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2012年  第33卷  第2期

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研究论文
The present study revisited the controversial taxonomic status of Petaurista yunanensis, P. philippensis, P.hainana, and P. petaurista by using a considerably extended set of morphometrical characters (26 cranial variables from 60 adult specimen skulls). The results revealed no sexual dimorphism in any of the four species but confirmed significant craniometric differences among the four species in both the principal components analysis (PCA) and discriminant function analysis (DFA), with the greatest distinction observed between P. petaurista and other Petaurista species. Both univariate and multivariate analysis indicated that the morphological differences between P. yunanensis and P.philippensis were less than that between P. philippensis and P. hainana. The morphometric results were concordant in geographic patterns with mtDNA data from previous studies and indicated that P. petaurista, P. hainana, P. philippensis,and P. yunanensis could be recognized as valid species.
Phrynocephalus przewalskii is one kind of lizards inhabiting inclusively in the desert that has controversial viewpoints on its phylogeny. Based on mitochondrial ND2 gene of 119 samples from 12 geographic populations, we analyzed the effects of environmental factors on the variation of genetic diversity, as well as its relationship to P. versicolor. The results showed that these populations clustered into three major lineages, with P.versicolor embedded within one lineage. The twelve populations had great genetic diversity variation, which was tightly linked with local altitude, annual precipitation, and variation of annual precipitation. High latitudes, increased annual precipitation and great variation in annual precipitations may all have resulted in the decrease of genetic diversity. It thus assumed that altitude can change the genetic diversity of different geographic populations of P. przewalskii resulting from the effects of different local annual precipitation.
The complete mitochondrial genome of the Parathyma sulpitia (Lepidoptera, Nymphalidae, Limenitidinae) was determined. The entire mitochondrial DNA (mtDNA) molecule was 15 268 bp in size. Its gene content and organization were the same as those of other lepidopteran species, except for the presence of the 121 bp long intergenic spacer between trnS1(AGN) and trnE. The 13 protein-coding genes (PCGs) started with the typical ATN codon, with the exception of the cox1 gene that used CGA as its initial codon. In addition, all protein-coding genes terminated at the common stop codon TAA, except the nad4 gene which used a single T as its terminating codon. All 22 tRNA genes possessed the typical clover leaf secondary structure except for trnS1(AGN), which had a simple loop with the absence of the DHU stem. Excluding the A+T-rich region, the mtDNA genome of P. sulpitia harbored 11 intergenic spacers, the longest of which was 121 bp long with the highest A+T content (100%), located between trnS1(AGN) and trnE. As in other lepidopteran species, there was an 18-bp poly-T stretch at the 3’-end of the A+T-rich region, and there were a few short microsatellite-like repeat regions without conspicuous macro-repeats in the A+T-rich region. The phylogenetic analyses of the published complete mt genomes from nine Nymphalidae species were conducted using the concatenated sequences of 13 PCGs with maximum likelihood and Bayesian inference methods. The results indicated that Limenitidinae was a sister to the Heliconiinae among the main Nymphalidae lineages in this study, strongly supporting the results of previous molecular data, while contradicting speculations based on morphological characters.
Human trefoil factor 2 (hTFF2) is considered as one of the most important initiators of mucosal healing in the gastrointestinal tract by promoting cell migration and suppressing apoptosis. However, it is hard to obtain hTFF2 from human tissue and many recombinant hTFF2 produced in vitro exist as fusion proteins. The purpose of the present study was to produce native hTFF2 while maintaining its biological activities. The open reading frame of hTFF2 was inserted into a pET-32a(+) expression vector, and hTFF2-TRX fusion protein was successfully expressed in Escherichia coli and purified by Nickel-nitrilotriacetic acid affinity chromatography and reverse-phase HPLC steps. The recombinant fusion protein (purity>95%) was cleaved by Factor Xa at 23 ℃ to release hTFF2. After removal of Factor Xa and undigested fusion proteins, hTFF2 was purified and identified by SDS-PAGE and Western blotting. The yield of recombinant hTFF2 was about 5 mg/L. The recombinant hTFF2 could promote IEC-6 cells migration and in vitro wound healing via the activation of ERK1/2. Recombinant hTFF2 could also inhibit apoptosis of HCT-116 cells induced by 50 μmol/L ceramide.In summary, our results showed that the recombinant hTFF2 was expressed in E. coli and successfully purified after cleavage with the fusion partner with high yield while maintaining its biological activities. Recombinant hTFF2 might be useful for investigating the molecular mechanism of hTFF2 and development of hTFF2-related drugs.
