Regenerating I messenger RNA and protein expression in the failing human testis: a potential molecular prognostic marker of seminoma.

Infertility has been stated as a risk factor for testicular cancer; but currently, there is no prognostic indicator of tumor development from the pathologic testis with impaired spermatogenesis. Regenerating proteins are expressed in many human tissues including the testis, and their role in carcinogenesis has been well documented. In the present work, regenerating I messenger RNA and protein expression and cellular protein localization were studied in testicular biopsies of patients with normal (obstructive azoospermia) or impaired spermatogenesis (nonobstructive azoospermia) and in seminoma testis by quantitative reverse transcriptase-polymerase chain reaction, Western blot, and immunofluorescence analyses. No significant differences in regenerating I transcripts were reported between the 3 groups studied. However, regenerating I protein was highly expressed in pure seminoma and in placental-like alkaline phosphatase-positive seminiferous tubules with in situ carcinoma. Regenerating I protein levels measured by Western blotting increased from the placental-like alkaline phosphatase-negative distal region of the seminoma to the pure placental-like alkaline phosphatase-positive tumoral region. Importantly, although cells localized in seminiferous tubules of obstructive azoospermic patients with normal spermatogenesis were very slightly labeled, persisting germ, Sertoli, and myoid cells and fibrous tissues were strongly regenerating I positive in seminiferous tubules of nonobstructive azoospermia. These results suggest the possibility to use regenerating I as a prognostic marker of tumoral development in the infertile testis.

A potential novel mechanism involving connexin 43 gap junction for control of sertoli cell proliferation by thyroid hormones.

There is strong evidence that thyroid hormones through triiodothyronine (T3) regulate Sertoli cell proliferation and differentiation in the neonatal testis. However, the mechanism(s) by which they are able to control Sertoli cell proliferation is unclear. In the present study in vivo approaches (PTU-induced neonatal hypothyroidism known to affect Sertoli cell proliferation) associated with in vitro experiments on a Sertoli cell line were developed to investigate this question. We demonstrated that the inhibitory effect of T3 on Sertoli cell growth, analyzed by evaluating DNA-incorporated [3H] thymidine, was associated with a time and dose-dependent increase in the levels of Cx43, a constitutive protein of gap junctions, known to participate in the control of cell proliferation and the most predominant Cx in the testis. These Cx43 changes were associated with increased gap junction communication measured by gap FRAP. Consistent with these results two specific inhibitors of gap junction coupling, AGA and oleamide, were able to significantly reverse the T3 inhibitory effect on Sertoli cell proliferation. The present data also revealed a nongenomic effect of T3 on Cx43 Sertoli cells that was evidenced by a rapid up-regulation of gap junction plaque number as identified in Cx43-GFP transfected cells exposed to the hormone. This process appears mediated through actin cytoskeleton since incubation of the cells with cytochalasin D totally reversed the T3 stimulatory effect on Cx43-GFP gap junction plaques. Based on these data, we propose a working hypothesis in which Cx43 could be an intermediate target for T3 inhibition of neonatal Sertoli cell proliferation.

Trophic status of the small intestine in young and aged rats: modulation by a yogurt-supplemented diet.

Among the multifactorial causes of undernutrition in old age, gastrointestinal mucosa altered function and resulting specific malabsorption are the most relevant. Despite numerous studies that have dealt with the effects of aging on the digestive tract of mammals, results showed discrepancies in terms of proliferation and biochemical aging small intestine events. However, the slowing-down of the maturation process and the poor adaptation of metabolism and intestinal function are obvious and there is evidence that protective mechanisms are impaired with age and contribute to affecting the trophic activity and related systemic homeostasis. Good prospects to improve gastrointestinal function in the elderly are essential and research on nutritional intervention to limit and counteract age-related impairments must be extensive. Probiotics are good candidates and fermented milks might be of great interest. In the present study we first show the main structural and functional variations between 3- and 23-month-old rat small intestines. The trophic consequences of aging and nutritional adaptation under basal conditions are also analyzed and discussed after 20 days of a yogurt-supplemented specific diet in both young and aged rats. The main variations that occur with aging and yogurt diet are located in the proximal small intestine. The present findings indicate a slight improvement of morphological trophic parameters in both young and aged rats by yogurt, whereas enzymatic changes are more discrete. Despite the obvious age-related decrease in trophicity, we suggest that assessment of probiotic potentials on trophicity requires a more altered model than normal, healthy aging animals.

Consequences of adrenalectomy on small intestine trophic parameters in aged and young rats: evidence of defective adaptation by aging and lack of corticoids.

Previous study pointed to an important role of adrenals and glucocorticoids in the trophic status of the adult small intestine mucosa, with possible implications during stress events. Small intestine morphological and biochemical consequences of 10-day bilateral adrenalectomy and also sham-related laparotomy were determined in 23-month-old Sprague-Dawley rats. As described in young rats, adrenalectomy in old rats leads to partial atrophy and disorganization of the proximal small intestine epithelium, with an increase in the number of Paneth cells and reduced crypt cell proliferation. We also observed a decrease of goblet cell number and a reduction of all enzyme activities including disaccharidases, in contrast with the specific induced response shown in young rats. A number of marked biochemical effects have also been noted in aged rats subjected to solely laparotomy, suggesting age-related adaptation impairments. In conclusion, adrenalectomy modified the differentiation processes of the small intestinal mucosa in both young and aged rats, and some parameters underlined that the lack of corticoid-mediated adaptive process are exacerbated by cumulative surgical stress (event) and aging.