The sexually dimorphic expression of glutamate transporters and their implication in pain after spinal cord injury

In addition to the loss of motor function,~60% of patients develop pain after spinal cord injury.The cellular-molecular mechanisms are not well understood,but the data suggests that plasticity within the rostral,epicenter,and caudal penumbra of the injury site initiates a cellularmolecular interplay that acts as a rewiring mechanism leading to central neuropathic pain.Sprouting can lead to the formation of new connections triggering abnormal sensory transmission.The excitatory glutamate transporters are responsible for the reuptake of extracellular glutamate which makes them a critical target to prevent neuronal hyperexcitability and excitotoxicity.Our previous studies showed a sexually dimorphic therapeutic window for spinal cord injury after treatment with the selective estrogen receptor modulator tamoxifen.In this study,we investigated the anti-allodynic effects of tamoxifen in male and female rats with spinal cord injury.We hypothesized that tamoxifen exerts anti-allodynic effects by increasing the expression of glutamate transporters,leading to reduced hyperexcitability of the secondary neuron or by decreasing aberrant sprouting.Male and female rats received a moderate contusion to the thoracic spinal cord followed by subcutaneous slow-release treatment of tamoxifen or matrix pellets as a control(placebo).We used von Frey monofilaments and the“up-down method”to evaluate mechanical allodynia.Tamoxifen treatment decreased allodynia only in female rats with spinal cord injury revealing a sexdependent effect.The expression profile of glutamatergic transporters(excitatory amino acid transporter 1/glutamate aspartate transporter and excitatory amino acid transporter 2/glutamate transporter-1)revealed a sexual dimorphism in the rostral,epicenter,and caudal areas of the spinal cord with a pattern of expression primarily on astrocytes.Female rodents showed a significantly higher level of excitatory amino acid transporter-1 expression while male rodents showed increased excitatory amino acid transporter-2 expression compared with female rodents.Analyses of peptidergic(calcitonin gene-related peptide-α)and non-peptidergic(isolectin B4)fibers outgrowth in the dorsal horn after spinal cord injury showed an increased calcitonin gene-related peptide-α/isolectin B4 ratio in comparison with sham,suggesting increased receptive fields in the dorsal horn.Although the behavioral assay shows decreased allodynia in tamoxifen-treated female rats,this was not associated with overexpression of glutamate transporters or alterations in the dorsal horn laminae fibers at 28 days post-injury.Our findings provide new evidence of the sexually dimorphic expression of glutamate transporters in the spinal cord.The dimorphic expression revealed in this study provides a therapeutic opportunity for treating chronic pain,an area with a critical need for treatment.

The Structure of Oryza Embryos and Their Dimorphic Cotyledons

It has been generally held in botany that Oryza sativa L. is a monocotyledon. Based on studies of rice embryo development we confirmed that rice embryo has two dimorphic cotyledons rather than just one cotyledon. In the present study we attempt to know if the morphology of embryos in other species of Oryza differs from O. sativa and if these embryos have dimorphic cotyledon. Two types of embryo structures were observed in 22 species and/or subspecies of genus Oryza under the scanning electron microscope. Type 1, the O.sativa type, which is characterized by ventral scale and lateral scales, was found in 16 species. Type 2, the O. meyeriana (Zoll. et Mor. ex Steud.) Baill. ssp. tuberculata W. C. Wu et Y. G. Lu, G. C. Wang type, with no ventral scale and lateral scales, was found in 6 species and subspecies. The embryogenic process of O.sativa and O.meyeriana sub. tuberculata showed that the scutellum primordium, coleorhiza primordium, coleoptile primordium and shoot apical meristem directly differentiate from proembryo. The former two later develop into the embryo envelope, which is the outside cotyledon; the coleoptile primordium develops into the coleoptile with the shape of inverted empty cone surrounding and covering the growth cone, which is the apical cotyledon. Both types of rice embryos have dimorphic cotyledons. The structural difference between them is that the scutellum primordium of the young embryo in type 2 does not differentiate ventral scale and lateral scales while the embryo of type 1 does. The dimorphic cotyledons of embryo of Oryza plants originate from the dorsiventrality of proembryo.

