A Comparative Study of Multiple: Acid Phosphatases in Cellular Fractions of Bean Hypocotyl

Six kinds of acid phosphatases were solubilized with TritonX-100 from the cell wall (W-I, W-II), mitochondrial (M-I, M-II,M-III) and microsomal (Ms) fractions of bean hypocotyl, andthey were partially purified by using Sephadex gel filtrationand DEAE-cellulose column chromatography. Acid phosphatasesfrom the soluble fraction were also fractionated into 12 isozymesby electrophoresis, and the properties of the isozymes werecompared. The soluble isozymes showed pH optima at 5·0,5·3 and 5·6; the isozymes possessed high affinityto p-nitrophenyl phosphate and ADP, and their mol. wts rangedfrom 30000 to 45000. Among the solubilized phosphatases, W-I,M-I and M-III showed maximum activity at pH 5·0 and theirmol. wts were between 50000 and 110000. W-I proved to have highaffinity to ATP and bis-p-nitrophenyl phosphate as M-I did top-nitrophenyl phosphate and M-III to phenyl phosphate. The characteristicsof these solubilized isozymes were different from those of thesoluble isozymes. On the contrary, W-II, M-II and Ms were quitesimilar to those of the solubles in pH optima, substrate specificity,K_m value, affinity to DEAE-cellulose and gel electrophoreticpatterns. These results suggest that W-II, M-II and Ms werederived from the soluble isozymes. Phaseolus vulgaris L., bean, hypocotyl, acid phosphatases, Michaelis constant     

Sex differences in the pharmacokinetics of pioglitazone in rats

Clinical studies have suggested that pioglitazone, an insulin sensitizer, has a stronger effect in women than in men. To determine the sex difference in the pharmacokinetics of pioglitazone, we examined the plasma and white adipose tissue levels of pioglitazone and its active metabolites (M-II, M-III and M-IV) in male and female rats treated with a single or repeated oral administration of pioglitazone (10 mg/kg). The AUCs of pioglitazone (149.6+/-22.6 vs. 103.3+/-14.0 microg.h/ml; P<0.01), M-III (31.4+/-8.1 vs. 20.2+/-4.7 microg.h/ml; P<0.05) and M-IV (41.9+/-15.5 vs. 14.1+/-1.6 microg.h/ml; P<0.01) were larger in female rats than in male rats, but the levels of M-II were similar. Any of the compounds did not accumulate in plasma after repeated administration. According to kinetic model analysis, the apparent elimination rate of pioglitazone and the formation rate of M-II were faster in male rats than in female rats. No significant sex difference was found in the tissue-to-plasma concentration ratios of pioglitazone or its active metabolites in white adipose tissue. These results suggest that there are sex differences in the plasma levels of pioglitazone and some of its active metabolites and that those differences are reflected in differences in white adipose tissue levels.     

Multiple forms of rat liver mitochondrial DNA polymerase.

Mitochondrial DNA polymerase (DNA polymerase mt) exists in two active forms. DNA polymerase present in crude extract (M-I) and ammonium sulfate precipitate (M-II) stages of purification sediments at 12.1S. The enzyme at the M-II stage of purification has a molecular weight of approximately 250,000 as determined by Sephadex G-200 chromatography in buffers of low ionic strength. In buffers containing 0.15 m NaCl, the enzyme sediments at 9.4S and has a molecular weight of approximately 190,000. When the enzyme is further purified on diethylaminoethyl cellulose (M-III stage of purification), the 9.4S activity predominates. Addition of a polymerase-free fraction from the M-III stage of purification changes the sedimentation coefficient of the enzyme from 9.4 to 12.1S.     

