The wave of the seminiferous epithelium in the rat

Perey, Bernard; Clermont, Y.; Leblond, C. P.

doi:10.1002/aja.1001080105

Cited by 321 publications

(143 citation statements)

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“…Little expression was observed in surrounding Leydig cells. During spermatogenesis in the adult mouse, germ-cell differentiation advances in highly ordered waves along the axis of the seminiferous tubule, and each cross-section of a tubule can represent one of 12 stages of spermatogenesis (35). The intensity of the RAP80 hybridization signal appears to be very similar between different seminiferous tubules suggesting that RAP80 is not regulated during spermatogenesis.…”

Section: Rap80mentioning

confidence: 89%

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RAP80, a Novel Nuclear Protein That Interacts with the Retinoid-related Testis-associated Receptor

Yan

Kim

Jetten

2002

Journal of Biological Chemistry

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In this study, we describe the characterization of a novel nuclear protein, referred to as RAP80. The RAP80 cDNA was cloned from a human testis cDNA library and encodes a 719-amino acid protein containing two potential CX 2 CX 11 HX 3 C-type zinc finger motifs at its carboxylterminal region. Analysis of its genomic structure revealed that the RAP80 gene covers more than 90 kb and consists of 15 exons and 14 introns. Fluorescence in situ hybridization mapped the RAP80 gene to human chromosome 5q35. RAP80 mRNA is expressed in many human tissues, but its expression is particularly high in testis. In situ hybridization showed that RAP80 is highly expressed in germ cells of mouse testis but is not differentially regulated during spermatogenesis. Confocal microscopy showed that RAP80 is localized to the nucleus, where it is distributed in a speckled pattern. Deletion analysis showed that a bipartite nuclear localization signal at the amino terminus is important in mediating nuclear transport of RAP80. Monohybrid analysis showed that RAP80 might function as an active repressor of transcription. Mammalian two-hybrid analysis demonstrated that RAP80 was able to interact with the retinoid-related testis-associated receptor (RTR), an orphan receptor that has been implicated in the control of embryonic development and spermatogenesis. Pulldown analysis showed that RAP80 and RTR physically interact in vitro. Deletion and point mutation analyses revealed that part of the hinge domain of RTR is required for this interaction. RAP80 is able to inhibit the interaction of RTR with the co-repressor N-CoR likely by competing with N-CoR for RTR binding. Our results suggest that RAP80 may be functioning as a modulator of RTR signaling.The nuclear receptor superfamily is composed of a large number of ligand-dependent transcription factors that include nuclear orphan receptors for which a ligand has not yet been identified (1-4). Nuclear receptors share a common domain structure that includes an amino-terminal domain, a DNAbinding domain (DBD), 1 hinge domain, and a ligand-binding domain (LBD). These domains are involved in the recognition of specific DNA response elements, receptor dimerization, nuclear localization, and ligand binding and contain repressor and transactivation functions. Repression and activation of transcription by nuclear receptors are mediated through interactions with co-repressor and co-activators, respectively. Nuclear receptors have been demonstrated to regulate many physiological processes, including embryonic development and cell growth and differentiation and have been implicated in a number of human diseases (5-8). Less, however, is known about the role of many nuclear orphan receptors, including the retinoidrelated, testis-associated receptor (RTR), also referred to as germ cell nuclear factor (9, 10). The orphan receptor RTR (named NR6A1 by the Nuclear Receptor Nomenclature Committee) has been cloned from several species, including mouse (9, 10), human (11-14), zebrafish (15), and Xenopus laevis (16). Duri...

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Section: Rap80mentioning

confidence: 89%

“…Fig. 8 shows that 35 S-labeled RAP80 was able to bind to GST-RTR but not to GST alone, suggesting that this binding is specific for RTR. No interaction was found between RAP80 and GST-ROR␥ (not shown).…”

Section: Figmentioning

confidence: 99%

RAP80, a Novel Nuclear Protein That Interacts with the Retinoid-related Testis-associated Receptor

Yan

Kim

Jetten

2002

Journal of Biological Chemistry

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“…ells to absorb out the binding activity of both XT-I XT-2 is detectable in the testis at 8 days, when the germ cells XT-II (Fig. 6b) (6), and then all cell types in the testis as spermatogenesis proceeds continadult (12,13 …”

mentioning

confidence: 99%

Recognition of differentiation antigens of spermatogenesis in the mouse by using antibodies from spleen cell-myeloma hybrids after syngeneic immunization.

