RESUMO
Axon guidance molecules are critical for neuronal pathfinding because they regulate directionality and growth pace during nervous system development. However, the molecular mechanisms coordinating proper axonal extension and turning are poorly understood. Here, metastasis suppressor 1 (Mtss1), a membrane protrusion protein, ensured axonal extension while sensitizing axons to the Semaphorin 3E (Sema3E)-Plexin-D1 repulsive cue. Sema3E-Plexin-D1 signaling enhanced Mtss1 expression in projecting striatonigral neurons. Mtss1 localized to the neurite axonal side and regulated neurite outgrowth in cultured neurons. Mtss1 also aided Plexin-D1 trafficking to the growth cone, where it signaled a repulsive cue to Sema3E. Mtss1 ablation reduced neurite extension and growth cone collapse in cultured neurons. Mtss1-knockout mice exhibited fewer striatonigral projections and irregular axonal routes, and these defects were recapitulated in Plxnd1- or Sema3e-knockout mice. These findings demonstrate that repulsive axon guidance activates an exquisite autoregulatory program coordinating both axonal extension and steering during neuronal pathfinding.
Assuntos
Moléculas de Adesão Celular , Glicoproteínas de Membrana , Proteínas do Tecido Nervoso , Semaforinas , Animais , Camundongos , Peptídeos e Proteínas de Sinalização Intracelular , Glicoproteínas de Membrana/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos Knockout , Semaforinas/genética , Semaforinas/metabolismoRESUMO
Misfolding of α-synuclein (α-Syn) in the brain causes cellular dysfunction, leading to cell death in a group of neurons, and consequently causes the progression of Parkinson's disease (PD). Although many studies have demonstrated the pathological connections between vascular dysfunction and neurodegenerative diseases, it remains unclear how neuronal accumulation of α-Syn affects the structural and functional aspects of the cerebrovasculature to accelerate early disease progression. Here, we demonstrated the effect of aberrant α-Syn expression on the brain vasculature using a PD mouse model expressing a familial mutant form of human α-Syn selectively in neuronal cells. We showed that young PD mice have an underdeveloped cerebrovasculature without significant α-Syn accumulation in the vasculature. During the early phase of PD, toxic α-Syn was selectively increased in neuronal cells, while endothelial cell proliferation was decreased in the absence of vascular cell death or neuroinflammation. Instead, we observed altered neuronal activation and minor changes in the activity-dependent gene expression in brain endothelial cells (ECs) in young PD mice. These findings demonstrated that neuronal expression of mutant α-Syn in the early stage of PD induces abnormal neuronal activity and contributes to vascular patterning defects, which could be associated with a reduced angiogenic potential of ECs.
Assuntos
Doenças Neurodegenerativas , Doença de Parkinson , Camundongos , Animais , Humanos , Doença de Parkinson/metabolismo , Células Endoteliais/metabolismo , alfa-Sinucleína/metabolismo , Neurônios/metabolismo , Doenças Neurodegenerativas/metabolismo , Células CultivadasRESUMO
In binocular animals that exhibit stereoscopic visual responses, the axons of retinal ganglion cells (RGCs) connect to brain areas bilaterally by forming a commissure called the optic chiasm (OC). Ventral anterior homeobox 1 (Vax1) contributes to the formation of the OC, acting endogenously in optic pathway cells and exogenously in growing RGC axons. Here, we generated Vax1AA/AA mice expressing the Vax1AA mutant, which is incapable of intercellular transfer. We found that RGC axons cannot take up Vax1AA protein from the Vax1AA/AA mouse optic stalk (OS) and grow slowly to arrive at the hypothalamus at a late stage. The RGC axons of Vax1AA/AA mice connect exclusively to ipsilateral brain areas after failing to access the midline, resulting in reduced visual acuity and abnormal oculomotor responses. Overall, our study provides physiological evidence for the necessity of intercellular transfer of Vax1 and the importance of the bilateral RGC axon projection in proper visuomotor responses.