Selection of reference genes for RT-qPCR analysis of Phenacoccus solenopsis(Hemiptera: Pseudococcidae) sex-dimorphic development

Mealybugs, such as Phenacoccus solenopsis, are highly sexually dimorphic. Winged adult males present such remarkable morphological differences from females that, to the untrained eye, conspecific adults of both sexes of P. solenopsis may be considered as two different insect species. A method to investigate sex-dimorphic mechanisms is by evaluating gene expression using RT-qPCR. However, the accuracy and consistency of this technique depend on the reference gene(s) selected. In this study, we analyzed the expression of 10 candidate reference genes in male and female P. solenopsis at different development stages, using common algorithms including the ?Ct method, NormFinder, geNorm, BestKeeper, and a web-based analysis tool, RefFinder. The results showed that EF1-β, RP-L32 and RP-18 S were selected as the most stable genes by both the ?Ct method and NormFinder; TUB-α was the most stable gene identified by BestKeeper; and RP-L40 and RP-L32 were the most stable genes ranked by geNorm. RefFinder, a comprehensive analysis software, ranked the ten genes and determined EF1-β and RP-L32 as the most suitable reference genes for the various developmental stages in male and female P. solenopsis. Furthermore, the two most suitable reference genes were validated by examining expression of the juvenile hormone acid O-methytransferase(JHAMT) gene. Results of the validation portion of the study showed that JHAMT expression was sex-biased towards males and exhibited a dynamic and classic expression pattern among the P. solenopsis developmental stages. The results can help further our knowledge on the molecular mechanisms underlying sexual dimorphic development in P. solenopsis.

Effect of salinity on seed germination, growth and ion content in dimorphic seeds of Salicornia europaea L. （Chenopodiaceae）

The halophyte Salicornia europaea L. is a widely distributed salt-tolerant plant species that produces numerous dimorphic seeds. We studied germination and recovery in dimorphic seeds of Central Asian S. europaea under various salinity conditions. We also tested the effects of various salts on Na＋ and K＋ accumulation during plant development from germination to anthesis under greenhouse conditions. We found good germination （close to control） of large seeds under NaCl between 0.5 and 2%, Na2SO4 and 2NaCl ＋ KCl ＋ CaCl between 0.5 and 3%, and 2Na2SO4 ＋ K2SO4 -- MgSO4 between 0.5 and 5%. For the small seeds, we found stimulating effects of chloride salts （both pure and mixed） under 0.5-1% concentrations, and sulfate salts under 0.5-3%. Both types of seeds showed high germination recovery potential. Salt tolerance limits of the two seed types during germination and at the later stages of development were very similar （4-5%）. During plant growth the optimal concentrations of mixed chloride and sulfate salts ranged from 0.5 to 2%. The mechanisms of salt tolerance in the two seed types of S. europaea appear to differ, but complement each other, improving overall adaptation of this species to high salinity.

Cellular Prion Protein and Sexual Dimorphic Areas in Rodents. Correlates with Alzheimer Disease

The cellular prion protein (PrPC) expression was analyzed by western-blot in the rat, in two different dimorphic brain areas such as the anterior hypothalamic and the preoptic areas. In both cases, the PrPC expression was increased in males, implying a sexual dimorphism for the PrPC protein. The study was also made in other two brain areas, frontal cortex and hyppocampus (a clearly dimorphic area);in this case, mice of different ages of both sexes were used. In both brain areas analyzed, although the PrPC expression was increased with age until the adult age (38 weeks), it was decreased in aged animals (56 weeks) in both sexes. The PrPC expression in mouse hippocampus was predominant in males in comparison to females. Moreover, the non-glycosylated band was increased with age and this increase was parallel with the increase observed for the glycolsylated band. The non-glycosylated band increases more in aged females. Altogether, these data suggest that PrP in rodents, in the brain areas analyzed, has a dimorphism role. As we discuss in the present study and in relation to previous studies of our group these data could be extrapolated to humans (specially in Alzhemer disease cases).

