Biomedical subjects

P Chieffi

Publications and source records attributed to P Chieffi.

24 records · Page 2Linked to original sources

PubMed1996

Regulation of avian precardiac mesoderm development by insulin and insulin-like growth factors.

Endoderm within the heart forming regions of vertebrate embryos has pronounced effects on myocardial cell development. Previous studies have suggested that these effects are mediated by soluble growth factors, in particular fibroblast growth factor 2 (FGF-2) and activin-A. Since both insulin and insulin-like growth factors (IGFs) are present in developing avian embryos at the time of heart formation, we have investigated the potential role of these molecules in promoting development of premyocardial cells in quail. Culture of precardiac mesoderm explants from stage 5 quail embryos in medium containing insulin, IGF-I, or IGF-II increased proliferation of premyocardial cells, with maximal stimulation observed at approximately 25 nM for each ligand. A direct comparison of the proliferative response of precardiac mesoderm to endoderm, fetal calf serum, insulin, IGF-I, IGF-II, activin-A, and FGF-2 showed that FGF-2 and activin-A increased proliferation of premyocardial cells approximately 2-fold, while insulin, IGF-I, and IGF-II stimulated proliferation approximately 3-fold. Insulin and IGF-I enhanced the rate of myocyte differentiation, similar to previously reported effects of endoderm. In contrast, exposure of precardiac mesoderm explants to transforming growth factor beta (TGF beta) reduced proliferation of premyocardial cells and moderated the proliferative effects of IGF-I. TGF beta did not block the differentiation of stage 5 premyocardial cells. Reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that mRNAs encoding insulin, IGF-II, insulin receptor, and IGF-I receptor were present in both precardiac mesoderm and endoderm, as well as in the forming heart at stage 8. Since premyocardial cells can survive and differentiate in a defined medium lacking these factors, precardiac mesoderm may produce IGF-II and insulin at levels that are sufficient to stimulate myocyte development. Taken together, these results suggest that insulin and/or IGF-II may promote cardiac development in vivo by both autocrine and paracrine mechanisms. Cardiogenesis may therefore be promoted by the combined action of several classes of growth factors.

Animals↗

PubMed1995

Detection of c-mos related products in the dogfish (Scyliorhinus canicula) testis.

The objective of the present paper was to do a comparative study to assess somatic versus germ cell localization of c-mos products in the testis. In mouse and amphibian oocytes, c-mos activity is necessary for meiotic maturation. Lack of c-mos expression has been reported in somatic cells of male and female gonads while transcripts have been found in germ cells of testis and ovary. Using a v-mos probe, we report here the detection of a c-mos related transcript (1.7 kb) in the dogfish Scyliorhinus canicula testis. Western blot analysis detects two proteins of 106 and 32 kDa. A specific immunostaining was exclusively localized in the interstitial tissue while the germinal compartment was completely negative. In conclusion, our results indicate for the first time the presence of c-mos products in an elasmobranch species and, moreover, their presence in somatic testicular cells rather than germ cells. Therefore, this finding in an ancient vertebrate indicates that c-mos activity does not have a direct universal role in the regulation of spermatogenesis.

Animals↗

PubMed1995

Ethane 1,2-dimethane sulfonate effects on the testis of the lizard, Podarcis s. sicula Raf: morphological and hormonal changes.

Ethane 1,2-dimethane sulfonate (EDS) destroys Leydig cells in the testis of some rodents (mice excluded), disrupts interstitial and germinal compartments in the frog, Rana esculenta, while it stimulates testicular activity in the teleost, Gobius paganellus. In the Japanese quail the toxin removes mature spermatozoa. There is no information on EDS effects in reptiles. The present study examines the effect of EDS treatment in the lizard Podarcis s. sicula Raf during two different periods of the testicular cycle (winter stasis and breeding season). Animals received a single EDS injection (100 mg/kg body wt) and were sacrificed at 0 and 24 hr and 3, 5, 7, 11, and 28 days after injection. Androgens were measured in plasma and right testes, while left testes were examined histologically. Plasma androgen levels decreased 5-7 days after EDS injection, alongside interstitial tissue destruction and mast cell appearance, with slight but significant increases on Days 11 and 28. Testicular androgen levels did not change. On Day 11 metaphases were present in the interstitial tissue which regenerated on Day 28. Between Days 5 and 7 some pycnotic nuclei of spermatocytes appeared, mitotic activity of spermatogonia was normal, but germ cell stages were disorganized and empty spaces appeared at the boundary of the tubule. These data show that a single EDS injection results in destruction and repopulation of the interstitial cells in a reptile. Moreover, the effects of EDS in the lizard suggest that P. s. sicula Raf testis responds to the toxin in a similar fashion to the rat testis.

