In vitro development of one-cell embryos from outbred mice: Influence of culture medium composition

Summary One-cell embryos from outbred mice (CF1, CD-1, and Dub:ICR) were cultured in various modifications of egg culture medium (ECM). The best development was observed in medium in which inorganic salts of modified T6 medium (mT6) replaced those of ECM. In this modification (TE), 66% of one-cell CF1 embryos developed into blastocysts, comared to 46 and 43% for ECM and mT6, respectively. Moreover, the cell numbers of blastocysts developing in TE (74.9±3.3) were higher than the cell numbers of those developing in ECM (55.1±2.4). The culture requirements of embryos varied between different stocks of mice: Fewer CF1 embryos developed to the blastocyst stage than either Dub:ICR embryos (90%) or CD-1 embryos (84%). Lowering the osmolarity of the medium from 300 to 280 mOsm, increasing the concentration of KC1 from 1.42 to 25 mM, or omitting lactate from the medium during Day 1 of culture did not further improve development of embryos, in contrast to previous reports. However, the time at which embryos were transferred to outgrowth medium influenced their postblastocyst development. The best development was observed when embryos were transferred on Day 4 of culture at the late morula-early blastocyst stage. This work was supported by the Office of Health and Environmental Research, U.S. Department of Energy, Washington, DC, contract DE-AC03-76-SF01012.     

杉木雌配子体与后胚发育过程中核酸、蛋白质和类脂的消长

在杉木胚胎分化期至成熟期,每个雌配子体总核酸和DNA,RNA含量在初期增加,后期则随着胚的分化发育逐渐下降,而蛋白质和类脂则一直上升。每胚总核酸、DNA,RNA则相应增加,而蛋白质、类脂的含量和干重亦逐渐增加。在胚分化早期RNA的迅速合成与细胞的分化及器官形成有关。但以胚干重为单位的DNA、RNA含量却随着胚的发育而有所减少;蛋白质含量先增加,至成熟后才下降。授粉前的胚珠,以及雌配子体、胚中都发现有凝集素存在。     

Functional morphology of genitalia of four species of julidan millipedes (Diplopoda: Nemasomatidae; Julidae)

The functional anatomy of the genitalia of Nemasoma varicorne (Nemasomatidae), Brachyiulus lusitanus, Unciger foehdus and Cylindroiulus boleti (Julidae) was investigated by shock freezing of animals in copula followed by serial semithin sectioning of the engaged genitalia. The species show conspicuous differences in the functional mechanism of their genitalia. In M varicorne and B. lusitanus the vulvae remain within the vulval sac during copulation while in U.foetidus and C. boleti parts of the gonopods (promerite and mesomerite) form clasper-like structures that pull out the vulvae from the vulval sacs. With the exception of C. boleti all investigated species have a 'central funnel' on the vulva which leads into the receptaculum seminis. The sperm-transferring part of the male gonopods (solenomerite) is introduced into this funnel during copulation. In B. lusitanus and C. boleti a projection of the posterior gonopods (end-projection, brachite) fits into a slit anterior to the openings of the receptacula. The results are discussed with regard to sexual selection theory and a hypothesis is proposed that explains the evolutionary change of millipede genitalia by a combination of female choice and sperm competition phenomena.     

Spectral sensitivity of photoreceptors mediating phase-shifts of circadian rhythms in retinally degenerate CBA/J (rd/rd) and normal CBA/N (+/+) mice

Light-dark cycles are the most important time cue for the circadian system to entrain the endogenous circadian clock to the environmental 24 h cycle. Although photic entrainment of circadian rhythms is mediated by the eye in mammals, photoreceptors implicated in circadian photoreception remain unknown. In our previous study, retinally degenerate CBA/J (rd/rd) mice were found to have lower circadian photo-sensitivity for phase-shifting the locomotor activity rhythms than normal CBA/N(+/+) mice. In the present study, the spectral sensitivity for phase-shifting the rhythms was examined in order to characterize the photopigments involved in circadian photoreception of these mice. The spectral sensitivity of CBA/J-rd/rd mice clearly fitted to the Dartnall nomogram for a retinal₁-based pigment with a maximum at 480 nm, while the best fitted nomogram had a maximum at 500 nm in CBA/N- +/+ mice. These results suggest that circadian photopigments involved in CBA/J-rd/rd and CBA/N- +/+ mice may be different.     

正常小鼠高频心电图时域值和功率谱的研究

本文用南京新博公司生产的ＮＨＥ－１０００型心电高频信息检测分析仪研究了正常小鼠（昆明种）高频心电图（ＨＦ－ＥＣＧ）的时域值和ＱＲＳ波群的功率谱。主要结果如下（以正导为例,－Ｘ±ＳＤ）：心率６０３±８８次／ｍｉｎ（ｎ＝７４）;Ｐ－Ｒ间期相对较长。为３４．９±４．７ｍｓ（ｎ＝５８）,占心动周期的３４．９±４．９％,这与人类有很大的不同;ＱＲＳ波宽９．２±１．２ｍｓ,占心动周期的９．２±１．４％（ｎ＝７４）,这一结果与以前的文献报道相差较大。Ｔ波宽１０．３±３．２ｍｓ,占心动周期的１０．３±３．２％;Ｑ－Ｔ间期１９．４±３．２ｍｓ,占心动周期的１９．５±３．６％;ＱＲＳ波群峰－峰值（Ｖｐ－ｐ）为１．４５６±０．４８０ｍＶ;Ｔ波高０．３３６±０．１１５ｍＶ;７３只动物Ⅱ导联高频切迹总数只有３个,扭挫２６个。Ⅱ导联ＱＲＳ波群的功率谱特点：０—８０Ｈｚ的相对能量为４５．４８±１５．３２％;８０—２００Ｈｚ为４３．９７±９．９５％;２００—３００Ｈｚ为８．８９±７．３８％;３００—１０００Ｈｚ为１．６６±２．７４％。     