Assuntos
Neuropeptídeos , Quiasma Óptico , Camundongos , Animais , Quiasma Óptico/metabolismo , Células Ganglionares da Retina , Encéfalo/metabolismo , Camundongos Endogâmicos C57BL , Neuropeptídeos/metabolismo , Proteínas de Homeodomínio/metabolismoRESUMO
Transcription factors (TFs) are transported from the cytoplasm to the nucleus and disappear from the nucleus after they regulate gene expression. Here, we discover an unconventional nuclear export of the TF, orthodenticle homeobox 2 (OTX2), in nuclear budding vesicles, which transport OTX2 to the lysosome. We further find that torsin1a (Tor1a) is responsible for scission of the inner nuclear vesicle, which captures OTX2 using the LINC complex. Consistent with this, in cells expressing an ATPase-inactive Tor1aΔE mutant and the LINC (linker of nucleoskeleton and cytoskeleton) breaker KASH2, OTX2 accumulated and formed aggregates in the nucleus. Consequently, in the mice expressing Tor1aΔE and KASH2, OTX2 could not be secreted from the choroid plexus for transfer to the visual cortex, leading to failed development of parvalbumin neurons and reduced visual acuity. Together, our results suggest that unconventional nuclear egress and secretion of OTX2 are necessary not only to induce functional changes in recipient cells but also to prevent aggregation in donor cells.
Assuntos
Núcleo Celular , Genes Homeobox , Animais , Camundongos , Lisossomos , Divisão Celular , Matriz Nuclear , VesículaRESUMO
Brain endothelial cells (BECs) are important conduits that deliver oxygen and nutrients, protect parenchyma cells from toxins, and drain wastes to maintain brain homeostasis. Impairment of BECs has been implicated in diverse neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease. Therefore, molecular analysis of BECs is important for understanding the molecular pathogenesis of these neurological diseases. Even though many transcriptome analyses for BECs have been developed, mRNA levels do not necessarily correlate with the levels of actively translated proteins. Translatome analysis using RiboTag mice, in which Rpl22, a ribosomal component, is tagged by the hemagglutinin epitope under Cre recombinase activation, could serve as an excellent tool that overcomes these caveats. However, implementation of this technique is limited by high noise-to-signal ratios as well as the low yield of mRNAs from BECs, which limits bulk gene expression analysis. In this study, we established a protocol to isolate highly pure mRNAs from BECs in the cortex of eight- to twelve-week-old male Tie2-Cre; Rpl22HA/HA mice by using a cell strainer to trap blood vessels prior to immunoprecipitation. According to the results of RT-PCR, the specificity of the mRNA pools isolated by our protocol was much higher than that of the pools isolated by the standard protocol. We were also able to generate a high-quality cDNA library for RNA-seq with the small amount of mRNA isolated with our protocol. Thus, this optimized method will be useful for future studies of BECs at the molecular level.
Assuntos
Células Endoteliais , Hemaglutininas , Animais , Encéfalo/metabolismo , Células Endoteliais/metabolismo , Epitopos/metabolismo , Hemaglutininas/metabolismo , Masculino , Camundongos , Oxigênio/metabolismo , RNA Mensageiro/metabolismoRESUMO
Background: Recent studies have reported that steamed and freeze-dried mature silkworms, also known as HongJam, have various health-promoting effects. Objective: The goal of this study was to elucidate changes in the various health-promoting effects of HongJam, after its digestion with a food-grade protease. Materials and Methods: We examined whether healthspan-promotion and rotenone-induced loss of motor-control prevention effects were enhanced in Drosophila fed with food-grade alkaline protease-digested HongJam compared to those fed with non-digested HongJam. The differences in mitochondrial functions, chemical susceptibilities, and activations of signal transduction pathways between Drosophila supplemented with various feed were examined to elucidate the molecular and biochemical basis of healthspan-promotion and locomotor-improvement effects of protease-digested HongJam. Results: We first found that the healthspan-promotion effect of HongJam digested with a food-grade protease was different depending on the silkworm variety used for its production. Digestion with food-grade protease into White-Jade HongJam (WJ) as prepared from the White-Jade silkworm variety that spins white cocoons did not enhance its functionality. However, compared to Golden-Silk HongJam (GS), a food-grade protease-digested Golden-Silk HongJam (GSD) produced from the Golden-Silk silkworm variety that spins yellow cocoons, it further promoted the healthspan in a Drosophila model. By conducting a series of studies to reveal the molecular and biochemical basis for healthspan-promoting effects, we found that GS and GSD similarly enhanced mitochondrial activity, but GSD activated autophagy signaling more than GS. In addition, GSD feed (GSDf)-, GSD supernatant feed (GSDsupf)-, and GSD precipitate feed (GSDprecf)-reared Drosophila were also found to have increased resistance to an autophagy inhibitor compared to that of normal feed- or GS feed-reared Drosophila. Furthermore, we found that the rotenone-induced loss of motor control prevention effect was superior for GSDsup compared to GS, GSD, or GSDprec. This result may have occurred because GSDsup has more phenolic compounds and antioxidant activities than other samples. Conclusion: GSDsup contained more digested small peptides and free phytochemicals than other samples due to the digestion of proteins with a food-grade protease. Thus, GSDsup leads to further healthspan-promoting and locomotor-improvement effects than GS, GSD, or GSDprec.