Androgens↗

PubMed1995

Changes in proto-oncogene activity in the testis of the frog, Rana esculenta, during the annual reproductive cycle.

Proto-oncogenes are said to influence the regulation of cellular growth and differentiation. Myc, Fos, Jun, and Mos protein localization has been studied by immunocytochemistry in the testis of the frog, Rana esculenta, during the annual reproductive cycle. Oncoproteins have been localized in the primary and secondary (I and II) spermatogonia (SPG). Myc and Mos also appear in I and II spermatocytes (SPC) while Jun appears in II SPC. Myc, Fos, and Jun in SPG translocate in the nucleus during the periods of active spermatogenesis. Myc, Fos, and Jun are also localized in Sertoli cells. Fos is present in interstitial cells during the period characterized by the androgen peak which precedes the sharp increase of estradiol. It is suggested that proto-oncogene activity exerts a regulatory role in steroidogenesis and spermatogenesis.

Androgens↗

PubMed1995

Chicken GnRH-II and salmon GnRH effects on plasma and testicular androgen concentrations in the male frog, Rana esculenta, during the annual reproductive cycle.

In the frog, Rana esculenta, two molecular forms of GnRH, coeluting with chicken (c) GnRH-II and salmon (s) GnRH, have been detected using HPLC and radioimmunoassay. Mammalian (m) GnRH seems to be also present. In amphibians the role of cGnRH-II seems to be primarily the involvement in the regulation of neuroendocrine processes and, while the mGnRH has been postulated to act as a neurotransmitter and/or neuromodulator, the activity of sGnRH-like material has not been investigated. Therefore, we have treated the frogs with single or multiple injections of cGnRH-II or sGnRH (6 micrograms) or both peptides (6 micrograms of each) to detect differences in the response measured as testicular or plasma androgen (testosterone plus 5 alpha-dihydrotestosterone) concentration during the annual reproductive cycle. The basal profile of testicular and plasma androgen shows that the spring peak disappeared in control animals given multiple injections and kept in short-term captivity. We show in the treatment with cGnRH-II and/or sGnRH that the effects of the peptides depend on the season, the experimental design, and the tissue in which androgen levels were measured. In particular, both peptides strongly stimulate androgen production during the autumn-winter period, the time of the greater response to the GnRHs when basal levels of steroids are highest.

Analysis of Variance↗

PubMed1995

The effects of testosterone and estradiol on mast cell number in the harderian gland of the frog, Rana esculenta.

The Harderian gland (HG) of the frog Rana esculenta contains mast cells in the interstitial tissue. The mast cell number (MCN) is influenced by sex hormones. Gonadectomy in both sexes provoked a decrease in MCN in January, while no effect was observed in September. Sex hormone-replacement therapy gave different results; estradiol treatment in castrated males and females always increased MCN, while testosterone did not. Acute estradiol treatment provoked an increase in MCN on days 2 and 4 of treatment and the morphology of the glandular compartment appeared normal. On days 8, 10 and 12 of treatment the MCN drastically decreased. The majority of glandular acini appeared strongly disorganized and the interstitial tissue became hypertrophic in concomitance with an increased vascularization. Our results suggest that estradiol acts by stimulating mast cells and acute estradiol treatment provokes proliferation of interstitial connective tissue together with glandular cells damage.

Animals↗