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PubMed2006

Sequential expression of type IV collagen networks: testis as a model and relevance to spermatogenesis.

The six alpha chains of type IV collagen are organized into three networks: alpha1/alpha2, alpha3/alpha4/alpha5, and alpha1/alpha2/alpha5/alpha6. A shift from the alpha1/alpha2 to the alpha3/alpha4/alpha5 network occurs in the developing glomerular basement membrane, but how the alpha1/alpha2/alpha5/alpha6 network fits into this sequence is less clear, because the three networks do not colocalize. Here, we studied the seminiferous tubule basement membrane of normal canine testis where all three networks do colocalize: the alpha1/alpha2 network is expressed from birth, the alpha1/alpha2/alpha5/alpha6 network by 5-6 weeks of age, and the alpha3/alpha4/alpha5 network by 2 months of age. A canine model of Alport syndrome allowed study of the absence of alpha3/alpha4/alpha5 and alpha1/alpha2/alpha5/alpha6 networks in testis. In Alport dogs, the seminiferous tubule basement membrane was thinner than in controls. Spermatogenesis began at the same time as with normal dogs; however, the number of mature sperm was significantly reduced in Alport dogs. Thus, it would appear that alpha3/alpha4/alpha5 and alpha1/alpha2/alpha5/alpha6 networks are not essential for onset of spermatogenesis, but long-term function may be compromised by the loss of one or both networks. This situation is analogous to the glomerular basement membrane in Alport syndrome. In conclusion, testis can serve as a model system to study the sequence of type IV collagen network expression.

Animals↗
PubMed1978

Fine structure and development of Sertoli junctions in human testis.

Complicated interdigitation-like junctions between the immature Sertoli cells were frequently observed in prepubertal testis (3--8-year-old). Tight or gap junctions could not be found. Subsurface cisternae appear in the testes of 7 and 8-year-old boys. In pubertal testis (11 to 13 years old), junctional specializations between Sertoli cells are composed of membrane fusions, bundles of microfilaments and associated cisternae. These tight junctions block the deep penetration of lanthanum into the seminiferous tubules. The lanthanum-filled Sertoli junctions show characteristic features of membrane fusions. In the seminiferous tubules spermatocytes were not present at 11 years of age but complete spermatogenesis was noted at 12-13 years of age. It is suggested that the human blood-testis barrier is estabilished shortly before or after the spermatogonia proliferate to give rise to primary spermatocytes.

Adolescent↗
PubMed1981

Acute cadmium intoxication: influence of cyproterone acetate on the testis and epididymis of the rat.

The changes resulting from treatment with cadmium were studied following the histological changes, the modification of both vascular permeability to vital dyes and of alkaline phosphatase activity in rat testis and epididymis. The testicular extravasation of acriflavine started 90 min following parenteral injection of cadmium and increased thereafter synchronous with an increase in testicular and epididymal weights due to edema. At 14 and 24 hr a striking decrease of interstitial fluorescence and tubular degeneration were noted in testis and caput epididymis due to thrombosis of the microvascular circulation. The barrier noted at 8 hr following cadmium injection. No changes of alkaline phosphatase activity was detected in testicular and epididymal blood vessels after cadmium injection. Previous treatment with cyproterone acetate accelerated the appearance of such alterations. The interstitial nuclear staining with acriflavine appeared in the testis at 1 hr and was diffuse at 90 and 120 min. cyproterone acetate seemed to accelerate the appearance of tubular degeneration at 8 hr after cadmium injection. The changes of the male rat gonad following cadmium treatment were characterized by an increased vascular permeability and generalized thrombosis. An inbalance of androgen stimulation seems to increase the blood vessels susceptibility to cadmium.

Acriflavine↗
PubMedDate not supplied

Cystic dysplasia of the testis: a report of three cases and review of the literature.

We describe the clinical and pathologic findings in three cases of cystic dysplasia of the testis and compare our findings to others in the English literature. The condition is characterized by dilatation of the rete testis with or without similarly affected adjacent structures. It is important to recognize this lesion for two reasons. First, it is associated with a high frequency of ipsilateral renal anomalies. Second, it is important to consider this condition in the differential diagnosis of cystic testicular mass in a neonate or a child. The management of this condition is quite different from the management of other testicular masses such as a hydrocele, testicular germ cell tumor, or infiltrative disorder (e.g., lymphoma, inflammation, torsion, or hematoma). Testicular cystic dysplasia is benign and the lesion can be removed surgically, leaving the rest of the testis intact. A discussion of the lesion's possible pathogenesis is also presented.

