PubMed Health⌕ Search

Biomedical subjects

G C Balboni

Publications and source records attributed to G C Balboni.

At least 19 recordsLinked to original sources

Sex steroids and odorants modulate gonadotropin-releasing hormone secretion in primary cultures of human olfactory cells.

Olfactory neurons and GnRH neurons share a common origin during development. In the nasal epithelia, GnRH neurons persist throughout fetal life and adulthood. The fate and function of these neurons in vivo have remained unknown. In a previous in vitro study, we isolated, cloned, and propagated primary long term cell cultures from the olfactory neuroepithelium of 8- to 12-week-old human fetuses. These cells expressed both neural proteins as well as olfactory genes and were responsive to odorant stimuli. We now report that these human olfactory cells also express the GnRH gene and protein. Combined HPLC and RIA studies have indicated that these cells release authentic GnRH in spent media. The release of GnRH was time dependent and was positively affected by sex steroids and odorants. Immunohistochemical data demonstrated the presence of sex steroid receptors in these cells. The presence of the alpha- and beta-subtypes of the estrogen receptor was also demonstrated by RT-PCR and Western blot analysis. When the cells were stimulated with increasing concentrations of 17beta-estradiol in the presence of a fixed concentration of progesterone (10(-7) mol/L), the combination of the two steroids induced a 3- to 4-fold increase in GnRH secretion. This stimulatory effect was completely blunted by tamoxifen. Neither 17beta-estradiol nor progesterone was effective when tested separately. Treatment with increasing concentrations of the odorant, l-carvone, induced a time- and dose-dependent dramatic increase in GnRH protein release (1000-fold increase) and gene expression. Repeated application of the stimulus resulted in a progressive lower responsiveness of the cells. To our knowledge, this is the first time that primary cell cultures from human fetal olfactory neuroepithelium have been shown to express and release GnRH. Our results also demonstrate that these cultures, which are sensitive to sex steroids and odorants, can be useful models in the study of the complex array of regulatory factors that finely tune GnRH secretion in humans.

Acyclic Monoterpenes↗

Alfa-inhibin and transferrin in human fetal testis.

Aim of this research was to investigate the immunohistochemical localization of alfa-inhibin and transferrin in the human fetal testis. In addition, an attempt was made to identify the Sertoli cells in primary cell cultures of fetal testes by the demonstration of the mRNA for transferrin and the presence of the FSH receptor. 15 fetal testes (8-12 weeks) were utilized. The results of this study show: 1. In sections, immunoreactivity for alfa-inhibin was present in the interstitial cells, whereas transferrin was localized in Sertoli cells; 2. After 20 days of primary culture: a) the percentage of immunoreactive cells for both the substances was about 15%; b) the presence of FSH receptor and the expression of transferrin mRNA were observed in 12-15% of cells. These findings suggest: 1. alfa-inhibin and transferrin are present in the human fetal testis; 2. the expression of transferrin mRNA and the presence of the FSH receptor permit the identification of the Sertoli cells in primary cultures of fetal testes (8-12 weeks).

Cells, Cultured↗

Neuroblast long-term cell cultures from human fetal olfactory epithelium respond to odors.

Primary cell cultures from human fetal olfactory neuroepithelium have been isolated, cloned, and propagated in continuous in vitro culture for approximately 1 year. The two clones we report here synthesize both neuronal proteins and olfactory-specific markers as well as the putative olfactory neurotransmitter, carnosine. In addition, patchclamp experiments reveal that these cells are electrically excitable. Following exposure to a panel of aromatic chemicals one of the cell cultures shows a specific increase in intracellular cAMP, indicating that some degree of functional maturity is expressed in vitro. The results suggest that these cells originate from the "stem cell" compartment that gives rise to mature olfactory receptor neurons. These long-term cell cultures represent models that will be useful in studying the mechanism(s) of olfaction and the regulation of olfactory neurogenesis and differentiation.

Abortion, Legal↗

The nasal respiratory epithelium in the human fetus: lectin histochemistry.

