Jong Yoon Bahk

Stage Specific Identification of the Expression of GnRH mRNA and Localization of the GnRH Receptor in Mature Rat and Adult Human Testis

PURPOSE To identify the expression of gonadotropin-releasing hormone (GnRH) messenger ribonuclueic acid (mRNA) and localize GnRH receptors in mature rat and adult human testes. MATERIALS AND METHODS In situ hybridization and enzymatic receptor binding localization were performed. RESULTS GnRH mRNA was expressed within the seminiferous tubules in both mature rat and adult human testis. In rats, expression of GnRH mRNA was identified in the Sertoli cells and spermatogenic cells of some seminiferous tubules, but the other tubules did not express any hybridization signal. In humans, expression of GnRH mRNA was identified only in some spermatogenic cells in some seminiferous tubules. The receptors for GnRH were localized to cells in the interstitial tissues of the testis, probably Leydig cells. CONCLUSION The authors believe that the mature rat and adult human seminiferous tubular cells produce GnRH at the same specific stage of the spermatogenic cycle and that GnRH produced within seminiferous tubules, including Sertoli cells and spermatogenic cells, reacts with neighboring GnRH receptors in interstitial cells, including Leydig cells. The GnRH produced from the Sertoli cells in seminiferous tubules would react with GnRH receptors in interstitial cells as a paracrine hormone.

Expression of gonadotropin-releasing hormone (GnRH) and GnRH receptor mRNA in prostate cancer cells and effect of GnRH on the proliferation of prostate cancer cells

Abstract The purpose of this study was to determine the production of gonadotropin-releasing hormone (GnRH), the co-occurrence of GnRH receptors in prostate cancer cells, and the effect of GnRH on prostate cancer cell proliferation. Four human prostate cancer cell lines were studied. LNCaP is an androgen sensitive prostate cancer cell line, DU-145 and PC-3 are androgen resistant, and TSU-Pr1 is uncharacterized. The expression of GnRH and GnRH receptor mRNAs were assessed by in situ hybridization and the effect of exogenous GnRH on proliferation of prostate cancer cells was measured by thymidine incorporation assay. GnRH mRNA expression, determined by in situ hybridization, was found in 83.48% of the LNCaP, 89.7% of the TSU-Pr1, 86.2% of the PC-3 and 95.3% of the DU-145. Signals of GnRH receptor mRNA were detected in more than 95% of the cells of all four cell lines. The proliferation of the prostate cancer cells grown in media supplemented with peptide hormone lacking charcoal-stripped serum was significantly (P < 0.05) suppressed. No significant effect of GnRH on the proliferation of all four prostate cancer cells was observed. In summary, prostate cancer cells produced GnRH and its receptors, and exogenous GnRH treatment did not affect the prostate cancer cell proliferation. The existence of GnRH and GnRH receptor mRNA in the same cell suggests that the role of GnRH produced by prostate cancer cells would be autocrine.

Chronic nicotine and smoking treatment increases dopamine transporter mRNA expression in the rat midbrain.

Previous pharmacokinetics and electrophysiological results indicated an important role of nicotine in the modulation of dopamine transporter (DAT). To elucidate the expression changes of DAT on chronic nicotine and smoke administration, the effects of nicotine and passive cigarette smoke on DAT mRNA expression in the ventral tegmental area (VTA) and the substantia nigra (SN) area were examined using in situ hybridization and RNase protection assay. The results showed that chronic nicotine and smoke exposure highly unregulated DAT mRNA in the VTA and SN areas, including the dorsal part of substantia nigra pars compacta. Smoke for 30 min showed the highest increasing effect, whereas nicotine and smoke for 10 min only had slightly increasing effects. However, smoke for 1 h showed an increasing effect to a lesser extent than 30 min. These results revealed a new aspect of nicotines modulation on the DAT, and may have important roles in neuropsychological disorders related to the midbrain abnormalities such as drugs addiction.