RESUMO
Post-stroke vascular remodeling, including angiogenesis, facilitates functional recovery. Proper vascular repair is important for efficient post-stroke recovery; however, the underlying mechanisms coordinating the diverse signaling pathways involved in vascular remodeling remain largely unknown. Recently, axon guidance molecules were revealed as key players in injured vessel remodeling. One such molecule, Semaphorin 3E (Sema3E), and its receptor, Plexin-D1, control vascular development by regulating vascular endothelial growth factor (VEGF) signaling. In this study, using a mouse model of transient brain infarction, we aimed to investigate whether Sema3E-Plexin-D1 signaling was involved in cerebrovascular remodeling after ischemic injury. We found that ischemic damage rapidly induced Sema3e expression in the neurons of peri-infarct regions, followed by Plexin-D1 upregulation in remodeling vessels. Interestingly, Plexin-D1 reemergence was concurrent with brain vessels entering an active angiogenic process. In line with this, Plxnd1 ablation worsened neurological deficits, infarct volume, neuronal survival rate, and blood flow recovery. Furthermore, reduced and abnormal vascular morphogenesis was caused by aberrantly increased VEGF signaling. In Plxnd1 knockout mice, we observed significant extravasation of intravenously administered tracers in the brain parenchyma, junctional protein downregulation, and mislocalization in regenerating vessels. This suggested that the absence of Sema3E-Plexin-D1 signaling is associated with blood-brain barrier (BBB) impairment. Finally, the abnormal behavioral performance, aberrant vascular phenotype, and BBB breakdown defects in Plxnd1 knockout mice were restored following the inhibition of VEGF signaling during vascular remodeling. These findings demonstrate that Sema3E-Plexin-D1 signaling can promote functional recovery by downregulating VEGF signaling in the injured adult brain.
Assuntos
Semaforinas , Acidente Vascular Cerebral , Animais , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular Neuronais/metabolismo , Regulação para Baixo , Infarto , Peptídeos e Proteínas de Sinalização Intracelular/genética , Glicoproteínas de Membrana/metabolismo , Camundongos , Semaforinas/genética , Semaforinas/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Remodelação VascularRESUMO
Coronavirus disease 2019 (COVID-19) has put hypertensive patients in densely populated cities at increased risk. Nurse-coordinated home blood pressure telemonitoring (NC-HBPT) may help address this. We screened studies published in English on three databases, from their inception to 30 November 2020. The effects of NC-HBPT were compared with in-person treatment. Outcomes included changes in blood pressure (BP) following the intervention and rate of BP target achievements before and during COVID-19. Of the 1916 articles identified, 27 comparisons were included in this review. In the intervention group, reductions of 5.731 mmHg (95% confidence interval: 4.120-7.341; p < 0.001) in systolic blood pressure (SBP) and 2.342 mmHg (1.482-3.202; p < 0.001) in diastolic blood pressure (DBP) were identified. The rate of target BP achievement was significant in the intervention group (risk ratio, RR = 1.261, 1.154-1.378; p < 0.001). The effects of intervention over time showed an SBP reduction of 3.000 mmHg (-5.999-11.999) before 2000 and 8.755 mmHg (5.177-12.334) in 2020. DBP reduced by 2.000 mmHg (-2.724-6.724) before 2000 and by 3.529 mmHg (1.221-5.838) in 2020. Analysis of the target BP ratio before 2010 (RR = 1.101, 1.013-1.198) and in 2020 (RR = 1.906, 1.462-2.487) suggested improved BP control during the pandemic. NC-HBPT more significantly improves office blood pressure than UC among urban hypertensive patients.
