The in vivo muscarinic receptor binding of antimuscarinic agents (oxybutynin, solifenacin, tolterodine, and imidafenacin) used to treat urinary dysfunction in patients with overactive bladder is reviewed. Transdermal administration of oxybutynin in rats leads to significant binding of muscarinic receptors in the bladder without long-term binding in the submaxillary gland and the abolishment of salivation evoked by oral oxybutynin. Oral solifenacin shows significant and long-lasting binding to muscarinic receptors in mouse tissues expressing the M3 subtype. Oral tolterodine binds more selectively to muscarinic receptors in the bladder than in the submaxillary gland in mice. The muscarinic receptor binding of oral imidafenacin in rats is more selective and longer-lasting in the bladder than in other tissues such as the submaxillary gland, heart, colon, lung, and brain, suggesting preferential muscarinic receptor binding in the bladder. In vivo quantitative autoradiography with (+)N-[11C]methyl-3-piperidyl benzilate in rats shows significant occupancy of brain muscarinic receptors with the intravenous injection of oxybutynin, solifenacin, and tolterodine. The estimated in vivo selectivity in brain is significantly greater for solifenacin and tolterodine than for oxybutynin. Imidafenacin occupies few brain muscarinic receptors. Similar findings for oral oxybutynin were observed with positron emission tomography in conscious rhesus monkeys with a significant disturbance of short-term memory. The newer generation of antimuscarinic agents may be advantageous in terms of bladder selectivity after systemic administration.
Administration, Cutaneous ; Animals ; Autoradiography ; Benzhydryl Compounds ; Brain ; Colon ; Cresols ; Heart ; Humans ; Imidazoles ; Injections, Intravenous ; Lung ; Macaca mulatta ; Mandelic Acids ; Memory, Short-Term ; Mice ; Muscarinic Antagonists ; Phenylpropanolamine ; Positron-Emission Tomography ; Quinuclidines ; Rats ; Receptors, Muscarinic ; Salivation ; Solifenacin Succinate ; Submandibular Gland ; Tetrahydroisoquinolines ; Tolterodine Tartrate ; Urinary Bladder ; Urinary Bladder, Overactive

OBJECTIVETo investigate the feasibility of using cartilage link protein of hyaluronic acid (HA-CLP) for defining the tumor boundary in a mouse model of lung carcinoma.

METHODSLung carcinoma was induced in KM mice by chemical carcinogenesis. HA-CLP separated from bovine cartilage and purified by affinity chromatography was labeled with (125)I for autoradiography. Immunohistochemical analysis and Western blotting were used to examine the efficiency of HA-CLP in defining the boundaries of the lung tumors.

RESULTSWith autoradiography, the clearest image of lung cancer was obtained at 2 h. With immunohistochemical method, the tumor boundary was the most clearly displayed at 2 h when the strongest signals of HA-CLP was detected; Western blotting also showed the clearest bands of HA-CLP at 2 h.

CONCLUSIONHA-CLP has the immunogenicity of HABP, and can efficiently indicate lung tumor boundary in autoradiography and immunohistochemistry.

Animals ; Autoradiography ; methods ; Extracellular Matrix Proteins ; metabolism ; pharmacology ; Female ; Hyaluronic Acid ; metabolism ; Immunohistochemistry ; Iodine Radioisotopes ; Lung Neoplasms ; radiotherapy ; Male ; Mice ; Proteoglycans ; metabolism ; pharmacology ; Radiotherapy, Image-Guided ; methods

OBJECTIVE: In humans, a single exposure to phencyclidine (PCP) can induce a schizophrenia-like psychosis which can persist for up to two weeks. In rats, an acute dose of PCP increases dopaminergic activity and causes changes in dopamine related behaviours some of which are sexually dimorphic. To better understand the effects of PCP on dopamine receptor adaptations in the short term we examined dopamine D1-like receptors (D1R) and D2-like receptors (D2R) in the mesolimbic and nigrostriatal dopamine pathways, 4 hours after exposure to PCP in female rats. METHODS: Animals received a single dose of 40 mg/kg PCP and were sacrificed 4 hours later. In vitro autoradiography was carried out using [3H] SCH 23390 and [3H] raclopride that target D1R and D2R respectively, in cryostat brain sections. RESULTS: Two way analysis of variance (ANOVA), revealed an overall effect of PCP treatment (F [1,63]=9.065; p=0.004) on D1R binding with an 18% decrease (p<0.01) in binding in the medial caudate putamen. PCP treatment also had an overall effect on D2R binding (F [1,47]=5.450; p=0.024) and a trend for an increase in D2R binding across all the brain regions examined. CONCLUSION: These results suggest opposing D1R and D2R adaptations in striatal subregions of female rats following acute exposure to PCP that may occur through indirect mechanisms.
Animals ; Autoradiography ; Benzazepines ; Brain ; Dopamine ; Female ; Humans ; Phencyclidine ; Psychotic Disorders ; Putamen ; Raclopride ; Rats ; Receptors, Dopamine