Isolation and identification of urinary metabolites of AF-2 (3-(5-nitro-2-furyl)-2-(2-furyl)acrylamide) in rabbits

After oral administration of AF-2 (3-(5-nitro-2-furyl)-2-(2-furyl)acrylamide) to rabbits, the two unique metabolites, M-I and M-II, were isolated from the urine. M-I, yellow needles of mp 117°, was identified as a new type metabolite of nitrofuran derivative, 2-(β-carboxypropionyl)-3-(5-methylthio-2-furyl) acrylamide by its mass, ir and nmr spectrometries. M-II, yellow solid, appears to be cis-trans isomer of M-I considering from its uv and mass spectral data, and the behavior on tlc.     

Carbon autonomy of reproductive shoots of Siberian alder (<Emphasis Type="Italic">Alnus hirsuta</Emphasis> var.<Emphasis Type="Italic"> sibirica</Emphasis>)

Carbon autonomy of current-year shoots in flowering, and of current-year shoots plus 1-year-old shoots (1-year-old shoot system) in fruiting of Siberian alder (Alnus hirsuta var. sibirica) was investigated using a stable isotope of carbon, ¹³C. The current-year shoot and 1-year-old shoot systems were fed ¹³CO₂ and the atom% excess of ¹³C in flowers and fruits was determined. The majority of photosynthate allocated to flower buds was originally assimilated in the leaves of the flowering current-year shoots. Of all the current-year shoots on fruiting 1-year-old shoots, only those nearest to the fruits allocated the assimilated photosynthate to fruit maturation. These results indicate that the current-year shoots and 1-year-old shoot systems are carbon-autonomous units for producing flowers and maturing fruits, respectively.     

Spindle dynamics in living mouse oocytes during meiotic maturation, ageing, cooling and overheating: a study by polarized light microscopy

A liquid crystal polarized light microscope (LC PolScope) was used to examine spindle dynamics in living mouse oocytes. Immature oocytes were cultured for 0-48 h and spindles were imaged with the PolScope at various time points of culture. Oocytes at metaphase I (M-I) and metaphase II (M-II) were also exposed to shifts of temperature from 25 to 41 degrees C to examine the effects of fluctuations of temperature on spindle dynamics. After examination with the PolScope, some oocytes were fixed and examined by immunocytochemical staining and confocal microscopy. After culturing for 6 h, 76% and 2% of the oocytes reached M-I and M-II stages and all oocytes had birefringent spindles. When the oocytes were cultured for 14-16 h, 88% and 6% of oocytes were at M-II and M-I stages respectively and all oocytes had birefringent spindles. However, when the oocytes were cultured for 22-48 h, the proportions of oocytes with birefringent spindles decreased as culture time was increased. Exposure of oocytes to 25 degrees C induced spindle disassembly within 10-20 min in both M-I and M-II oocytes. Most (93-100%) oocytes reassembled spindles after warming at 37 degrees C. Furthermore, exposure of oocytes at M-I stage but not at M-II stage, to 30 degrees C also induced significant microtubule disassembly. However, exposure of oocytes to 38-41 degrees C did not obviously change the quantity of microtubules in the spindles, which was measured by retardance. This study indicates that the PolScope can be used to examine spindle dynamics in living oocytes, and it has the advantage over the routine fluorescence microscope in that images can be obtained in the same individual oocyte and the quantity of microtubules can be measured by retardance in living oocytes. These results also indicate that the M-II spindle in mouse oocytes is sensitive to oocyte ageing and cooling, but not heating, and M-I spindle is more sensitive to temperature decline than M-II spindle.     