Osteoblast-restricted Disruption of the Growth Hormone Receptor in Mice Results in Sexually Dimorphic Skeletal Phenotypes

Growth hormone （GH） exerts profound anabolic actions during postnatal skeletal development, in part, through stimulating the production of insulin-like growth factor-1 （IGF-1） in liver and skeletal tissues. To examine the requirement for the GH receptor （GHR） in osteoblast function in bone, we used Cre-LoxP methods to disrupt the GHR from osteoblasts, both in vitro and in vivo. Disruption of GHR from primary calvarial osteoblasts in vitro abolished GH-induced signaling, as assessed by JAK2/STAT5 phosphorylation, and abrogated GH-induced proliferative and anti-apoptotic actions. Osteoblasts lacking GHR exhibited reduced IGF-l-induced Erk and Akt phosphorylation and attenuated IGF-1-induced proliferation and anti-apoptotic action. In addition, differentiation was modestly impaired in osteoblasts lacking GHR, as demonstrated by reduced alkaline phosphatase staining and calcium deposition. In order to determine the requirement for the GHR in bone in vivo, we generated mice lacking the GHR specifically in osteoblasts （△GHR）, which were born at the expected Mendelian frequency, had a normal life span and were of normal size. Three week-old, female AGHR mice had significantly reduced osteoblast numbers, consistent with the in vitro data. By six weeks of age however, female AGHR mice demonstrated a marked increase in osteoblasts, although mineralization was impaired; a phenotype similar to that observed previously in mice lacking IGF-1R specifically in osteoblasts. The most striking phenotype occurred in male mice however, where disruption of the GHR from osteoblasts resulted in a ＂feminization＂ of bone geometry in 16 week-old mice, as observed by faCT. These results demonstrate that the GHR is required for normal postnatal bone development in both sexes. GH appears to serve a primary function in modulating local IGF-1 action. However, the changes in bone geometry observed in male AGHR mice suggest that, in addition to facilitating IGF-1 action, GH may function to a greater extent than previously appreciated in establishing the sexual dimorphism of the skeleton.

Gestational heat stress alters skeletal muscle gene expression profiles and vascularity in fetal pigs in a sexually dimorphic manner

Background: There is evidence that sow heat stress(HS) during gestation affects fetal development with implications for impaired muscle growth. We have previously demonstrated that maternal HS during early to midgestation compromised muscle fibre hyperplasia in developing fetal pigs. Thus, we hypothesised these phenotypic changes are associated with a change in expression of genes regulating fetal skeletal muscle development and metabolism. To test this, at d 60 of gestation, RNA sequencing and immunohistochemistry were performed on fetal longissimus dorsi(LD) muscle biopsies collected from pregnant gilts that had experienced either thermoneutral control(CON, 20 ℃, n = 7 gilts, 18 LD samples) or controlled HS(cyclic 28 to 33 ℃, n = 8 gilts, 23 LD samples)conditions for 3 weeks.Results: A total of 282 genes were differentially expressed between the HS and CON groups in female LD muscles(false discovery rate(FDR) ≤ 0.05), whereas no differentially expressed genes were detected in male LD muscles between the two groups(FDR > 0.05). Gestational HS increased the expression of genes associated with transcription corepressor activity, adipogenesis cascades, negative regulation of angiogenesis and pro-inflammatory signalling in female LD muscles. Immunohistochemical analyses revealed a decreased muscle vascularity density in fetuses from HS group for both sexes compared to those from the CON group(P = 0.004).Conclusions: These results reveal gilt HS during early to mid-gestation altered gene expression profiles in fetal LD muscles in a sexually dimorphic manner. The molecular responses, including transcription and angiogenesis repressions and enhanced adipogenesis cascades, were exclusively observed in females. However, the associated reductions in muscle vascularity were observed independently of sexes. Collectively this may indicate female fetal pigs are more adaptive to gestational HS in terms of gene expression changes, and/or there may be sexually dimorphic differences with respect to the timing of muscle molecular responses to gestational HS.