OBJECTIVE: To observe whether bFGF could cross the blood-labyrinth barrier (BLB) after intra-abdominal injection and to establish an experimental basis for its clinical applications. METHOD: Thirty guinea pigs were divided into three groups. Animals in group 1 were administered o I-bFGF, while animals in group 2 and 3 were administered 125 and saline, respectively, via intra-abdominal injection. The both cochlea, blood, liver, brain, thyroid gland and kidney were collected and weighted. A radioimmunoassay analyzer was employed to measure counts per minute (CPM) of each sample, and autoradiography was performed on both cochlea. RESULT: The CPM value of organ samples in the 125I group was higher than that in other groups, and radioactive grain was observed in cochlear samples of this group. In the 125I-bFGF group, blood demonstrated the highest CPM value, while cochlea and brain demonstrated the lowest CPM value, with no radioactive grain observed in cochlear samples. CONCLUSION bFGF has some difficulties in getting across BLB, so the way of bFGF application in clinics need further study.
Animals ; Autoradiography ; Cochlea ; cytology ; metabolism ; Fibroblast Growth Factor 2 ; administration & dosage ; Guinea Pigs ; Injections, Intraperitoneal ; Iodine Radioisotopes ; administration & dosage

PURPOSE: Hydrodynamic-based procedure is a simple and effective gene delivery method to lead a high gene expression in liver tissue. Non-invasive imaging reporter gene system has been used widely with herpes simplex virus type 1 thymidine kinase (HSV1-tk) and its various substrates. In the present study, we investigated to image the expression of HSV1-tk gene with 5-(2-iodovinyl)-2'-deoxyuridine (IVDU) in mouse liver by the hydrodynamicbased procedure. MATERIALS AND METHODS: HSV1-tk or enhanced green fluorescence protein (EGFP) encoded plasmid DNA was transferred into the mouse liver by hydrodynaminc injection. At 24 h post-injection, RT-PCR, biodistribution, fluorescence imaging, nuclear imaging and digital wholebody autoradiography (DWBA) were performed to confirm transferred gene expression. RESULTS: In RT-PCR assay using mRNA from the mouse liver, specific bands of HSV1-tk and EGFP gene were observed in HSV1-tk and EGFP expressing plasmid injected mouse, respectively. Higher uptake of radiolabeled IVDU was exhibited in liver of HSV1-tk gene transferred mouse by biodistribution study. In fluorescence imaging, the liver showed specific fluorescence signal in EGFP gene transferred mouse. Gamma-camera image and DWBA results showed that radiolabeled IVDU was accumulated in the liver of HSV1-tk gene transferred mouse. CONCLUSION: In this study, hydrodynamic-based procedure was effective in liver-specific gene delivery and it could be quantified with molecular imaging methods. Therefore, co-expression of HSV1-tk reporter gene and target gene by hydrodynamic-based procedure is expected to be a useful method for the evaluation of the target gene expression level with radiolabeled IVDU.
Animals ; Autoradiography ; DNA ; Fluorescence ; Gene Expression ; Genes, Reporter ; Herpes Simplex ; Herpesvirus 1, Human ; Idoxuridine ; Liver ; Methylmethacrylates ; Mice ; Molecular Imaging ; Optical Imaging ; Plasmids ; Polystyrenes ; RNA, Messenger ; Simplexvirus ; Thymidine Kinase

AIMTo study the molecular mechanism of epididymal protease inhibitor (Eppin) modulating the process of prostate specific antigen (PSA) digesting semenogelin (Sg).

