The effects of an in utero exposure to 2,3,7,8-tetrachloro-dibenzo-p-dioxin on male reproductive function: identification of Ccl5 as a potential marker.

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and dioxin-like compounds are widely encountered toxic substances suspected of interfering with the endocrine systems of humans and wildlife, and of contributing to the loss of fertility. In this study, we determined the changes in testicular gene expression caused by in utero exposure to TCDD along with the intra-testicular testosterone levels, epididymal sperm reserves, daily sperm production (DSP) and testis histology. To this purpose, female pregnant Sprague-Dawley rats orally received TCDD (10, 100 or 200 ng/kg body weight) or vehicle at embryonic day 15, and the offspring was killed throughout development.

Lack of effect on rat testicular organogenesis after in utero exposure to 3-monochloropropane-1,2-diol (3-MCPD).

3-Monochloropropane-1,2-diol (3-MCPD) is a food-born contaminant known to display toxic effects on male reproduction, producing infertility in rats and humans. Using the rat as a model, we investigated whether or not testicular organogenesis, which, in the rat species, occurs during the second half of gestation, was at particular risk regarding 3-MCPD toxicity. Pregnant rats were given daily doses of 5, 10 or 25 mg/kg BW of 3-MCPD from days 11.

Diethylstilbestrol inhibits the expression of the steroidogenic acute regulatory protein in mouse fetal testis.

This study investigated the early deleterious effects of an in-utero exposure to diethylstilbestrol (DES) on mouse testicular development. To that purpose, pregnant mice were injected daily with up to 100 microg/kg DES from 10.5 to 17.

Evidence for similar expression of protein C inhibitor and the urokinase-type plasminogen activator system during mouse testis development.

Plasminogen activators (PAs) and their inhibitors (PAIs) are predicted to be involved in the restructuring events that characterize the testis throughout development. We here demonstrate that PAI-3 or protein C (PC) inhibitor (PCI) was expressed in a sexually dimorphic fashion during mouse gonad genesis, whereas PAI-1 and -2 exhibited no sex differences. PCI transcripts accumulated rapidly in the male gonad, from 12.

Differential expression of tissue inhibitor of metalloproteinases type 1 (TIMP-1) during mouse gonad development.

In mammals, the gene Sry initiates signaling pathways triggering the differentiation of a testis from a sexually indifferent gonad. Assuming that these morphogenetic events may alter the proteolytic balance, the expression of matrix metalloproteinases (MMPs) and inhibitors (TIMPs) was investigated in gonads from 11.5 days postcoitum (dpc) onward, when testicular organogenesis occurs.

Overexpression of a dominant-negative type II TGFbeta receptor tagged with green fluorescent protein inhibits the effects of TGFbeta on cell growth and gene expression of mouse adrenal tumor cell line Y-1 and enhances cell tumorigenicity.

Transforming growth factor beta (TGFbeta) has been reported to be a potent growth inhibitor of epithelial cells. The purpose of the present work was to study in vitro and in vivo the effects of overexpression of a dominant-negative type II TGFbeta receptor on the proliferation and differentiation of Y-1 cells. Stable transfections were performed with a mutant TbetaRII (TbetaRII-KR) fused with the Enhanced Fluorescent Green Protein (EGFP).

Antisense oligonucleotide targeting the transforming growth factor beta1 increases expression of specific genes and functions of Leydig cells.

Transforming growth factor beta1 (TGFbeta1) has been reported to be a potent inhibitor of differentiated functions of many steroidogenic cells. Porcine Leydig cells (LC), as well as Sertoli cells (SC), express TGFbeta1 mRNA and secrete this peptide, suggesting that it might play an autocrine role. Moreover, many studies have suggested a possible paracrine regulation of LC by SC-secreted factors.

Sorbin in the porcine gastrointestinal tract and pancreas: an immunocytochemical analysis.

Sorbin is a 153-amino acid peptide that was initially discovered in the porcine duodenum. We have reported previously that this peptide regulates intestinal electrolyte transport and have described accumulation sites in the rat digestive tract. In the present study, we investigated the anatomical distribution and the site(s) of sorbin production in the porcine digestive tract using immunocytochemistry.

Repression of transforming growth factor beta 1 protein by antisense oligonucleotide-induced increase of adrenal cell differentiated functions.