Sex-dimorphic adverse drug reactions to immune suppressive agents in inflammatory bowel disease

AIM:To analyze sex differences in adverse drug reactions(ADR) to the immune suppressive medication in inflammatory bowel disease(IBD) patients.METHODS:All IBD patients attending the IBD outpatient clinic of a referral hospital were identified through the electronic diagnosis registration system.The electronic medical records of IBD patients were reviewed and the files of those patients who have used immune suppressive therapy for IBD,i.e.,thiopurines,methotrexate,cyclosporine,tacrolimus and anti-tumor necrosis factor agents(anti-TNF);infliximab(IFX),adalimumab(ADA) and/or certolizumab,were further analyzed.The reported ADR to immune suppressive drugs were noted.The general definition of ADR used in clinical practice comprised the occurrence of the ADR in the temporal relationship with its disappearance upon discontinuation of the medication.Patients for whom the required information on drug use and ADR was not available in the electronic medical record and patients with only one registered contact and no further followup at the outpatient clinic were excluded.The difference in the incidence and type of ADR between male and female IBD patients were analyzed statistically by χ 2 test.RESULTS:In total,1009 IBD patients were identified in the electronic diagnosis registration system.Out of these 1009 patients,843 patients were eligible for further analysis.There were 386 males(46%),mean age 42 years(range:16-87 years) with a mean duration of the disease of 14 years(range:0-54 years);578 patients with Crohn's disease,244 with ulcerative colitis and 21 with unclassified colitis.Seventy percent(586 pts) of patients used any kind of immune suppressive agents at a certain point of the disease course,the majority of the patients(546 pts,65%) used thiopurines,176 pts(21%) methotrexate,46 pts(5%) cyclosporine and one patient tacrolimus.One third(240 pts,28%) of patients were treated with anti-TNF,the majority of patients(227 pts,27%) used IFX,99(12%) used ADA and five patients certolizumab.There were no differences between male and female patients in the use of immune suppressive agents.With regards to ADR,no differences between males and females were observed in the incidence of ADR to thiopurines,methotrexate and cyclosporine.Among 77 pts who developed ADR to one or more anti-TNF agents,significantly more females(54 pts,39% of all anti-TNF treated women) than males(23 pts,23% of all antiTNF treated men) experienced ADR to an anti-TNF agent [P = 0.011;odds ratio(OR) 2.2,95%CI 1.2-3.8].The most frequent ADR to both anti-TNF agents,IFX and ADA,were allergic reactions(15% of all IFX users and 7% of all patients treated with ADA) and for both agents a significantly higher rate of allergic reactions in females compared with males was observed.As a result of ADR,36 patients(15% of all patients using anti-TNF) stopped the treatment,with significantly higher stopping rate among females(27 females,19% vs 9 males,9%,P = 0.024).CONCLUSION:Treatment with anti-TNF antibodies is accompanied by sexual dimorphic profile of ADR with female patients being more at risk for allergic reactions and subsequent discontinuation of the treatment.

Association between specific proteins and dimorphic growth of penicillium marneffei

Objective： To investigate the difference of proteome between yeast form and mould form of Penicillium mameffei, and to investigate the association of specific proteins expressed with biochemical properties, susceptibility of antifungal agent with dimorphic growth. Methods： Biochemistry identity plates were used to test the assimilation of carbohydrates and E-test strips were used to detect the minimum inhibitory concentration （MIC） of mould form and yeast form 16 P. mameffei. Surface enhanced laser desorption/ionization （SELDI） mass spectrometry with ProteinChip WCX2 was performed to compare the expressed proteins in yeast form and mould form. Protein profiles were read by PBSⅡ proteinchip reader and the proteome database was analyzed by proteincbip software 3.2.0. Results： Mould form assimilated lactose, melibiose significantly stronger （ P 〈0.01）, while yeast form assimilated sorbinose significantly stronger （ P 〈 0.05）. The mean MIC of fluconazole against mould form increased significantly （ P 〈 0.01 ） compared with yeast form. Seventy-five distinct proteins were found in yeast form and mould form of P. mameffei, in which proteins of 2900Da and 3151Da were specifically expressed in yeast form and other two proteins of 13151Da and 13285Da were specifically expressed in mould form （ P 〈 0.01 ）. Conclusion： The assimilation of carbohydrates and drug susceptibility of P. mameffei may change partly due to the morphogenetic conversion and different texture. Specific proteins may he involved in the regulation, the change of biochemical reaction and drug susceptibility during dimorpbic growth.

