Expression of pro-opiomelanocortin-like gene in the testis and epididymis.

Adrenocorticotropin (ACTH), beta-endorphin, and the melanocyte-stimulating hormones (MSHs), which are products of a common precursor, pro-opiomelanocortin (POMC), are present in a variety of tissues other than pituitary. The recent detection of immunoreactive POMC-derived peptides in the male reproductive tract raised the possibility that these hormones might regulate reproductive function. To determine whether the low concentrations of POMC-derived peptides in the male reproductive tract are synthesized locally and are not contaminants from blood, we have demonstrated POMC-like gene expression in both testis and epididymis. The identification of cells in testis capable of synthesizing POMC mRNA was established by showing the presence of this mRNA in mouse Leydig cell lines (TM3 and I10A). The hybridizing species of POMC-like mRNA in the testis, epididymis, and Leydig cell lines (TM3 and I10A) were approximately 150 bases shorter than those in the pituitary or hypothalamus but were similar in size to that in the amygdaloid nucleus of rat brain. The concentration of POMC-like mRNA in the testis is almost as high as that in the hypothalamus. This finding is quite unexpected because the concentrations of POMC-derived peptides in the testis were 2-3 orders of magnitude lower than those in the hypothalamus. The demonstration of a POMC-like gene expression in male reproductive tissues suggests that POMC-derived peptides are synthesized in Leydig cells and epididymis. These observations are consistent with the postulate that POMC-derived peptides may exert paracrine and/or autocrine effects in these organs.     

Expression and regulation of kit ligand in the ovary of the hen

The Kit system, composed of Kit ligand (KL) and its tyrosine kinase receptor, cKit, has been well characterized in mammals. Studies have shown that it is involved in signaling between the oocyte and somatic cells during the process of follicle maturation. We characterized KL mRNA expression during follicle maturation in the domestic hen, examined regulation of KL and a possible function of the Kit system. KL mRNA expression was assessed using quantitative PCR (n=4 replicates) in follicles of various sizes (1, 3, 5, 6-12mm, F1). Expression of KL mRNA decreased significantly (p<0.01) with follicle development and was highest in <1mm follicles, which contained the theca as well as granulosa layers, with high levels also found in the granulosa layer of 3mm follicles and ovarian stroma. To study regulation of KL mRNA, granulosa cells from 6-8mm follicles (n=4 replicates) were plated in M199 plus 0.1% BSA in the presence of various treatments including: oocyte conditioned medium (OCM), Vitamin D(3), FSH, estradiol, progesterone and testosterone. OCM caused a dose-related increase (p<0.05) in expression of KL mRNA; Vitamin D(3) increased and FSH decreased expression of KL mRNA. cKit was detected (at the expected size) in the theca layer of 3-5mm follicles and in a lysate of whole <1mm follicles. Culture of granulosa cells in the presence of OCM resulted in a decrease of P4 secretion, an effect blocked by pre-incubation of OCM with cKit antibody. Although OCM caused a dose-related increase in E2 secretion from theca, this was not blocked by cKit antibody.     

Expression of preproenkephalin-like mRNA and its peptide products in mammalian testis and ovary.

The distribution of preproenkephalin mRNA and proenkephalin-derived peptides have been examined in gonadal tissues from rats, hamsters, and cattle. A preproenkephalin mRNA band was detected in the ovaries of all three species and in hamster testis that is identical in size to the 1450-nucleotide mRNA typically found in tissues that express proenkephalin. Rat testis, on the other hand, expresses at least one preproenkephalin-like mRNA that is substantially greater in size (1900 nucleotides). [Met]enkephalin-containing peptides were also detected in each of the gonadal tissues examined. Although the abundance of preproenkephalin-like mRNA in rat testis was comparable to that in rat brain, the testicular content of proenkephalin-derived [Met]enkephalin sequences was less than 4% of the rat brain content. Together these data suggest that preproenkephalin-like mRNA in rat testis is not efficiently translated, proenkephalin-derived peptides undergo rapid turnover in this tissue, or the mRNA in rat testis has a frameshift resulting in an altered coding sequence.     