Antigens, Neoplasm↗
PubMedDate not supplied

Structure of rete testis, vas efferens, epididymis and vas deferens of langur monkey (Presbytis entellus entellus Dufresne).

Morphological and histological features of rete testis, vas efferens, epididymis and vas deferens were studied in the langur monkey. Tubular extensions of rete were located towards lateral side of the testis. Its epithelium comprises mostly of cuboidal cells with hyaline cytoplasm. Three to nine bundles of vas efferens, emerging below the cranial pole of the testis, were observed. Vas efferens epithelium comprises of ciliated and nonciliated cells. Epididymis could be divided into six zones on the basis of cytological features. Principal cells, basal cells, apical cells and intraepithelial lymphocytes were observed in the epididymal epithelium, but their number, shape, size and location of nuclei varied in different zones. Vas deferens epithelium comprises of principal cells, basal cells, apical cells and few intraepithelial lymphocytes. Epithelium is surrounded by lamina propria, longitudinal, circular and longitudinal muscle layers.

Animals↗
PubMed1985

Lesions of the rete testis in mice exposed prenatally to diethylstilbestrol.

Adenocarcinoma of the rete testis is an exceptionally rare and malignant testicular neoplasm. Although treatment of pregnant women with diethylstilbestrol (DES) results in reproductive tract abnormalities in their male offspring, increased incidence of testicular tumors has not been verified. However, recently three cases of seminoma have been described in men prenatally exposed to DES, suggesting an association of prenatal DES treatment and the subsequent development of testicular tumors. This report describes the treatment of outbred pregnant CD-1 mice with DES (100 micrograms/kg) on Days 9 through 16 of gestation and its effects on their male offspring. In addition to nonmalignant abnormalities such as retained testes which have been reported in men exposed prenatally to DES, lesions resembling adenocarcinoma of the rete testis were seen in prenatally DES-treated mice at 10 to 18 mo of age (11 of 233; 5%). No comparable lesions were seen in 96 age-matched control male mice. These results suggest an association of prenatal DES exposure and the subsequent development of testicular lesions in the rete testis of mice.

Adenocarcinoma↗
PubMed1994

[Effectiveness of blood-testis and blood-epididymis barriers for lead].

Over the period of the last 30 years there was a marked decrease in the number of spermatozoa produced by the gonads of men. It is felt that this observation is due to the influence of environmental pollution, wherein the lead plays quite an important role. In sperms of men, who are professionally exposed to lead compounds, oligo-, asteno-, and teratospermia is disclosed. Studies were performed on sexually mature male rats after 9-month-long exposure to lead acetate (II) as well as a group of animals after 3-month-long interval in the exposure. The changes provoked by the lead were evaluated by employing a number of study techniques, namely: morphological examination of testes with taking into account the stages of seminiferous epithelial cycle, and epididymis, giving due consideration to zones; electron microscopic examination of seminiferous cells and interstitial tissue, as well as the cells in the wall of epididymis and spermatozoa; X-ray microanalysis determining the presence and type of elements on ultrathin section; spectrophotometrical determination of Pb content in blood, testes and epididymides; determination of testosterone concentration (T) in blood serum. It has been revealed that the blood-testis barrier protects seminiferous epithelium against the toxic action of the lead. No deposits of Pb were observed either in germinal cells or Sertoli cells. The endocrine testicular cells outside the barrier had also unchanged ultrastructure, and contained no Pb. That finding was expressed by normal T level in blood serum. The only cells in the area of the testis, in whose cytoplasm there was Pb confirmed by X-ray microanalysis, were macrophages in the interstitial tissue of the testis. Apart from that, it has been disclosed that the blood-epididymis barrier does not provide a barrier against this element. Pb deposits were seen in smooth myocytes, epithelial cells and in the lumen of epididymal duct. That correlated with a marked decrease in the number of epididymal spermatozoa and numerous damages involving their ultrastructure. It has been shown that the lead, when passing to the duct lumen of epididymis through cells and structures constructing the wall of that organ, is being excreted from the male genital system with the sperm.