Cut-off Value of Testes Volume in Young Adults and Correlation Among Testes Volume, Body Mass Index, Hormonal Level, and Seminal Profiles

OBJECTIVES To set a population-based cut-off value of normal adult and to determine correlations of testicular volume with body mass index (BMI), seminal profiles, and hormone levels. Testicular volume is an index of male fertility but cut-off values of normal adult has not been reported. METHODS During 54 months from January 2004, 1139 normal young men, 19-27 years old in military service were enrolled. Testicular volumes were measured by ultrasonometry. Height, body weight, and BMI were measured and semen analysis and hormone assay (follicle-stimulating hormone [FSH], luteinizing hormone [LH], and testosterone) were performed. RESULTS The mean age was 23.52+/-2.74. The mean testicular volume was 18.37+/-3.62 cm3 in left, and 18.13+/-3.85 cm3 in right. The mean body weight was 67.4+/-7.91 kg, the mean height was 176.2+/-6.64 cm, and mean BMI was 22.49+/-2.02 kg/m2. Testicular volumes had significant but weak correlations with height, body weight, and BMI. The semen analyses showed a mean pH of 7.63+/-0.74, volume of 2.49+/-1.12 mL, count of 68.63+/-13.62x10(6), motility of 69.93%+/-10.28%, and morphology of 68.62%+/-7.48%. Sperm counts and motility had positive correlations with testicular volume. The mean hormonal levels of FSH, LH, and testosterone were 7.31+/-2.42 mIU/mL, 7.81+/-2.49 mIU/mL, and 6.23+/-1.69 ng/mL, respectively. Testicular volume was negatively correlated with FSH and LH and positively with testosterone. CONCLUSIONS In this population-based study, we conclude that the cut-off testicular volume in normal young adults is around 18 mL and that testicular volume is positively correlated with height, body weight, BMI, semen profile, and testosterone, and negatively correlated with FSH and LH.

Dopamine D1 and D2 receptor mRNA up-regulation in the caudate-putamen and nucleus accumbens of rat brains by smoking.

Nicotine, the toxic substance that is exclusively absorbed from smoking, produces a wide array of behavior and collectively propels drug-seeking behaviors when abused. The neurotransmitter dopamine (DA) is important in the reward and reinforcing properties of many addictive drugs: however, the effect of nicotine by cigarette smoking itself on the expression of DA receptors in the caudate-putamen (CPu), nucleus accumbens (NAc) and olfactory tubercle (OTu) has not been elucidated completely. Hence, the effects of smoking and nicotine on DA receptors need to be defined. In this research, the effect of smoking and nicotine addiction on the DA D1 and D2 receptors in the rat CPu, NAc and OTu were studied. Adult male Spraque-Dawley (S.D., n=50) rats were administered with cigarette smoke (passive inhaled for 10 min and 1 h, 500 ml x 3 times/day, 4 weeks) and nicotine (oral, 3 mg/day). DA D1 and D2 receptor mRNA levels were determined by in situ hybridization and RNase protection assay (RPA). In the smoking groups (10 min and 1 h), DA D1 and D2 receptor mRNA greatly increased in the CPu and NAc, and most of all in the NAc. The nicotine treated group showed increased expression of DA D1 and D2 receptor mRNA too, but statistically less than in the smoking group. In the smoking group, DA D1 and D2 receptor mRNA levels were significantly higher in the CPu and NAc than in the nicotine group (P<.01). These results suggest that smoking and nicotine administration both influence DA receptor mRNAs in the CPu, NAc and OTu, in terms of up-regulation. The up-regulation was much more evident in the smoking group than in the nicotine group. In conclusion, we believe that smoking up-regulate the DA receptor mRNA expression significantly higher in rat CPu and NAc than nicotine but only a little bit higher in OTu.