Transforming growth factor beta 1 (TGF beta 1) is a potent inhibitor of several differentiated functions in bovine adrenal fasciculata cells (BAC). In addition, these cells express and secrete this factor. To determine whether this peptide plays an autocrine role in BAC, cells were transfected with 10 microM unmodified sense (SON) or antisense (AON) oligonucleotide complementary to the translation initiation region of the TGF beta 1 mRNA in an attempt to inhibit TGF beta 1 protein synthesis.

Expression and regulation of transforming growth factor-beta 1 messenger ribonucleic acid and protein in cultured porcine Leydig and Sertoli cells.

In the present work, the expression and secretion of transforming growth factor-beta 1 (TGF beta 1) by immature pig Leydig and Sertoli cells were investigated. Both cell types express two TGF beta 1 mRNA transcripts of 2.5 and 3.

Differential expression of the two nonallelic proinsulin genes in the developing mouse embryo.

In the mouse, insulin is produced from two similar but nonallelic genes that encode proinsulins I and II. We have investigated expression of these two genes during mouse embryonic development, using a PCR to detect the two gene transcripts and immunocytochemistry to visualize the two corresponding proteins. At appearance of the dorsal pancreatic anlage at day 9.

Insulin, glucagon and somatostatin in fetal rat islets maintained free-floating in culture: immunocytochemistry and radioimmunoassay.

Fetal rat islets maintained free-floating in tissue culture represent a source of B-cells. Because we recently noted the occurrence of other cell types during long-term tissue culture, this in vitro model was used to examine the possible development of non B-cells. The changes in the numbers and percentages of B, A and D-cells in vitro were estimated by counting the hormone-positive cells after immunocytochemical staining.

ACTH and angiotensin II regulation of insulin-like growth factor-I and its binding proteins in cultured bovine adrenal cells.

Insulin-like growth factor-I (IGF-I) is required for the maintenance of differentiated functions of bovine adrenal fasciculata cells in culture. We have investigated, by immunocytochemistry, the presence of IGF-I in cells cultured in the absence or presence of ACTH and angiotensin II (AII), as well as the secretion of IGF-I and its binding proteins (IGFBPs). In control cultures, very few cells were specifically stained with the anti-IGF-I serum.

Polyclonal origin of pancreatic islets in aggregation mouse chimaeras.

In the present study, we have examined the origin and growth pattern of the beta cells in pancreatic islets, to determine whether a single progenitor cell gave rise to all the precursors of the islets, or if each of a few progenitor cells is the founder of a different islet, or if each islet is a mixture of cells originating from a pool of progenitor cells. Aggregation mouse chimaeras where the pancreatic beta cells derived from each embryo can be identified in the islets on histological sections were analyzed. In two chimaeras, all the islets contained cells from both the aggregated embryo.

Light and electron microscopic immunocytochemical evidence that delta sleep-inducing peptide and gonadotropin-releasing hormone are coexpressed in the same nerve structures in the guinea pig median eminence.

The relationships between delta sleep-inducing peptide (DSIP) and GnRH immunoreactivity within the guinea pig median eminence are investigated by light and electron microscopic immunocytochemistry. Indirect immunofluorescence and elution-restaining experiments show that at the light microscopic level the distribution patterns of DSIP and GnRH immunoreactivity are indistinguishable. This suggested the possible coexistence of both immunoreactivities within the same fibers and neurosecretory endings.

Anatomical distribution of substance P-immunoreactive neurons in human brainstem during the first postnatal year.

The localization of substance P (SP)-immunoreactive structures in the infant brainstem was investigated by immunohistochemistry using the peroxidase antiperoxidase technique. SP-immunoreactive structures are widely distributed throughout the brainstem region. SP-immunoreactive cell bodies are prominent in the superior colliculis, the central grey, the nucleus tractus solitarius and the reticular formation.

[Demonstration of the secretion of IGF-I and of its binding proteins by bovine adrenal fasciculata cells in culture. Immunocytochemistry, radioimmunoassay and ligand blotting].

Insulin-like growth factor I (IGF-I) is required for the maintenance of differentiated functions of bovine adrenal fasciculata cells in culture. We have investigated, by immunocytochemistry, the presence of IGF-I in cells cultured in the absence or presence of corticotropin (ACTH) and angiotensin II (A-II), as well as the secretion of IFG-I and its binding proteins (IGF-BP). In control cultures, very few cells were specifically stained with the anti-IGF-I serum.

Anatomical distribution of LHRH-immunoreactive neurons in the human infant hypothalamus and extrahypothalamic regions.

