Pregnancy and delivery after intracytoplasmic injection of an immobilized, killed spermatozoon into an oocyte.
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Adult↗
Biomedical subjects
H Yazawa
Publications and source records attributed to H Yazawa.
Characterization of muscarinic cholinoceptor in primary culture of smooth muscle cells from human prostate.
We obtained a primary culture of prostatic cells by an explant method from patients with benign prostatic hypertrophy (BPH). Ultrastructural morphology and growth characteristics of these cells conformed to those reported for smooth muscle cells isolated from vascular and visceral tissue sources. The cells retained their original character including the presence of androgen receptor, acid phosphatase and normal chromosomal number. [3H]-methyl-quinuclidinyl benzilate (QNB) saturation experiments showed the existence of a homogeneous population of binding sites with a high affinity and low capacity (KD = 0.17 +/- 0.05 nM., Bmax = 15,000 sites per cell). Inhibition of [3H]-methyl-QNB binding by nonlabelled compounds showed these [3H]-methyl-QNB binding sites to be M2 muscarinic cholinoceptors. cAMP formation induced by forskolin and isoproterenol was inhibited by carbamoyl choline and oxotremorine. These results suggest that prostatic smooth muscle cells contain M2 muscarinic cholinoceptors and that these cholinoceptors couple adenylate cyclase inhibition.
Cells, Cultured↗
Pronuclear formation and cleavage of mammalian eggs after microsurgical injection of freeze-dried sperm nuclei.
A study was conducted to evaluate the ability of mammalian oocytes to develop to the pronuclear stage and beyond if injected with freeze-dried sperm nuclei. The rate of development of hamster eggs to the pronuclear stage after microinjection of freeze-dried hamster sperm nuclei was 90%. The pronuclear formation rate of hamster eggs injected with freeze-dried human sperm nuclei was 85%. On the other hand, the rates for eggs injected with untreated sperm nuclei were 84% and 89% respectively. The survival rate of rabbit eggs microinjected with freeze-dried rabbit sperm nuclei was 64%, the fertilisation rate 56% and the cleavage rate 38%. The survival, fertilisation and cleavage rates of eggs injected with non-freeze-dried sperm nuclei were 78%, 45% and 34%, respectively. There was no difference between eggs injected with freeze-dried sperm nuclei and those injected with sperm nuclei that had not been freeze-dried. Of the viable rabbit eggs injected with freeze-dried rabbit sperm nuclei, 6% developed to the 6- to 8-cell stage 62 h after injection.
Animals↗
Effect of thioltransferase on the cystamine-activated fructose 1,6-bisphosphatase by its redox regulation.
Fructose 1,6-bisphosphatase [EC. 3.1.3.11] is activated by the treatment with 0.1 mM cystamine up to about 400% compared to its original activity (dithiothreitol-reduced form). Thiol compounds (0.1 mM of cysteamine and dithiothreitol) can restore its activity effectively. Reduced glutathione, at 0.2 mM, also restores fructose 1,6-bisphosphatase activity only in the presence of cystamine. When excess cystamine is removed, the addition of 1.0 U/ml thioltransferase is able to restore FBPase activity very efficiently coexistence with 0.2 mM reduced glutathione though reduced glutathione alone does not work.
Animals↗
[The results of microinsemination by intracytoplasmic sperm injection].
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Calcimycin↗
The M3-muscarinic cholinoceptor subtype in rat prostate and its down regulation by aging.