Gonadotropin-Releasing Hormone (GnRH) and GnRH Receptor in Bladder Cancer Epithelia and GnRH Effect on Bladder Cancer Cell Proliferation

Introduction: Gonadotropin-releasing hormone (GnRH) is the pivotal hormone in the hypothalamopituitary gonadal axis. Recently, localization of extrahypothalamic GnRHs and GnRH receptors has been made in several organs but not in the bladder. We studied the extrahypothalamic GnRH and GnRH receptor in bladder cancer epithelia, and the role of GnRH on bladder cancer cell proliferation. Material and Methods: Normal human bladder mucosa, human transitional bladder cancer tissue, and 4 bladder cancer cell lines were used. For culture media, normal or charcoal-stripped serum with exogenous GnRH of different concentrations (0, 10–3, 10–5, and 10–7M) were supplemented. For detection of GnRH and the GnRH receptor and its mRNAs, RNase protection assay, in situ hybridization, and immunocytochemistry were performed. Cellular proliferation was measured by hemocytometer and the cell cycle by flow cytometer. Results: GnRH and GnRH receptor mRNA expression were detected in all cancer cell lines and human bladder cancer tissues. GnRH and GnRH receptors were localized in bladder cancer epithelial cells, but the density was not even. There were no significant differences (p > 0.05) in proliferation, and no significant change (p > 0.05) in cell cycle with GnRH supplements compared to controls. Conclusion: The bladder epithelia produce GnRH and the GnRH receptor in both normal and malignant tissues, but GnRH does not induce proliferation or cell cycle change of bladder cancer cells.

Chronic nicotine and smoking exposure decreases GABAB1 receptor expression in the rat hippocampus

Nicotine and smoking have long been proved to play an important role in cognition and memory in the hippocampus. This effect is closely related to the gamma-aminobutyric acid (GABA)ergic system. Previous research has focused on functional and pharmacological aspects of nicotines modulation activity. In this study, the effects of nicotine and different doses of smoking on GABA(B1) expression in the rat hippocampus have been examined using in situ hybridization and RNase protection assay. GABA(B1) receptor mRNAs were intensely expressed in the CA1, CA2, CA3, and dentate gyrus areas of the hippocampus. Nicotine and smoking doses dependently decreased GABA(B1) receptor expression in the hippocampus. These results revealed new aspects of nicotines modulation on GABA(B) receptor, and on learning and memory.

CONCENTRATION OF OFLOXACIN IN CANINE PROSTATE TISSUE AND PROSTATE FLUID AFTER INTRAPROSTATIC INJECTION OF BIODEGRADABLE SUSTAINED-RELEASING MICROSPHERES CONTAINING OFLOXACIN