蛹虫草线粒体基因型快速检测体系的构建及线粒体遗传稳定性的初步研究

本研究的目的是建立一种快速确定蛹虫草菌株线粒体基因型的技术体系,并探讨蛹虫草连续传代培养后线粒体的遗传稳定性。从已知线粒体基因组的蛹虫草菌株中扩增线粒体内含子位点,将扩增产物混合并制作出两套DNA分子量标准,即在8个内含子位点分别具有内含子的8条扩增条带组成的M-I和在6个内含子位点分别缺失内含子的6条扩增条带组成的M-II。从待检测的蛹虫草菌株（包括3个已知和2个未知线粒体基因组的菌株）中扩增同样的（假定）内含子位点,然后通过琼脂糖凝胶电泳分别与制备好的两个DNA分子量标准进行比较,能够准确判断蛹虫草菌株的线粒体内含子分布模式,从而验证了所构建的线粒体基因型快速检测体系的有效性。选择10个蛹虫草组织分离菌株和8个单分生孢子菌株连续转接培养15代,没有发现线粒体内含子分布模式发生改变。本研究成功构建了快速检测蛹虫草线粒体基因型的技术体系,并发现蛹虫草线粒体具有很高的遗传稳定性,为开展蛹虫草线粒体遗传规律的研究奠定了基础。     

Quantitation of a new potent angiotensin II receptor antagonist,TCV-116, and its metabolites in human serum and urine

A sensitive high-performance liquid chromatographic (HPLC) method is described for the determination of a new potent antihypertensive agent, TCV-116, and its two metabolites (M-I and M-II) in human serum or urine. After pre-treatment of the specimens, the analytes were determined using a column switching technique, except for the metabolites in urine which were determined by gradient elution mode HPLC. The quantitation limits for TCV-116, M-I and M-II were all 0.5 ng/ml in serum, and 0.5, 10 and 10 ng/ml in urine, respectively. The methods were applied to clinical trials of TCV-116.     

Genotypic differences in branching pattern and fruiting habit in common walnut (Juglans regia L.)

Architectural analysis of 840 Slovenian walnut (Juglans regia L.) genotypes was performed to determine the most typical and frequent morphological types and to evaluate their vegetative and generative potential. Four branching and fruiting patterns (I-IV) were detected. A 3-year-old fruiting branch, consisting of a 3-year-old shoot plus corresponding 2-year-old and 1-year-old shoots, was used as a structural unit for quantitative analysis. In the intermediate fruit-bearing types with mesotonic and acrotonic branching pattern (types II and III), the total lengths of 3-, 2- and 1-year-old shoots were 385 and 380 cm, respectively, compared with 275 and 253 cm in the terminal and lateral-fruiting types (types I and IV). In type I, 1-year-old shoots had significantly fewer nodes than in other types. In addition, they had a thinner basal diameter than types III and IV, and their angles were the most erect (39 degrees ). Only 0.4 out of 3.6 1-year-old shoots were flowering with one mixed bud with 1.9 female flowers. In type IV, 2-year-old shoots had significantly more nodes and a larger basal diameter than other types. One-year-old shoots in type IV are thicker than those in other types. Ratios between the number of flowering and the total number of 1-year-old shoots were 0.7 in type IV, 0.6 in type III, 0.5 in type II and 0.1 in type I. On 1-year-old shoots in type IV, 1.7 mixed buds with a mean of three female inflorescences per bud were counted. Consequently, the generative potential is highest in type IV and lowest in type I. In types II and III, growth and the ability to bear fruits are more balanced.     

Intra-oocyte localization of MAD2 and its relationship with kinetochores, microtubules, and chromosomes in rat oocytes during meiosis