Animals↗
PubMed1999

D-type cyclins in adult human testis and testicular cancer: relation to cell type, proliferation, differentiation, and malignancy.

D-type cyclins are proto-oncogenic components of the 'RB pathway', a G1/S regulatory mechanism centred around the retinoblastoma tumour suppressor (pRB) implicated in key cellular decisions that control cell proliferation, cell-cycle arrest, quiescence, and differentiation. This study focused on immunohistochemical and immunochemical analysis of human adult testis and 32 testicular tumours to examine the differential expression and abundance of cyclins D1, D2, and D3 in relation to cell type, proliferation, differentiation, and malignancy. In normal testis, the cell type-restricted expression patterns were dominated by high levels of cyclin D3 in quiescent Leydig cells and the lack of any D-type cyclin in the germ cells, the latter possibly representing the only example of normal mammalian cells proliferating in the absence of these cyclins. Most carcinoma-in-situ lesions appeared to gain expression of cyclin D2 but not D1 or D3, while the invasive testicular tumours showed variable positivity for cyclins D2 and D3, but rarely D1. An unexpected correlation with differentiation rather than proliferation was found particularly for cyclin D3 in teratomas, a conceptually significant observation confirmed by massive up-regulation of cyclin D3 in the human teratocarcinoma cell line NTera2/D1 induced to differentiate along the neuronal lineage. These results suggest a possible involvement of cyclin D2 in the early stages of testicular oncogenesis and the striking examples of proliferation-independent expression point to potential dual or multiple roles of the D-type cyclins, particularly of cyclin D3. These findings extend current concepts of the biology of the cyclin D subfamily, as well as of the biology and oncopathology of the human adult testis. Apart from practical implications for the assessment of proliferation and oncogenic aberrations in human tissues and tumours, this study may inspire further research into the emerging role of the cyclin D proteins in the establishment and/or maintenance of the differentiated phenotypes.

Adult↗
PubMed1996

Localization of GnRH molecular forms in the brain, pituitary, and testis of the frog, Rana esculenta.

In the amphibian brain four molecular forms of GnRH have been identified so far: mammalian GnRH (m- and hydroxyproline9m-), chicken II GnRH (cII), and a salmon (s) GnRH-like peptide. In Rana esculenta, cII- and s-GnRH-like molecules have been partially characterized in the brain extracts using HPLC combined with radioimmunoassay. Moreover, since cII-GnRH-like material has been detected in Rana esculenta testis, the present study describes the localization of the above peptides in the brain and testis of the frog. Immunoreactive cII-GnRH and m-GnRH neurons and fibers were identified in the anterior preoptic area (APOA) and in the median septal area (MSA). A population of cells located on the dorsal side of the caudal preoptic region was also stained. Immunopositive fibers were seen to overlap the median eminence before ending within the pars nervosa. Moreover, densely packed fibers made close contact with the vascular complex in the median eminence. Conversely, immunoreactive s-GnRH-like material was absent in APOA and MSA, but weakly scattered elements were detected by the anti-s-GnRH serum in the dorsal side of the caudal preoptic region. Using m-GnRH antiserum, a strong immunopositivity was observed in the median eminence but not within the pars nervosa, indicating that, besides cII-GnRH and s-GnRH-like material, also m-GnRH-like material is present in Rana esculenta brain. In the testis, cells of the interstitial and germinal compartment were detected by anti-cII-GnRH during different periods of the annual cycle. In particular, in October and February interstitial tissue was intensely stained, coinciding with periods of increased androgen production and the onset of the new spermatogenic wave, respectively.

Animals↗
PubMed1999

Temporal difference between testis and ovary determinations with possible involvement of testosterone and aromatase in gonadal differentiation in TSD lacking lizard, Calotes versicolor.