Expression of the growth hormone variant gene in human placenta

Besides the hGH-N gene, which codes for the pituitary 22 and 20K GH variants, the human genome contains a second GH gene, namely the GH-V, which has been thought to be silent. We recently discovered a placental variant of human growth hormone (hPGH), which appears in maternal serum at mid-pregnancy and which rises in concentration thereafter to term. As hPGH and GH-V proteins display very similar characteristics, including a high affinity for hepatic GH receptors, they could be identical. To verify this hypothesis, we sought hGH-V mRNA in placenta. Hybridization experiments were performed between dot-blotted mRNA originating either from placenta or from one pituitary hGH secreting adenoma and synthetic polynucleotide probes corresponding to specific portions of the hGH-V or hGH-N gene sequences. The results indicate that the V gene is indeed expressed in the placenta and, at a very low level, in the pituitary adenoma. Therefore hPGH is most likely the expression product of the hGH-V gene.     

Expression and regulation of adiponectin and receptor in human and rat placenta

CONTEXT: Adiponectin is an adipocyte hormone involved in glucose and lipid metabolism. Recently, two receptors of this protein, called adiponectin receptor 1 (Adipo-R1) and Adipo-R2, have been cloned. OBJECTIVE: The aim of this study was to examine whether adiponectin and its receptors are expressed in human and rat placentas and to evaluate the regulation of these factors by gestational age and nutritional status. RESULTS: Our results demonstrate that adiponectin and Adipo-R2 are localized in both human and rat placentas. Human adiponectin and Adipo-R2 are presented in cytotrophoblast and syncytiotrophoblast cells. However, rat adiponectin and Adipo-R2 change their specific cell type immunostaining during gestation. Furthermore, placental adiponectin mRNA expression is increased during pregnancy in the rat, whereas Adipo-R2 has the contrary pattern. We also assessed the effect of food restriction (30%) during gestation, and we observed that adiponectin mRNA levels decrease after 16 d of undernutrition. In contrast, placental Adipo-R2 mRNA is unchanged by undernutrition. Finally, treatment with adiponectin during gestation decreases Adipo-R2, glucose transporter 3, lipoprotein lipase, and TGF-beta mRNA expression. CONCLUSION: Taken together, our results suggest that, at least in rodents, adiponectin may be involved in the regulation of several placental functions.     

POTE,a highly homologous gene family located on numerous chromosomes and expressed in prostate,ovary, testis,placenta, and prostate cancer

We have identified a gene located on chromosomes 21 that is expressed in normal and neoplastic prostate, and in normal testis, ovary, and placenta. We name this gene POTE (expressed in prostate, ovary, testis, and placenta). The POTE gene has 11 exons and 10 introns and spans approximately equal 32 kb of chromosome 21q11.2 region. The 1.83-kb mRNA of POTE encodes a protein of 66 kDa. Ten paralogs of the gene have been found dispersed among eight different chromosomes (2, 8, 13, 14, 15, 18, 21, and 22) with preservation of ORFs and splice junctions. The synonymous:nonsynonymous ratio indicates that the genes were duplicated rather recently but are diverging at a rate faster than the average for other paralogous genes. In prostate and in testis, at least five different paralogs are expressed. In situ hybridization shows that POTE is expressed in basal and terminal cells of normal prostate epithelium. It is also expressed in some prostate cancers and in the LnCAP prostate cancer cell line. The POTE protein contains seven ankyrin repeats between amino acids 140 and 380. Expression of POTE in prostate cancer and its undetectable expression in normal essential tissues make POTE a candidate for the immunotherapy of prostate cancer. The existence of a large number of closely related but rapidly diverging members, their location on multiple chromosomes and their limited expression pattern suggest an important role for the POTE gene family in reproductive processes.     

Expression of ER-alpha and ER-beta in the hamster ovary: differential regulation by gonadotropins and ovarian steroid hormones

Spatiotemporal expression patterns of ER-alpha and ER-beta protein and mRNA in hamster ovarian cells during the estrous cycle and following hypophysectomy and selective hormone replacement were evaluated by immunofluorescence, immunoblotting and in situ hybridization analyses. Whereas ER-beta mRNA and protein expression predominated in granulosa cells and ER-alpha expression was in interstitial and thecal cells, overlap in receptor subtype expression across cell types was evident. Both ER subtypes were present from primordial follicle stage onward. ER-alpha mRNA levels and immunoreactivity started increasing from D3:0900 h in interstitial and granulosa cells and peaked on the proestrous (D4:0900 h). Regionalized higher expression of ER-alpha in granulosa cells in and around the forming antrum was evident. Surface epithelial cells were also positive. ER-beta mRNA and protein expression increased markedly in granulosa and interstitial cells on D2:0900 h, reached a peak on D3:0900 h, and then declined sharply on D4:0900 h. No change in ER expression occurred following the preovulatory gonadotropin surge. Whereas FSH or human CG stimulated ER-alpha mRNA and protein expression in hypophysectomized hamsters, only FSH could stimulate ER-beta mRNA and protein, and the effect was significantly attenuated by human CG. ER expression was stimulated by estrogen, but progesterone strongly inhibited estrogen action. These results indicate that ER expression is cell type specific to the larger extent and is critically regulated by reproductive hormones.     