The present study was designed to investigate subcellular localization of MAD2 in rat oocytes during meiotic maturation and its relationship with kinetochores, chromosomes, and microtubules. Oocytes at germinal vesicle (GV), prometaphase I (ProM-I), metaphase I (M-I), anaphase I (A-I), telophase I (T-I), and metaphase II (M-II) were fixed and immunostained for MAD2, kinetochores, microtubules and chromosomes. The stained oocytes were examined by confocal microscopy. Some oocytes from GV to M-II stages were treated by a microtubule disassembly drug, nocodazole, or treated by a microtubule stabilizer, Taxol, before examination. Anti-MAD2 antibody was also injected into the oocytes at GV stage and the injected oocytes were cultured for 6 h for examination of chromosome alignment and spindle formation. It was found that MAD2 was at the kinetochores in the oocytes at GV and ProM-I stages. Once the oocytes reached M-I stage in which an intact spindle was formed and all chromosomes were aligned at the equator of the spindle, MAD2 disappeared. However, when oocytes from GV to M-II stages were treated by nocodazole, spindles were destroyed and MAD2 was observed in all treated oocytes. When nocodazole-treated oocytes at M-I and M-II stages were washed and cultured for spindle recovery, it was found that, once the relationship between microtubules and chromosomes was established, MAD2 disappeared in the oocytes even though some chromosomes were not aligned at the equator of the spindle. On the other hand, when oocytes were treated with Taxol, MAD2 localization was not changed and was the same as that in the control. However, immunoblotting of MAD2 indicated that MAD2 was present in the oocytes at all stages; nocodazole and Taxol treatment did not influence the quantity of MAD2 in the cytoplasm. Significantly higher proportions of anti-MAD2 antibody-injected oocytes proceeded to premature A-I stage and more oocytes had misaligned chromosomes in the spindles. The present study indicates that MAD2 is a spindle checkpoint protein in rat oocytes during meiosis. When the spindle was destroyed by nocodazole, MAD2 was reactivated in the oocytes to overlook the attachment between chromosomes and microtubules. However, in this case, MAD2 could not check unaligned chromosomes in the recovered spindles, suggesting that a normal chromosome alignment is maintained only in the oocytes without any microtubule damages during maturation.     

The heterogeneity of the polymeric intracellular hemoglobin of Glycera dibranchiata and the cDNA-derived amino acid sequence of one component

The erythrocytes of the marine polychaete Glycera dibranchiata contain a number of different, single-chain hemoglobins, some of which self-associate into a 'polymeric' fraction. An oligodeoxynucleotide probe was synthesized based on partial amino acid sequences determined by chemical methods, and used to screen a cDNA library constructed from the poly(A+)mRNA of Glycera erythrocytes (Simons, P.C. and Satterlee, J.D. (1989) Biochemistry 28, 8525-8530). The longest positive inserts found were sequenced using the dideoxy nucleotide chain termination method. One complete clone was obtained: clone 5A, 816 bases long, contained 59 bases of 5'-untranslated RNA, an open reading frame of 441 bases coding for 147 amino acids and a 3'-untranslated region of 316 bases. The derived amino acid sequence of Glycera globin P1 was in agreement with the partial amino acid sequences obtained by chemical methods. Three additional inserts obtained in the screening were also sequenced: the inferred amino acid sequences proved to be partial globin sequences which were different from each other and from the sequence of P1. Thus, the 'polymeric' fraction of the intracellular hemoglobin of Glycera probably consists of at least four different globin chains much like the 'monomeric' fraction. Comparison of the 'polymeric' sequence with the two known 'monomeric' sequences, M-II and M-IV, shows that they share 54 identical residues. At 74 positions, the identical residues in M-II and M-IV differ from the corresponding residue in P1, including at E-7, where P1 has a distal His, in contrast to Leu in M-II and M-IV. The alignment of Bashford et al. ((1987) J. Mol. Biol. 196, 199-216) and their templates were used to examine the principal differences between the two types of Glycera globin sequences. They appear to consist of uncommon surface amino acid residues at positions C6 (Phe vs. Ala), E10 (Val vs. Lys), E17 (Lys vs. Val), G1 (Arg vs. Lys), G10 (Met vs. Ala) and H5 (Arg vs. Lys). One or more of these residues could be responsible for the self-association exhibited by the 'polymeric' Glycera globins.     