Contribution of Estrogen to Sex Dimorphic Expression of Cardiac Natriuretic Peptide and Nitric Oxide Synthase Systems in ANP Gene-Disrupted Mice

Background: Sex dimorphism in the prevalence, onset, development and progression of cardiovascular disease (CVD) is well recognized, but the mechanisms whereby sex hormones are believed to confer cardioprotection are still not fully understood. Objective: This study more closely delineates the effect of 17β-Estradiol (E2) on the expression and signaling of the cardiac NP and NOS systems, well-known cardioprotective modulators of the cardiac hypertrophy (CH) response, that both contribute to downstream production of cyclic guanosine 3’,5’-monophosphate (cGMP). Materials and Methods: Ovariectomized (OVX) female ANP+/+ and ANP-/- mice, 6 - 7 weeks old, were subjected to a five-week treatment with E2 (100 μg/100 μL/day) or vehicle (VEH). Left ventricle from these treatment groups, along with that from age-matched male ANP+/+ and ANP-/- mice was used to assess expression of these systems by real-time quantitative PCR (qPCR). Left ventricle tissue and plasma cGMP were measured by enzyme immunoassay to assess alterations in resultant downstream signaling. Results: NP system expression was unchanged across genotype, sex and E2 treatment. Sex-specific differences in NOS system expression were observed;female mice showed an increased expression of NOS system genes that were significantly elevated in all but one of the E2 treatment groups. Left ventricle tissue cGMP remained unchanged across genotype, sex and E2 treatment. Plasma cGMP levels were unchanged in ANP+/+ treatment groups. In ANP-/- treatment groups, plasma cGMP in the female OVX-E2 mice was significantly higher compared to male and female OVX-VEH mice. Conclusion: These findings demonstrate that in the absence of ANP, E2 upregulates cardiac NOS system expression to produce cGMP. This study confirms the importance of the cardiac NOS system in females;this particular system may be a promising future target for sex-specific treatments and therapies for CVD in women.

Sex-Dimorphic Kidneyy-Brain Connectivity Map of Mice

The kidneys are essential organs that help maintain homeostasis,and their function is regulated by the neural system.Despite the anatomical multi-synaptic connection between the central autonomic nuclei and the kidneys,it remains unclear whether there are any variations in neural connections between the nervous systems and the renal cortex and medulla in male and female mice.Here,we used the pseudorabies virus to map the central innervation network of the renal cortex and medulla in both sexes.The data revealed that specific brain regions displayed either a contralateral-bias or ipsilateral-bias pattern while kidney-innervating neurons distributed symmetrically in the midbrain and hindbrain.Sex differences were observed in the distribution of neurons connected to the left kidney,as well as those connected to the renal cortex and medulla.Our findings provide a comprehensive understanding of the brain-kidney network in both males and females and may help shed light on gender differences in kidney function and disease susceptibility in humans.

Molecular characterization, tissue and developmental expression profiles of cryptochrome genes in wing dimorphic brown planthoppers, Nilaparvata lugens

Cryptochromes （CRYs） are blue and UV light photoreceptors, known to play key roles in circadian rhythms and in the light-dependent magnetosensitivity of insects. Two novel cryptochrome genes were cloned from the brown planthopper, and were given the designations of Nlcryl and Nlcry2, with the accession numbers KM108578 and KM108579 in GenBank. The complementary DNA sequences ofNlcryl andNlcry2 are 1935 bp and 2463 bp in length, and they contain an open reading frame of 1629 bp and 1872 bp, encoding amino acids of 542 and 623, with a predicted molecular weight of 62.53 kDa and 70.60 kDa, respectively. Well-conserved motifs such as DNA-photolyase and FAD-binding-7 domains were observed in Nlcry1 and Nlcry2. Phylogenetic analysis demonstrated the proteins of Nlcry1 and Nlcry2 to be clustered into the insect＇s cryptochrome 1 and cryptochrome 2, respectively. Quantitative polymerase chain reaction showed that the daily oscillations of messenger RNA （mRNA） expression in the head of the brown planthopper were mild for Nlcryl, and modest for Nlcry2. Throughout all developmental stages, Nlcryl and Nlcry2 exhibited extreme fluctuations and distinctive expression profiles. Cryptochrome mRNA expression peaked immediately after adult emergence and then decreased subsequently. The tissue expression profiles of newly emerged brown planthopper adults showed higher expression levels of CRYs in the head than in the thorax or abdomen, as well as significantly higher levels of CRYs in the heads of the macropterous strain than in the heads of the brachypterous strain. Taken together, the results of our study suggest that the two cryptochrome genes characterized in the brown planthopper might be associated with developmental physiology and migration.

