Ua tsaug rau koj tuaj xyuas Nature.com.Qhov browser version koj siv tau txwv CSS kev txhawb nqa.Rau qhov kev paub zoo tshaj plaws, peb xav kom koj siv qhov browser tshiab (lossis lov tes taw Compatibility Hom hauv Internet Explorer).Nyob rau lub sijhawm no, txhawm rau ua kom muaj kev txhawb nqa txuas ntxiv, peb yuav ua rau lub xaib tsis muaj qauv thiab JavaScript.
Fertility ntawm cov noog nyob ntawm lawv lub peev xwm khaws cov phev txaus rau lub sijhawm ntev hauv cov phev cia tubules (SST).Qhov tseeb mechanism uas spermatozoa nkag mus, nyob hauv, thiab tawm hauv SST tseem muaj teeb meem.Cov phev ntawm sharkasi hens pom muaj kev nyiam rau agglutination, tsim cov pob khoom txawb uas muaj ntau lub hlwb.Vim qhov nyuaj ntawm kev soj ntsuam kev txav mus los thiab kev coj tus cwj pwm ntawm cov spermatozoa hauv lub raj opaque fallopian, peb siv cov khoom siv microfluidic nrog microchannel hla ntu zoo ib yam li cov spermatozoa los kawm cov spermatozoa agglutination thiab motility.Txoj kev tshawb no tham txog seb cov phev bundles tsim li cas, lawv txav mus los li cas, thiab lawv lub luag haujlwm tseem ceeb hauv kev nthuav dav cov phev nyob hauv SST.Peb tshawb xyuas cov phev tshaj tawm thiab tus cwj pwm rheological thaum cov kua dej ntws tawm hauv lub microfluidic channel los ntawm hydrostatic siab (ntws tus nqi = 33 µm / s).Cov spermatozoa nyiam ua luam dej tawm tsam tam sim no (zoo rheology) thiab qhov nrawm ntawm cov phev spermatozoa tau txo qis dua piv rau ib leeg spermatozoa.Cov phev bundles tau raug soj ntsuam kom txav mus rau hauv ib lub kauv thiab nce qhov ntev thiab tuab raws li ntau cov phev raug xaiv. Cov phev bundles tau pom los ze thiab ua raws li cov phab ntsa ntawm cov kab hauv microfluidic kom tsis txhob raug hws nrog cov kua dej nrawm tshaj> 33 µm / s. Cov phev bundles tau pom los ze thiab ua raws li cov phab ntsa ntawm cov kab hauv microfluidic kom tsis txhob raug hws nrog cov kua dej nrawm tshaj> 33 µm / s. Было замечено, что пучми сперматозоидов приближаются и прилипают к боковым стенкам микрофлюньтных, ания со скоростью потока жидкости > 33 мкм / с. Cov phev bundles tau raug soj ntsuam kom mus txog thiab ua raws li cov phab ntsa ntawm cov microfluidic channel kom tsis txhob raug cheb tawm ntawm cov kua dej ntws> 33 µm / s.观察到精子束接近并粘附在微流体通道的侧壁上，以避免被流体流速> 33 µm/s 扫过。33 µm/s. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрожидкость стенкам микрожидкостьного метания потоком жидкости со скоростью > 33 мкм/с. Cov phev bundles tau raug soj ntsuam kom ze thiab ua raws li cov phab ntsa ntawm cov microfluidic channel kom tsis txhob raug cheb tawm los ntawm cov kua dej ntws ntawm> 33 µm / s.Kev kuaj xyuas thiab kev xa hluav taws xob hluav taws xob microscopy qhia tias cov phev bundles tau txais kev txhawb nqa los ntawm cov khoom siv ntom ntom.Cov ntaub ntawv tau qhia pom qhov txawv txav ntawm Sharkazi qaib spermatozoa, nrog rau lub peev xwm ntawm cov spermatozoa rau agglutinate thiab tsim cov pob khoom txawb, uas ua rau muaj kev nkag siab zoo dua ntawm kev cia ntev ntawm cov spermatozoa hauv SMT.
Yuav kom ua tiav fertilization nyob rau hauv tib neeg thiab tsiaj feem ntau, phev thiab qe yuav tsum tuaj txog ntawm qhov chaw fertilization ntawm lub sijhawm.Yog li ntawd, mating yuav tsum tshwm sim ua ntej los yog thaum lub sij hawm ntawm ovulation.Ntawm qhov tod tes, qee cov tsiaj txhu, xws li dev, nrog rau cov tsiaj uas tsis yog tsiaj txhu, xws li kab, ntses, tsiaj reptiles, thiab noog, khaws cov phev hauv lawv lub cev xeeb tub rau lub sijhawm ntev mus txog thaum lawv cov qe npaj rau fertilization (asynchronous fertilization 1).Noog muaj peev xwm tswj tau cov spermatozoa muaj peev xwm fertilizing qe rau 2-10 lub lis piam2.
Qhov no yog qhov tshwj xeeb tshwj xeeb uas txawv cov noog los ntawm lwm cov tsiaj, vim nws muab qhov muaj feem ntau ntawm fertilization tom qab ib zaug insemination rau ob peb lub lis piam yam tsis muaj ib txhij mating thiab ovulation.Lub ntsiab phev cia lub cev, hu ua phev cia tubule (SST), nyob rau hauv lub internal mucosal folds ntawm lub uterovaginal hlws ris.Txog niaj hnub no, cov txheej txheem uas cov phev nkag mus, nyob, thiab tawm ntawm cov phev bank tsis nkag siab tag nrho.Raws li cov kev tshawb fawb yav dhau los, ntau qhov kev xav tau muab tso rau tom ntej, tab sis tsis muaj leej twg tau lees paub.
Forman4 xav tias spermatozoa tswj lawv qhov chaw nyob hauv SST kab noj hniav los ntawm kev txav mus tas li oscillatory tawm tsam cov kev taw qhia ntawm cov kua dej ntws los ntawm cov protein raws nyob rau ntawm SST epithelial hlwb (rheology).ATP yog depleted vim qhov kev ua haujlwm tas li uas yuav tsum tau ua kom cov phev nyob rau hauv SST lumen thiab motility nws thiaj li poob mus txog rau thaum cov phev tau nqa tawm ntawm cov phev bank los ntawm cov kua dej ntws thiab pib ib tug tshiab txoj kev mus rau lub ascending fallopian raj rau fertilize cov phev.Qe (Forman4).Cov qauv ntawm cov phev khaws cia no tau txais kev txhawb nqa los ntawm kev kuaj pom los ntawm immunocytochemistry ntawm aquaporins 2, 3 thiab 9 tam sim no hauv SST epithelial hlwb.Txog niaj hnub no, kev tshawb fawb txog qaib cov phev rheology thiab nws lub luag haujlwm hauv SST cia, kev xaiv cov phev ntawm qhov chaw mos, thiab kev sib tw phev tsis muaj.Hauv cov qaib, cov phev nkag mus rau hauv qhov chaw mos tom qab mating, tab sis ntau dua 80% ntawm cov phev raug tshem tawm ntawm qhov chaw mos sai tom qab mating.Qhov no qhia tias qhov chaw mos yog qhov chaw tseem ceeb rau kev xaiv phev hauv cov noog.Tsis tas li ntawd, nws tau tshaj tawm tias tsawg dua 1% ntawm spermatozoa fertilized nyob rau hauv qhov chaw mos mus rau hauv SSTs2.Hauv kev tsim cov menyuam yaus hauv qhov chaw mos, cov spermatozoa ncav cuag SST nyhav nce 24 teev tom qab insemination.Txog tam sim no, cov txheej txheem ntawm kev xaiv cov phev thaum tus txheej txheem no tsis meej, thiab phev motility yuav ua lub luag haujlwm tseem ceeb hauv SST phev uptake.Vim yog cov phab ntsa tuab thiab opaque ntawm cov hlab ntsha, nws yog ib qho nyuaj rau saib xyuas cov phev motility hauv cov hlab ntsha ntawm cov noog.Yog li ntawd, peb tsis muaj kev paub txog yuav ua li cas spermatozoa hloov mus rau SST tom qab fertilization.
