Silver staining of nucleolus organizer regions during human spermatogenesis

Schmid, Michael; Müller, Hj.; Stasch, S.; Engel, Wolfgang

doi:10.1007/bf00292368

Cited by 31 publications

(11 citation statements)

References 39 publications

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“…and postmeiotic stages of mouse spermatogenesis, there is total agreement between the present re sults obtained by immunocytoehemical methods and those of sil ver staining (Hofgarlner et al. 1979b;Schmid et al. 1983).…”

Section: Discussionsupporting

confidence: 65%

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Immunocytogenetics: localization of transcriptionally active rRNA genes in nucleoli and nucleolus organizer regions by use of human autoantibodies to RNA polymerase I

Haaf¹,

Reimer²,

Schmid³

1988

Cytogenet Genome Res

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Cytological staining with silver nitrate (AgN0₃) has proved useful for the localization of nucleoli in interphase nuclei, as well as of nucleolus organizer regions (NORs) in metaphase chromosomes. The affinity of interphase nucleoli and chromosomal NORs to silver is a direct measure of the ongoing transcriptional activity of the rRNA genes or their activity during the preceding interphase, respectively. Correspondingly, human autoantibodies directed against chromatin-associated RNA polymerase I (RPI) should also be of value in the investigation of transcribed rRNA genes. Indirect immunofluorescence using the anti-RPI antibody as a probe has been employed successfully to visualize the chromosomal distribution of NORs in various mammalian species, as well as in human tumor cells. Immunofluorescence staining even permits the identification of heteromorphisms and small aberrations of the chromosomal NORs. The fluorescent intensity of interphase nucleoli is correlated with the different stages of nucleolar activation. In male gametogenesis, RPI-positive granules are present during meiotic prophase up to pachytene, as well as during the early and middle spermatid stages.

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Section: Discussionsupporting

confidence: 65%

“…mouse cmbryogenesis (Engel et al. 1977), and malegametogenesis (Hofgartnerel al.. 1979b: Schmid et al. 1983.…”

mentioning

confidence: 99%

Immunocytogenetics: localization of transcriptionally active rRNA genes in nucleoli and nucleolus organizer regions by use of human autoantibodies to RNA polymerase I

Haaf¹,

Reimer²,

Schmid³

1988

Cytogenet Genome Res

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“…The finding that total cell nucleolar size at diplotene is significantly larger than that at leptotene indicates that some nucleolar material might be synthesized during meiotic prophase I, in close resemblance to chordates, where nucleolar activity reaches its maximum at pachytene (Schmid et al 1982, 1983, Wachtler & Stahl 1993. However, we cannot rule out that the observed differences in NA between leptotene and diplotene might be an artefact yielded by higher squashing ease at diplotene because of the lack of a nuclear membrane which is, on the other hand, present at leptotene.…”

Section: Nucleolus Size Variation During Meiosismentioning

confidence: 95%

“…During meiosis, in chordates, there is vigorous nucleolar activity during prophase I, with a maximum at pachytene (Schmid et al 1982, 1983, Wachtler & Stahl 1993. Ribosomal RNA synthesis ceases after pachytene in spermatogenesis but continues until diplotene in oogenesis.…”

Section: Introductionmentioning

confidence: 99%

Nucleolus size variation during meiosis and NOR activity of a B chromosome in the grasshopper Eyprepocnemis plorans