Morph-specific differences in metabolism related to flight in the wing-dimorphic Aphis gossypii

The cotton aphid, Aphis gossypii Glover, is a wing-dimorphic species, which causes globally important agricultural losses. In this present study, we compared the biochemical basis of wing polymorphism in A. gossypii with respect to trade-off of energy resources, including glycogen, trehalose, lipids （total lipid, triglyceride and phospholipid）, free fatty acids, and soluble protein between dispersal and reproduction morphs during the wing-bud nymph and adulthood. Total lipid, triglyceride and free fatty acids were significantly higher in winged versus wingless morphs at 12 h of adulthood, the period during which alates are able to fly. By contrast, the wingless morph contained more glycogen than the winged morph from the 4th nymphal stage to adulthood. Trehalose content in the wingless morph was also higher than that in the winged morph during the 3rd and 4th nymphal stages, but vice versa at 12 h of adulthood. Finally, soluble protein content increased from nymphs to adults and was higher during adulthood in aptera versus alate. Whole-body water content in 12-h adults was significantly higher in apterae than that in alatae. These results indicate significant physiological differences between morphs related to specialization for flight.

Variation in sexual dimorphism and assortative mating do not predict genetic divergence in the sexually dimorphic Goodeid fish Girardinichthys multiradiatus

Sexual dimorphism is often used as a proxy for the intensity of sexual selection in comparative studies of sexual selection and diversification. The Mexican Goodeinae are a group of livebearing freshwater fishes with large variation between species in sexual dimorphism in body shape. Previously we found an association between variation in morphological sexual dimorphism between species and the amount of gene flow within populations in the Goodeinae. Here we have examined if mor- phological differentiation within a single dimorphic species is related to assortative mating or gene flow between populations. In the Amarillo fish Girardinichthys multiradiatus studies have shown that exaggerated male fins are targets of female preferences. We find that populations of the species differ in the level of sexual dimorphism displayed due to faster evolution of differences in male than female morphology. However, this does not predict variation in assortative mating tests in the laboratory; in fact dif- ferences in male morphology are negatively correlated with assortative mating. Microsatellite markers reveal significant genetic differences between populations. However, gene flow is not predicted by either morphological differences or assortative mating. Rather, it demonstrates a pattern of isolation by distance with greater differentiation between watersheds. We discuss the caveats of predicting behavioural and genetic divergence from so-called proxies of sexual selection [Current Zoology 58 （3）： 440-452, 2012].

Sexual Dimorphism and Female Reproductive Characteristics in the Hainan Water Skink(Tropidophorus hainanus)from Hainan,South China

We studied sexual dimorphism in body size and shape and female reproductive characteristics in the Hainan Water Skink(Tropidophorus hainanus)from Hainan,South China.In our sample the largest female and male were 61.1 and 55.2 mm snout-vent length(SVL),respectively.The mean SVL was larger in adult females(52.0 mm)than in adult males(48.3 mm).Juveniles were sexually dimorphic in head length but not in other examined morphological variables.Adult males were longer in head length and shorter in abdomen length and fore-limb length than adult females of the same SVL.Ontogenetic shifts in sexual dimorphism in body shape were evident,as revealed by the fact that morphological differences between the sexes were more pronounced in adults than in juveniles.Females produced a single litter of 3–6 offspring per season from early August to early September.Litter size,litter mass and offspring(neonate)mass were positively related to female SVL.Neonate mass was independent of relative fecundity.From the above findings we draw three main conclusions.First,females are the larger sex in T.hainanus,and sexual dimorphism in body shape is more pronounced in adults than in juveniles.Second,larger female T.hainanus produce more and larger offspring and thus heavier litters than smaller ones.Third,the offspring size-number trade-off does is not evident in T.hainanus.