Shoot preformation in clones of Fraxinus pennsylvanica in relation to site and year of bud formation

Summary Shoot preformation was investigated in buds of four clones of Fraxinus pennsylvanica var. subintegerrima (Vahl) Fern. at two sites in Manitoba in the second (1988) and third (1989) growing seasons after grafting. More preformed primordia were produced in terminal buds in 1989 compared to 1988 at each site. Both terminal and lateral buds at Morden contained significantly more primordia than those at Winnipeg. The numbers of preformed primordia were significantly different among clones. Clone 3 produced the most and clone 1 the fewest primordia in terminal buds. Despite quantitative variation, the pattern was similar among clones for terminal buds at each site and in each year. A similar pattern was evident for lateral buds at the Winnipeg site in 1989 but at Morden, clones 4 and 1 had the largest number of preformed primordia. Data from 1989 revealed that numbers of primordia were correlated with bud dimensions, parent shoot length, diameter and number of leaves, and location of the bud on the parent. Shoot dry weight was also related to these variables and revealed a non-linear increase in dry weight with shoot length. Multiple regression, with parent shoot length and location of buds along the parent axis as independent variables provided a reliable indicator of preformation in the crown. Although there is a genotypic component to preformation, variation between sites, years and crown locations suggests plasticity in bud development.     

Disbudding treatments on pistachio trees cv. mateur: dry matter accumulation and distribution within fruiting and non-fruiting branches under dry climate

Key message

After applying disbudding treatments, removal of fifty percent of flower bud each year improves dry matter accumulation in fruiting and non-fruiting branches of pistachio trees, which could minimize alternate bearing.

Abstract

Dry matter accumulation and distribution within branches of pistachio trees were investigated during 2005-2008 to determine the effects of fruiting on shoot growth under rain-fed conditions in arid climate. Four treatments were applied: T ₀ normal alternation cycle, T ₁ trees disbudded for 1 year, T ₂ trees disbudded for two successive years, and T ₃ removal of 50 % of all floral buds for each year. Consecutive disbudded treatment (T ₂) allowed a higher growth potential of pistachio trees with reference to normal biennial cycle (T ₀). Individual current shoot of T ₂ accumulated 44 % as much dry matter cm^?1 as those of ‘On’ trees, which have the highest yield. Removal of 50 % of floral buds (T ₃) significantly increased the dry matter accumulation in fruiting branch to reach 57 g in postharvest compared to 42.6 g for the control T ₀. Trees disbudded for 2 years (T ₂) had increased dry matter accumulation in the non-fruiting branch from 3.3 to 16.3 g. Leaves, current shoot, 1-year-old wood and inflorescence buds represented, respectively, 87, 5.3, 5 and 2.7 % of the total dry matter of individual branch of T ₂. In fruiting branches, nuts consisted of 83–87 %, leaves 7–10 %, rachises 4 %, 1-year-old wood 1.6–2 % and current shoot 0.8–1.3 % of the total dry matter. One-year-old wood played a major role as sources and sinks for developing current year shoot, leaves, inflorescence buds and nuts. Removal of 50 % of flower bud (T ₃) improves the dry matter accumulation in fruiting and non-fruiting branches. Thus, under rain-fed conditions, annual pruning could be used to minimize alternate bearing of pistachio.     

Double column-switching high-performance liquid chromatographic method for the determination of TAK-603 and its metabolites in human serum

A double column-switching high-performance liquid chromatographic (HPLC) method for the determination of concentrations for TAK-603 (T) and its metabolites, T-72258 (M-I) and T-72294 (M-III), in human serum was developed. The analytes were extracted with ethyl acetate from human serum samples treated with triethylamine and injected into the HPLC system. Separation of the analytes was performed on the HPLC system with double column-switching technique. The mobile phases A and B for the first column and the mobile phase C for the second column used were a mixture of methanol–10 mM aqueous ammonium acetate solution (1:1, v/v), methanol and a mixture of methanol–10 mM aqueous ammonium acetate solution (11:9, v/v), respectively. The eluate was monitored with a UV detector at a wavelength of 253 nm. The work-up procedure was reproducible and more than 90% of the analytes could be recovered from human serum. The lower limits of quantitation were all 1 ng/ml for the analytes when 0.5 ml of human serum was used. Standard curves were linear with a correlation coefficient (R) of more than 0.999 in the range of 1–500 ng/ml for T, M-I and M-III in human serum. The intra- and inter-day precision of the method for the various analytes were below 4.8%. The accuracy was good with the deviations between spiked and calculated concentrations of the analytes being within 11.0%. The method was successfully applied to analyze serum samples after an oral administration of T to healthy male volunteers.     