METHODSHuman Sg cDNA (nucleotides 82-849) and Eppin cDNA (nucleotides 70-723) were generated by polymerase chain reaction (PCR) and cloned into pET-100D/TOPO. Recombinant Eppin and Sg (rEppin and rSg) were produced by BL21 (DE3). The association of Eppin with Sg was studied by far-western immunoblot and radioautography. In vitro the digestion of rSg by PSA in the presence or absence of rEppin was studied. The effect of anti-Q20E (N-terminal) and C-terminal of Eppin on Eppin-Sg binding was monitored.

RESULTSEppin binds Sg on the surface of human spermatozoa with the C-terminal of Eppin (amino acids 75-133). rSg was digested with PSA and many low molecular weight fragments were produced. When rEppin is bound to rSg, then digested by PSA, incomplete digestion and a 15-kDa fragment results. Antibody binding to the N-terminal of rEppin did not affect rSg digestion. Addition of antibodies to the C-terminal of rEppin inhibited the modulating effect of rEppin.

CONCLUSIONEppin protects a 15-kDa fragment of rSg from hydrolysis by PSA.

Animals ; Antibodies ; pharmacology ; Autoradiography ; Humans ; Hydrolysis ; Male ; Prostate-Specific Antigen ; metabolism ; Proteinase Inhibitory Proteins, Secretory ; genetics ; immunology ; metabolism ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Semen ; cytology ; metabolism ; Seminal Vesicle Secretory Proteins ; metabolism ; Spermatozoa ; metabolism

BACKGROUND AND OBJECTIVES: Reperfusion of ischemic myocardium is necessary to salvage tissue from eventual death. However, new pathophysiological changes are initiated after reperfusion. The aim of this study was to investigate one of the mechanisms of ischemia/reperfusion (I/R) injury, and we focused on transferrin. MATERIALS AND METHODS: Male Spragre-Dawley (SD) rats were used for the I/R model. Myocardial ischemia was produced by occlusion of the left anterior descending coronary artery for 30 minutes. 99mTc Transferrin-Chitosan-hydrazino nicotinate hydrochloride (HYNIC) (Tfc) (/37 MBq/mL) was injected once after the reperfusion was finished. Autoradiography, hematoxylin and eosin (H & E) staining and determination of the tissue myeloperoxidase (MPO) activity were performed. RESULTS: Autoradiography showed remarkable 99mTc-Tfc uptake in the left ventricular myocardium at the reperfusion period from 0 to 1.5 hours, whereas no uptake was demonstrated at 3 hours. The uptake was increased again at 6 and 24 hours. Western blotting showed that the transferrin receptor (TfR) proteins were increased at 0 to 1.5 hours compared with that of the control; this expression of TfR disappeared at 3 hours, and it showed up for the second time at 6 and 24 hours. The MPO activity only at 24 hours was significantly higher than that of the control and those MPO activities at 0 to 6 hours (p=0.001). CONCLUSION: In the rodent model of 30 minutes occlusion and reperfusion, our study revealed, with using 99mTc-Tfc, that the TfR expression increased in the myocardium till 3 hours after reperfusion. TfR-mediated entry of iron into the cardiomyocytes may represent that this process plays a role in the I/R injury during the early reperfusion period.
Animals ; Autoradiography ; Blotting, Western ; Coronary Vessels ; Eosine Yellowish-(YS) ; Hematoxylin ; Humans ; Iron ; Male ; Myocardial Ischemia ; Myocardium ; Myocytes, Cardiac ; Niacin ; Peroxidase ; Pilot Projects ; Proteins ; Rats ; Receptors, Transferrin ; Reperfusion ; Reperfusion Injury ; Rodentia ; Transferrin

OBJECTIVETo explore the feasibility of tracing mesenchymal stem cells in vivo with scintigraphy.

METHODSTransferrin receptor expression of cultured mesenchymal stem cells (hMSCs) was quantified with radioligand-receptor binding assay before the cells were transplanted into the spinal cord of rabbits. (131)I-labeled transferrin was then administered into the subarachnoid space of the rabbits, and scintigraphic images were acquired with a gamma camera at different time points after the administration. In the control experiments, (131)I-labeled human serum albumin was used in stead of (131)I-transferrin as the tracer, or only PBS was injected without stem cell transplantation. The images were semi-quantitatively analyzed with region of interest (ROI) techniques, and the phosphor imaging on the spinal sections were performed.