Flower development and sexual dimorphism in Vernicia montana

The mu oil tree(Vernicia montana Lour.) is a dioecious species, but the genetic mechanisms underlying its phenotypic sexual dimorphism are unclear. In this study, we determined two pivotal phases of sex differentiation of mu oil tree via morphological and histological analyses of unisexual flowers:(Ⅰ) differentiation of male or female primordia to produce staminate flowers(SFs) or transient hermaphrodite flowers(HFs),and(Ⅱ) complete abortion of stamens in transient HFs to generate pistillate flowers(PFs). A total of 1621 sex-biased genes were identified by comparative transcriptome analysis which exhibited elevated rates of protein evolution than unbiased genes. The female-biased genes were enriched in the production of defense compounds while male-biased genes were focused on the production of viable pollens. Transcriptomebased analysis revealed that the differentially expressed genes(DEGs) between PFs and SFs in phase Ⅰ involved in abscisic acid(ABA), auxin(AUX), cytokinin(CK), ethylene(ET), and gibberellin(GA) biosynthesis and signaling showed higher expression levels in males than in females in general, whereas the DEGs involved in jasmonic acid(JA) and salicylic acid(SA) pathways displayed opposite expression patterns. Moreover,differentially expressed endogenous ABA, AUX, GAs, JA, and SA exhibited consistent biased expression patterns with the DEGs by UPLC-MSbased analysis. Exogenous application of an anti-ethylene plant growth regulator could promote the development of stamens in PFs and generated HFs. Comparative transcriptomic and hormonal analyses of PFs and SFs in phase Ⅱ indicated an increase in ET concentration when abortion of stamens in PFs occurred. This study suggested that phytohormones play key roles in sex dimorphism and ET may determine the development of stamens in PFs of mu oil tree, which provides an insight into plant sex differentiation mechanisms.

Osteology of Leptobrachella bijie(Anura,Megophryidae)

The Asian leaf litter toad genus Leptobrachella contains more than 100 species widely distributed in southwestern Asia.However,the systematic profiles of this group remain unresolved.Osteological morphology is important for taxonomic and phylogenetic studies.However,few studies have focused on the osteology of the genus.Herein,we comprehensively described the osteological features of a representative species L.bijie based on micro-CT scanning and double-staining methods.The results show that the skull of adult L.bijie is well-ossified,exhibiting sexual dimorphism and minimal intraspecific variation.The skull length is slightly greater than the width,with the maxilla slightly overlapping with the quadratojugal.The nasal connects with the sphenethmoid in males,but only part or not connects with the sphenethmoid in females.The transverse processes of the sacrum are robust and symmetrically butterfly-shaped,and the presacral vertebrae are procoelous.The pectoral girdle is arciferous.The phalangeal formula is 2-2-3-3 for the hand,and 2-2-3-4-3 for the foot.This study provides the first detailed and comprehensive osteological accounts of the genus Leptobrachella.

Male Morphological Dimorphism in the Genji Firefly Nipponoluciola cruciata in Central Japan

The Japanese Genji firefly Nipponoluciola cruciata is famous for geographic variation in flash pattern. Such geographic variation has also been well studied in terms of ecology and molecular phylogeny. However, there are very few studies on geographic variation in morphology of this species. This study focused on morphological characteristics of males of this species in Yamanashi and Gunma Prefectures separated by the Kanto Mountains in central Japan. Previous studies have shown that this species in the two prefectures exhibits different genetic features as well as different flash patterns. This study aimed to explore morphological characteristics of males of this species and performed multivariate analysis using measurements previously published in the literature. As a result, allometric analysis between body length and elytra width suggested that this species exhibits different allometric patterns between the two prefectures. Moreover, cluster analysis of measurements of genitalia suggested that this species is completely separated into two different groups between the two prefectures. These results suggest the importance of further morphological studies to clarify the divergence of this species.