Auto-regulation of thyroid hormone receptors in the goldfish ovary and testis

Thyroid hormones, acting via their cognate thyroid receptors (TRs) act as mediators and modulators of several physiological processes and homeostasis. A clear role for the TRs in reproduction has not yet been established although several lines of recent evidence suggest that they are involved in the regulation of reproduction. To further study the role of TRs in control of reproduction, we investigated homologous regulation of TR subtypes in the gonads of goldfish, in vivo and in vitro. It was found that tri-iodothyronine (T₃) down-regulates the traditional TRs (TRα-1 and TRβ) and up-regulates a dominant-negative form, TRα-t. This indicates a ‘feedback’ mechanism whereby an acute treatment with T₃ down regulates further T₃ mediated response. The results provide novel information on auto-regulation of TRs in the goldfish ovary and testis, and support the hypothesis that thyroid hormones are involved in the control of reproduction.     

PET imaging in the management of tumors of testis and ovary: current thinking and future directions

The role of fluoro-D-deoxyglucose positron-emission tomography (FDG-PET) in testicular malignancies has been examined in various studies primarily in three specific settings: 1) differentiation of active disease from fibrosis/mature teratoma in patients with residual mass following chemotherapy and evaluation of the response to treatment; 2) initial staging and disease assessment after orchidectomy identification of suspected recurrences in the context of elevated circulating serum markers; and 3) predicting response to treatment. Of these, the area where FDG-PET imaging has been examined the most in testicular tumors is the evaluation of postchemotherapy residual mass in both seminoma and nonseminomatous germ cell tumors (NSGCT) of the testis, a critical step in determining the subsequent management approach of these tumors that vary amongst various centers. From the available data, this should be the test of choice for the assessment of a computed tomography (CT)-visualized residual mass following chemotherapy. In patients with residual masses or raised marker levels following therapy, positron-emission tomography (PET) appears sensitive and specific for detecting recurrent disease, at suspected and unsuspected sites. Fewer studies are available investigating its usefulness for staging at diagnosis and this requires further investigation to determine its eventual place as an imaging modality in this setting. Its precise role in disease prognostification is yet to be clearly defined in this malignancy but the initial results are promising. With regard to its role in ovarian carcinoma, it appears to be particularly useful for the diagnosis of recurrence when CA125 levels are rising and conventional imaging is inconclusive or negative. The role of fluoro-D-deoxyglucose (FDG)-PET/CT for the detection of recurrent ovarian cancer appears very promising and has the potential to replace the current surveillance techniques in detecting recurrent disease.     

Expression of the epidermal growth factor gene in mouse lachrymal gland: comparison with that in the submandibular gland and kidney

Expression of the epidermal growth factor (EGF) gene in mouse lachrymal gland was studied. The lachrymal gland of adult male and female mice contained immunoreactive EGF at a concentration of 0.013 +/- 0.002 (S.E.M.) and 0.014 +/- 0.003 ng/mg wet tissue respectively. Northern blot analysis of RNA from the lachrymal gland revealed the presence of 4.1-4.3 kb preproEGF mRNA, which was smaller in size than the 4.7-4.8 kb preproEGF mRNA in the submandibular gland and kidney. There was no sex difference in the mRNA level in the lachrymal gland, whereas the mRNA levels in the submandibular gland and kidney of adult male mice were 42-fold higher and twofold lower respectively than those of female animals. Administration of testosterone propionate or L-thyroxine to female mice increased submandibular gland levels of preproEGF mRNA by 35-fold or 12-fold respectively, but caused no change in lachrymal gland levels of preproEGF mRNA. The level of mRNA in the kidney was decreased by administration of androgen to one-third of that in control animals. The mRNA in the lachrymal gland was detected as early as 2 weeks of age and thereafter it remained at a constant level throughout adulthood, while that in the submandibular gland increased greatly during the pubertal stage. These results indicate differential hormonal and developmental regulation of EGF gene expression in the lachrymal gland, submandibular gland and kidney.