Sertoli cells synthesize and secrete transferrin-like protein

One of the major proteins secreted by Sertoli cells (Band 3) has a molecular weight on sodium dodecyl sulfate-polyacrylamide gels which corresponds to rat serum transferrin. The preparation of Sertoli cell-secreted proteins contains an iron-binding protein with electrophoretic properties on native polyacrylamide gels similar to serum transferrin. Antibodies against Sertoli cell-secreted proteins contain a component which will immunoprecipitate serum transferrin. Antibodies to serum transferrin cross-react with and immunoprecipitate a major component (Band 3) of [35S]-methionine-labeled Sertoli cell-secreted proteins. We propose that testicular transferrin is a major secretory product of Sertoli cells in culture. In addition to transferrin, antiserum to rat serum proteins will immunoprecipitate one other polypeptide from preparations of Sertoli cell-secreted proteins

X-rays from T Tau: A test case for accreting T Tauri stars

We test models for the generation of X-rays in accreting T Tauri stars (TTS), using X-ray data from the classical TTS T Tau. High-resolution spectroscopy from the Reflection Grating Spectrometers on XMM-Newton is used to infer electron densities, element abundances and the thermal structure of the X-ray source. We also discuss the ultraviolet light curve obtained by the Optical Monitor, and complementary ground-based photometry. A high-resolution image from Chandra constrains contributions from the two companions of T Tau N. The X-ray grating spectrum is rich in emission lines, but shows an unusual mixture of features from very hot (~30 MK) and very cool (1-3 MK) plasma, both emitted by similar amounts of emission measure. The cool plasma confirms the picture of a soft excess in the form of an enhanced OVII/OVIII Lya flux ratio, similar to that previously reported for other accreting TTS. Diagnostics from lines formed by this plasma indicate low electron densities (<~ 1E10 cm-3). The Ne/Fe abundance ratio is consistent with a trend in pre-main sequence stars in which this ratio depends on spectral type, but not on accretion. On the basis of line density diagnostics, we conclude that the density of the cool ``soft-excess'' plasma is orders of magnitude below that predicted for an accretion shock, assuming previously determined accretion rates of (3-6)E-8 M_sun/y. We argue that loading of magnetic field lines with infalling material suppresses the heating process in a part of the corona. We thus suggest that the X-ray production of T Tau is influenced by the accretion process although the X-rays may not form in the bulk of the accretion footpoints.Comment: 12 pages, 7 figures, A&A style. Accepted by A&A, to appear in a special section/issue dedicated to the XMM-Newton Extended Survey of the Taurus Molecular Cloud (XEST). See also http://www.issibern.ch/teams/Taurus/papers.htm

Role of the Transcriptional Coactivator CBP/p300 in Linking Basic Helix-Loop-Helix and CREB Responses for Follicle-Stimulating Hormone-Mediated Activation of the Transferrin Promoter in Sertoli Cells

Sertoli cells are the epithelial cells responsible for the onset of pubertal development and the maintenance of spermatogenesis in the adult. Transferrin is one of the major secretory products expressed by differentiated Sertoli cells. Investigation of the transcriptional control of transferrin gene expression provides insight regarding the regulation of Sertoli cell differentiation. The optimal activation of the mouse transferrin promoter (mTf) by FSH requires the synergistic actions of the cAMP response element-binding protein (CREB) binding to the cAMP response element-like proximal region II (PRII) and the basic helix-loop-helix (bHLH) binding to the E-box. Proximal region II alone is sufficient for cAMP-mediated activation. The proximity of the PRII and E-box (220 base pairs apart) suggests the possibility of interaction between CREB and bHLH proteins. Such an interaction can be mediated by transcriptional integrators such as CREB-binding protein (CBP) and/or p300 and may stabilize the binding of trans-acting factors to their respective cis-elements. Such an interaction may also provide a mechanism for cell-specific promoter activation. The hypothesis tested in this study was that CBP/p300 is required for the synergistic activation of the transferrin promoter involving PRII and E-box through the formation of a ternary complex. In the Sertoli cells, both CBP and p300 proteins are expressed. The effect of CBP/p300 on transferrin promoter activation and, hence, Sertoli cell function was studied by using antisense oligonucleotides (AS-oligo). In the presence of CBP/p300 AS-oligo, activity of the FSH-induced mTf-chloramphenicol acetyl transferase (CAT) was significantly lower as compared to the respective controls. Interestingly, AS-oligo had no effect on cAMP-induced activation of the transferrin promoter reporter construct (mTf-CAT). Mutations in the E-box (EB*) significantly reduced the FSH response. The presence of AS-oligo had no further effect on the FSH-mediated activation of the EB*-mTf-CAT construct but reduced cAMP-mediated activation. Mutations in the CRE-like PRII (PRII*) also significantly reduced the FSH response. Activation of the PRII*-mTf-CAT in response to cAMP was completely abolished. The presence of AS-oligo had no further effect on the FSH- or cAMP-mediated activation of the PRII*-mTf-CAT construct. In Sertoli cells, CBP/p300 was coimmunoprecipitated with CREB and the bHLH protein E47. These observations suggest that CBP/p300 appears to be involved in regulating FSH-mediated activation of the transferrin promoter by linking bHLH and CREB activities