Geographic Differences in Allometric Patterns of Males of the Japanese Firefly Luciola parvula

It has been well known for many years that males of Luciola parvula (Coleoptera, Lampyridae) have two morphs, namely small and large morphs in Japan. This article performed a statistical reanalysis of their morphological measurements based on previously published data and compared them. As a result, the two morphs were found to have different allometric patterns in the Hakone and Nagano areas, Japan. In Hakone, the relationships between their pronotum width and length were expressed by a common allometric line and a shift in location along the line. However, in Nagano, the relationships between their pronotum width and body length were expressed by two parallel allometric lines (a common slope and different intercepts). The allometric pattern of males of this species may vary among different parts of the body or among different areas. Therefore, this article recommends future studies to measure the same parts of the body as previous studies to clarify geographic variation in allometric relationships.

Embryogenesis,Germination,Structure and Cotyledon Dimorphism of Zea mays Embryo

A series of new cognitions on the morphogenesis of maize ( Zea mays L.) embryo have been obtained with scanning electron microscopy and semi-thin section techniques. 1. The proembryo. The proembryo from zygotic cell divisions may be divided into three parts: proper, hypoblast and suspensor. The suspensor is short and small, and only exists transiently. As to the hypoblast there is a growth belt, which promotes elongation of the hypoblast. Eventually the upper portion of the hypoblast contributes to the formation of the coleorhiza and the remainder dries up, sticking to the end of the coleorhiza. 2. The maize embryo possesses dorsiventrality and cotyledon dimorphism. During early proembryo stage, the dorsiventrality appears in the proper of the embryo. On the ventral side, the cells are small with dense cytoplasm and few vacuoles. On the dorsal side, the cells are larger with lower cytoplasmic density and have more vacuoles. During later proembryo stage, the proper develops into two parts: the ventrum and the dorsurn. The ventrum rises up from the center of the ventral side. The dorsurn is composed of the marginal area of the ventral side and the whole dorsal side of the proper. During young embryo development, the ventrum differentiates into the coleoptile, apical meristem, hypocotyl, radicle and the main part of the coleorhiza. What is more important, the emergence of coleoptile primordium and radicular initials occur at the axis of the proper, then the coleoptile primordium expands from its two ends toward left and right to form a ring, and the endogenous radicular initials expand in all directions to form a conical radicular tip. All these morphogenetic activities of the ventrum follow a bilateral symmetrical pattern. The dorsurn forms the scutellum. primordium. Then the scutellum primordium, expands rapidly toward the left, right, front and back, while thickening itself, so as to make all components originating from the ventrum become hidden in the longitudinal groove of the scutellum. Lastly, the left and right lateral scales emerge from the edges of the longitudinal groove and expand toward the central line of the axis. As a consequence, morphologically, the bilateral symmetry of the ventral side of the embryo is revealed entirely. Morphogenetically, the coleoptile primordium and apical meristem in maize are similar to the coleoptile (apical cotyledon) and apex formation of the nice embryo, so the coleoptile of the maize embryo can also be considered as an apical cotyledon. The scutellum is a lateral cotyledon. These dimorphic cotyledons of the maize embryo originate from the dorsiventrality of the proper. 3. The true morphological structure of the maize embryo is recognized and its developmental stages are established. A maize embryo is a hypocotyl, in which the apical part is the shoot apex (or plumule) with the coleoptile, the central part consists mainly of the hypocotyl with a lateral cotyledon (scutellum), and the basal part is the radicle with coleorhiza. The left and right lateral scales derived from the scutellum overlap at the ventral side, leaving only two little pores at both ends of the seam from which the coleoptile and coleorhiza can be seen. The four sequential stages of maize embryonic development are as follows: (1) proembryo, stage. This stage covers a period from zygotic cell division to the appearance of the dorsum and ventrum. (2) ventrum rapid differentiation stage. (3) scutellum rapid expansion stage. (4) lateral scale development stage (or embryonic envelope formation stage). 4. To obtain a median longitudinal section perpendicular to the ventral surface is crucial for recognizing the genuine morphological structure of the maize embryo.