In the garden lizard, Calotes versicolor, which lacks identifiable sex chromosomes, incubation temperature also does not have a deterministic effect on the gender. However, the embryos reared at high temperature (33-35 degrees C) have a shorter duration of incubation as well as gonadal differentiation. In contrast, exogenous application of the male hormone testosterone to embryos at ambient temperature (28 degrees C) results in almost all individuals with only testis. Thus the testosterone treatment reverts genic females to males and accelerates the differentiation of testis, a feature similar to the high-temperature treatment. Treatment of eggs with estradiol shows no difference from that seen in the untreated eggs. The present series of experiments was done to establish the "window" of testosterone sensitivity and to understand the interaction between sex hormones and high temperature on gonadal differentiation. The period between day 5 and 15 of embryonic development was the window period of testosterone sensitivity for sex reversal. This period coincided with the formation of the genital ridge and its differentiation into cortex and medulla. Treatment of the 33 degrees C-reared embryos with testosterone resulted in hatchlings of both the sexes, in contrast to only males at the ambient temperature. In contrast, at the same temperature (33 degrees C), all the dihydrotestosterone (nonaromatisable testosterone)-treated embryos hatched into males. However, those given estradiol showed no sex bias regardless of the day of application and the concentration of drug. Eggs were also treated with aromatase inhibitor, CGS 16949 A, at ambient temperature and at 33 degrees C. All the 33 degrees C eggs to which the drug was given on day 25 hatched into males. These results suggest that though high temperature has no direct effect on sex determination in this species, it may have a stimulatory effect on aromatase activity, leading to the conversion of the exogenously applied testosterone into estradiol and permitting ovarian differentiation in the genic females. It also follows from the present report that the pathway of testis formation in Calotes versicolor is triggered much earlier, and irreversibly, than that for the ovary.

Animals↗
PubMed1997

The expression of mouse gene P1A in testis does not prevent safe induction of cytolytic T cells against a P1A-encoded tumor antigen.

Tumor antigen P815AB is recognized by cytolytic T lymphocytes (CTL) on mouse mastocytoma P815. This antigen is encoded by P1A, a gene activated in several tumors but silent in normal tissues except for testis and placenta. Notwithstanding the expression of P1A in testis, we found that male mice mounted P815AB-specific CTL responses as efficiently as females. The responding males remained fertile and no autoimmune lesions were observed in their testes. By immunohistochemistry with a rabbit antiserum directed against the P1A protein, we identified spermatogonia as the testicular cells expressing P1A. The absence of MHC class-I molecules on spermatogonia could be one of the mechanisms of protection against testicular autoimmunity, as the antigenic peptide should not be displayed at the cell surface. Human genes MAGE, BAGE and GAGE, which also code for tumor antigens recognized by autologous CTL, are not expressed in normal tissues other than testis. The results obtained in mice with antigen P815AB suggest that immunization of human males with such antigens will not generate autoimmune side-effects. Although P1A is strongly expressed in placenta, we also found that gestation did not prevent generation of CTL responses against antigen P815AB, and that such CTL responses did not affect gestation outcome. We identified labyrinthine trophoblasts as the placental cells expressing P1A. Again, the absence of MHC class-I molecules on these cells provides a plausible explanation for placental protection, although other mechanisms may also play a role.

Animals↗
PubMed1998

Several novel transcripts of glyceraldehyde-3-phosphate dehydrogenase expressed in adult chicken testis.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), in addition to being a classic glycolytic enzyme, is a multifunctional protein involved in relevant cell functions such as DNA replication, DNA repair, translational control of gene expression, and apoptosis. Although the multifunctional nature of GAPDH suggests versatility in the mechanisms regulating its expression, no major qualitative changes and few quantitative changes in the GAPDH transcripts have been reported. While studying the expression of GAPDH during spermatogenesis, we detected alternative initiations to TATA box and alternative splicings in the 5' region of the pre-mRNA, resulting in at least six different types of mRNAs. The amount and the polyadenylation of the GAPDH transcripts increased in mature testis in relation to immature testis and further increased when cell suspensions from mature testis were exposed to heat shock. These results suggest that alternative initiation, alternative splicing, and polyadenylation could provide the necessary versatility to the regulation of the expression of this multifunctional protein during spermatogenesis.

Alternative Splicing↗
PubMed1996

Murine interleukin-11 (IL-11) is expressed at high levels in the hippocampus and expression is developmentally regulated in the testis.

