearly showed that spermatogenesis can be induced up to the elongating spermatid stage even when the STs were cut into short segments and cultured in isolation. In addition, we demonstrated that lowered oxygen tension was favorable for spermatogenesis both for meiotic progression and for producing elongating spermatids in isolated STs. Culturing isolated STs rather than tissue masses is advantageous for explicitly assessing the various environmental parameters that influence the progression of spermatogenesis.
Topics: Male; Mice; Animals; Spermatogonia; Semen; Seminiferous Tubules; Spermatogenesis; Testis; Spermatids; Mice, Transgenic
PubMed: 37023052
DOI: 10.1371/journal.pone.0283773 -
Genetics and Molecular Research : GMR Mar 2015The current study investigated the relationship between the level of expression and tyrosine phosphorylation of the sperm protein 32 (sp32) and the activation of the...
The current study investigated the relationship between the level of expression and tyrosine phosphorylation of the sperm protein 32 (sp32) and the activation of the boar proacrosin/acrosin system. The acrosomal membrane proteins of boar sperm for use in different treatment experiments (i.e., fresh sperm, freezing and thawing, capacitation, and acrosome reaction) were separated, stained by Coomassie brilliant blue, and analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot. The results showed that there were differences in the expression level of sp32 among capacitated, frozen-thawed, and post acrosomal reaction sperms. sp32 expression was higher and significantly higher in capacitated and post-acrosomal reaction sperms than in frozen-thawed sperms and fresh semen, respectively. The level of sp32 tyrosine phosphorylation was significantly different between the frozen-thawed sperms and sperms in the other experimental groups. However, bands with molecular masses of 38 to 170 ku in the fresh semen group were more noticeable, indicating that large acrosomal membrane proteins underwent modification and degradation during capacitation and the acrosomal reaction. As a proacrosin binding protein, sp32 shows upregulated expression and increase in tyrosine phosphorylation levels during the activation of the boar proacrosin/acrosin system.
Topics: Acrosin; Acrosome; Acrosome Reaction; Animals; Blotting, Western; Carrier Proteins; Cryopreservation; Enzyme Precursors; Male; Phosphorylation; Semen Preservation; Sperm Capacitation; Spermatozoa; Swine; Tyrosine
PubMed: 25867384
DOI: 10.4238/2015.March.27.23 -
The International Journal of... Feb 1996Spermatogenesis is a complex developmental process which involves amplification of germinal stem cells, their differentiation into spermatocytes, meiotic division and... (Review)
Review
Spermatogenesis is a complex developmental process which involves amplification of germinal stem cells, their differentiation into spermatocytes, meiotic division and finally transformation into mature spermatozoa. Therefore, spermatogenesis provides an interesting system for examining the regulation of gene expression during development and differentiation. The genes expressed during spermatogenesis can be divided into two main groups: diploid and haploid expressed genes. In this review, we report about the regulation of expression of a diploid expressed gene, namely the proacrosin gene, and that of a haploid expressed gene, the transition protein 2 gene.
Topics: Acrosin; Animals; Base Sequence; Chromosomal Proteins, Non-Histone; DNA, Complementary; Diploidy; Enzyme Precursors; Gene Expression Regulation, Developmental; Haploidy; Humans; Male; Protamines; Protein Biosynthesis; Spermatogenesis
PubMed: 8735951
DOI: No ID Found -
Cell Proliferation Feb 2016Previous studies have shown that germ-like cells can be induced from human umbilical cord mesenchymal stem cell (hUC-MSCs) in vitro. However, induction efficiency was...
OBJECTIVES
Previous studies have shown that germ-like cells can be induced from human umbilical cord mesenchymal stem cell (hUC-MSCs) in vitro. However, induction efficiency was low and a stable system had not been built. CD61, also called integrin-β3, plays a significant role in cell differentiation, in that CD61-positive-cell-derived pluripotent stem cells easily differentiate into primordial germ-like cells (PGC). Here, we have explored whether overexpression of CD61 would promote hUC-MSC differentiation into PGC and male germ-like cells.
MATERIALS AND METHODS
hUC-MSCs were cultured and transduced using pCD61-CAGG-TRIP-pur (oCD61) and pTRIP-CAGG plasmid (Control), and hUC-MSCs overexpressed CD61 were induced by bone morphogenetic protein 4 (BMP4, 12.5 ng/ml), to differentiate into PGC and male germ cells. Quantitative real-time PCR (RT-qPCR), western blotting and immunofluorescence staining were used to examine PGC- and germ cell-specific markers.
RESULTS
High expression levels of PGC-specific markers were detected in oCD61 hUC-MSCs compared to controls. After BMP4 induction, expression levels of male germ cell markers such as Acrosin (ACR), Prm1 and meiotic markers including Stra8, Scp3 in oCD61 were significantly higher than those of the Control group.
CONCLUSIONS
Under induction of BMP4, CD61-overexpressing hUC-MSCs, which had turned into PGC-like cells, could be further differentiated into male germ-like cells. Thus, a simple and efficient approach to study male germ cell development by using hUC-MSCs has been established.