Actions of extracellular matrix on Sertoli cell morphology and function

Sertoli cells were isolated and cultured in the absence or presence of extracellular matrix (ECM) to determine whether ECM may influence Sertoli cell function on a molecular level. As previously described, a morphological analysis of the cells indicated that ECM allows the expression of a columnar histotype and the formation of junctional complexes. The combined actions of ECM and hormones were found to have a profound effect in promoting the expression of a polarized Sertoli cell morphology. In our investigation of the effects of ECM on Sertoli cells, we used transferrin and androgen-binding protein (ABP) production as biochemical markers of Sertoli cell function. The presence of ECM was found to cause a 25% increase in the basal level of transferrin production; however, ECM had no effect on the basal level of ABP production by Sertoli cells. Regulatory agents such as follicle-stimulating hormone (FSH) and a combination of FSH, insulin, retinol, and testosterone stimulated the production of both transferrin and ABP. The ability of hormones to stimulate these Sertoli cell functions was not influenced by the presence of ECM. Similar results were obtained with 2-microns- or 50-microns-thick ECM and with a seminiferous tubule biomatrix preparation. ECM was found to increase the maintenance of long-term Sertoli cell cultures; however, the decline in Sertoli cell functional integrity, which occurs during cell culture, was not affected by the presence of ECM. An additional functional parameter examined was the radiolabeled proteins secreted by Sertoli cells. ECM did not promote the production or affect the electrophoretic profile of Sertoli cell-secreted proteins under basal or hormonally stimulated conditions. Combined results indicated that although ECM allowed the expression of a normal Sertoli cell histotype, ECM had no major effects on the Sertoli cell functions analyzed nor on the hormonal regulation of these functions. The inability of ECM to affect Sertoli cell function on a molecular level is discussed with regard to environmental as opposed to regulatory cellular interactions. Our observations imply that dramatic effects of ECM on cell morphology do not necessarily correlate to subsequent effects on cellular function

Sertoli Cells Produce Vitamin-Binding Proteins

The seminiferous tubule in the rat testis is composed primarily of three cell types, Sertoli cells, peritubular myoid cells, and germinal cells in various stages of development. Tight junctions near the basal surfaces of adjacent Sertoli cells contribute to a blood-testis barrier, which creates a unique microenvironment within the tubule. The barrier effectively blocks passage of small and large molecules from plasma into the seminiferous tubule compartment formed by Sertoli cells.' Consequently, plasma constituents required for most stages of germinal cell development apparently must first pass through Sertoli cells. One approach to understanding the role of Sertoli cells in the maintenance and control of spermato-genesis is to determine the function of proteins secreted by the cell and then to monitor or to manipulate their function under various conditions. For example, Sertoli cells produce testicular transferrid and testicular ceruloplasmin,3 which specifically bind components (iron and copper, respectively) that are believed essential for germinal cell development. Vitamins are required for proper metabolic function and consequently germi-nal cell development. Vitamin deficiencies can have dramatic effects on the morphology of the seminiferous tubule and a negative effect on spermat~genesis.~ Vitamins that reach the inside of cells from plasma are quickly bound to proteins. Free vitamins would therefore be unlikely to exist in high concentrations in Sertoli cell cytosol and could not readily diffuse through the plasma membrane to developing germ cells. Further, the blood-testis barrier probably prevents diffusion of vitamins directly from the bloodstream to developing germinal cells. A logical, testable, hypothesis is that Sertoli cells synthesize and secrete proteins that bind and transport vitamins to germinal cells. Preliminary results are reported that suggest that Sertoli cells in serum-free primary monoculture secrete a substance that binds riboflavin with a high affinity. Twenty-day-old rats were killed by cervical dislocation, the testes were removed , and Sertoli cells were ~ r e p a r e d

Transforming growth factor-beta (beta 1, beta 2, and beta 3) gene expression and action during pubertal development of the seminiferous tubule: potential role at the onset of spermatogenesis

The potential role of transforming growth factor-beta (TGF beta) as a mediator of cell-cell interactions during the pubertal development of the seminiferous tubule was examined. Mesenchymal-derived peritubular cells and epithelial-like Sertoli cells were isolated from prepubertal, midpubertal, and late pubertal rat testes. The developmental expression of the multiple forms of TGF beta (TGF beta 1, -beta 2, and -beta 3) in whole testis and isolated somatic cell types was determined using a nuclease protection analysis. TGF beta 1 and TGF beta 2 mRNA expression was predominant in the immature testis and decreased at the onset of puberty. TGF beta 3 mRNA expression, the most abundant form of TGF beta present, peaked at an early pubertal stage, coincident with the initiation of spermatogenesis. Peritubular and Sertoli cells expressed each isoform of TGF beta during development. Peritubular cell mRNA expression of TGF beta 1, -beta 2, and -beta 3 decreased during pubertal development upon differentiation of this cell type. Sertoli cell expression of TGF beta 1 increased slightly and plateaued during pubertal development. TGF beta 2 mRNA expression was evident only in immature prepubertal Sertoli cells. Sertoli cell mRNA expression of TGF beta 3 increased transiently at the onset of puberty, corresponding with the peak of expression observed during the analysis of whole testicular development. Immunoblot analysis indicated that both cultured peritubular and Sertoli cells can produce the proteins for TGF beta 1, -beta 2, and -beta 3. Analysis of the hormonal regulation of TGF beta expression revealed that FSH caused a dramatic decrease in Sertoli cell TGF beta 2 expression while having no effect on TGF beta 1 or TGF beta 3 expression. Potential actions of TGF beta in the seminiferous tubule were also examined. TGF beta 1 inhibited TGF alpha-induced [3H]thymidine incorporation into peritubular cell DNA with cells from each developmental stage examined. TGF beta 1 had no effect on Sertoli cell proliferation. Previously, germinal cells have been shown to be responsive to TGF beta. This study demonstrates the potential of having a unique hormone-dependent pattern of TGF beta isoform expression during postnatal organ development. Observations demonstrate that the suppression of TGF beta 2 expression, in part in response to FSH, and the transient increase in TGF beta 3 expression correlate with the onset of puberty and the induction of spermatogenesis