Increase in the number of detectable preoptic glutamic acid decarboxylase 67-immunoreactive cells in immature male rats

Gonadotropin-releasing hormone (GnRH) neurons are mainly located in the anterior preoptic area (aPOA) and gamma-aminobutyric acid (GABA) is known as a potent regulator of the GnRH neurons. To examine the development of the GABAergic system in the aPOA, immunocytochemistry of glutamic acid decarboxylase 67 (GAD(67)) was performed in immature (postnatal d16, d25 and d30) and mature (postnatal 10 weeks) male rats. All immunocytochemical procedures were simultaneously performed. In the lateral part of the aPOA, the detectable number of GAD(67)-immunoreactive cells was small in the d16 group, but significantly increased in the d25, d30 and mature groups, up to 2.7, 4.8 and 5.7 times the number in the d16 group, respectively. In the diagonal band of Broca (DBB), the number was also small in the d16 group, and significantly increased in the d25, d30 and mature groups upto 1.8, 2.2 and 2.8 times the number in the d16 group, respectively. However, in the cingulate cortex, no significant developmental change was observed. These results suggest that the development of the GABAergic system in the lateral aPOA and the DBB occurs before sexual maturation of male rats.     

Effects of estrogen and progesterone on the expression of connexin-36 mRNA in the suprachiasmatic nucleus of female rats

In rat, helium pressures induce locomotor and motor activity which requires dopaminergic and N-methyl-D-aspartate (NMDA) receptor activities at striatal level. However, biochemical studies have suggested that pressure exposure may increase striatal glutamate level. We used microdialysis technique to study the effects of pressure on glutamate level in the striatum and the effects of local administration of D1 (SCH23390) or D2 (sulpiride) on these changes. Pressures increase both glutamate and glutamine levels in striatal microdialysates. Administration of sulpiride (1 microM) or SCH23390 (1 microM) by reverse microdialysis did not affect significantly pressure induced glutamate increase. So, protective effects of D1 and D2 antagonists against locomotor and motor hyperactivity (LMA) are probably independent of the processes involved in the striatal glutamate increase evoked by pressure.     

N-Glycosylation contributes to the limited cross-reactivity between hemagglutinin neuraminidase proteins of human parainfluenza virus type 4A and 4B

cDNAs encoding human parainfluenza virus type 4B (hPIV-4B) hemagglutinin neuraminidase (HN) protein were cloned and the nucleotide sequences were determined. A high degree of identity (81.4%) was observed between the nucleotide sequences of hPIV-4A and -4B HN proteins, and an 87.3% identity was found between the deduced amino acid sequences. This degree of identity is considered to be greater than immunological similarity between hPIV-4A and -4B HN proteins determined using monoclonal antibodies. To elucidate the causes of the antigenic difference between HN proteins of hPIV-4A and -4B, we constructed three cDNAs of hPIV-4B HN whose potential N-glycosylation sites were partially or completely the same as in hPIV-4A HN cDNA. We compared the antigenicity of the expressed wild-type and mutant proteins, and found that the antigenicities of the mutant hPIV-4B HN proteins were more similar to the hPIV-4A HN protein than to the non-mutant hPIV-4B HN protein. This study indicated that the antigenic diversity between hPIV-4A and -4B was partly caused by deletion or creation of glycosylation sites, showing that the point mutations resulting in deletion or creation of glycosylation sites is one of the initial steps leading to the division of virus into subtypes. Received: 21 January 2000     

Quantitative analysis for soluble elastin in circulation and cell culture fluids using monoclonal antibody-based sandwich immunoassay

We have newly established 3 distinct murine monoclonal antibodies (MoAbs) against human soluble elastin by using chemically denatured immunogen isolated from human aorta; they are designated as HASG-2, HASG-30, and HASG-61-1. All of these MoAbs were highly reactive with soluble forms of native elastin in normal human serum. HASG-2 and HASG-61-1 MoAbs can recognize soluble bovine elastin as well as human antigen, but HASG-30 cannot. The sandwich enzyme-linked immunosorbent assay (ELISA) for human soluble elastin was developed with HASG-61-1 labeled with peroxidase and HASG-30 immobilized on the microplates. The circulating levels of soluble elastin in human healthy subjects (mean +/- SD; 42.9 +/- 19.9ng/mL; n = 85) could be measured with full accuracy and reproducibility, and gradually increased with aging. The positive correlation between the levels and ages was statistically significant (r = 0.581, p < 0.0001). In addition, we could also determine the concentration of tropoelastin secreted from cultured human dermal fibroblasts accurately by this ELISA. This simple assay can be utilized for the routine clinical laboratory screening of patients with arteriosclerotic vascular diseases or to accurately determine the concentrations of tropoelastin secreted from cultured human cells.     

Mechanomyography of the human quadriceps muscle during incremental cycle ergometry

The mechanical activity of the human quadriceps muscle during maximal incremental cycle ergometry was investigated by mechanomyography (MMG). MMG and surface electromyography (EMG) recordings of vastus lateralis muscle activity were obtained from nine males. Cycle ergometry was performed at 60?rev/min and work load was incremented step wise by 20?W (3.2?Nm) every minute until volitional fatigue. The mean amplitudes of MMG (mMMG) and EMG (mEMG) during the contraction phase were calculated from the last six contractions in each load. The duration, load and work rate of exercise at exhaustion were 13.3 (1.6)?min, 44.1 (5.5)?Nm, 276.7 (34.7)?W, respectively. A linear relationship between mMMG and load was evident in each subject (r?=?0.868–0.995), while mEMG seemed to dissociate as the load became greater. In the grouped mean data, mMMG was linearly related to load whether aligned to the absolute (r?=?0.995) or maximal (r?=?0.995) load. Involvement of the noise component was further investigated by studying passive cycling by four subjects. Pedals were rotated passively for the first half of each stage (PAS) and the subject then pushed the pedals for the second half (ACT). In the lighter load region, the mMMG of ACT was as small as that of PAS. However, the change in the mMMG of PAS was very small compared with that of ACT. In conclusion, this study demonstrates a linear relationship between the mMMG of the quadriceps muscle and work load during maximal incremental cycle ergometry. The effect of movement noise was thought to be small and stable.     