Lectin binding histochemistry was used to investigate the glycoconjugate saccharidic moieties in the respiratory epithelium of human fetuses ranging in age from the 8th to the 12th week of gestation. The distribution and changes in the sugar residues of the glycoconjugates at the level of the basal cells, the bathyprismatic cells and the bathyprismatic ciliated cells are reported. PNA and SBA allowed the identification of some 'elongated cells', which were characterized by lectin-reactive supranuclear granules. These epithelial cells, which have not previously been identified using conventional histochemical methods, might be the precursors of secretory cells.

Embryonic and Fetal Development↗

Immunolocalization of inhibin alpha-subunit in the human testis. A light- and electron-microscopy study.

The localization of inhibin alpha-subunit in the human testis was studied at the light- and electron-microscope level with immunostaining techniques. Antibodies against specific fragments of porcine and human inhibin alpha-subunits were utilized. At light microscopy, inhibin alpha-subunit immunoreactivity was detected in Sertoli cells, spermatocytes and in some Leydig cells. At electron microscopy, gold labeling was found in the cisternae of the Golgi apparatus and in the endoplasmic reticulum of Sertoli and Leydig cells. Gold labeling for inhibin was also found in coated vesicles in the cytoplasm of Sertoli cells as well as in coated pits and coated vesicles in the cytoplasm of some spermatocytes. The results of the present study suggest that, in the human testis, inhibin is produced by Sertoli and Leydig cells and is taken up by spermatocytes, on which it might act in a paracrine manner.

Coated Pits, Cell-Membrane↗

Sertoli-germ cells interactions in the human testis.

In previous histoimmunochemical studies we reported that transferrin (TF) and insulin-like growth factor I (IGF-I) are present in the cytoplasm of the Sertoli cells of the adult human testis. Receptors for TF were found mainly in adluminal germ cells and type I receptors for IGF-I both in Sertoli and germ cells. Using electron microscopy, evidence of transfer of both TF and IGF-I from the Sertoli to the germ cells through a receptor-mediated endocytosis mechanism was also found. In this paper we report the results of the histoimmunochemical localization of alpha inhibin in the human fetal, prepubertal and adult testis. In 8- to 14-week-old fetal testes a positive immunostaining was found mainly in the interstitial cells, whereas no staining was found in the germ cords. In the prepubertal testis the immunostaining was present in the Sertoli cells but not in the interstitial cells. In the adult human testis the immunostaining was present not only in the Sertoli cells but also in the spermatocytes and in several Leydig cells. Using electron microscopy and immunogold labeling the presence of alpha inhibin immunoreactivity was found in the rough endoplasmic reticulum and in the Golgi cisternae of both Sertoli and Leydig cells. Moreover we found evidence of transfer of alpha inhibin from the Sertoli to the germ cells through receptor-mediated endocytosis.

Adult↗

Identification of sugar residues in human fetal olfactory epithelium using lectin histochemistry.

Lectin-binding histochemistry was used to investigate the distribution and the changes of the glycoconjugate saccharidic moieties in the olfactory epithelium of human fetuses ranging from 8 to 12 weeks of gestation. It was found that the basal cells, the sustentacular cells and the olfactory neurons exhibit differences in oligosaccharide cellular content and distribution. Differences in lectin binding was also demonstrated at the dendrite, cell body and axon of the receptor cells. From the 11th week onwards, Ulex europaeus agglutinin I was found to be a marker of the olfactory neurons.

Embryonic and Fetal Development↗

Plasminogen activator and inhibin in the human testis. An immunohistochemical investigation.

It is generally accepted that Sertoli cells are the main source of active proteins in the human testis, these proteins can act by autocrine and/or paracrine mechanism, modulating both the gametogenic and the endocrine function of the male gonad. As to the Plasminogen Activators (T-PA, U-PA) the immunocytochemical reactivity was detectable in the Sertoli cells and in some Leydig cells. A positivity in spermatocytes and early spermatides appeared to be in relation to the stage of the cycle of the seminiferous epithelium. Inhibin was studied in foetal as well in prepubertal and adult life. The positivity for alpha human and alpha porcine subunits was intense only in the interstitial cells at 12-15 weeks of foetal life. Afterwards the staining was detectable in the Sertoli cells while it decreased in the interstitial cells. In prepubertal life, only the Sertoli cells were stained. In adult life, at the light microscopic level the Sertoli and Leydig cells were positive as well as the spermatocytes. At the electron microscopic level, coated pits and vesicles containing labelled golden particles were shown in both Sertoli cells and spermatocytes, suggesting a paracrine activity of the substance.