Muscarinic cholinoceptor subtypes in the rat prostatic membrane were characterized by using [3H]-methyl-quinuclidinyl benzilate (QNB) in ligand binding studies. [3H]-Methyl-QNB saturation experiments showed the existence of a homogeneous population of binding sites with a high affinity (KD value) of 0.24 +/- 0.04 nM and a maximum binding site number (Bmax) of 219 +/- 65 fmol/mg protein. Inhibition of [3H]-methyl-QNB binding by nonlabelled compounds was in the following order of effectiveness in rat prostate: atropine > 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) > hexahydro-sila-difenidol hydrochloride, p-fluoroanalog (p-F-HHSiD) > pirenzepine > methoctramine > [11-((2-((dimethylamino)-methyl)-1-piperidinyl)acetyl)-5,11-dihydro-6H- pyrido(2,3-b)(1,4)benzodiazepine-6-one] (AF-DX 116). This ranking order was similar to that for the salivary gland (M3 subtype), but not for the brain (M1 subtype) or the heart (M2 subtype). These results indicate that the muscarinic cholinoceptors in the rat prostate belong mainly to the M3 subtype. Furthermore, Bmax values for muscarinic cholinoceptors in the aged rat prostate (approximately 1-year-old) were smaller than those in the young rat prostate (6- to 8-week-old) (87 +/- 13 vs. 183 +/- 32 fmol/mg protein). However, KD values for muscarinic cholinoceptors, and Bmax and KD values for beta-adrenoceptors showed no change. These results suggest that the number of prostatic muscarinic cholinoceptors decreases with aging.
Aging↗
Alpha 1-adrenoceptor subtype in the rat prostate is preferentially the alpha 1A type.
alpha 1-Adrenoceptors in the rat prostate were characterized by a binding assay using the newly synthesized radioligand [3H]-YM617 (5-[2-[[2[ethoxyring(n)-3H](o-ethoxyphenoxy)ethyl]amino]prop yl]-2-methoxybenzenesulfonamide HCl) and an in vitro assay. Specific [3H]-YM617 binding in the rat prostate was saturable and of high affinity (KD = 61.5 +/- 5.9 pM) with 23.2 +/- 6.9 fmol/mg of protein as the maximal number of binding sites (Bmax). alpha-Adrenoceptor agonists and antagonists inhibited the binding of the radioligand with the following order of effectiveness: YM617 > prazosin = bunazosin > WB4101 > 5-methylurapidil = phenoxybenzamine > phentolamine > S(+)-isomer of YM617 > yohimbine > norepinephrine > phenylephrine > methoxamine. alpha 1-Adrenoceptors in the rat prostate preferred the R(-)-isomer of YM617 to the S(+)-isomer. Preincubation with chlorethylclonidine (CEC; 10(-5) M, 10 min) just slightly changed the Bmax value for [3H]-YM617 without changing the KD value in the prostate; however, CEC reduced the Bmax in the aorta. In the isolated tissue, pretreatment with CEC (10(-5) M, 10 and 30 min) time-dependently shifted to the right the dose-response curve for phenylephrine and decreased the maximal contraction of aortas induced by phenylephrine, but did not shift or decrease those of prostates. The present results indicate that the alpha 1-adrenoceptors in the rat prostate are mainly CEC-insensitive (alpha 1A), whereas those in the aorta are CEC-sensitive (alpha 1B).
Adrenergic alpha-Agonists↗
Human aortic smooth muscle cells containing angiotensin II type 1 receptors.
We characterized the angiotensin II (AII) receptor in human aortic smooth muscle cells (HASMCs). This receptor binds [125I]Sar1,Ile8-angiotensin II with a high affinity of 0.20 +/- 0.04 nM and a low capacity of 5.3 +/- 0.4 fmol/mg protein (230 +/- 17 sites/cell). Based on the Ki values, the ranking order of [125I]Sar1,Ile8-AII binding inhibition was as follows: Sar1,Ile8-AII > AII > Dup 753 > AII > AI >> PD 123319. The addition of AII to HASMCs induced a rapid, transient increase in intracellular free Ca2+ concentration followed by a lower, sustained phase. When extracellular Ca2+ was removed by adding 3 mM EGTA, this initial transient increase was not changed, but the sustained phase was abolished. These results revealed AII receptors in HASMCs to be of the type 1 receptor subtype, which induce Ca2+ mobilization mainly from intracellular Ca2+ stores.
Angiotensin II↗
Characterization of the endothelin receptor in primary cultures of human aortic smooth muscle cells.