IL-11, derived from a bone marrow stromal cell line, has pleiotropic effects on both hematopoietic cells and nonhematopoietic cells. However, no previous studies have systematically addressed expression of IL-11 in primary tissues in vivo and the relationship of IL-11 tissue specific gene expression and function of IL-11 is not clear. In the present study, we examined constitutive IL-11 expression in various murine adult tissues in vivo. IL-11 mRNA is expressed in a wide range of normal tissues (including hematopoietic organs) at levels only detected by RT-PCR. IL-11 protein was detected in brain and testis by Western blot analysis. The in vivo cellular distribution of IL-11 expression was examined by in situ hybridization. In brain, IL-11 message is distributed in granular layer dentate gyrus and pyramidal cell layers of hippocampus. IL-11 is also expressed in anterior horn cells and lateral column neuronal cells of the spinal cord. In testis, IL-11 mRNA is expressed in round spermatids at stage VI-IX seminiferous tubules. IL-11 expression in testis is restricted to developing spermatogonia and is developmentally regulated, since no expression is seen in mice genetically deficient in germ cells and in mice prior to sexual maturation. These expression data correlate with functional data demonstrating that IL-11 stimulates proliferation in vitro of a hippocampus neuronal progenitor cell line and administration of IL-11 in vivo accelerates recovery of spermatogenesis after cytotoxic therapy. These studies suggest that IL-11 may be an important regulator in neural and testicular function.

Animals↗
PubMed1996

Testis-specific expression of mRNAs for a unique human type 1 hexokinase lacking the porin-binding domain.

Several enzymes in the glycolytic pathway are reported to have spermatogenic cell-specific isozymes. We reported recently the cloning of cDNAs representing three unique type 1 hexokinase mRNAs (mHk1-sa, mHk1-sb, and mHk1-sc) present only in mouse spermatogenic cells and the patterns of expression of these mRNAs (Mori et al., 1993: Biol Reprod 49:191-203). The mRNAs contain a spermatogenic cell-specific sequence, but lack the sequence for the porin-binding domain that somatic cell hexokinases use to bind to a pore-forming protein in the outer mitochondrial membrane. We now report the cloning of cDNAs representing three unique human type 1 hexokinase mRNAs (hHK1-ta, hHK1-tb, and hHK1-tc) expressed in testis, but not detected by Northern analysis in other human tissues. These mRNAs also contain a testis-specific sequence not present in somatic cell type 1 hexokinase, but lack the sequence for the porin-binding domain. The hHK1-tb and hHK1-tc mRNAs each contain an additional unique sequence. The testis-specific sequence of the human mRNAs is similar to the spermatogenic cell-specific sequence of the mouse mRNAs. Furthermore, Northern analysis of RNA from mouse, hamster, guinea pig, rabbit, ram, human, and rat demonstrated expression of type 1 hexokinase mRNAs lacking the porin-binding domain in the testes of these mammals. These results suggest that hexokinase may have unique structural or functional features in spermatogenic cells and support a model proposed by others for hexokinase gene evolution in mammals.

Amino Acid Sequence↗
PubMed1996

Creatine kinase in trout male germ cells: purification, gene expression, and localization in the testis.

High creatine kinase (CK) activity (16.5 +/- 7.6 IU/mg) is present in trout spermatozoa. In order to partly characterize the CK isozyme predominantly present in sperm and to study the expression of this protein in spermatogenesis, we purified to homogeneity a CK (s-CK) from trout sperm, by nitrogen cavitation followed by two chromatography steps (DEAE-Trisacryl and Blue Sepharose). Specific antisera to 5-CK were developed. A cDNA encoding for a CK named TCK1, and whose transcript shows enhanced testicular expression, was previously isolated from trout testis (Garber et al., 1990: Biochim Biophys Acta 1087:256-258). A CK subunit expressed in vitro by this cDNA cross-reacts with anti-s-CK. A 21-amino-acid residue sequence near the N-terminus of s-CK is identical to the cDNA-derived sequence of TCK1, which is unlike any previously reported CK sequence. Using in situ hybridization, the TCK1 mRNA was detectable in primary and secondary spermatocytes and in early spermatids. Immunohistochemical staining of testis and various organs revealed that s-CK was confined to testis and, in this organ, to late spermatids and spermatozoa. In gill, some cells exhibited a positive signal, but another study rules out the presence of s-CK in this organ (Garber et al., 1990: Biochim Biophys Acta 1087:256-258). These results demonstrate that s-CK/TCK1 is a germ cell-specific protein, the transcription of which starts in meiotic germ cells, while translation starts in late spermatids.