PURPOSE Excellent treatment results in chronic prostatitis by direct intra-prostatic injection of antibiotic were reported several decades ago with only minimal scientific background. We examined the distribution, in prostatic tissue and fluid, of the antibiotic in canines after intra-prostatic injection of biodegradable sustained-releasing microspheres containing 12 mg. of ofloxacin. MATERIALS AND METHODS A total of 36 male dogs, 12 controls and 24 experimental, older than 2 years, were used. Experimental dogs were given biodegradable sustained releasing microspheres containing ofloxacin 12 mg. and poly(D,L-lactic) acid 28 mg., designed to release over more than a 4 week period. The 12 control animals were divided into 2 groups, and oral ofloxacin 100 mg. was given twice a day for 2 and 4 weeks. The 24 experimental animals were divided into 4 subgroups of 6 dogs each, 4 for prostatic tissue and 2 for prostatic fluid level of ofloxacin determination. Anesthesia was initiated with ketamine HCl and xylazine, and maintained with intermittent ketamine HCl. In the experimental groups, 1 ml. of resolved formula was injected into one lobe of surgically exposed prostates. The concentration of ofloxacin was measured by high performance liquid chromatography (HPLC) of blood, prostatic tissue and prostatic fluid. Pilocarpine 0.5 mg./kg. was used for the collection of the prostatic fluid. RESULTS The total ofloxacin of controls were 2,800 (2 weeks) and 5,600 (4 weeks) mg. In control groups, tissue concentrations of ofloxacin were relatively even at all segments of prostate, 7.4 +/- 0.8 (2 weeks) and 9.2 +/- 1.1 microg./ml. (4 weeks). The blood level ranged between 3.6 to 5.1 microg./ml. The prostatic fluid level ranged from 3.1 to 5.7 microg. /ml. In the experimental groups, the tissue levels of ofloxacin were 10.5 +/- 3.0 (1 week), 13.8 +/- 4.5 (2 weeks), 7.1 +/- 0.9 (3 weeks) and 7.7 +/- 3.0 microg./ml. (4 weeks) in the injected lobe. The opposite lobes were 8.0 +/- 1.1 (1 week), 10.2 +/- 4.2 (2 weeks), 5. 1 +/- 1.4 (3 weeks) and 7.6 +/- 0.8 (4 weeks) microg./ml. The blood level in the experimental groups ranged between 0.16 to 0.59 microg./ml. The prostate fluid level ranged from 2.9 to 6.1 microg./ml. in 8 dogs. Upon pathologic examination, the microspheres were interposed between prostate stroma and their size was reduced over time. CONCLUSIONS Our study indicates that there is communication between the right and left prostate lobes. Direct injection of biodegradable sustained releasing ofloxacin formula into the prostate may be a substitute for long term antibiotic medication in humans for chronic prostatitis in the future without hurting the minimal inhibitory concentration(MIC)90.

Ethanol modulates GABAB receptor expression in cortex and hippocampus of the adult rat brain

Using in situ hybridization, RNase protection assay and Western blot, we studied the effects of ethanol on the expression levels of GABA B receptor mRNA and protein in the cortex and hippocampus from adult rat brain. The results showed that ethanol significantly increased GABA B1 and GABA B2 receptor protein expression in the cortex, whereas only GABA B2 was increased in the hippocampus. GABA B receptor agonist baclofen could partially reverse the effect of ethanol. Further studies of the mRNA levels defined that GABA B1 mRNA levels were significantly increased in the hippocampus, with no significant changes of GABA B2 mRNA levels. Moreover, GABA B1 and GABA B2 receptor mRNA levels were increased on 3-week ethanol treatment. Finally, GABA B agonist baclofen and antagonist phaclofen showed significant decreasing effects on GABA B1 receptor mRNA levels in the cortex, but not in the hippocampus. These results were further confirmed by in situ hybridization. Thus, the present results showed the effects of ethanol on GABA B receptors in the cortex and hippocampus, implying the possible role of GABA B receptor in ethanol effects. The effects of GABA B receptor agonist and antagonist suggested that the possible mechanisms underlying that GABA B receptor modulated the behavioral effect induced by ethanol.

Ethanol modulates the expression of GABAB receptor mRNAs in the prenatal rat brain in an age and area dependent manner

Prenatal ethanol exposure has various deleterious effects on neuronal development. As GABA(B) receptor is known to play an important role during the development of the CNS, we now focused on its mRNA expression pattern in the rat brain during the late gestational days (GD) from 15.5 to GD 21.5. Ethanols effect was also observed from GD 11.5 to GD 21.5. GABA(B1) receptor mRNA showed a high expression level in GD 15.5 and 19.5, while GABA(B2) receptor mRNA did in GD 15.5 and 21.5. The mRNAs levels depended on age and area during development. Ethanol exposure decreased GABA(B1) receptor from GD 11.5 to GD 19.5 with slight increases in GD 21.5. The decreasing effects were area dependent, with the highest effects in the forebrain including cortex, whereas slight effects were observed in the midbrain and hindbrain. The present results suggest an important role of GABA(B) receptor in the effects of ethanol on prenatal brain developmental processes.