Promiscuous Expression of α-Tubulin II in Maturing Male and Female Plasmodium falciparum Gametocytes 英文参考文献.docVIP

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Promiscuous Expression of α-Tubulin II in Maturing Male and Female Plasmodium falciparum Gametocytes 英文参考文献.doc

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Promiscuous Expression of α-Tubulin II in Maturing Male and Female Plasmodium falciparum Gametocytes 英文参考文献

PromiscuousExpressionofa-TubulinIIinMaturingMale andFemalePlasmodiumfalciparumGametocytes SamanaSchwank,ColinJ.Sutherland*,ChrisJ.Drakeley DepartmentofImmunologyandInfection,FacultyofInfectiousandTropicalDiseases,LondonSchoolofHygieneandTropicalMedicine,London,UnitedKingdom Abstract Background:AntimalarialinterventionsdesignedtoimpactonthetransmissiblesexualstagesofPlasmodiumfalciparumare evaluatedbymeasurementofperipheralgametocytecarriageinvivoandinfectivitytomosquitoes.Drugorvaccine-elicited effectsmaydifferentiallyaffecttherelativeabundanceofmaturemaleandfemalesexualforms,andthiscanbemeasured byestimationofsexratiosbeforeandafterinterventioninvivoandinvitro.Measuringtheimpactofanti-gametocytedrugs onsexualcommitmentofimmaturegametocytestagesinvitroisnotcurrentlypossibleasmaleandfemaleparasitescannot bedistinguishedbymorphologyalonepriortostageIV. Methodology/Principal Findings: We have modified an existing immunofluorescence-based approach for distinguishing male and female gametocytes during development in vitro, by using highly synchronised magnetically-enriched gametocytepreparationsatdifferentstagesofmaturity.Antibodiesrecognisinga-tubulinII(males)andPfg377(females) wereused toattempttodiscriminatethesexes.Transcriptionofthesetwoproteins wasnotcoordinatedduring invitro development, with pfg377 transcripts accumulating only late in development, immediately prior to immunofluorescent signals from the PfG377 protein appearing in stage IV gametocytes. Contrary to previous descriptions of this protein as male-specificinP.falciparum,a-tubulinIIrecognisedbothmaleandfemalegametocytesatstagesItoIV,butevidenceof differential expression levels of this protein in late stage male and female gametocytes was found. Using antibodies recognisingPfG377astheprimarymarkeranda-tubulinIIasasecondarymarker,robustestimatesofsexratioininvitro cultureswereobtainedforgametocytesatstageIVorlater,andvalidatedbylightmicroscopiccounts.However,sexratio estimationwasnotpossiblefor