Compensatory growth in shoot populations of young white pine trees

Summary White pine (Pinus strobus L.) trees have shoot populations composed of subpopulations of terminal and lateral shoots. I tested whether the subpopulations would show compensatory (increased) growth when separated from each other. Ten-year-old white pine (Pinus strobus L.) trees growing under an oak (Quercus) overstory were untreated or treated in winter by removing either all terminal, or all lateral buds (10 trees per treatment). Growth was compared between control and treated shoot subpopulations. In the 1st year, shoot-length frequency distributions were similar between control and treated subpopulations. There was significant compensatory shoot elongation (mean of 1.5 cm per shoot) in both treated subpopulations. In the 2nd year each subpopulation produced both terminal and lateral shoots. Shoot-length frequency distributions were similar, but shifted toward longer shoots in treated populations. Shoot number, mean length and total shoot length were greater in treated populations. The increased growth in treated subpopulations was due both to differences in parent shoot length and to compensatory shoot production and elongation.     

Development to the blastocyst stage of immature pig oocytes arrested before the metaphase-II stage and fertilized in vitro

In vitro fertilization (IVF) and embryonic development of mature and meiotically arrested porcine oocytes were compared in the present study. After in vitro maturation (IVM) of cumulus-oocyte complexes for 48 h, 75.4% of them extruded a visible polar body (PB). Most of the oocytes with a first polar body (PB+ group) were at the metaphase-II (M-II) stage (91.4%). Most of the oocytes without a visible polar body (PB− group) appeared to be arrested at the germinal vesicle (GV) (41.6%) and metaphase-I (M-I) (34.0%) stages. After IVF of oocytes (day of IVF = Day 0), there was no difference between PB⁺ and PB⁻ groups in rates of sperm penetration, mono-spermy, however oocyte activation rate after penetration was greater in the PB+ than in the PB− group (P < 0.05). On Day 2, there was no difference between rates of embryos cleaved at the 2–4 cell stages in PB+ and PB− groups (42.1 ± 48.8% and 33.6 ± 2.1%, respectively). On Day 4, the rate of PB+ embryos developing beyond the 4-cell stage was greater than that of PB− embryos (P < 0.05, 31.7 ± 3.9% and 14.1 ± 1.5%, respectively), and PB+ embryos had more cells than the PB− embryos (P < 0.05, 8.3 ± 0.4 and 6.0 ± 0.8 cells, respectively). On Day 6, a greater proportion of PB+ embryos developed to the blastocyst stage than did PB− embryos (P < 0.05, 34.6 ± 2.4% and 20.7 ± 2.8%, respectively). However, when the GV oocytes of the PB− group were not included in recalculations, there was no difference in blastocyst rates between M-I arrested and M-II oocytes (35.3 and 34.6%, respectively). The number of blastomere nuclei in embryos obtained from the PB+ group (52.0 ± 2.5) was greater than that from the PB− group (P < 0.05, 29.1 ± 2.8). The proportion of degenerated parts in the blastocysts, as determined by morphological appearance, was the same in the PB+ and PB− groups. Although the quality of PB+ embryos was enhanced as compared with that of the PB− group, the proportion of inner cell mass and trophectoderm cells in PB+ and PB− blastocysts did not differ (1:1.9 and 1:2.2, respectively). Chromosome analysis revealed that PB+ blastocysts had more diploidy (P < 0.05, 69.7%) than did PB− blastocysts (44.0%), whereas PB− blastocysts had more triploid cells (P < 0.05, 34.0%) than did PB+ oocytes (8.4%). These results indicate that pig oocytes arrested before the M-II stage (M-I oocytes) undergo cytoplasmic maturation during maturation culture and have the same ability to develop to blastocysts after IVF as M-II oocytes, but some of them resulted in degeneration or delayed development with poor embryo quality.     