Assuntos
COVID-19 , Hipertensão , Pressão Sanguínea , Monitorização Ambulatorial da Pressão Arterial , Humanos , Hipertensão/epidemiologia , SARS-CoV-2RESUMO
Genome editing by clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR associated protein (Cas)9, a third-generation gene scissors, and molecular breeding at the genome level are attracting considerable attention as future breeding techniques. In the present study, genetic and phenotypic analyses were conducted to examine the molecular breeding of Bombyx mori through CRISPR/Cas9-mediated editing of the kynurenine 3-monooxygenase (KMO) gene. The synthesized guide RNAs (gRNAs) were analyzed using T7 endonuclease I after introduction into the BM-N silkworm cell line. To edit the silkworm gene, K1P gRNA, and Cas9 complexes were microinjected into silkworm embryos. After microinjection, the hatching rate and the incidence of mutation were determined as 18.1% and 60%, respectively. Gene mutation was verified in the heterozygous G0 generation, but no phenotypic change was observed; however, certain embryos and moths produced through sib-mating had significant differences compared to the wild-type. In successive generations, a distinct phenotypic change was also observed by continuous mating. Thus, although there are limitations in the phenotypic expression in breeding through the induction of deletion mutations, as in the present study, the process is believed to yield successful results within a shorter period compared to traditional breeding and is safer than transgenic technology.
Assuntos
Bombyx/genética , Embaralhamento de DNA/métodos , Edição de Genes/métodos , Quinurenina 3-Mono-Oxigenase/genética , Animais , Sistemas CRISPR-Cas/genéticaRESUMO
The homeodomain is found in hundreds of transcription factors that play roles in fate determination via cell-autonomous regulation of gene expression. However, some homeodomain-containing proteins (HPs) are thought to be secreted and penetrate neighboring cells to affect the recipient cell fate. To determine whether this is a general characteristic of HPs, we carried out a large-scale validation for intercellular transfer of HPs. Our screening reveals that intercellular transfer is a general feature of HPs, but it occurs in a cell-context-sensitive manner. We also found the secretion is not solely a function of the homeodomain, but it is supported by external motifs containing hydrophobic residues. Thus, mutations of hydrophobic residues of HPs abrogate secretion and consequently interfere with HP function in recipient cells. Collectively, our study proposes that HP transfer is an intercellular communication method that couples the functions of interacting cells.
Assuntos
Comunicação Celular/genética , Proteínas de Homeodomínio/metabolismo , Transporte Proteico/genética , Motivos de Aminoácidos/genética , Animais , Encéfalo/embriologia , Encéfalo/metabolismo , Linhagem Celular , Feminino , Ensaios de Triagem em Larga Escala , Proteínas de Homeodomínio/genética , Humanos , Interações Hidrofóbicas e Hidrofílicas , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mutação , Gravidez , Retina/metabolismoRESUMO
The non-cell autonomous transfer of OTX2 homeoprotein transcription factor into juvenile mouse cerebral cortex regulates parvalbumin interneuron maturation and critical period timing. By analyzing gene expression in primary visual cortex of wild-type and Otx2+/GFP mice at plastic and nonplastic ages, we identified several putative genes implicated in Otx2-dependent visual cortex plasticity for ocular dominance. Cortical OTX2 infusion in juvenile mice induced Gadd45b/g expression through direct regulation of transcription. Intriguingly, a reverse effect was found in the adult, where reducing cortical OTX2 resulted in Gadd45b/g upregulation. Viral expression of Gadd45b in adult visual cortex directly induced ocular dominance plasticity with concomitant changes in MeCP2 foci within parvalbumin interneurons and in methylation states of several plasticity gene promoters, suggesting epigenetic regulation. This interaction provides a molecular mechanism for OTX2 to trigger critical period plasticity yet suppress adult plasticity.