We characterized the endothelin receptor subtypes in primary cultures of human aortic smooth muscle cells (HASMCs) by binding studies. [125I]-Endothelin (ET)-1 saturation experiments showed the existence of a homogeneous population of binding sites with the high affinity (KD value) of 97 +/- 37 pM and maximum number of binding sites (Bmax) of 54 +/- 10 fmol/mg protein. However, almost no specific [125I]-ET-3 binding was observed. Inhibition of [125I]-ET-1 binding in the HASMCs membrane by nonlabeled compounds showed the following order of effectiveness: ET-1 = ET-2 = FR139317 >> ET-3. These results suggest that the endothelin receptor of HASMCs is of the ETA type. We also studied the effect of ET-1 on the cytosolic [Ca2+]i in HASMCs loaded with fura-2/AM. In 1.3 mM Ca2+, ET-1 produced a dose-dependent, biphasic increase in signal with a maximal effect at 10 nM. At this concentration, ET-1 produced a transient increase in [Ca2+]i that reached a peak at 1 min, which was followed by a slow but sustained increase in [Ca2+]i. This second phase was attenuated in Ca(2+)-deficient medium. Furthermore, ET-1 increased inositol 1,4,5-triphosphate in a time- and dose-dependent manner. These results suggest that the endothelin receptors of HASMCs are of the ETA type, which couple with Ca2+ channels.
Aorta↗
[Application of DNA fingerprinting to investigation of genetic relationships between laboratory rabbit strains].
It is well known that laboratory rabbits are not controlled genetically like laboratory mice and rats. In order to test the usefulness of DNA fingerprinting in investigation of genetic uniformity of the laboratory rabbits strains and their relationships, we applied DNA fingerprinting using bacteriophage M13 probe to five strains (2 inbreds (JWY-NIBS and DuY-NIBS) and 3 outbreds (JW-NIBS, Icl:JW and WHHL)). DNA fingerprints of 2 inbred strains showed the same banding patterns within each strain but the strain-specific patterns. Although there were no rabbits showing the same banding patterns in 3 outbred strains, average percent differences (APD) were 13.7 to 18.6. A dendrogram based on APD of DNA fingerprints was constructed by 2 large clusters, JW group and DuY. The dendrogram was essentially similar to that based on rabbit mandible measurements. These results suggest that DNA fingerprinting is available not only for the genetic monitoring of the laboratory rabbit strains but also for the investigation of their genetic relationships.
Animals↗
[Short intertrial interval as a phrasing cue in the reinforcement-pattern learning in rats].
The effect of short intertrial interval as a phrasing cue on the reinforcement-pattern learning was investigated in a rat experiment using a runway. In the acquisition session, 20 rats were trained for NNR sequence in which two nonreinforced trials were followed by a reinforced one with equal ITI of 30 min. In the sequence-addition session, a new sequence with consisting of three nonreinforced trials with 30-min ITI was added 30 min and 30 s after the original sequence of trials for Group L-ITI and for Group S-INT, respectively. Rats of Group S-INT responded on trials 4-6 with the similar pattern of running speed as they showed on Trials 1-3 from the second day. In Group L-ITI, the running speed on Trial 4 did not decrease. This suggests that Group S-INT rats learn to use the ITI of 30 s as a phrasing cue not on the first day but from the second day on. The phrasing effect of short ITI (30 s) was discussed in comparison with that of longer ITI (30 min).
Animals↗
[Effect of interrun interval on serial pattern learning in rats].
The effect of interrun interval (IRI) on serial pattern learning was investigated in a runway. One of two serial sequences, a monotonic decreasing sequence (14-7-3-1-0 food pellets) or a nonmonotonic sequence (14-1-3-7-0), was combined factorially with a short (30 s) or a long (30 min) IRI. Following 28 acquisition trials, a short (or long) IRI was transferred to the same sequence with a long (or short) IRI. It was found that anticipation of the O-pellet developed more rapidly in the group receiving a monotonic sequence with a 30 s IRI than that with a 30 min IRI in acquisition phase. The O-pellet was not anticipated by either of the two nonmonotonic groups. Anticipation was eliminated by the increase in IRI, whereas it was developed by the decrease in IRI in the monotonic sequence. These results suggest that time-related factors such as the decay of the memory of numbers of pellets are needed for Capaldi's memory discrimination theory.
Animals↗
Characterization of [3H]YM617, R-(-)-5-[2-[[2[ethoxyring(n)-3H](o-ethoxyphenoxy)ethyl]amino]- propyl]-2-methoxybenzenesulfonamide HCl, a potent and selective alpha-1 adrenoceptor radioligand.