Utilization of Iron Sources and Its Possible Roles in the Pathogenesis of Vibrio parahaemolyticus

Vibrio parahaemolyticus is an important enteropathogen in Japan, Taiwan and other coastal regions. The influence of the regulation of iron on the pathogenesis of this pathogen has not been well characterized. The growth of pathogenic and non-pathogenic strains of V. parahaemolyticus on iron-limited agar plates was stimulated by ferritin, lactoferrin and transferrin at 30 μM , and also by hemin, hemoglobin and ferric ammonium citrate at 100 μM . Spontaneous iron-utilizing mutant strains (mutants) were derived from a clinical strain, ST550. Compared with the parent strain, lowered virulence was demonstrated for these mutants, as assayed by adult mouse and suckling mouse models. The in vivo growth and enterotoxigenicity of these mutants were also lower in the suckling mice. Adherence of the mutants to excised mouse intestine was lower as demonstrated by scanning electron microscopy. The iron-regulated outer membrane protein profile also changed in selected mutants. These results indicate that iron-regulated outer membrane proteins and other unknown factors associated with iron utilization may have profound influences, besides iron acquisition, on the pathogenesis of V. parahaemolyticus.     

The 21-aminosteroid U-74389F increases the number of glial fibrillary acidic protein-expressing astrocytes in the spinal cord of control and wobbler mice

Summary 1. Wobbler mice suffer an autosomal recessive mutation producing severe motoneuron degeneration and dense astrogliosis, with increased levels of glial fibrillary acidic protein (GFAP) in the spinal cord and brain stem. They have been considered animal models of amyotrophic lateral sclerosis and infantile spinal muscular atrophy. 2. Using Wobbler mice and normal littermates, we investigated the effects of the membrane-active steroid Lazaroid U-74389F on the number of GFAP-expressing astrocytes and glucocorticoid receptors (GR). Lazaroids are inhibitors of oxygen radical-induced lipid peroxidation, and proved beneficial in cases of CNS injury and ischemia. 3. Four days after pellet implantation of U-74389F into Wobbler mice, hyperplasia and hypertophy of GFAP-expressing astrocytes were apparent in the spinal cord ventral and dorsal horn, areas showing already intense astrogliosis in untreated Wobbler mice. In control mice, U-74389F also produced astrocyte hyperplasia and hypertophy in the dorsal horn and hyperplasia in the ventral-lateral funiculi of the cord. 4. Givenin vivo U-74389F did not change GR in spinal cord of Wobbler or control mice, in line with the concept that it is active in membranes but does not bind to GR. Besides, U-74390F did not compete for [³H]dexamethasone binding when addedin vitro. 5. The results suggest that stimulation of proliferation and size of GFAP-expressing astrocytes by U-74389F may be a novel mechanism of action of this compound. The Wobbler mouse may be a valuable animal model for further pharmacological testing of glucocorticoid and nonglucocorticoid steroids in neurodegenerative diseases.     

细小病毒非结构蛋白转基因小鼠模型建立

将小鼠乳腺癌病毒启动子控制的细小病毒非结构蛋白基因(长5.7kb)氯化铯超速离心,纯化透析后用显微注射法导入C57BL/SJL F_1小鼠受精卵雄核,植入假孕母鼠输卵管,得成活小鼠15只。抽取鼠尾DNA,对其中10只小鼠作PCR和southern blot鉴定,其中4只(40％)整合有目的基因。对首建者B_6()的8只子代小鼠鉴定,3只(37.5％)整合有目的基因。说明导入的目的基因能传代。     

Female wing spreading as acceptance signal in theDrosophila virilis group of species

Females of manyDrosophila species spread apart their wings prior to copulation. In the present study we found female wing spreading to provoke male copulation attempts inDrosophila virilis-group species, helping the males to attempt copulation when the female is ready to mate. The males of most species, however, rarely responded to female wing spreading by copulation attempt without licking the female genitalia before and/or after female wing spreading bout. Blocking the female genitalia (D. virilis, D. novamexicana) reduces males' tendency to attempt copulation after female wing spreading. In these, and most other species of the group, female wing spreading seems to be an efficient signal only when combined with stimuli from female genitalia.     

The effect of variation in parameters of the male calling song of the katydid,Amblycorypha parvipennis (Orthoptera: Tettigoniidae), on female phonotaxis and phonoresponse

After being placed equidistant from playbacks of two computer-generated male calls, females of the katydidAmblycorypha parvipennis preferentially moved toward call phrases that were louder, longer, and leading (initiated without being overlapped by other call phrase endings). Because earlier tests with calling males had indicated that male mating success was related to these call parameters, we suggest that mating success is partly a result of female choice. Females also preferentially responded to the initial, rather than the latter, half of male phrases. Results of other tests suggested that females were responding to increased phonatome rate characteristic of the first half of the phrase. Males may compete to lead in order to prevent jamming of initial phrase information. Females also preferentially phonoresponded (ticked) in response to longer phrases. In earlier studies of male mating success and female phonotaxis using live males, male weight, sound level, and leading were intercorrelated; however, none of these parameters were correlated with phrase length. We suggest that females may respond to different call parameters under different environmental conditions.