et al. 2007

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The number of nucleoli and nucleolar area were measured in meiotic cells from males of the grasshopper Eyprepocnemis plorans collected in three natural populations. Number of nucleoli per cell showed no significant correlation among cells in different meiotic stages, but there was strong positive correlation for nucleolar area between leptotene and interkinesis cells in individuals from distant populations (Salobreña in Spain, and Smir in Morocco). No correlation was, however, observed for both parameters between the meiotic stages analysed in individuals from the population of Torrox (Spain). The number of nucleoli at leptotene was about double the number at interkinesis, as expected from the double ploidy level at leptotene and the corresponding double number of rDNA clusters. Leptotene nucleolar area, however, was about fourfold that in interkinesis, presumably due to higher requirements for ribosome biogenesis in meiosis I than meiosis II. In Torrox, diplotene cells showed a lower number of nucleoli but larger nucleolar area than in leptotene cells, suggesting an increase in nucleolus size during prophase I. Significant differences were found among populations for nucleolar area but not for number of nucleoli, the smallest nucleolar area being observed in Torrox, which is the population harbouring the most parasitic B chromosome variant. No clear effects on nucleolar area or number of nucleoli were associated with the B-chromosome number. However, B-chromosome effects on the nucleolar area were apparent in the Torrox population when data were analysed with respect to a B-chromosome odd-even pattern in leptotene and interkinesis cells. However, in diplotene cells no odd-even pattern was observed for both nucleolar parameters, suggesting that the increase in nucleolar size from leptotene to diplotene dilutes the leptotene odd-even pattern. The rDNA distally located in the B chromosome was associated with a nucleolus in 6.5% out of the 247 diplotene cells analysed. The implications of these findings are discussed in the context of B chromosomes as stress-causing genome parasites and the nucleolus as a sensor of stress.

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“…Using the newer fixation method, spirochetes and sperm cells were visible in semen samples and showed distinct morphology ( Figure 1C). Sperm cells are known to stain with silver stains ( Pathak et al , 1979; Schmid et al , 1983). Sperm cells were seen in all semen samples except for Patients 2 and 6, who had vasectomies (data not shown).…”

Section: Resultsmentioning

confidence: 99%

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions

Middelveen¹,

Burke

Sapi

et al. 2015

F1000Res

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Background: Recent reports indicate that more than 300,000 cases of Lyme disease are diagnosed yearly in the USA. Preliminary clinical, epidemiological and immunological studies suggest that infection with the Lyme disease spirochete Borrelia burgdorferi (Bb) could be transferred from person to person via intimate human contact without a tick vector. Failure to detect viable Borrelia spirochetes in vaginal and seminal secretions would argue against this hypothesis. Methods: Patients with and without a history of Lyme disease were selected for the study after informed consent was obtained. Serological testing for Bb was performed on all subjects. Semen or vaginal secretions were inoculated into BSK-H medium and cultured for four weeks. Examination of genital cultures and culture concentrates for the presence of spirochetes was performed using light and darkfield microscopy, and spirochete concentrates were subjected to Dieterle silver staining, anti-Bb immunohistochemical staining, molecular hybridization and PCR analysis for further characterization. Immunohistochemical and molecular testing was performed in three independent laboratories in a blinded fashion. Positive and negative controls were included in all experiments. Results: Control subjects who were asymptomatic and seronegative for Bb had no detectable spirochetes in genital secretions by PCR analysis. In contrast, spirochetes were observed in cultures of genital secretions from 11 of 13 subjects diagnosed with Lyme disease, and motile spirochetes were detected in genital culture concentrates from 12 of 13 Lyme disease patients using light and darkfield microscopy. Morphological features of spirochetes were confirmed by Dieterle silver staining and immunohistochemical staining of culture concentrates. Molecular hybridization and PCR testing confirmed that the spirochetes isolated from semen and vaginal secretions were strains of Borrelia, and all cultures were negative for treponemal spirochetes. PCR sequencing of cultured spirochetes from three couples having unprotected sex indicated that two couples had identical strains of Bb sensu stricto in their semen and vaginal secretions, while the third couple had identical strains of B. hermsii detected in their genital secretions. Conclusions: The culture of viable Borrelia spirochetes in genital secretions suggests that Lyme disease could be transmitted by intimate contact from person to person. Further studies are needed to evaluate this hypothesis.

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Silver staining of nucleolus organizer regions during human spermatogenesis

Cited by 31 publications

References 39 publications

Immunocytogenetics: localization of transcriptionally active rRNA genes in nucleoli and nucleolus organizer regions by use of human autoantibodies to RNA polymerase I

Immunocytogenetics: localization of transcriptionally active rRNA genes in nucleoli and nucleolus organizer regions by use of human autoantibodies to RNA polymerase I

Nucleolus size variation during meiosis and NOR activity of a B chromosome in the grasshopper Eyprepocnemis plorans

Culture and identification of Borrelia spirochetes in human vaginal and seminal secretions

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