The binding properties of a new radioligand, R(-)-5-[2-[[2[ethoxyring(n)-3H](o- ethoxyphenoxy)ethyl]amino]propyl]-2-methoxybenzenesulfonamide++ + HCl ([3H]YM617), were studied in membranes of the rat hippocampus and spleen. [3H]YM617 rapidly associated with its binding sites in both membranes and reached steady state by 20 min at 25 degrees C. The specific binding of [3H]YM617 appeared to be saturable, and Scatchard analysis revealed a linear plot, suggesting a single population of binding sites with a dissociation constant of 0.170 +/- 0.016 nM (n = 6) in the hippocampus and 0.195 +/- 0.036 (n = 4) in the spleen. The maximal binding sites in the hippocampus and spleen were 203.0 +/- 43.2 (n = 6) and 72.4 +/- 17.0 (n = 4) fmol/mg protein, respectively. Chlorethylclonidine (10(-5) M for 10 min) treatment reduced the Bmax values of [3H]YM617 and [3H]prazosin to a similar degree in the rat hippocampus (10-15%) and spleen (40-50%). Alpha adrenoceptor agonists and antagonists competed with [3H]YM617 for binding sites in the following order: YM617 > prazosin > WB4101 > bunazosin > 5-methylurapidil > S(+)-isomer of YM617 > phentolamine > yohimbine > norepinephrine = phenylephrine > methoxamine in the hippocampus, and prazosin > YM617 > bunazosin > WB4101 > 5-methylurapidil > phentolamine > S(+)-isomer of YM617 > yohimbine > norepinephrine > phenylephrine > methoxamine in the spleen. In the hippocampus, prazosin and bunazosin produced biphasic displacement of [3H]YM617, but not [3H]prazosin binding. In contrast, only monophasic curves were obtained against either radioligand in the spleen.(ABSTRACT TRUNCATED AT 250 WORDS)
Adrenergic alpha-Agonists↗
[A study of relation between phrasing and intertrial interval in reinforcement pattern learning in rats].
Effects of the temporal interval on reinforcement pattern learning were investigated in four experiments using a runway. The reinforced trial was always the first trial for R5N sequence (Experiments 1a and 1b), and the fourth trial for 3NR2N sequence (Experiments 2a and 2b). Group S-ITI received the given sequence at 30-s ITI, Group L-INT received the same procedure as Group S-ITI except for a 30-min ITI inserted between Trials 3 and 4 (Experiments 1a and 2a). Group L-ITI received at 30-min ITI, Group S-INT did as Group L-ITI except for the ITI between Trials 3 and 4 was 30 seconds (Experiments 1b and 2b). It was found that the running speed on each of Trials 1 and 4 was faster than any other trials under R5N and 3NR2N sequences for Group L-INT. That is, the running pattern for Trials 1-3 was similar to that for Trials 4-6. On the other hand, the running speed on Trial 4 under R5N sequence and that on Trial 1 under 3NR2N sequence did not increase for Group S-INT. These results suggest that a longer or shorter ITI plays the roles of both phrasing cue and discriminative stimulus. For Group S-ITI and Group L-ITI, there was little evidence that sequences were phrased. Therefore, when trials are separated by equal ITIs, neither 30-s ITI nor 30-min ITI becomes a phrasing cue.
Animals↗
[Interval effects of added sequences on reinforcement pattern learning in rats].
Three experiments examined how intervals of added sequences affected rat's learning of the reinforcement pattern. Animals were trained for runway performance corresponding to the series of one reinforcement trial (R) and two nonreinforcement trials (N) run with 30-s ITIs. The series was NNR, RNN, NRN for Experiments 1, 2, 3, respectively. Following this acquisition training, a second series of three nonreinforced trials with 30-s ITIs was added to the first series. Animals were assigned to two groups matched for performance level and they were given an added series with either long or short inter-session intervals. Subjects in Group S-ITI were given totally six trials with 30-s ITIs, while subjects in Group L-INT were given 30-min between the first and second series. Running speed for the first series differed with structure of the series (reinforcement pattern). The pattern of running speed for added series (Trials 4-6) was similar to that for original series (Trials 1-3) in Group L-INT, while running speed was kept at a low level for added series of nonreinforced trials in Group S-ITI. As is suggested by these findings, the events regularly occurred with equal ITIs can be remembered as one series, even when a new series of events is added to already experienced one with the same ITIs. However, when the second series is temporally separated from the first one by long intervals, the memory system may be reset so that events can be segregated as two sequences.