Rheology tsis ntev los no tau lees paub tias yog ib qho tseem ceeb ntawm kev tswj cov phev thauj mus rau hauv cov tsiaj txhu ntawm qhov chaw mos.Raws li lub peev xwm ntawm motile spermatozoa migrate countercurrently, Zaferani et al8 tau siv lub corra microfluidic system los cais cov spermatozoa motile los ntawm cov qauv phev.Hom phev sorting no yog ib qho tseem ceeb rau kev kho mob infertility thiab kev tshawb fawb soj ntsuam, thiab yog nyiam tshaj li ib txwm txoj kev uas yog lub sij hawm thiab kev ua hauj lwm hnyav thiab muaj peev xwm cuam tshuam cov phev morphology thiab structural integrity.Txawm li cas los xij, txog niaj hnub no, tsis muaj kev tshawb fawb tau tshawb fawb txog qhov cuam tshuam ntawm kev zais ntawm qhov chaw mos ntawm cov qaib ntawm cov phev motility.
Tsis hais txog ntawm cov txheej txheem tswj cov phev khaws cia hauv SST, ntau tus neeg tshawb xyuas tau pom tias cov neeg nyob hauv cov spermatozoa agglutinate taub hau-rau-lub taub hau hauv SST ntawm qaib 9, 10, quails 2, thiab qaib ntxhw 11 los tsim cov phev agglutinated bundles.Cov kws sau ntawv qhia tias muaj kev sib txuas ntawm qhov kev sib txuam no thiab kev khaws cia ntev ntev ntawm cov spermatozoa hauv SST.
Tingari thiab Lake12 tau tshaj tawm txog kev sib raug zoo ntawm cov spermatozoa nyob rau hauv cov phev-ceev caj pas ntawm cov qaib thiab nug seb avian spermatozoa agglutinate zoo ib yam li cov spermatozoa mammalian.Lawv ntseeg tias qhov sib sib zog nqus sib txuas ntawm cov phev hauv vas deferens tuaj yeem yog vim muaj kev ntxhov siab los ntawm qhov muaj cov phev ntau hauv qhov chaw me me.
Thaum soj ntsuam tus cwj pwm ntawm cov spermatozoa ntawm cov iav dai tshiab, cov cim ntu ntu ntawm agglutination tuaj yeem pom, tshwj xeeb tshaj yog nyob ntawm cov npoo ntawm cov phev tee dej.Txawm li cas los xij, agglutination feem ntau cuam tshuam los ntawm kev ua haujlwm sib hloov cuam tshuam nrog kev txav mus tas li, uas piav qhia txog qhov xwm txheej ntawm qhov tshwm sim no.Cov kws tshawb fawb kuj pom tias thaum cov kua dej ntxiv rau hauv cov phev, elongated "xov zoo li" cell aggregates tshwm.
Thaum ntxov sim ua raws li cov phev spermatozoon tau ua los ntawm kev tshem cov hlau nyias los ntawm kev dai, uas ua rau cov phev elongated zoo li vesicle protruding los ntawm cov phev poob.Cov spermatozoa tam sim ntawd lined nyob rau hauv ib tug sib npaug zos zam nyob rau hauv lub vesicle, tab sis tag nrho cov chav tsev sai sai ploj vim 3D txwv.Yog li ntawd, txhawm rau kawm cov spermatozoa agglutination, nws yog ib qho tsim nyog yuav tsum tau soj ntsuam kev txav mus los thiab kev coj tus cwj pwm ntawm cov spermatozoa ncaj qha rau hauv cov phev cia cov tubules, uas yog qhov nyuaj rau kev ua tiav.Yog li ntawd, nws yog ib qho tsim nyog los tsim ib qho cuab yeej uas ua rau cov spermatozoa los txhawb kev tshawb fawb ntawm phev motility thiab agglutination cwj pwm.Brillard thiab al13 tau tshaj tawm tias qhov nruab nrab ntev ntawm cov phev cia tubules hauv cov menyuam yaus hnub nyoog yog 400-600 µm, tab sis qee qhov SSTs tuaj yeem ntev li 2000 µm.Mero thiab Ogasawara14 tau muab faib cov qog seminiferous rau hauv cov phev loj thiab cov phev tsis loj, ob qho tib si ntawm qhov ntev (~ 500 µm) thiab caj dab dav (~ 38 µm), tab sis qhov nruab nrab lumen txoj kab uas hla ntawm cov tubules yog 56.6 thiab 56.6 µm..11.2 μm, feem.Hauv txoj kev tshawb fawb tam sim no, peb siv cov khoom siv microfluidic nrog lub channel loj ntawm 200 µm × 20 µm (W × H), uas nws qhov hla yog me ntsis ze rau ntawm amplified SST.Tsis tas li ntawd, peb tau soj ntsuam cov phev motility thiab agglutination tus cwj pwm nyob rau hauv cov kua ntws, uas yog raws li Foreman lub hypothesis hais tias cov kua uas tsim los ntawm SST epithelial hlwb khaws cov phev nyob rau hauv lub lumen nyob rau hauv ib tug countercurrent (rheological) kev taw qhia.
Lub hom phiaj ntawm txoj kev tshawb no yog kom kov yeej cov teeb meem ntawm kev soj ntsuam spermatozoa motility nyob rau hauv lub fallopian raj thiab kom tsis txhob muaj teeb meem ntawm kev kawm rheology thiab tus cwj pwm ntawm spermatozoa nyob rau hauv ib tug dynamic ib puag ncig.Cov khoom siv microfluidic tau siv los ua kom muaj zog hydrostatic los simulate phev motility hauv qhov chaw mos ntawm tus qaib.