RESULTSRadioligand-receptor binding assay showed 10 770 binding sites with high affinity (KD=0.982 nmol/L) for Fe saturated transferrin on each human mesenchymal cell. Visible accumulation of radioactivity at the cell transplantation sites was observed 16 h and 24 h after intrathecal injection of (131)I-transferrin tracer, but not in two control groups. ROI analysis showed that the difference between (131)I-transferrin and the control groups was statistically significant (P<0.05). Phosphor imaging further verified that it was the specific coupling of transferrin to the implanted cells that resulted in radioactivity accumulation at the transplantation sites.

CONCLUSIONSTransferrin receptor imaging is capable of in vivo tracing of the implanted stem cells, and has the potential for use in non-invasive monitoring for stem cell transplantation therapy after further technical improvements.

Animals ; Autoradiography ; Cell Survival ; Feasibility Studies ; Female ; Gene Expression Regulation ; Humans ; Iodine Radioisotopes ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Molecular Imaging ; methods ; Rabbits ; Receptors, Transferrin ; metabolism ; Spinal Cord ; diagnostic imaging ; metabolism ; Tomography, Emission-Computed, Single-Photon ; Transferrin ; chemistry ; metabolism

OBJECTIVEIn recent years, abnormal changes in the endocannabinoid system have been found in schizophrenia. The superior temporal gyrus (STG) is strongly implicated in the pathophysiology of schizophrenia, particularly with regards to auditory hallucinations. In this study, we investigated the binding density of cannabinoid CB1 receptors in the STG of schizophrenia patients compared to control subjects.

METHODSQuantitative autoradiography was used to investigate the binding densities of [(3)H]SR141716A (a selective antagonist) and [(3)H]CP-55940 (an agonist) to the CB1 receptors in the STG. Post-mortem brain tissue was obtained from the NSW Tissue Resource Centre (Australia).

RESULTSContrasting to previous findings in the alterations of CB1 receptor densities in the prefrontal, anterior and posterior cingulate cortex of schizophrenia, which were suggested to be associated to impairment of cognition function, no significant difference was found between the schizophrenia and control cases in both [(3)H]SR141716A and [(3)H]CP-55940 binding.

CONCLUSIONWe suggest that CB1 receptors in the STG are not involved in the pathology of schizophrenia and the auditory hallucination symptom of this disease.

Adult ; Aged ; Autoradiography ; Case-Control Studies ; Humans ; Middle Aged ; Receptor, Cannabinoid, CB1 ; agonists ; antagonists & inhibitors ; metabolism ; Reference Values ; Schizophrenia ; metabolism ; physiopathology ; Temporal Lobe ; metabolism

OBJECTIVETo investigate the approach of basic fibroblast growth factor(bFGF) entering inner ear, as well as its the protective mechanism to inner ear and nerve tissue in pathological situation.

METHODS125I-bFGF was injected into guinea pigs body via the lateral ventricle and muscle under physical situation as well as pathological situation. Then the per minute gamma-radioactive in blood, liver, thyroid gland, brain, cochlear and perilymph fluid was counted, and the distribution and metabolism of bFGF in the inner ear and autoradiography of the cochlea were also observed.

RESULTSGamma-radioactive cpm of blood and liver increased significantly, while it did not change in brain, cochlea and perilymph after 125I-bFGF intramuscular injections. Gamma-radioactive cpm in blood, liver, brain, perilymph and cochlea had increased and autoradiography granules was found in the cochlea in 30 min after 125I-bFGF injected into CSF. In brain, perilymph and cochlea, a maximal value of gamma-radioactive cpm was obtained between 2 h and 4 h, while that in 8 h decreased significantly. Autoradiography granules still were seen in 8 h. gamma-radioactive cpm in 12 h was still higher than that in control group, but autoradiography granules can't be seen. The result in 24 h was similar to that in control group. The time course of cpm in the blood, cochlea and perilymph always parallel changed.

CONCLUSIONSbFGF has some difficulties in getting across blood-labyrinth barrier (BLB) and blood-brain barrier (BBB) under physical and pathological situation, such as acute anoxia, aminoglycoside-induced deafness. bFGF can reach inner ear, perilymph fluid, brain tissue and blood rapidly when it is injected into CSF and excreted slowly in those tissues. Permeability of BBB and BLB to bFGF is similar and has orientation.

Animals ; Autoradiography ; Blood-Brain Barrier ; metabolism ; Ear, Inner ; metabolism ; Fibroblast Growth Factor 2 ; administration & dosage ; pharmacokinetics ; Guinea Pigs ; Iodine Radioisotopes ; administration & dosage