A novel presenilin 1 mutation (Y154N) in a patient with early onset Alzheimer's disease with spastic paraparesis

Early onset familial Alzheimer's disease with spastic paraparesis (FAD-SP) has been associated with mutations of the presenilin 1 gene (PSEN1). We report a pedigree of FAD-SP due to a novel missense mutation of PSEN1 (Y154N). The symptoms of the proband were characterized by presenile dementia in her 40s, preceded by spastic paraparesis in her 30s, whereas the mother of the proband presented with spastic paraparesis in her 40s, followed by symptoms of dementia in her mid 60s. The mutation was found only in the proband, and not in a normal family member, normal Japanese control subjects, patients with sporadic Alzheimer's disease or patients with familial spastic paraparesis without dementia. Thus, Y154N is a novel PSEN1 mutation responsible for FAD-SP of Japanese origin.     

Scanning and transmission electron microscopic observations of the acute morphological response of the mouse urinary bladder to 4-ethylsulfonylnaphthalene-1-sulfonamide

A total of 75 BALB/cStCrlfC3H/Nctr male weanling mice were administered either 0 or 250 ppm of 4 ethylsulfonylnaphthalene-1-sulfonamide (ENS) in the diet for periods up to 14 days to evaluate the early morphological changes of the transitional epithelium of the urinary bladder with scanning (SEM) and transmission (TEM) electron microscopy. Primary TEM changes included hyperplasia of the epithelium, loosening of the intercellular junctions, autophagic vacuoles and electron dense granules in the mitochondria. Primary SEM changes included sloughing of epithelial cells, irregularity in the size and shape of the transitional epithelial cells and the presence of microvilli. Although pleomorphic microvilli were present after only three days of treatment with ENS, it appears that they are a transient observation in a series of morphological changes. The reversibility or transient nature of the pleomorphic microvilli may indicate that they are an acute toxic response and may not necessarily indicate a preneoplastic change.     

Effects of gap junction blocker on vasopressin and vasoactive intestinal polypeptide rhythms in the rat suprachiasmatic nucleus in vitro

We examined effects of gap junction blockers, octanol and halothane, on circadian rhythms in the release of arginine-vasopressin (AVP) and vasoactive intestinal polypeptide (VIP) in suprachiasmatic nucleus (SCN) slice cultures of the rat. Circadian rhythms in AVP and VIP release maintained when the SCN culture was treated with octanol for 42 h. However, the release of AVP and VIP showed no circadian rhythms after 7 days incubation with octanol or halothane. Circadian rhythmicity in the two peptide rhythms appeared after the removal of the drug from the culture medium. These findings suggested that the gap junction communication may be involved in intercellular coupling within each subpopulation of AVP or VIP neurons in the SCN.     

Detection of skeletal muscle fatigue using an accelerometer in dynamic cardiomyoplasty

 An animal experiment was done using six mongrel dogs that weighed 28 ± 3 kg to show that an accelerometer could detect skeletal muscle fatigue in dynamic cardiomyoplasty. Through left-side thoracotomy, the heart was exposed and an electrode to sense the heartbeat was positioned on the left ventricle. A left latissimus dorsi muscle flap (LDMF) was inserted into the left chest cavity and rolled around the heart. An accelerometer was put on the rolled LDMF to sense the ventricular acceleration by contraction of the LDMF and the heart. The LDMF was stimulated under these settings: pulse width, 210 μs; stimulation output, 6 V; burst frequency, 30 Hz; burst duration, 200 ms; synchronous ratio, 1 : 4; and synchronous delay, 66 ms. Output voltage from the accelerometer was recorded 1, 3, 5, 10, and 15 min after the beginning of stimulation. Percentages of the amplitude in all dogs after 3, 5, 10, and 15 min were 81 ± 10%, 63 ± 12%, 48 ± 11%, and 45 ± 14% of the values after 1 min, respectively. Significant differences were found between the values after 1 min and those after 3 min, between the values after 3 min and those after 5 min, and between the values after 5 min and those after 10 min. This study suggests that muscle fatigue is detectable with an accelerometer in actual dynamic cardiomyoplasty. Received: May 11, 2001 / Accepted: September 10, 2002 Acknowledgments This work was financially supported in a part by a Grant in Aid for Scientific Research (05671113) from the Ministry of Education, Science, and Culture of Japan. Correspondence to:H. Kuroda     

Intracerebroventricular injection of arginine-vasopressin V1 receptor antagonist attenuates the surge of luteinizing hormone and prolactin secretion in proestrous rats

Metamorphosis in the fly (Musca domestica) involves extensive lysis, neurogenesis and reorganization of neural tissue. Despite this, two experiments are reported which show aspects of larval memory persist into the adult form. Experimental larvae were reared in sawdust scented with aversive odours (mint or geraniol). Control larvae were reared on plain sawdust. In blind post-metamorphosis testing using a Y maze, adult flies showed a preference for the specific odour to which the larvae had been exposed. Control flies found both test odours aversive. A second experiment explores the possible role of cell survival through metamorphosis as a mechanism of the persistence of memory. Cells from trained donor flies when grafted into untrained recipients produced specific changes in recipient odour preference reflecting the donor larvae training.