Thaum ib tug poob ntawm ib tug diluted phev qauv (1:40) twb loaded rau hauv lub microchannel ntaus ntawv, ob hom phev motility yuav raug txheeb xyuas (kab phev cais thiab khi phev).Tsis tas li ntawd, spermatozoa nyiam ua luam dej tawm tsam tam sim no (zoo rheology; video 1, 2). Txawm hais tias cov phev bundles muaj qis tshaj tshaj li cov phev kho siab (p <0.001), lawv tau nce qhov feem pua ​​​​ntawm cov phev uas pom zoo rheotaxis (p <0.001; Table 2). Txawm hais tias cov phev bundles muaj qis tshaj tshaj li cov phev kho siab (p <0.001), lawv tau nce qhov feem pua ​​​​ntawm cov phev uas pom zoo rheotaxis (p <0.001; Table 2). ISки ппи сперматоззотоззотли иотовериолериолериколерикоть, Орость снинооидтоззоидов (P <0,001), ои увели уротент рматозовеозв, демонстрирующительный реотакс (P <0,001; Txawm hais tias cov spermatozoa bundles muaj qis tshaj tawm tshaj li ntawm ib leeg spermatozoa (p <0.001), lawv tau nce qhov feem pua ​​​​ntawm cov spermatozoa uas qhia qhov zoo rheotaxis (p <0.001; Table 2).尽管精子束的速度低于孤独精子的速度（p < 0.001），但它们增加了明孤独精子的速度（p < 0.001），但它们增加了明孤独精子的速度（p < 0.001），但它们增加了明孤独精子的篔岔成。 ； 2).尽管精子束的速度低于孤独的速度（p <0.001），但增加了 显示阳性流徆0。 01; 2 . . . . . . . . . Хотя скорость пучков сперматозоидов была ниже, чем у одиночных сперматозоидов (p < 0,001), они вони увелиричи ва с положительной реологией (p < 0,001; таблица 2). Txawm hais tias qhov ceev ntawm cov phev bundles yog qis dua li ntawm ib leeg spermatozoa (p <0.001), lawv tau nce qhov feem pua ​​​​ntawm cov phev nrog cov rheology zoo (p <0.001; Table 2).Qhov zoo rheology rau ib leeg spermatozoa thiab tufts yog kwv yees li ntawm 53% thiab 85%, feem.
Nws tau raug pom tias spermatozoa ntawm sharkasi qaib tam sim ntawd tom qab ejaculation daim ntawv linear bundles, muaj ntau ntawm cov neeg.Cov tufts no nce nyob rau hauv ntev thiab tuab lub sij hawm thiab tej zaum yuav nyob twj ywm nyob rau hauv vitro rau ob peb teev ua ntej dissipating (video 3).Cov pob khoom filamentous no zoo li zoo li echidna spermatozoa uas tsim thaum kawg ntawm lub epididymis.Sharkashi hen phev tau pom tias muaj lub siab nyiam rau agglutinate thiab tsim ib pob reticulate hauv tsawg dua ib feeb tom qab sau.Cov kab teeb no muaj zog thiab muaj peev xwm lo rau tej phab ntsa nyob ze lossis cov khoom zoo li qub.Txawm hais tias phev bundles txo qhov ceev ntawm cov phev hlwb, nws yog tseeb hais tias macroscopically lawv nce lawv linearity.Qhov ntev ntawm cov pob khoom sib txawv nyob ntawm seb muaj pes tsawg tus phev sau hauv cov pob.Ob feem ntawm cov pob khoom raug cais tawm: thawj feem, suav nrog lub taub hau dawb ntawm cov phev agglutinated, thiab lub davhlau ya nyob twg, suav nrog tus Tsov tus tw thiab tag nrho qhov kawg ntawm cov phev.Siv lub koob yees duab ceev ceev (950 fps), lub taub hau dawb ntawm agglutinated spermatozoa tau pom nyob rau hauv thawj feem ntawm lub nras, lub luag haujlwm rau kev txav ntawm lub nras vim lawv txoj kev oscillatory, rub cov seem rau hauv lub nras nrog cov lus tsa suab (Video 4).Txawm li cas los xij, nyob rau hauv cov tufts ntev, nws tau pom tias qee cov phev dawb lub taub hau adhered rau lub cev thiab lub davhlau ya nyob twg ntawm cov tuft ua raws li cov vanes los pab txhawb lub plab.
Thaum nyob rau hauv ib qho qeeb ntawm cov kua dej, cov phev bundles txav mus rau ib leeg, txawm li cas los xij, lawv pib sib tshooj thiab lo rau txhua yam uas tseem nyob, kom tsis txhob raug ntxuav tawm los ntawm cov dej ntws tam sim no raws li qhov khiav ceev nce.Cov bundles tsim thaum ib txhais tes ntawm cov phev hlwb mus rau ib leeg, lawv pib txav mus rau hauv synchrony thiab qhwv ib ncig, thiab tom qab ntawd lo rau ib yam khoom nplaum.Daim duab 1 thiab 2 qhia tias cov phev mus rau ib leeg, tsim kev sib tshuam raws li cov tails qhwv ib ncig.
Cov kws tshawb fawb tau siv hydrostatic siab los tsim cov dej ntws hauv microchannel los kawm cov phev rheology.Lub microchannel nrog qhov loj ntawm 200 µm × 20 µm (W × H) thiab qhov ntev ntawm 3.6 µm tau siv.Siv cov microchannels nruab nrab ntawm cov ntim nrog cov koob txhaj tshuaj haum rau qhov kawg.Cov xim zaub mov tau siv los ua kom cov channel pom ntau dua.
Txuas cov kab txuas txuas thiab cov khoom siv rau ntawm phab ntsa.Cov vis dis aus tau coj nrog lub tshuab ntsuas qhov sib piv theem.Nrog rau txhua daim duab, theem sib piv microscopy thiab daim duab kos duab tau nthuav tawm.(A) Kev sib txuas ntawm ob lub kwj dej tiv thaiv qhov ntws vim yog helical motion (xub xub liab).(B) Kev sib txuas ntawm lub raj txuas thiab cov phab ntsa channel (cov xub liab), tib lub sij hawm lawv txuas nrog ob pawg (cov xub daj).(C) Cov phev bundles nyob rau hauv lub microfluidic channel pib sib txuas nrog ib leeg (cov xub liab), ua ib tug mesh ntawm phev bundles.(D) Tsim kom muaj ib lub network ntawm phev bundles.
Thaum ib tug poob ntawm cov phev diluted loaded mus rau hauv lub microfluidic ntaus ntawv thiab ib tug txaus tsim, cov phev beam raug soj ntsuam kom txav tawm tsam cov kev taw qhia ntawm cov ntws.Cov bundles haum snugly tiv thaiv phab ntsa ntawm microchannels, thiab lub taub hau dawb nyob rau hauv thawj feem ntawm cov bundles haum snugly tiv thaiv lawv (video 5).Lawv kuj lo rau txhua qhov chaw nyob ruaj khov hauv lawv txoj kev, xws li cov khib nyiab, kom tiv thaiv tau los ntawm qhov tam sim no.Thaum lub sij hawm, cov tufts no ua ntev filaments cuab lwm yam spermatozoa thiab luv luv tufts (Video 6).Raws li qhov ntws pib qeeb, cov kab phev ntev pib tsim ib lub network ntawm cov phev kab (Video 7; Daim duab 2).
Ntawm qhov nrawm nrawm (V> 33 µm / s), cov kauv txav ntawm cov xov tau nce ntxiv raws li kev sim ntes ntau tus phev ua cov pob khoom zoo dua tiv thaiv lub zog ntawm qhov ntws. Ntawm qhov nrawm nrawm (V> 33 µm / s), cov kauv txav ntawm cov xov tau nce ntxiv raws li kev sim ntes ntau tus phev ua cov pob khoom zoo dua tiv thaiv lub zog ntawm qhov ntws. При высокой скорости потока (V > 33 мкм/с) спиралевидные движения нитей усиливаются, поскольку они сет поскольку они ятат ельных сперматозоидов, образующих пучки, которые лучше противостоят дрейфующей силе потока. Ntawm qhov siab txaus (V> 33 µm / s), helical txav ntawm cov strands nce thaum lawv sim ntes ntau tus neeg phev ua cov pob uas zoo dua tuaj yeem tiv thaiv lub zog ntawm qhov ntws.在高流速(V > 33 µm/s) 时，螺纹的螺旋运动增加，以试图捕捉许多形成束的单个精匐，以试图捕捉许多形成束漂移力.在高流速 (v> 33 µm/s) 时，的螺旋运动增加，以试图 许多形成束单个精子漂移力 . . . . . . . . При высоких скоростях потока (V > 33 мкм/с) спиральное движение нитей увеличивается в попытке захватит матозоидов, образующих пучки, чтобы лучше сопротивляться силам дрейфа потока. Ntawm qhov siab txaus (V> 33 µm / s), helical txav ntawm cov filaments nce nyob rau hauv ib qho kev sim mus ntes ntau tus spermatozoa ua bundles kom zoo dua tiv thaiv drift quab yuam ntawm cov ntws.Lawv kuj tau sim txuas cov microchannels rau ntawm phab ntsa.