Assuntos
Antígenos de Diferenciação/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Plasticidade Neuronal/fisiologia , Fatores de Transcrição Otx/metabolismo , Córtex Visual/fisiologia , Animais , Dominância Ocular/fisiologia , Epigênese Genética , Regulação da Expressão Gênica , Interneurônios/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Parvalbuminas/metabolismoRESUMO
Retinal ganglion cell (RGC) axons of binocular animals cross the midline at the optic chiasm (OC) to grow toward their synaptic targets in the contralateral brain. Ventral anterior homeobox 1 (Vax1) plays an essential role in the development of the OC by regulating RGC axon growth in a non-cell autonomous manner. In this study, we identify an unexpected function of Vax1 that is secreted from ventral hypothalamic cells and diffuses to RGC axons, where it promotes axonal growth independent of its transcription factor activity. We demonstrate that Vax1 binds to extracellular sugar groups of the heparan sulfate proteoglycans (HSPGs) located in RGC axons. Both Vax1 binding to HSPGs and subsequent penetration into the axoplasm, where Vax1 activates local protein synthesis, are required for RGC axonal growth. Together, our findings demonstrate that Vax1 possesses a novel RGC axon growth factor activity that is critical for the development of the mammalian binocular visual system.
Assuntos
Axônios/metabolismo , Proteínas de Homeodomínio/metabolismo , Neuropeptídeos/metabolismo , Células Ganglionares da Retina/metabolismo , Animais , Drosophila melanogaster/metabolismo , Espaço Extracelular/metabolismo , Proteoglicanas de Heparan Sulfato/metabolismo , Discos Imaginais/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Camundongos , Modelos Biológicos , Ligação Proteica , Biossíntese de Proteínas , Imagem com Lapso de Tempo , Asas de Animais/metabolismoRESUMO
Plasticity in the visual cerebral cortex is regulated by the internalization of Otx2 homeoprotein into parvalbumin neurons in cortical layers II/III and IV. However the Otx2 locus is not active in these neurons and the protein is imported from external sources, including the choroid plexus. Because Otx1 and Otx2 may have redundant functions, we wanted to verify if part of the staining in parvalbumin neurons corresponds to Otx1 transported from cortical layer V neurons. It is demonstrated here that Otx staining in layer IV cells is maintained in Otx1-null mice. The immunoprecipitation of extracts from finely dissected granular and supragranular cortex (layers I-IV) gave immunoblots with a band corresponding to Otx2 and not Otx1. Moreover, high-resolution mass spectrometry analysis after immunoprecipitation identifies two peptides within the Otx2 homeodomain. One of these peptides is specific for Otx2 and is not found in Otx1. These results unambiguously establish that the staining in parvalbumin neurons revealed with the anti-Otx2 antibodies used in our previous studies identifies non-cell autonomous Otx2.
RESUMO
This paper proposes a simple indicator to measure the exposure to natural disasters for the poor and non-poor population, in order to assess the global and regional trend of natural hazard and poverty. Globally, poor people are two times more exposed to natural disasters than the non-poor in the twenty-first century. The time trend varies across regions, with poor people in East Asia and Pacific being most exposed to natural disasters, followed by those in South Asia and Sub-Saharan Africa. The change of exposure measure over time is decomposed into two factors: a pure exposure change, which could be fuelled by climate change; and a concentration component. The result shows that the total net increase of exposure between the 1970s and the 2000s is driven significantly by the increased concentration of the poor (26 per cent) in disaster-prone areas, whereas the contribution of that factor remains very small for the non-poor (six per cent).
Assuntos
Desastres , Saúde Global , Áreas de Pobreza , África Subsaariana , Ásia , Ásia Oriental , Humanos , Ilhas do Pacífico , Medição de RiscoRESUMO
Retinal pigment epithelium (RPE) plays a critical role in vertebrate vision by providing functional and structural support to the retina. Degeneration of RPE by cumulative oxidative stresses or acute injury frequently results in retinal degenerative diseases, notably age-related macular degeneration (AMD). Moreover, it has been shown that phosphorylation-mediated inactivation of PTEN (phosphatase and tensin homolog) in RPE is closely linked to AMD-like retinal degeneration in mice [1]. In this study, we used AMD mouse models, in which chemokine (C-C motif) ligand 2 (Ccl2) or chemokine (C-C motif) receptor 2 (Ccr2) were genetically ablated, to examine mechanisms linking reactive oxygen species (ROS) to phosphorylation/inactivation of PTEN in RPE. We found that ROS levels were increased in these RPE cells in association with phosphorylation/inactivation of PTEN. Both PTEN phosphorylation/inactivation and consequent Akt activation in the RPE of AMD model mice were inhibited by antioxidant treatment, indicating a functional role for elevated intracellular ROS. We further discovered that PTEN phosphorylation in oxidatively stressed RPE was repressed by a phosphoinositide 3-kinase (PI3K) inhibitor, but not by an Akt inhibitor. Taken together, these results suggest that ROS-activated PI3K potentiates AMD-related RPE pathogenesis through phosphorylation/inactivation of PTEN.