Amino Acid Sequence↗
PubMed1997

Distribution of gelsolin in human testis.

Gelsolin, an actin-binding and severing protein present in many mammalian cells, was characterized in human testis. Although abundant in testicular extracts, gelsolin was not detected in purified spermatogenic cells by immunoblot analysis. Immunofluorescence studies of testis sections showed that gelsolin has two main localizations: peritubular cells and the seminiferous epithelium. In peritubular cells, gelsolin was present together with alpha-SM actin, in agreement with the myoid cell characteristics of these cells. In a large proportion of the tubules, gelsolin was found mainly, together with actin, in the apical part of the seminiferous epithelium. This localization of gelsolin also was observed in seminiferous tubules with a partial or complete absence of germinal cells, which evokes a presence of gelsolin at the apex of Sertoli cells. However, in normal testis, a complex pattern of gelsolin labeling was also present, mostly in the apical third of the epithelium, around cells or groups of cells, mainly spermatids, and, less frequently, in various other localizations from the apical to the basal part of the seminiferous epithelium. Taken together, these observations suggest that gelsolin may play different functions in the seminiferous epithelium: (1) regulation of the dynamic alterations of the actin cytoskeleton in the apical cytoplasm of Sertoli cells, and (2) modification of actin filaments assemblies in specific structures at germ cell-Sertoli cell contacts. Thereby, the actin-modulating properties of gelsolin are probably involved in reorganization of the seminiferous epithelium related to germ cell differentiation.

Animals↗
PubMed1998

The identification and characterization of expression of Pftaire-1, a novel Cdk family member, suggest its function in the mouse testis and nervous system.

We have isolated a murine cDNA encoding for a novel putative Cdk-related protein kinase, which has been named Pftaire-1, by screening a testis cDNA library for new serine/threonine kinases. Pftaire-1 showed 50% and 49% amino acid identity with Cdk5 and Pctaire-3, respectively, and contains the eleven subdomains characteristic of the protein kinases. By northern blot analysis we detected two transcripts of approximately 5.5 and 4.9 kb in size. These transcripts were expressed at low level in all murine tissues tested, except in the brain, testis and embryo, where high expression was detected. Cellular localization of the mRNAs by in situ hybridization analysis shows that Pftaire-1 is expressed in late pachytene spermatocytes in the testis and in post mitotic neuronal cells both in the brain and the embryo, suggesting a role of Pftaire-1 both in the process of meiosis as well as neuron differentiation and/or function.

Amino Acid Sequence↗
PubMed1998

Expression of the human antigen SPAG2 in the testis and localization to the outer dense fibers in spermatozoa.

ABSTRACT Antisperm antibodies (ASAs) have been implicated in some instances of infertility. To characterize sperm antigens relevant to immunologic and immunocontraceptive development, SPAG2 (sperm-associated antigen 2) was identified by screening a human testis cDNA library with human sera positive for ASAs. Subsequently, two isoforms, SPAG2-1 and SPAG2-2, were identified in testis and placenta libraries, respectively. In the current study, Southern analysis of human genomic DNA with a probe common to the two SPAG2 isoforms indicated a single SPAG2 gene; therefore, alternative splicing is a likely mechanism for production of variant mRNAs. In situ hybridization of human testis sections demonstrated the expression of SPAG2 in primary spermatocytes, with decreased or arrested expression in postmeiotic cells. Immunofluorescence of Triton X-100-extracted spermatozoa with an anti-SPAG2 peptide antiserum indicate that SPAG2 is an intracellular component of the sperm flagellum. Electron microscopy refined this localization to the outer dense fibers (ODFs), structural filaments associated with the mammalian sperm axoneme. The ODFs have been reported to be composed of keratin-like intermediate filament proteins. However, SPAG2 does not exhibit the molecular characteristics of such proteins, nor does SPAG2 demonstrate sequence homology with previously characterized ODF proteins. Therefore, SPAG2 represents a novel protein of human sperm ODFs. Characterization of SPAG2 will further our understanding of ODF function in normal sperm motility and of flagellar abnormalities that lead to male infertility.

Antigens, Surface↗