Cov kab phev tau raug txheeb xyuas raws li pawg ntawm cov phev taub hau thiab curling tails siv lub teeb microscopy (LM).Cov phev bundles nrog ntau cov aggregates kuj tau txheeb xyuas tias yog twisted taub hau thiab flagellar aggregates, ntau fused phev tails, phev taub hau txuas rau tus Tsov tus tw, thiab cov phev taub hau nrog khoov nuclei li ntau fused nuclei.kis tau tus mob electron microscopy (TEM).Scanning electron microscopy (SEM) qhia tau hais tias cov phev bundles tau sheathed aggregates ntawm cov phev taub hau thiab cov phev aggregates pom ib tug txuas network ntawm qhwv tails.
Lub morphology thiab ultrastructure ntawm spermatozoa, tsim cov spermatozoa bundles tau kawm siv lub teeb microscopy (ib nrab seem), scanning electron microscopy (SEM) thiab kis electron microscopy (TEM), phev smears tau stained nrog acridine txiv kab ntxwv thiab tshuaj xyuas siv epicopyes.
Cov phev smear staining nrog acridine txiv kab ntxwv (Daim duab 3B) qhia tias cov phev taub hau tau daig ua ke thiab npog nrog cov ntaub ntawv secretory, uas ua rau tsim cov tufts loj (Fig. 3D).Cov phev bundles muaj cov phev aggregates nrog ib tug network ntawm txuas tails (Fig. 4A-C).Cov phev bundles yog tsim los ntawm cov tails ntawm ntau spermatozoa daig ua ke (Fig. 4D).Secrets (Fig. 4E, F) npog lub taub hau ntawm spermatozoa bundles.
Tsim cov spermatozoa bundle Siv theem sib piv microscopy thiab phev smears stained nrog acridine txiv kab ntxwv, qhia tias lub taub hau ntawm spermatozoa lo ua ke.(A) Kev tsim cov phev thaum ntxov pib nrog cov phev (lub voj voog dawb) thiab peb cov phev (lub voj voog daj), nrog cov kauv pib ntawm tus Tsov tus tw thiab xaus ntawm lub taub hau.(B) Photomicrograph ntawm ib tug phev smear stained nrog acridine txiv kab ntxwv qhia adherent phev taub hau ( xub).Qhov tso tawm npog lub taub hau.Magnification × 1000. (C) Kev loj hlob ntawm cov nqaj loj thauj los ntawm kev ntws hauv microfluidic channel (siv lub koob yees duab ceev ntawm 950 fps).(D) Micrograph ntawm cov phev smear stained nrog acridine txiv kab ntxwv uas qhia loj tufts ( xub xub).Loj: × 200.
Scanning electron micrograph ntawm cov phev kab thiab cov phev smear stained nrog acridine txiv kab ntxwv.(A, B, D, E) yog cov xim digital scanning electron micrographs ntawm spermatozoa, thiab C thiab F yog micrographs ntawm acridine txiv kab ntxwv stained phev smears uas qhia txuas ntawm ntau cov spermatozoa wrapping lub vas sab caudal.(AC) Sperm aggregates yog qhia raws li lub network ntawm cov tails txuas (xub xub).(D) Adhesion ntawm ob peb spermatozoa (nrog rau cov khoom nplaum, cov duab paj yeeb, xub) qhwv ib ncig ntawm tus Tsov tus tw.(E thiab F) Sperm head aggregates (pointers) covered with adhesive material (pointers).Cov spermatozoa tsim cov pob khoom nrog ntau lub vortex zoo li cov qauv (F).(C) × 400 thiab (F) × 200 magnification.
Siv cov khoom siv hluav taws xob xa hluav taws xob, peb pom tias cov phev bundles tau txuas cov tails (Fig. 6A, C), taub hau txuas rau cov tails (Fig. 6B), los yog lub taub hau txuas rau tails (Fig. 6D).Lub taub hau ntawm cov spermatozoa nyob rau hauv cov pob khoom yog nkhaus, nthuav tawm nyob rau hauv seem ob thaj tsam nuclear (Fig. 6D).Nyob rau hauv cov pob qij txha, cov spermatozoa muaj lub taub hau twisted nrog ob thaj tsam nuclear thiab ntau thaj tsam flagellar (Fig. 5A).
Cov xim hluav taws xob digital micrograph qhia txog kev sib txuas ntawm cov tails hauv cov phev pob thiab cov khoom siv agglutinating txuas cov phev taub hau.(A) Txuas tus Tsov tus tw ntawm ntau tus spermatozoa.Daim ntawv ceeb toom yuav ua li cas tus Tsov tus tw zoo li hauv ob qho tib si portrait ( xub xub) thiab toj roob hauv pes ( xub) qhov projections.(B) Lub taub hau ( xub xub) ntawm cov phev txuas nrog tus Tsov tus tw ( xub xub).(C) Ob peb phev tails ( xub) txuas.(D) Agglutination khoom (AS, xiav) txuas plaub lub phev taub hau (ntsuab).
Kev tshuaj ntsuam xyuas electron microscopy tau siv los kuaj xyuas cov phev taub hau hauv cov phev bundles uas npog nrog cov kua nplaum los yog daim nyias nyias (Daim duab 6B), qhia tias cov phev bundles tau anchored los ntawm extracellular khoom.Cov khoom siv agglutinated tau tsom mus rau hauv cov phev taub hau (lub taub hau zoo li jellyfish sib dhos; Fig. 5B) thiab nthuav dav distally, muab lub ntsej muag daj zoo nkauj nyob rau hauv fluorescence microscopy thaum stained nrog acridine txiv kab ntxwv (Fig. 6C).Cov tshuaj no pom meej meej nyob rau hauv lub tshuab kuaj kab mob thiab suav tias yog ib qho kev sib khi.Semi-thin seem (Fig. 5C) thiab cov phev smears stained nrog acridine txiv kab ntxwv pom cov phev bundles uas muaj densely packed taub hau thiab curled tails (Fig. 5D).
Ntau yam photomicrographs qhia txog kev sib sau ntawm cov phev taub hau thiab folded tails siv ntau txoj kev.(A) Hla-sectional digital xim kis tau tus mob electron micrograph ntawm cov phev pob uas qhia lub taub hau phev coiled nrog ob feem ntawm cov nucleus (xiav) thiab ntau qhov chaw flagellar (ntsuab).(B) Cov xim digital scanning electron micrograph qhia ib pawg ntawm jellyfish zoo li phev taub hau (xub xub) uas tshwm sim los npog.(C) Semi-thin seem uas qhia cov phev taub hau (cov xub) thiab curled tails ( xub).(D) Micrograph ntawm cov phev smear stained nrog acridine txiv kab ntxwv qhia aggregates ntawm phev taub hau ( xub) thiab curled adherent tails ( xub).Nco ntsoov tias ib yam khoom nplaum (S) npog lub taub hau ntawm cov phev spermatozoon.(D) × 1000 magnification.