Assuntos
Degeneração Macular/enzimologia , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Epitélio Pigmentado da Retina/enzimologia , Animais , Linhagem Celular , Quimiocina CCL2/genética , Modelos Animais de Doenças , Humanos , Degeneração Macular/genética , Degeneração Macular/patologia , Camundongos , Camundongos Knockout , Estresse Oxidativo , PTEN Fosfo-Hidrolase/genética , Fosforilação , Espécies Reativas de Oxigênio/metabolismo , Receptores CCR2/genética , Epitélio Pigmentado da Retina/patologiaRESUMO
This study was conducted to confirm the possible use of female Yangwonjam as a host for synnemata production of Isaria tenuipes in eight local areas in Korea. Silkworm pupation rate, infection rate and synnemata characteristics of I. tenuipes were examined. Normal silkworms had a higher pupation rate than silkworms inoculated with I. tenuipes. The pupae survival percentage of normal silkworm in cocoons was 92.5~97.6%, whereas it ranged from 91.1~95.6% in silkworms sprayed with I. tenuipes. Female Yangwonjam showed the highest survival percentage at 97.6% among the silkworm varieties tested. I. tenuipes infection rate of larvae of 5th instar newly-exuviated silkworms was 89.2~90.7% in the spring rearing season and 98.2~99.3% in the autumn rearing season. Synnemata production of I. tenuipes was excellent in female Yangwonjam with an incidence rate of 98.0% followed by male Yangwonjam (94.1%) and Baegokjam (93.3%) in the spring rearing season. Synnemata living weight ranged from 1.44~0.94 g in the spring rearing season. The female Yangwonjam had the heaviest synnemata weight (1.44 g) in the spring rearing season. The synnemata of I. tenuipes produced on pupae were white or milky-white in color, and were similar in shape and color to wild synnemata collected in Korea.
RESUMO
Honey production from approximately 1.6 million colonies owned by about 199,000 Korean beekeepers was almost 23,000 metric tons in 2009. Nosema causes significant losses in honey production and the virus decreases population size. We initiated a survey of honey bee colonies on the blooming period of Acacia to determine the prevalence of Nosema and virus in 2011. Most Korean beekeepers have moved from the south to north of Korea to get Acacia nectar for 2 mon. This provided a valuable opportunity to sample bees originating from diverse areas in one location. Twenty hives owned by 18 beekeepers were sampled in this year. Nosema spore counts ranged from zero to 1,710,000 spores per bee. The average number of nosema spores per bee was 580,000. Approximately 95% of the colonies were infected with Nosema, based on the presence of spores in the flowering period of Acacia. This indicates that Nosema is the predominant species affecting honeybee colonies. Also, the seven most important honeybee viruses were investigated by reverse transcription-PCR. Among them, four different viruses were detected in samples. Black queen cell virus was present in all samples. Chronic bee paralysis virus was detected in 10% of samples. Deformed wing virus was present in only 5% of the samples. Prevalence of Sacbrood virus was 15%. However, Cloudy wing virus, Israel acute paralysis virus and kashmir bee virus were not detected in any of samples.
RESUMO
Injection inoculation protocols for fruit body formation of Cordyceps militaris (C. militaris) were investigated to improve the incidence of infection in the silkworm species Bombyx mori (B. mori). Injection, with suspensions of C. militaris hyphal bodies into living silkworm pupae, was used to test for fruit body production. Use of Daeseungjam rather than Baegokjam or Keumokjam varieties of B. mori is thought to be suitable for infection by C. militaris. From mounting, nine-day-old to 11-day-old pupae showed the best incidence of infection with a 100 µL injection volume. Silkworm pupae injected with a hyphal suspension concentration of more than 2 × 10(5) colony-forming unit (cfu) recorded a greater than 96% incidence of infection. Also, fruit bodies of C. militaris were induced and produced at a light intensity between 500 and 1,000 lx.