Effects of MULTIFOLIATE-PINNA, AFILA, TENDRIL-LESS and UNIFOLIATA genes on leafblade architecture in Pisum sativum

In order to dissect the genetic regulation of leafblade morphogenesis, 16 genotypes of pea, constructed by combining the wild-type and mutant alleles of MFP, AF, TL and UNI genes, were quantitatively phenotyped. The morphological features of the three domains of leafblades of four genotypes, unknown earlier, were described. All the genotypes were found to differ in leafblade morphology. It was evident that MFP and TL functions acted as repressor of pinna ramification, in the distal domain. These functions, with and without interaction with UNI, also repressed the ramification of proximal pinnae in the absence of AF function. The expression of MFP and TL required UNI function. AF function was found to control leafblade architecture multifariously. The earlier identified role of AF as a repressor of UNI in the proximal domain was confirmed. Negative control of AF on the UNI-dependent pinna ramification in the distal domain was revealed. It was found that AF establishes a boundary between proximal and distal domains and activates formation of leaflet pinnae in the proximal domain.     

Localization of mitotic arrest deficient 1 (MAD1) in mouse oocytes during the first meiosis and its functions as a spindle checkpoint protein

The present study was designed to investigate the localization of mitotic arrest deficient 1 (MAD1) in mouse oocytes during meiotic maturation and its relationship with kinetochores, chromosomes, and microtubules. Oocytes at various stages during the first meiosis were fixed and immunostained for MAD1, kinetochores, microtubules, and chromosomes. The stained oocytes were examined by confocal microscopy. Some oocytes were treated with nocodazole or Taxol before examination. The anti-MAD1 antibody was injected into the oocytes at the germinal vesicle (GV) stage for examination of chromosome alignment and spindle formation. It was found that MAD1 was present in the oocytes from the GV to prometaphase I stages around the nuclei. When the oocytes reached the metaphase I (M-I) to metaphase II (M-II) stages, MAD1 was mainly localized at the spindle poles. However, MAD1 relocated to the vicinity of the chromosomes when spindles were disassembled by nocodazole or cooling, and the relocated MAD1 moved back to the spindle poles during spindle recovery. Taxol treatment did not affect the MAD1 localization. Although anti-MAD1 antibody injection did not affect nuclear maturation, significantly higher proportions of injected oocytes had misaligned chromosomes when the oocytes reached the M-I to M-II stages. The results of the present study indicate that MAD1 is present in mouse oocytes at all stages during the first meiosis and that it participates in spindle checkpoint during meiosis. However, MAD1 could not check misaligned chromosomes during spindle recovery after the spindles were destroyed by drug or cooling, which caused some chromosomes to scatter in the oocytes.     

Allosomal and Autosomal Control of Sex Ratio in Phh and Phl Mice

When male mice from inbred PHH (sex ratio .535) and PHL (.435) are mated to females of various inbred lines, the sex ratio follows the male parent. The sex ratios from litters sired by reciprocal cross F1 males (letting A represent a set of autosomes) are 0.510 from AH/AL, XL/YH and 0.469 from AH/AL, XH/YL. The difference is statistically significant but only half the difference between pure strains. The paternal effect, presumably due to the Y, persists in progeny of the two kinds of F2 males. In backcrosses to the female parent, resulting finally in AH/AH, XH/YL and AL/AL, XL/YH, and in outcrosses, the effect of the Y chromosome does not persist, indicating that neither the Y alone, nor the autosomes alone, will cause the sex ratio to depart significantly from equality of sexes. When pairs of males in all possible combinations were presented with C57BL/6 females mating success gave the following ranking: AH-YL, AL-YL, AH-YH, AL-YH. The combination of autosomes from PHH and Y-chromosome from PHL seems to confer the greatest competitive advantage.