Siv kev xa xov hluav taws xob microscopy (Fig. 7A), nws kuj tau muab sau tseg tias cov phev taub hau tau twisted thiab cov nuclei muaj ib tug kauv, raws li tau lees paub los ntawm cov phev smears stained nrog acridine txiv kab ntxwv thiab tshuaj xyuas siv fluorescence microscopy (Fig. 7B).
(A) Cov xim digital kis tau tus mob electron micrograph thiab (B) Acridine txiv kab ntxwv stained phev smear qhia coiled hau thiab txuas ntawm cov phev taub hau thiab tails ( xub).(B) × 1000 magnification.
Qhov kev tshawb pom nthuav yog tias Sharkazi cov phev sib sau ua ke los tsim cov pob khoom txawb.Cov khoom ntawm cov pob khoom no tso cai rau peb nkag siab txog lawv lub luag haujlwm hauv kev nqus thiab khaws cia cov spermatozoa hauv SST.
Tom qab mating, cov phev nkag mus rau hauv qhov chaw mos thiab raug kev xaiv khaus, ua rau tsuas yog cov phev txwv nkag mus rau SST15,16.Txog niaj hnub no, cov txheej txheem uas cov phev nkag mus thiab tawm ntawm SST tsis meej.Hauv cov nqaij qaib, cov spermatozoa khaws cia rau hauv SST rau lub sijhawm ncua ntawm 2 txog 10 lub lis piam, nyob ntawm hom 6.Kev tsis sib haum xeeb tseem nyob ntawm qhov xwm txheej ntawm cov phev thaum khaws cia hauv SST.Puas yog lawv nyob hauv kev txav los yog so?Hauv lwm lo lus, cov phev hlwb ua li cas tswj lawv txoj haujlwm hauv SST kom ntev?
Forman4 qhia tias SST qhov chaw nyob thiab ejection tuaj yeem piav qhia txog cov phev motility.Cov kws sau ntawv xav tias cov phev tswj lawv txoj haujlwm los ntawm kev ua luam dej tawm tsam cov kua dej tsim los ntawm SST epithelium thiab cov phev raug tshem tawm ntawm SST thaum lawv qhov nrawm poob qis dua qhov uas lawv pib txav rov qab vim tsis muaj zog.Zaniboni5 tau lees paub tias muaj aquaporins 2, 3, thiab 9 nyob rau hauv apical feem ntawm SST epithelial hlwb, uas tej zaum yuav indirectly txhawb Foreman tus phev cia qauv.Hauv txoj kev tshawb fawb tam sim no, peb pom tias yuav luag ib nrab ntawm Sharkashi's spermatozoa qhia qhov zoo rheology nyob rau hauv cov kua dej ntws, thiab cov phev agglutinated ua rau cov phev spermatozoa qhia qhov zoo rheology, txawm tias agglutination slows lawv.Yuav ua li cas cov phev hlwb mus txog tus noog lub fallopian raj mus rau qhov chaw ntawm fertilization yog tsis to taub tag nrho.Hauv cov tsiaj txhu, cov kua dej follicular chemoattracts spermatozoa.Txawm li cas los xij, chemoattractants tau ntseeg tias yuav coj cov spermatozoa mus rau qhov deb7.Yog li ntawd, lwm cov txheej txheem yog lub luag haujlwm rau kev thauj cov phev.Lub peev xwm ntawm cov phev mus rau taw qhia thiab ntws tawm tsam cov kua dej hauv lub raj tso tawm tom qab kev sib tw tau raug tshaj tawm tias yog qhov tseem ceeb hauv kev tsom cov phev hauv cov nas.Parker 17 qhia tias spermatozoa hla lub oviducts los ntawm kev ua luam dej tawm tsam ciliary tam sim no hauv cov noog thiab cov tsiaj reptiles.Txawm hais tias nws tsis tau sim ua kom pom hauv cov noog, Adolphi18 yog thawj zaug pom tias cov phev avian muab cov txiaj ntsig zoo thaum cov txheej txheem nyias nyias ntawm cov ntaub npog thiab cov swb yog tsim nrog ib daim ntaub lim dej.Rheology.Hino thiab Yanagimachi [19] muab tso rau nas zes qe menyuam-tubal-uterine complex nyob rau hauv lub nplhaib perfusion thiab txhaj 1 µl ntawm tus number case rau hauv isthmus kom pom cov kua dej ntws hauv cov hlab ntsha.Lawv pom muaj zog heev ntawm contraction thiab so nyob rau hauv lub fallopian raj, nyob rau hauv uas tag nrho cov number case balls tsis tu ncua mus rau lub ampulla ntawm lub fallopian raj.Cov kws sau ntawv hais txog qhov tseem ceeb ntawm cov kua dej ntws los ntawm qhov qis mus rau sab sauv qaum rau cov phev uplift thiab fertilization.Brillard20 tau tshaj tawm tias nyob rau hauv qaib thiab qaib ntxhw, spermatozoa tsiv teb tsaws los ntawm kev txav mus los ntawm qhov chaw mos nkag, qhov chaw uas lawv khaws cia, mus rau qhov chaw mos ntawm lub tsev menyuam, qhov chaw uas lawv khaws cia.Txawm li cas los xij, qhov kev txav no tsis tas yuav tsum tau nyob nruab nrab ntawm lub tsev menyuam thiab cov infundibulum vim tias cov spermatozoa raug thauj los ntawm kev txav mus los.Paub txog cov lus pom zoo dhau los no thiab cov txiaj ntsig tau txais hauv txoj kev tshawb fawb tam sim no, nws tuaj yeem xav tias lub peev xwm ntawm cov phev tuaj yeem txav mus rau sab saud (rheology) yog ib qho ntawm cov khoom ntawm cov txheej txheem xaiv raws li.Qhov no txiav txim siab qhov kev tso ntawm spermatozoa los ntawm qhov chaw mos thiab lawv nkag mus rau hauv CCT rau kev cia.Raws li Forman4 tau hais tseg, qhov no kuj tseem yuav pab txhawb cov txheej txheem ntawm cov phev nkag mus rau SST thiab nws qhov chaw nyob rau lub sijhawm thiab tom qab ntawd tawm thaum lawv qhov nrawm pib qeeb.
Ntawm qhov tod tes, Matsuzaki thiab Sasanami 21 tau tawm tswv yim tias avian spermatozoa tau hloov pauv hloov ntawm dormancy mus rau motility hauv cov txiv neej thiab poj niam cov kab mob.Inhibition ntawm cov phev motility nyob rau hauv lub SST tau npaj los piav qhia lub sij hawm ntev cia ntawm phev thiab ces rejuvenation tom qab tawm hauv SST.Nyob rau hauv cov xwm txheej hypoxic, Matsuzaki li al.1 tau tshaj tawm ntau lawm thiab tso tawm lactate hauv SST, uas yuav ua rau inhibition ntawm cov phev motility.Nyob rau hauv cov ntaub ntawv no, qhov tseem ceeb ntawm phev rheology yog xav txog nyob rau hauv kev xaiv thiab nqus cov spermatozoa, thiab tsis nyob rau hauv lawv cia.