Adult↗

[Sugar residues of glycoconjugates in the olfactory epithelium of the human fetus: histochemical study using peroxidase-conjugated lectins].

The oligosaccharide content of glycoconjugates was studied at the olfactory epithelium of human fetuses ranging from 8 to 12 weeks of gestation by mean of peroxidase labelled lectins (DBA, PNA, WGA, SBA, ConA, LTA, UEA I). The main results demonstrated that: 1-The olfactory epithelium (olfactory cells, supporting cells and basal cells) was generally characterized by different amount of a-D-mannose, a-D-galactosamine, a-D-glucose, D-galactose-(beta 1-3)-N-acetyl-galactosamine, sialic acid and a-L-fucose. 2-At the 11th-12th week of gestation the largest amount of sugar residues was detected at the olfactory cells and at some basal cells. 3-At the 12th week of gestation, UEA I may be considered a specific marker of the olfactory cells in different stages of development.

Carbohydrate Sequence↗

Insulin-like growth factor-I receptors in nonfunctioning thyroid nodules.

We have recently demonstrated the production of insulin-like growth factor-I (IGF-I) as well as the presence of type I IGF receptors in human thyroid cells in primary culture. The role of IGF-I in the control of thyroid cell growth has been well established. In order to investigate the involvement of IGF-I in abnormal thyroid growth, the density of IGF-I receptors in solitary, cold, micro- and macro-follicular thyroid adenomas, and in extranodular histological normal tissue was studied. Forty-three euthyroid patients with isolated cold nodules were selected for the study. In 30 patients the presence of IGF-I receptors was evaluated by using quantitative immunohistochemistry; in 10 patients by using radioligand binding studies, and in 3 patients by using affinity labeling. Cross-linking and binding studies clearly demonstrated the presence of a homogeneous class of binding sites for type I IGF receptors. Furthermore, radioligand studies did not show any significant differences in receptor density between the 2 types of thyroidal tissues. Conversely, the computerized analysis of 900 fields of nodular and normal thyroid tissues immunostained with the monoclonal antibody alpha-IR3, strongly indicated that higher concentrations of IGF-I receptors were present in the epithelial cells of non-functioning thyroid nodules than in the adjacent extranodular thyroid tissues. These studies strongly suggest that the same type I IGF receptor is present in thyroid follicular adenomas as in histological normal thyroid tissue removed from the same patient. The higher concentration of IGF-I receptors as documented by immunostaining in the adenomas suggests that IGF-I may contribute to the abnormal growth of the neoplasms.

Adenoma↗

[Growth factors in human tooth development].

Our research concerns the immunohistochemical localization of EGF and IGF-I receptors in the tooth germ, using monoclonal antibodies. The results show that in the early phases of human tooth development EGF and IGF-I receptors are present. At bud stage both receptors are localized at dental laminae level, in some epithelial cells of the tooth bud and in some mesenchymal cells. At cap stage the receptors are present in the outer and inner enamel epithelium, and in some cells of stellate reticulum. As far as concerns the mesenchymal cells, some cells of dental papilla in contact with enamel organ, are intensely positive. The immunopositivity is present also in some mesenchymal cells at follicular level. At late cap stage and at early bell stage receptors are not present at inner enamel epithelium level but they can be detectable in the mesenchyma of dental papilla and in some cells of the follicle. On the basis of these results it may be hypothesized that EGF and IGF-I can act as growth factors in the modulation of cellular proliferation and differentiation during the human tooth morphogenesis. Moreover, it is possible that these substances can play a role in the mesenchymal-epithelial interaction in the developing human tooth.

Cell Differentiation↗

Sertoli cell proteins in the human seminiferous tubule.