The morphological features and distribution of luteinizing hormone-releasing hormone (LHRH)-immunoreactive cell bodies and fibers of the hypothalamic and the neighboring mesencephalic regions were studied in the normal newborn infant by immunohistochemistry. Within the hypothalamus, numerous LHRH-immunoreactive like (IL) cell bodies were found mainly in the ventral portion of the infundibular nucleus close to the median eminence and at a lower extent in the medial preoptic area. In addition, sparse immunoreactive cell bodies were displayed in the paraventricular and medial mammillary nuclei.

Processing of thyrotropin-releasing hormone prohormone (pro-TRH) in the adult rat pancreas: identification and localization of pro-TRH-related peptides in beta-cells of pancreatic islets.

Rat TRH prohormone (pro-TRH) contains five separate copies of the TRH progenitor sequence, Gln-His-Pro-Gly. All five sequences are flanked by paired basic amino acid cleavage sites and linked together by connecting sequences. RIAs to synthetic TRH and prepro-TRH-(178-199) were used to investigate pro-TRH processing in the endocrine pancreas of adult rats.

Immunohistochemical distribution of somatostatin in the infant hypothalamus.

Somatostatin (SS)-containing neurons were mapped in the normal infant hypothalamus with immunohistochemistry, using the peroxidase anti-peroxidase technique. Neurons displaying SS immunoreactivity show a widespread distribution throughout the hypothalamic region. Principal SS-immunoreactive like (SS-IL) perikarya are located in the paraventricular, infundibular and posterior nuclei and in the preoptic region.

Immunocytochemical evidence for the presence of S-100 protein in insulin-containing cells of cultured fetal rat islets.

S-100 protein was long considered to be specific to glial and Schwann cells, but was subsequently proved to be present in various organs. In particular, S-100 proteinimmunoreactivity was demonstrated in the parathyroid gland, adenohypophysis and endocrine pancreas. In the present study cultured fetal rat islets were investigated in view of the possible presence of S-100 protein immunoreactivity in their cells.

Anatomical distribution of somatostatin immunoreactivity in the infant brainstem.

The distribution of somatostatin-immunoreactive structures in the infant brainstem was investigated using the peroxidase-antiperoxidase technique. A wide distribution of somatostatin-immunoreactive cell bodies and fibers was observed throughout the brainstem. Numerous somatostatin-immunoreactive cell bodies and fibers were present in several areas of the brainstem including the substantia grisea centralis and the reticular formation.

Coexistence of thyrotropin-releasing hormone and insulin in cultured fetal rat islets: a light and electron microscopic immunocytochemical study during islet neoformation.

Pancreatic thyrotropin-releasing hormone (TRH) is without doubt localized in the insulin-containing beta-cells. A previous study reported cellular continuity between beta-cells and ducts in cultured fetal rat islets, but it is not known whether these insulin-containing beta-cells form a cell type that is different from the TRH cells producing insulin. On the other hand, the subcellular coexistence of both peptides as yet remains unresolved.

Persistence of peptidylglycine alpha-amidating monooxygenase activity and elevated thyrotropin-releasing hormone concentrations in fetal rat islets in culture.

Two experimental systems were used to investigate the relationship between TRH content and peptidylglycine alpha-amidating monooxygenase (PAMase) activity of the neonatal rat pancreatic islets: freshly isolated islets from rats aged 1-14 days, and fetal islets maintained in culture for 3 weeks. TRH was present in freshly isolated islets and in newly formed fetal islets in culture. However, while the TRH concentration in freshly isolated islets measured by RIA followed the same ontogenic pattern as that in the total pancreas, peaking during the first week of life (78 pg/micrograms DNA on day 4) and decreasing thereafter to reach 4 pg/micrograms DNA on day 14, the TRH content of fetal islets in culture did not decrease with time (65 pg/micrograms DNA on day 1 and 80 pg/micrograms DNA after 3 weeks in culture).

Localization of thyrotropin-releasing hormone- and insulin-immunoreactivity in the pancreas of neonatal rats after injection of streptozotocin at birth.

Streptozotocin treatment at birth induces, in the pancreas of rats, first depletion of insulin and thyrotropin-releasing hormone and then early regeneration of beta cells and insulin, but not TRH. This study was undertaken to investigate whether the reduction in pancreatic TRH content can be associated with changes in the intensity and the distribution of TRH-immunoreactivity, and to follow the pattern of regeneration of beta cells through insulin- and TRH-immunoreactivity. In control animals, strong TRH-immunoreactivity was seen in insulin-containing cells on days 1-4 after birth.