C9, a component of the membrane attack complex, participates in the final stage of the complement cascade which lyses foreign organisms by disrupting the integrity of their cell membranes. In the present study, a full-length ayu C9 (aC9) cDNA was cloned which contains 2,125 nucleotides and encodes a protein of 592 amino acids.A signal peptide was deposited in the N-terminal 22 residues. The deduced amino acid sequence of aC9 showed 56.8% identity to the C9 of rainbow trout, and 40.9% to 53.8% identity to the C9 of other teleosts. RT-PCR analysis demonstrated that the mRNA of aC9 was expressed in the liver, spleen, intestine, gill and muscle of healthy ayu fish with the highest level in the liver. Quantitative RT-PCR analysis showed that aC9 transcripts were significantly up-regulated in the liver at 4 h post Listonella anguillarum infection, peaked at 16 h post injection. Western blotting analysis revealed that serum aC9 significantly increased in Listonella anguillarum infected ayu fish. Our results suggested that aC9 may play an important role in fish immune response of anti-bacteria.
Tbx3, a member of the TBX2 subfamily of T-box gene family, encodes a transcription factor with a highly conserved DNA-binding domain, which called T-domain. Tbx3 is involved in morphogenesis and organogenesis in vertebrates, such as limb development, heart remodeling, and neural placode differentiation. In the present study, a full-length 2 908 bp Tbx3 cDNA from Acipenser baerii (AbTbx3) was obtained using RT-PCR and RACE technique,which includes a 2 166 bp complete open reading frame encoding a putative peptide of 721 amino acids. AbTbx3 shares 73.5% identity with its human homolog. Particularly, the DNA-binding domain of AbTbx3 shared 95.2% identity with human Tbx3. Phylogenetic analysis revealed that AbTbx3 was grouped with Tbx3s in other vertebrates, which were clustered with Tbx2s and separated from Tbx4/5s. The predicted secondary and three-dimensional structures of the T-domain of AbTbx3 were remarkably similar to human Tbx3. Through semi-quantity RT-PCR, the expression of AbTbx3 was first detected at blastula stage during Siberian sturgeon embryonic development, increased gradually, reached its peak at early tail-bud stage and then decreased slightly. In adult sturgeon, AbTbx3 was strongly expressed in eye, brain, gill,intestines, pectoral fin and pelvic fin, but not in liver, blood, heart, kidney and muscle. The whole mount in situ hybridization showed that AbTbx3 was mainly expressed in the otic vesicle, hindbrain, dorsal notochord, pineal organ and dorsal fin bud in the larvae of stage 37 and 43.
The olfactory system may play a pivotal role in drug addiction. To clarify the issues, we investigated the morphine dependence and psychological craving in morphine addicted mice using the conditioned place preference (CPP)paradigm by taking an only odor cue as the conditioned stimulus (CS). The results showed that by pairing morphine with odor, the CPP could be induced in mice. When the morphine addicted mice were exposed to a novel environment during morphine withdrawal, they spent significantly longer time in the chamber with morphine-paired odor than in the control chamber. The effects of odor cue on the morphine CPP were blocked by the administration of dopamine D1 or D2 antagonists. The studies indicated that olfactory system plays an important role in drug addiction.