Cov phev agglutination qauv yog suav hais tias yog ib qho kev piav qhia rau lub sijhawm ntev ntawm cov phev khaws cia hauv SST, vim qhov no yog ib qho qauv ntawm cov phev khaws cia hauv cov nqaij qaib2,22,23.Bakst et al.2 tau pom tias feem ntau cov spermatozoa adhered rau ib leeg, tsim fascicular aggregates, thiab ib leeg spermatozoa tsis tshua pom nyob rau hauv quail CCM.Ntawm qhov tod tes, Wen et al.24 pom cov spermatozoa tawg ntau dua thiab tsawg dua spermatozoa tufts hauv SST lumen hauv qaib.Raws li cov kev soj ntsuam no, nws tuaj yeem xav tias qhov kev xav tau rau cov phev agglutination txawv ntawm cov noog thiab nruab nrab ntawm cov spermatozoa hauv tib lub ejaculate.Tsis tas li ntawd, Van Krey et al.9 qhia tias kev sib cais tsis sib xws ntawm agglutinated spermatozoa yog lub luag haujlwm rau kev nkag mus ntawm cov spermatozoa maj mam mus rau hauv lub lumen ntawm lub raj mis.Raws li qhov kev xav no, spermatozoa nrog qis agglutination peev xwm yuav tsum raug tshem tawm ntawm SST ua ntej.Hauv cov ntsiab lus no, lub peev xwm ntawm spermatozoa rau agglutinate tuaj yeem yog qhov cuam tshuam rau qhov tshwm sim ntawm kev sib tw phev hauv cov noog qias neeg.Tsis tas li ntawd, qhov ntev ntawm cov phev agglutinated dissociates, ntev fertility yog khaws cia.
Txawm hais tias spermatozoa aggregation thiab aggregation rau hauv pob khoom tau pom nyob rau hauv ntau qhov kev tshawb fawb 2,22,24, lawv tsis tau piav qhia meej vim qhov nyuaj ntawm lawv qhov kev soj ntsuam kinematic hauv SST.Ntau qhov kev sim tau ua los kawm cov phev agglutination hauv vitro.Kev nthuav dav tab sis ntu ntu tau pom thaum cov hlau nyias raug tshem tawm ntawm cov dangling noob poob.Qhov no ua rau lub fact tias ib tug elongated npuas protrudes los ntawm qhov poob, ua raws li lub caj pas seminal.Vim muaj kev txwv 3D thiab lub sij hawm luv luv kom qhuav, tag nrho cov block sai poob rau hauv disrepair9.Hauv txoj kev tshawb fawb tam sim no, siv Sharkashi qaib thiab microfluidic chips, peb tuaj yeem piav qhia seb cov tufts no ua li cas thiab lawv txav mus li cas.Sperm bundles tsim tam sim ntawd tom qab sau cov phev thiab pom tau txav mus rau hauv ib lub kauv, qhia qhov zoo rheology thaum tam sim no ntws.Tsis tas li ntawd, thaum saib macroscopically, cov phev bundles tau pom los ua kom cov linearity ntawm motility piv rau cov spermatozoa cais.Qhov no qhia tias phev agglutination yuav tshwm sim ua ntej SST nkag mus thiab cov phev ntau lawm tsis txwv rau ib cheeb tsam me me vim muaj kev ntxhov siab raws li tau hais dhau los (Tingari thiab Lake12).Thaum lub sij hawm tuft tsim, cov spermatozoa ua luam dej nyob rau hauv synchrony kom txog rau thaum lawv tsim ib qho kev sib tshuam, ces lawv cov tails qhwv ib ncig thiab lub taub hau ntawm cov spermatozoon tseem dawb, tab sis tus Tsov tus tw thiab distal ib feem ntawm cov spermatozoon stick ua ke nrog ib tug nplaum.Yog li ntawd, lub taub hau dawb ntawm ligament yog lub luag haujlwm rau kev txav, rub tus so ntawm ligament.Kev kuaj xyuas electron microscopy ntawm cov phev bundles pom cov phev taub hau uas txuas nrog ntau cov khoom nplaum, qhia tias cov phev taub hau tau txuas rau hauv cov khoom so, uas yuav tshwm sim tom qab mus txog qhov chaw cia (SST).
Thaum cov phev smear yog stained nrog acridine txiv kab ntxwv, extracellular nplaum cov ntaub ntawv nyob ib ncig ntawm cov phev hlwb yuav pom nyob rau hauv ib tug fluorescent microscope.Cov tshuaj no tso cai rau cov phev bundles ua raws li thiab cling rau tej qhov chaw nyob ib puag ncig los yog hais kom lawv tsis drift nrog cov nyob ib ncig ntawm ntws.Yog li, peb cov kev soj ntsuam qhia lub luag haujlwm ntawm spermatozoa adhesion nyob rau hauv daim ntawv ntawm mobile bundles.Lawv lub peev xwm ua luam dej tawm tsam tam sim no thiab lo rau qhov chaw nyob ze tso cai rau cov phev nyob ntev dua hauv SST.
Rothschild25 siv lub koob yees duab hemocytometry los kawm txog kev faib tawm ntawm cov phev bovine hauv qhov poob ntawm kev ncua, noj photomicrographs los ntawm lub koob yees duab nrog ob txoj kab ntsug thiab kab rov tav optical axis ntawm lub tshuab tsom.Cov txiaj ntsig tau pom tias cov spermatozoa tau nyiam mus rau saum lub chamber.Cov kws sau ntawv qhia tias tej zaum yuav muaj kev cuam tshuam hydrodynamic ntawm cov phev thiab qhov chaw.Ua raws li qhov no rau hauv tus account, nrog rau lub peev xwm ntawm Sharkashi qaib cov phev los ua cov nplaum nplaum, nws yuav ua rau kom cov phev yuav ua raws li phab ntsa SST thiab khaws cia rau lub sijhawm ntev.
Bccetti thiab Afzeliu26 qhia tias cov phev glycocalyx yuav tsum tau rau gamete paub thiab agglutination.Forman10 pom tias hydrolysis ntawm α-glycosidic bonds hauv glycoprotein-glycolipid txheej los ntawm kev kho avian phev nrog neuraminidase ua rau txo qis fertility yam tsis muaj kev cuam tshuam cov phev motility.Cov kws sau ntawv qhia tias cov nyhuv ntawm neuraminidase ntawm glycocalyx cuam tshuam cov phev sequestration ntawm lub tsev menyuam-qhov chaw mos, yog li txo qis fertility.Lawv cov kev soj ntsuam tsis tuaj yeem tsis quav ntsej txog qhov ua tau tias kev kho neuraminidase tuaj yeem txo cov phev thiab oocyte paub.Forman thiab Engel10 pom tias kev xeeb tub tau txo qis thaum cov hens tau inseminated intravaginally nrog phev kho nrog neuraminidase.Txawm li cas los xij, IVF nrog neuraminidase kho phev tsis cuam tshuam rau fertility piv rau kev tswj qaib.Cov kws sau ntawv tau xaus lus tias kev hloov pauv ntawm glycoprotein-glycolipid txheej nyob ib ncig ntawm cov phev membrane txo cov phev kom fertilize los ntawm impairing sequestration ntawm cov phev nyob rau hauv lub utero-vaginal junction, uas nyob rau hauv lem nce phev poob vim lub ceev ntawm lub qe ntawm lub tsev me nyuam hlws ris, tab sis tsis cuam tshuam.