Animals↗
[Pathophysiological analysis of rapidly progressive periodontitis].
Pathophysiological features were studied on 7 patients with rapidly progressive periodontitis but without any evidence of systemic disease, to analyse the clinical pathogenesis. The patients consisted of 5 females, 2 males, between the ages of 32 and 42 years. All patients had severe and rapid alveolar bone destruction on the basis of radiographic measurement. Abnormal serum levels of IgG and IgM were detected in some patients. Higher IgG level was found in 4 patients and higher IgM level was found in 2 patients. The proportion of lymphocyte subsets was calculated in mononuclear cells from peripheral blood of patients. Higher OKT4/OKT8 ratio was found in all patients. The percentage of OKT4 positive cells in 2 patients was higher than that in normal subjects while the percentage of OKT8 positive cells in 4 patients was lower than that in the healthy controls. Microorganisms from periodontal pockets were examined in 5 patients. Bacteriodes was isolated in all 5 patients and Haemophilus actinomycetemcomitans in 2 patients.
Adult↗
Morphometrical observations on the mandible of five strains of rabbits and strain identification using mandible measurements.
In order to clarify the relationships between five strains of rabbits and to identify the strains, principal component and discriminant analyses were carried out using 12 mandibular measurements of three inbred strains (JW-NIBS/Y (JW/Y), NW-NIBS/Y (NW/Y) and Dutch-NIBS/Y (D/Y)) and two outbred strains (JW-NIBS (JW) and NW-NIBS (NW] which were maintained at the Nippon Institute for Biological Science. The results obtained were as follows. (1) Principal component analysis revealed that in the males the mandible of JW was the largest but with considerable variation. D/Y was the smallest of all strains examined. The mandibles of NW and NW/Y were similar to the JW mandibles but had a shape which was shorter and higher. In the females the mandible of NW was the largest of all strains and, as in the males, D/Y was the smallest and JW varied markedly. (2) Discriminant analysis showed the probability of erroneous discrimination to be 14.8% (34/229) when the inbred and outbred strains were combined. In both sexes erroneous discrimination mostly occurred between NW and NW/Y, which have the same origin, and between JW and NW, which have a common ancestor. However, when the inbreds and outbreds were identified separately by discriminant analysis, the probability of erroneous discrimination was low in both cases (4.5% (5/112) and 7.7% (9/117) respectively). These results indicate that strain differences are found in the size and shape of rabbit mandibles and that mandible analysis is effective for strain identification of laboratory rabbits.
Animals↗
Effects of glucocorticoid and cycloheximide on the activity and amount of RNA polymerase I in nuclei of rat liver.
The activity of the template-engaged form of RNA polymerase I from livers of adrenalectomized rats was about 50-60% of that of normal control rats, and increased about 2-fold at 6 h after the administration of dexamethasone. However, no change was found in the activity of the 'free' form of RNA polymerase I or the template-engaged form of RNA polymerase II. Immunochemical studies using guinea-pig anti-(RNA polymerase I) serum disclosed that the total number of RNA polymerase I molecules did not vary during the treatment with dexamethasone. Cycloheximide caused a rapid decrease in the template-engaged form of RNA polymerase I activity in normal rats and in dexamethasone-treated (6 h) adrenalectomized rats, to the value in adrenalectomized rats, but affected it only slightly in adrenalectomized rats. The elongation rate of rRNA-precursor synthesis in liver nuclei was not affected by a change in the concentration of circulating dexamethasone. From these results, it is concluded that about half the rRNA-precursor synthesis in rat liver is regulated by glucocorticoids, probably through the synthesis of short-lived protein(s) which may play a role in conversion of the 'dormant' form of RNA polymerase I into the 'engaged' form.
Adrenalectomy↗