研究报告
Habitat selection of four sympatric pheasants (Syrmaticus ellioti, Lophura nycthemera, Pucrasia macrolopha and Bumbusicola thoracica) was studied in Guanshan National Nature Reserve, Jiangxi, China from October 2009 to November 2010. We measured seventeen ecological factors across 388 sites used by the four pheasants by direct observation in six transects. The results show that the pheasants preferred broad-leaved forest and coniferous forest and a sunny or semi-shaded slope. There were differences in elevation, degree of slope, arbor coverage, shrub coverage, shrub quantity, shrub height, herbage cover, herbage species, herbage quantity, leaf litter coverage, distance to water and arbor species between the habitat selected by the four pheasants. Canonical scores indicated that the four pheasants differed in habitat selection to some extent. A stepwise discriminant analysis revealed that elevation, herbage quantity, shrub height,distance to water, shrub cover, arbor cover, degree of slope, leaf litter cover, shrub quantity and herbage cover were the critical factors distinguishing habitat selection between the species. Predicted classification accuracy of the model was 74.7%.
The morphology and diet characteristics of three Myotis species roosting in the same cave were studied in Anlong County, Guizhou from September to November 2005. The three mouse-eared bats were the Chinese water myotis (Myotis laniger) “body mass: (4.46±0.53) g, forearm: (34.63±1.45 mm)”, fringed long-footed myotis (Myotis fimbriatus) “body mass: (5.15±1.76) g, forearm: (35.20±1.07) mm” and szechwan myotis (Myotis altarium) “body mass: (10.94±0.87) g, forearm: (45.21±1.15) mm”. There were significant differences in the body masses of the three species. The forearm length of M. altarium was significantly longer than the other two species. M. laniger preyed mostly on dipsters and their larvae (79.7% in volume and 100% in frequency, Diptera). M. fimbriatus preyed mostly on dipsters and small beetles (59.6% and 91.3%, Diptera; 28.8% and 80.1%, Coleoptera). The prey of M. altarium was mostly ground-dwelling beetles (80.8% and 100%, Carabidae and Silphidae, Coleoptera). These dietary examinations indicate that the three species are highly adapted to different foraging habitats. Our work suggests that the spatial differences in foraging niches and trophic resource partitioning represent the major mechanism behind the levels of co-existence seen in this particular bat community.
In addition to pollinator fig wasps, there are several non-pollinating fig wasps associated with monoecious Ficus sp. In order to understand how pollinator fig wasps and non-pollinating fig wasps are distributed across the same syconium, the spatial distribution of fig wasps associated with Ficus altissima and F. benjamina were compared using the pedicle lengths of galls containing each species. The results indicate that in Ficus altissima, the average pedicel length of galls containing Eupristina sp. is longer than that containing E. altissima. Average pedicel length of galls containing Sycobia sp., Micranisa ralianga and Sycoscapter sp. two did not show significant difference. The range of pedicel lengths of galls containing Sycobia sp., M. ralianga or Sycoscapter sp. two is narrower than that of galls containing E. altissima,indicating these non-pollinating fig wasps and pollinator have partially separated spatial niches. In F. benjamina, E.koningsbergeri was distributed in galls from the outer layer to inner layer, while most Walkerella sp. were found in outer layer galls, indicating E. koningsbergeri and Walkerella sp. have partially separated spatial niches.
A new spider species, Sudharmia tridenticula sp. nov., is described from primary evergreen forests of Sumatra, Indonesia.