Bechtol¹,

Brown²,

Kennett³

1979

Proc. Natl. Acad. Sci. U.S.A.

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Two differentiation antigens of spermatogenesis in the mouse are defined by monoclonal antibodies from hybridomas produced between the myeloma P3-X63Ag8 and spleen lymphocytes immunized with syngeneic testis cells. 4 and 5) between immune lymphocytes and a myeloma cell line. The resulting monoclonal antibodies derived from individual hybrid lines then provide highly specific reagents, which can be used to dissect the membrane composition of individual cell types and the cell type composition of a complex mixture.In spermatogenesis, because of the "blood-testis barrier" syngeneic immunizations are possible, and they produce a generally tissue-specific response. Thus, hybridomas produced from mice immunized syngeneically with testis cells produce testis-binding activity which in most cases is testis-specific. We present here an analysis of two differentiation antigens of mouse spermatogenesis that are recognized by monoclonal antibodies from hybrids between P3-X63Ag8 (X63) and syngeneically immunized antitestis spleen cells (hybrids XT-I and XT-II). The antigens recognized, XT-1 and XT-2, respectively, are testis specific, and also species, but not strain specific. Both are trypsin sensitive and collagenase insensitive. In addition, on the basis of their development during the onset of spermatogenesis in juvenile mice, the two determinants appear to be expressed at different but overlapping times during spermatogenesis.MATERIALS AND METHODS Animals. 129/Sv, C3H/HeSn, B10.6R (a noninbred line of T/t6 mice), and Fisher rats were bred and maintained at the Immunization and Fusion. A 2-month-old strain 129 female mouse was immunized by intraperitoneal injection of 5 X 106 testis cells, in phosphate-buffered saline, from 4-month-old strain 129 males. Three weeks later 107 testis cells in buffered saline from 26-day-old strain 129 males were injected intravenously. Because of the age of this second group of donors, the cell preparation contained differentiating germ cells only through middle spermatids-i.e., no spermatozoa were present (ref. 6; our unpublished observations on strain 129 mice). Four days after the intravenous injection, the immune splenic lymphocytes were fused with X63 by using the method of Kennett et al. (7). The final fused cell suspension was distributed one drop per well into six 96-well flat-bottom tissue culture plates (Falcon).Subclones. Positive wells were subcloned in soft SeaKem agarose (Microbiological Associates) over a feeder layer of fibroblasts. When visible subclones appeared, approximately 30 were picked for each hybrid. All of these were found to be producing binding activity. One subelone was then chosen from each original well, and these were used as the source of all antibody-containing culture supernatant used in characterization of the recognized antigens.Quantitative Radioimmunoassay. Antibody binding was quantitated by a two-step binding assay modified from Morris and Williams (8). F(ab')2 rabbit anti-mouse IgG, a gift of A. F.Abbreviations: X63 = the myeloma line P3...

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“…Its composition probably reflects the conditions within the adluminal compartment of the germinal epithelium in which meiosis is completed (Setchell, 1970(Setchell, , 1978(Setchell, , 1980Setchell & Waites, 1975). In addition, a possible involvement of fluid in the entrainment of the spermatogenic wave has been suggested (Perey, Clermont & Leblond, 1961) and flow of fluid may be important in the regulation of spermatogenesis. Suoranta (1971) selectively damaged single seminiferous tubules of rats by touching the exposed testicular capsule for about two seconds with a hot tungsten wire.…”

Section: Introductionmentioning

confidence: 99%

Effects of obstruction of the flow of seminiferous tubule fluid on the germinal epithelium in the rat

Pilsworth¹,

Hinton²,

Setchell³

1981

Reproduction

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The wave of the seminiferous epithelium in the rat

Cited by 321 publications

References 14 publications

RAP80, a Novel Nuclear Protein That Interacts with the Retinoid-related Testis-associated Receptor

RAP80, a Novel Nuclear Protein That Interacts with the Retinoid-related Testis-associated Receptor

Recognition of differentiation antigens of spermatogenesis in the mouse by using antibodies from spleen cell-myeloma hybrids after syngeneic immunization.

Effects of obstruction of the flow of seminiferous tubule fluid on the germinal epithelium in the rat

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