Hauv qaib cov txwv Bakst thiab Bauchan 11 pom cov hlwv me me thiab cov kab mob me me hauv cov lumen ntawm SST thiab pom tias qee cov granules tau fused nrog cov phev membrane.Cov kws sau ntawv qhia tias cov kev sib raug zoo no tuaj yeem ua rau lub sijhawm ntev ntawm cov spermatozoa hauv SST.Txawm li cas los xij, cov kws tshawb fawb tsis tau qhia meej txog qhov chaw ntawm cov khoom no, txawm tias lawv tau zais los ntawm CCT epithelial hlwb, tsim thiab tso tawm los ntawm cov txiv neej kev ua me nyuam, lossis tsim los ntawm cov phev nws tus kheej.Tsis tas li ntawd, cov khoom no yog lub luag haujlwm rau agglutination.Grützner et al27 tau tshaj tawm tias cov kab mob epididymal epithelial tsim thiab tso tawm cov protein tshwj xeeb uas yuav tsum tau tsim los ntawm ib qho pore seminal tracts.Cov kws sau ntawv kuj tau tshaj tawm tias qhov kev tawg ntawm cov pob khoom no nyob ntawm kev sib cuam tshuam ntawm cov proteins epididymal.Nixon et al28 pom tias adnexa secrete protein, acidic cysteine-nplua nuj osteonectin;SPARC koom nrog hauv kev tsim cov phev tufts hauv luv-beaked echidnas thiab platypuses.Kev tawg ntawm cov kab teeb no cuam tshuam nrog kev poob ntawm cov protein no.
Nyob rau hauv txoj kev tshawb no tam sim no, kev tsom xam ultrastructural siv electron microscopy pom tias cov spermatozoa adhered mus rau ib tug loj npaum li cas ntawm cov khoom ntom.Cov tshuaj no tau xav tias yog lub luag haujlwm rau lub agglutination uas condenses ntawm thiab nyob ib ncig ntawm lub taub hau adherent, tab sis nyob rau hauv qis concentrations nyob rau hauv lub Tail cheeb tsam.Peb xav tias cov tshuaj agglutinating no tau tawm los ntawm cov txiv neej lub cev xeeb tub (epididymis lossis vas deferens) nrog rau cov phev, txij li thaum peb feem ntau pom cov phev cais tawm ntawm cov qog ntshav thiab cov ntshav ntshav thaum lub sijhawm ejaculation.Nws tau raug tshaj tawm tias raws li avian spermatozoa hla dhau ntawm epididymis thiab vas deferens, lawv tau txais kev hloov pauv ntsig txog kev loj hlob uas txhawb nqa lawv lub peev xwm los khi cov proteins thiab tau txais cov ntshav plasma lemma-koom nrog glycoproteins.Qhov kev pheej hmoo ntawm cov proteins no ntawm cov phev hauv cov phev hauv SST qhia tias cov proteins no tuaj yeem cuam tshuam rau kev tau txais cov phev membrane ruaj khov 30 thiab txiav txim siab lawv fertility 31 .Ahammad et al32 tau tshaj tawm tias cov spermatozoa tau txais los ntawm ntau qhov chaw ntawm cov txiv neej kev ua me nyuam (los ntawm cov qog mus rau qhov distal vas deferens) tau pom tias muaj kev vam meej nyob rau hauv cov kua dej cia, tsis hais qhov kub thiab txias, thiab kev muaj sia nyob hauv qaib kuj nce hauv cov hlab ntsha tom qab kev sib txuas ntxiv.
Sharkashi qaib phev tufts muaj cov yam ntxwv thiab kev ua haujlwm ntau dua li lwm hom tsiaj xws li echidnas, platypuses, ntoo nas, mos lwj nas, thiab guinea npua.Hauv cov qaib sharkasi, tsim cov spermatozoa bundles txo lawv cov kev ua luam dej nrawm piv rau ib leeg spermatozoa.Txawm li cas los xij, cov pob no tau nce qhov feem pua ​​​​ntawm rheologically zoo spermatozoa thiab nce lub peev xwm ntawm spermatozoa kom stabilize lawv tus kheej hauv ib puag ncig dynamic.Yog li, peb cov txiaj ntsig tau lees paub cov lus qhia yav dhau los uas cov phev agglutination hauv SST yog txuam nrog cov phev cia ntev.Peb kuj xav tias qhov kev xav ntawm cov phev los ua cov tufts tuaj yeem tswj cov phev poob hauv SST, uas tuaj yeem hloov cov txiaj ntsig ntawm kev sib tw phev.Raws li qhov kev xav no, spermatozoa nrog qis agglutination muaj peev xwm tso tawm SST ua ntej, thaum spermatozoa nrog siab agglutination muaj peev xwm tsim tau feem ntau ntawm cov xeeb ntxwv.Kev tsim cov phev ib leeg-pore phev bundles muaj txiaj ntsig thiab cuam tshuam rau niam txiv-me nyuam piv, tab sis siv cov txheej txheem sib txawv.Hauv echidnas thiab platypuses, cov spermatozoa tau teem rau tib lub sijhawm kom nce qhov nrawm ntawm lub nqaj.Cov pob khoom ntawm echidnas txav mus txog peb zaug sai dua li cov phev spermatozoa.Nws ntseeg tau tias qhov tsim ntawm cov phev tufts hauv echidnas yog ib qho kev hloov pauv hloov mus rau kev tswj hwm, txij li cov poj niam yog promiscuous thiab feem ntau yog khub nrog ob peb txiv neej.Yog li ntawd, spermatozoa los ntawm txawv ejaculates sib tw hnyav rau fertilization ntawm lub qe.
Agglutinated spermatozoa ntawm sharkasi qaib yog ib qho yooj yim rau kev pom kev siv theem sib piv microscopy, uas yog suav tias yog qhov zoo vim nws tso cai yooj yim kawm txog tus cwj pwm ntawm spermatozoa hauv vitro.Cov txheej txheem los ntawm kev tsim cov phev tuft txhawb kev yug me nyuam hauv sharkasi qaib kuj txawv ntawm qhov uas pom nyob rau hauv ib co placental mammals sawv cev rau kev koom tes phev cwj pwm xws li ntoo nas, qhov twg ib co spermatozoa ncav cuag lub qe, pab lwm tus neeg muaj feem xyuam mus txog thiab puas lawv cov qe.los ua pov thawj koj tus kheej.altruistic tus cwj pwm.Self-fertilization 34. Lwm qhov piv txwv ntawm kev sib koom tes ntawm cov spermatozoa tau pom nyob rau hauv cov nas mos mos, qhov chaw spermatozoa muaj peev xwm txheeb xyuas thiab ua ke nrog cov genetic spermatozoa feem ntau thiab tsim cov pab pawg sib koom ua ke kom lawv nrawm dua piv rau cov spermatozoa tsis sib xws.