The complete mitochondrial genome of Apatura ilia (GenBank accession no. JF437925) was determined as a circular DNA molecule of 15 242 bp, with common genes of 13 putative proteins, 2 rRNAs, and 22 tRNAs and of the same gene arrangement as in other sequenced lepidopterans. All protein-coding genes had the typical start codon ATN,except for the COI’s using CGA as its start codon as previously demonstrated in other lepidopteran species. The comparison of the nucleotide sequences of the A. ilia mitogenome with ten other Nymphalidae species showed nearly identical gene orientation and arrangement, with only a few alterations in non-coding fragments. The nucleotide composition and codon frequency all fell into the range estimated for the order Lepidoptera. The A. ilia mitochondrial genome had the canonical set of 22 tRNA genes folded in the typical cloverleaf structure, with an unique exception of tRNASer(AGN). The mitochondrial genes from A. ilia were overlapped in a total of 33 bp at 9 locations, as well as interleaved with a total of 155 bp intergenic spacers, spread over 12 regions with the size ranging from 1 to 49 bp.Furthermore, the spacer between ND6 and Cyt b harbored a microsatellite-like repeat (TA)23 not found in other completely sequenced nymphalid genomes. The 403 bp AT-rich region harbored two conserved motifs (ATAGA,ATTTA), a 21 bp polyT stretch, a 10 bp poly-A region, along with two microsatellite-like repeats ((TA)10and (TA)7), as detected in other nymphalid butterflies
Firstly, RAPD was conducted to analyze genetic diversity of Trachidermus fasciatus in the Fuchun River population (FR), Yellow River population (YR), Luan River population (LR), and Yalu River population (YL), with 32 polymorphic 10-bp random primers selected from 294 ones. Thirty wild individuals were detected in each population.The results indicated that the genetic diversity of T. fasciatus was relatively rich. The major results were as the following:1) Altogether, 591 bands were detected and 515 of them were polymorphic, accounted for 87.14%. The range of proportion of polymorphic loci (P) was: FR(89.17%)>YR(87.99%)>YL(86.63%)>LR(83.25%). 2) The Shannon’s information index(IT) and Nei’s genetic diversity(HT) among populations were 0.3393 0.3566 and 0.2157 0.2279,respectively. Compare to other three populations, LR population had relative lower values. If took the populations as a whole, the total Nei’s genetic diversity(HT) and Shannon’s information index(IT) was 0.2336±0.1643 and 0.3710±0.2153,respectively. 3) The value of gene flow (Nm) (5.76103 19.84497) were high, indicating certain gene exchange existed among the four populations. But the AMOVA results exhibited significantly differentiation (P<0.05 or P<0.01) among the populations. 4) In the UPGMA tree constructed according to genetic distance, YL and YR populations clustered firstly, then with FR population, and finally they joined to LR population. Obviously, the YL, YR and FR populations had relatively close relationship according to their geographic distance, whereas LR population showed clear divergence to the other three populations. Secondly, out of the five special RAPD bands (S1225525bp, S1225605bp, S1225841bp, S1345695bp and S1345825bp), SCAR maker SCAR01560bp and SCAR02443bpwere successfully transformed from S1225605bp and S1225841bp,respectively. After large samples examination of the two markers, we found the highest frequency (96.67% and 93.33%)in the YL population, higher frequency (83.33% and 90%) in the FR population, high frequency (56.67% and 66.67%) in the YR population, and the lowest frequency (13.33% and 20 %) in the LR population. Therefore, SCAR01560bpand SCAR02443bpcan be used as special molecular markers for the population identification between LR and other three populations.
The strep avidin-biotin-peroxidase complex (SABC) immunohistochemical methods were applied to investigate the localization and semi-quantitative distribution of IFN-γ, IL-1α, NGF-β and TNF-α-immunoreactive cells in the cerebellar cortex of Western Anhui white goose at embryonic day 13, 19, 24, 28 (E13, E19, E24, E28) and postnatal day 7, 15 (P7, P15). The possible roles of IFN-γ、IL-1α、NGF-β and TNF-α in the development of cerebellar cortex were discussed. The results indicated that in the external granular layer, there were IFN-γ and TNF-αpositive cells at E13, E19, E24, E28, P7, IL-1α positive cells at E13, E19, E24, E28 and NGF-β positive cells at E13,E19, E24. The expression levels of these four cytokines all reached peaks at E19 of the six tested periods in this study. In the Purkinje cell layer, there were IFN-γ, IL-1α and TNF-α positive cells at E13, E19, E24, E28, P7, P15 and NGF-βpositive cells at E13, E19, E24, E28, P7. In the internal granular layer, there were IFN-γ positive cells at E13, E19, E24,E28, P7, P15, IL-1α and TNF-α positive cells at E13, E19, E24, E28, P7 and NGF-β positive cells at E13, E19, E24, E28.These results showed that E19 might be the “critical stage” in the cerebellar cortex development of Western Anhui white goose. IFN-γ, IL-1α and TNF-α might be synthesized by cerebellar cortex itself, and NGF-β could be transported from regions which project to Purkinje cells. IFN-γ may interfer the transfer of granular cells, and NGF-β may have neurotrophic functions that are beneficial to the growth and development of Purkinje cells.