It is well known that rat Sertoli cells in culture secrete both testis-specific proteins, such as inhibin and androgen binding protein (ABP), and proteins which are very similar, if not identical, to serum proteins, such as transferrin (TF), ceruloplasmin, and IGF-I. It is also well known that very few data have been reported about the secretory activity and the hormonal regulation of the Sertoli cell in man, mainly because of the difficulties associated with the isolation of pure cell populations from human tissue. Using histoimmunochemical techniques we tried to localize, with specific antisera, Sertoli cell proteins and, when possible, their receptors in the human testis. The results obtained with our Light Microscopy studies suggest that: (1) human Sertoli cells produce and/or store transferrin (TF), IGF-I, an albumin-like protein and ABP; (2) TF receptors are localized in spermatocytes and early spermatids and are absent in spermatogonia, in the cytoplasm of Sertoli cells and in differentiated spermatids; (3) IGF-I type I receptors are localized in the same germ cells and in the cytoplasm of Sertoli cells. The results obtained with our Electron Microscopy studies suggest that TF and IGF-I are internalized through a receptor mediated endocytosis mechanism both in Sertoli cells (basal compartment) and in germ cells (spermatocytes and early spermatids).

Androgen-Binding Protein↗

Insulin-like growth factor-I (IGF-I) and IGF-I receptor in human testis: an immunohistochemical study.

Using specific monoclonal antibodies, the authors studied the distribution of insulin growth factor-I (IGF-I) and its receptor in normal human testis by immunostaining techniques. IGF-I was preferentially localized in Sertoli cells. Less evident positivity was found in primary spermatocytes. In the interstitium some Leydig cells were positive for IGF-I. The major positivity for the IGF-I receptor was found in secondary spermatocytes and early spermatids, whereas Sertoli cells were less positive. An intense positivity was found in some Leydig cells.

Antibodies, Monoclonal↗

Immunologically reactive albumin-like protein in human testis and seminal plasma.

An immunologically reactive albumin-like protein (albumin) was localized, by an immunostaining technique, in the testis of infertile men (normal spermatogenesis, obstructive azoospermia) at the level of the Sertoli cells and in some cells of the germinal epithelium (secondary spermatocytes and early spermatids). No positive reaction was detectable in prepubertal testis. In vasectomized men, mean seminal albumin values were drastically reduced (by about 80%) in comparison to fertile controls, indicating a probable testicular origin. Mean seminal albumin values were also decreased in patients affected by azoospermia due to a seminiferous tubular lesion (about 40%) and in oligozoospermic patients (about 30%). In the same seminal samples transferrin, an index of Sertoli cell function, was also measured. Albumin and transferrin results were well correlated in the seminal plasma of each group (with the exception of vasectomized subjects), including a group of men with abnormally high concentrations of seminal transferrin. A weak correlation was found between seminal albumin and sperm count. We suggest that the presence of albumin in the human adult testis and in seminal plasma could be related to its ability to transport androgens.

Albumins↗

Transferrin and somatomedin C receptors in the human ovarian follicles.

Transferrin and somatomedin C receptors (TFr and SMCr) were studied in human ovaries by immunostaining using monoclonal antisera. The oocyte of evoluting follicles was intensely positive for both types of receptors, but this positivity decreases in large follicles. An evident positivity for both TFr and SMCr was shown in granulosa cells of evoluting follicles. However, not all of these cells were equally immunoreactive. An intense positivity was present in the developing thecal cells of early cavitary follicles as well as in thecal cells of follicles of medium and large size (greater than 6 mm) and in some cells of involuting follicles in initial atresia. These results seem to demonstrate that TFr and SMCr are present in different cellular components of the human developing and early involuting follicles.

Adolescent↗

Immunostaining of transferrin and transferrin receptor in human seminiferous tubules.

Transferrin (TF) and transferrin receptor (TFr) were studied in human testicular biopsy specimens with the use of immunostaining techniques. A polyclonal antibody to human TF (obtained in goat), a murine monoclonal antibody (B3/25) to human TFr, and antisera antigoat IgG and antimouse IgG, both labeled with peroxidase, were used. In seminiferous tubules of subjects with normal spermatogenesis, TF was found mainly in Sertoli cells and, in lesser amounts (probably related to the presence of receptor-TF complexes), in spermatocytes and early spermatids. TFrs were found only in spermatocytes and early spermatids. In patients with spermatogenetic disorders, TF was always found in Sertoli cells, whereas TFrs were found in spermatocytes only when they were present. These results seem to demonstrate that in human seminiferous tubules, Sertoli cells are devoted to the production and/or storage of TF, whereas spermatocytes and early spermatids use TF.

Cytoplasm↗