Topics: Amino Acid Motifs; Amino Acid Sequence; Base Sequence; Biomarkers; Bone Morphogenetic Protein 4; Cell Differentiation; Cloning, Molecular; Codon; Germ Cells; Humans; Integrin beta3; Male; Mesenchymal Stem Cells; Models, Biological; Molecular Sequence Data; Protein Structure, Secondary; Sequence Analysis, Protein; Sequence Homology, Nucleic Acid; Umbilical Cord
PubMed: 26840189
DOI: 10.1111/cpr.12236 -
Activation and maturation mechanisms of boar acrosin zymogen based on the deduced primary structure.The Journal of Biological Chemistry Jul 1989We have isolated cDNA clones encoding boar acrosin, a serine protease participating in the initial stage of fertilization, from boar testis lambda gt11 cDNA libraries.... (Comparative Study)
Comparative Study
We have isolated cDNA clones encoding boar acrosin, a serine protease participating in the initial stage of fertilization, from boar testis lambda gt11 cDNA libraries. Nucleotide sequencing of the overlapping clones indicates that the composite cDNA inserts contain 1,391 base pairs coding for a 5'-untranslated region, an open reading frame, a stop codon, a 3'-untranslated region, and a poly(A)+ tail. A polyadenylation signal, AATAAA, is located 33 bases upstream from the start of the poly(A)+ tail. The amino acid sequence deduced from the cDNAs shows that boar acrosin is initially synthesized as a prepro-protein with a 16-residue signal peptide at the NH2 terminus. This signal sequence is followed by a 399-residue sequence corresponding to the acrosin zymogen. COOH-terminal sequence analysis of boar sperm 55-kDa proacrosin and its processed forms indicates that the mature acrosin molecule contains 322 amino acid residues in two polypeptide chains, a 23-residue light chain and a 299-residue heavy chain, with a combined molecular mass of 35,735 Da, and that the 55-kDa proacrosin molecule has 14-, 18-, and 43-residue segments as COOH-terminal extensions that are removed during proacrosin maturation. The COOH-terminal 43-residue segment is rich in proline residues, including an unusual repeat of 23 consecutive prolines. The deduced amino acid sequence of boar acrosin shows a high degree of identity with major portions of other serine proteases, including the active site region and the location of cysteine residues. We conclude that boar acrosin is synthesized as a single-chain polypeptide with the regions corresponding to the light and heavy chains covalently connected by two disulfide bonds, and that the single-chain molecule is autoactivated by cleavage of the Arg23-Val24 bond after removal of the COOH-terminal 14-residue segment, resulting in the formation of the light and heavy chains. This two-chain molecule is then converted to the mature enzyme by removal of the COOH-terminal 18- and 43-residue segments.
Topics: Acrosin; Amino Acid Sequence; Animals; Base Sequence; Blotting, Northern; Enzyme Precursors; Female; Male; Molecular Sequence Data; Nucleic Acid Hybridization; Protein Conformation; Rabbits; Restriction Mapping; Serine Endopeptidases; Swine; Testis
PubMed: 2745422
DOI: No ID Found -
Theriogenology Nov 1997Acrosin amidase activity of spermatozoa has been been associated with in vitro fertilization success in humans and has been proposed as an additional method for...
Acrosin amidase activity of spermatozoa has been been associated with in vitro fertilization success in humans and has been proposed as an additional method for assessing sperm function in vitro. In this study, acrosin amidase activity was determined in equine spermatozoa by the hydrolysis of an arginine amide substrate. This assay includes a detergent to release acrosomal enzymes into a medium of basic pH to activate proacrosin to acrosin, which subsequently hydrolyses N-alpha-benzoyl-DL-arginine para-nitroanilide-HCl (BAPNA) to a chromogenic product. Spermatozoa (n = 3 ejaculates from each of 4 stallions) were washed free from seminal plasma by centrifugation through Ficoll and incubated with a detergent-substrate mixture (BAPNA in triton X-100; pH = 8.0) at room temperature for 3 h in the dark. At the end of the 3-h incubation, benzamidine was added to test samples to stop the reaction, and samples were centrifuged to remove spermatozoa. Absorbance at 410 nm was measured to determine acrosin amidase activity (microIU acrosin/10(6) sperm). Acrosin amidase activity increased with sperm concentration (P < 0.001; r(2) = 0.75), and there were significant effects (P < 0.001) of stallion and ejaculate within stallion on acrosin activity. Acrosin activity detectable in equine seminal plasma was 312 +/- 49 microU/ml (n = 3 ejaculates). Addition of a cryopreservation medium containing egg yolk, skim-milk, glycerol and sucrose to equine spermatozoa and subsequent cryopreservation significantly (P < 0.05) increased acrosin amidase activity compared with spermatozoa from raw semen. This result is in contrast to that previously reported for frozen-thawed human spermatozoa. Determination of acrosin amidase activity in equine spermatozoa may provide an alternative method for assessing sperm function in vitro; however, further studies are needed to determine the relationship between acrosin activity and fertility in the horse.
PubMed: 16728208
DOI: 10.1016/s0093-691x(97)00352-x