Psychophysical studies suggest that lateral extrastriate visual cortical areas in cats may mediate the sparing of vision largely by network reorganization following lesions of early visual cortical areas. To date, however, there is little direct physiological evidence to support this hypothesis. Using in vivo single-unit recording techniques, we examined the response of neurons in areas 19, 21, and 20 to different types of visual stimulation in cats with or without acute bilateral lesions in areas 17 and 18. Our results showed that, relative to the controls, acute lesions inactivated the response of 99.3% of neurons to moving gratings and 93% of neurons to flickering square stimuli in areas 19, 21, and 20. These results indicated that acute lesions of primary visual areas in adult cats may impair most visual abilities. Sparing of vision in cats with neonatal lesions in early visual cortical areas may result largely from a postoperative reorganization of visual pathways from subcortical nucleus to extrastriate visual cortical areas.
Glaucoma is a typical irreversible blind neurodegenerative disease for which there is no effective treatment for halting visual deterioration. The recent development of neural stem cells studies sheds light on a potential resolution for this disease. As a result, an appropriate glaucoma modeling method for stem cell transplantation study is needed. In the present study, Dexamethasone was injected unilaterally into the conjunctiva of New Zealand rabbit at the dose of 2.5 mg (5 mg/mL), three times a week. After eight weeks, the eye ground photography showed that the optic nerve head of the treated eye was expanded, and the blood vessel was geniculate compared to the control eye, while the ocular media remained transparent. The hematoxylin-eosin (HE) stain of the retinal nerve fiber layer (RNFL) sections showed optic neuron death in the treated eye. The Heidelberg Retina Tomography (HRT) results showed optic disk morphological changes consistent with the pathophysiology of glaucoma in the treated eye, including a decrease in the rim area (1.10?0.88) mm2 and mean RNFL thickness (0.44?0.31) mm, and an increase in the cup/disk ratio 0.17?0.13.Then neural stem cells were injected into the vitreous body of the treated eye. After five months, surviving transplanted cells were observed. These results suggest a simple and reproducible chronic glaucoma model, which is appropriate for neural stem cell transplant research, has been successfully developed.
We investigated the contribution of locomotor activity and learning and memory abilities to Alzheimer’s disease induced by metal neurotoxicity. The metal neurotoxicity model was established by using locomotor activity tests and activity-avoidance conditioning paradigms, and observing changes in zebrafish behaviors after exposure to 50 μg/L of AlCl3 with different pH values (pH 7.8, pH 6.8 and pH 5.8) for 24 hours and 96 hours. The results showed Alzheimer’s disease-like behavior in locomotor activity tests and activity-avoidance conditioning paradigms by zebrafish receiving AlCl3in pH 5.8 significantly decreased compared to the control group and groups receiving AlCl3in pH 7.8 and pH 6.8 for 96 hours and 24 hours. These results provide evidences that exposure to AlCl3in an acid environment can influence locomotor activity and learning and memory abilities in zebrafish. In addition, exposure time and concentration of Al induced neurotoxicity and damaged the brain area of memory, resulting in Alzheimer’s disease-like behavior.
In this study, the 5 d and 7 d training paradigms were adopted to investigate the influences of different training procedures on the performance of spatial learning and memory of the hippocampal injured rats. The results showed that during the hidden-platform acquisition training, similar spatial learning dysfunctions were indicated in those two training paradigms. Whereas, when the spatial memories have been evaluated, compared with the 5 d training groups,the rats under 7 d training procedure not only crossed the platform location less, but also preferred to spend less time in the target quadrant.