Cov txiaj ntsig tau txais hauv qhov kev tshawb fawb no tsis cuam tshuam nrog Foman txoj kev xav ntawm kev cia cov spermatozoa mus ntev hauv SWS.Cov kws tshawb fawb tau tshaj tawm tias cov phev hlwb txuas ntxiv mus rau hauv cov dej ntws ntawm cov hlwb epithelial nyob rau hauv SST rau lub sijhawm ntev, thiab tom qab qee lub sijhawm, cov khw muag khoom hluav taws xob ntawm cov phev hlwb tau ploj mus, ua rau txo qis, uas tso cai rau kev tshem tawm cov khoom hnyav molecular.lub zog ntawm spermatozoa nrog cov kua dej ntws los ntawm lub lumen ntawm SST Cov kab noj hniav ntawm cov hlab ntsha fallopian.Hauv txoj kev tshawb fawb tam sim no, peb tau pom tias ib nrab ntawm cov phev ib leeg tau pom tias muaj peev xwm ua luam dej tawm tsam cov dej ntws, thiab lawv qhov adhesion hauv cov pob no tau nce lawv lub peev xwm los qhia qhov zoo rheology.Tsis tas li ntawd, peb cov ntaub ntawv zoo ib yam nrog cov Matsuzaki li al.1 leej twg tau tshaj tawm tias kev tso kua mis ntau ntxiv hauv SST tuaj yeem cuam tshuam cov phev motility.Txawm li cas los xij, peb cov txiaj ntsig tau piav qhia txog kev tsim cov phev motile ligaments thiab lawv tus cwj pwm rheological nyob rau hauv lub xub ntiag ntawm ib puag ncig dynamic nyob rau hauv ib tug microchannel nyob rau hauv ib qho kev sim los qhia lawv tus cwj pwm hauv SST.Kev tshawb fawb yav tom ntej yuav tsom mus rau kev txiav txim siab cov tshuaj muaj pes tsawg leeg thiab keeb kwm ntawm tus neeg sawv cev agglutinating, uas yuav pab tau cov kws tshawb fawb tsim txoj hauv kev tshiab los khaws cov kua phev thiab ua kom lub sijhawm fertility.
Kaum tsib 30 lub lis piam-hnub liab qab-caj dab txiv neej sharkasi (homozygous dominant; Na Na) raug xaiv los ua cov phev pub dawb hauv txoj kev tshawb no.Cov noog tau raug txhawb nqa ntawm Kev Tshawb Fawb Poultry Farm ntawm Kws Qhia Txog Kev Ua Liaj Ua Teb, Ashit University, Ashit Governorate, Egypt.Cov noog tau nyob hauv ib lub tawb (30 x 40 x 40 cm), raug rau qhov kev pab cuam lub teeb (16 teev ntawm lub teeb thiab 8 teev tsaus ntuj) thiab noj cov zaub mov uas muaj 160 g ntawm cov protein crude, 2800 kcal ntawm metabolizable zog, 35 g ntawm calcium txhua.5 grams muaj phosphorus ib kilogram ntawm cov khoom noj.
Raws li cov ntaub ntawv 36, 37, cov phev tau sau los ntawm cov txiv neej los ntawm kev zaws plab.Tag nrho ntawm 45 cov qauv phev tau sau los ntawm 15 tus txiv neej dhau 3 hnub.Cov phev (n = 15 / hnub) tau diluted tam sim ntawd 1: 1 (v: v) nrog Belsville Poultry Semen Diluent, uas muaj poov tshuaj diphosphate (1.27 g), monosodium glutamate monohydrate (0.867 g), fructose (0.5 d) anhydrous sodium.acetate (0.43 g), tris (hydroxymethyl) aminomethane (0.195 g), potassium citrate monohydrate (0.064 g), potassium monophosphate (0.065 g), magnesium chloride (0.034 g) thiab H2O (100 ml), pH = 7, 5, osmolarity / 3.Cov phev diluted tau kuaj thawj zaug hauv lub teeb tsom iav kom paub tseeb tias cov phev zoo (nqus dej) thiab tom qab ntawd muab tso rau hauv dej da dej ntawm 37 ° C kom txog thaum siv li ib nrab ib teev tom qab sau.
Lub kinematics thiab rheology ntawm spermatozoa tau piav qhia siv lub kaw lus ntawm cov khoom siv microfluidic.Cov qauv phev tau ntxiv mus rau 1:40 hauv Beltsville Avian Semen Diluent, thauj khoom rau hauv cov khoom siv microfluidic (saib hauv qab), thiab cov kev ntsuas kinetic tau txiav txim siab siv Computerized Semen Analysis (CASA) system yav dhau los tsim rau microfluidics characterization.ntawm kev txav mus los ntawm cov spermatozoa hauv cov kua dej (Department of Mechanical Engineering, Kws qhia ntawv ntawm Engineering, Assiut University, Egypt).Lub plugin tuaj yeem rub tawm ntawm: http://www.assiutmicrofluidics.com/research/casa39.Curve tshaj tawm (VCL, μm / s), linear tshaj tawm (VSL, μm / s) thiab nruab nrab trajectory tshaj tawm (VAP, μm / s) tau ntsuas.Cov yeeb yaj kiab ntawm spermatozoa tau raug coj los siv qhov hloov pauv ntawm Optika XDS-3 theem sib piv lub tshuab ntsuas (nrog 40x lub hom phiaj) txuas nrog lub koob yees duab Tucson ISH1000 ntawm 30 fps rau 3 s.Siv CASA software los kawm tsawg kawg peb thaj chaw thiab 500 phev trajectories ib tus qauv.Cov yeeb yaj kiab kaw tau ua tiav siv CASA hauv tsev.Lub ntsiab lus ntawm motility nyob rau hauv CASA plug-in yog raws li nyob rau hauv tus da dej ceev ntawm cov phev piv rau cov ndlwg tus nqi, thiab tsis suav nrog rau lwm yam tsis xws li ib sab-rau-sab zog, raws li qhov no tau pom tias yuav txhim khu kev qha nyob rau hauv cov kua.Rheological motion yog piav raws li kev txav ntawm cov phev hlwb tawm tsam cov kev taw qhia ntawm cov kua dej.Spermatozoa nrog rheological zog tau muab faib los ntawm tus naj npawb ntawm motile spermatozoa;spermatozoa uas nyob so thiab convectively txav spermatozoa raug tshem tawm ntawm suav.
Tag nrho cov tshuaj siv tau los ntawm Elgomhoria Pharmaceuticals (Cairo, Egypt) tshwj tsis yog sau tseg.Cov cuab yeej tau tsim tawm raws li tau piav qhia los ntawm El-sherry li al.40 nrog qee qhov kev hloov kho.Cov ntaub ntawv siv los tsim cov microchannels suav nrog iav phaj (Howard Glass, Worcester, MA), SU-8-25 tsis zoo tiv thaiv (MicroChem, Newton, CA), diacetone cawv (Sigma Aldrich, Steinheim, Lub teb chaws Yelemees), thiab polyacetone.-184, Dow Corning, Midland, Michigan).Microchannels yog fabricated siv lithography mos.Ua ntej, lub ntsej muag tiv thaiv lub ntsej muag meej nrog qhov xav tau microchannel tsim tau luam tawm ntawm lub tshuab luam ntawv siab (Prismatic, Cairo, Egypt thiab Pacific Arts thiab Tsim, Markham, ON).Cov masters tau tsim siv iav daim hlau ua substrates.Cov phaj raug ntxuav hauv acetone, isopropanol thiab deionized dej thiab tom qab ntawd coated nrog 20 µm txheej ntawm SU8-25 los ntawm cov txheej txheej (3000 rpm, 1 min).Cov txheej SU-8 ces maj mam qhuav (65 ° C, 2 min thiab 95 ° C, 10 min) thiab raug UV hluav taws xob rau 50 s.Tom qab raug ci ntawm 65 ° C thiab 95 ° C rau 1 min thiab 4 min rau crosslink nthuav tawm SU-8 txheej, ua raws li kev loj hlob hauv diacetone cawv rau 6.5 min.Hard ci lub waffles (200 ° C rau 15 min) kom ntxiv dag zog rau SU-8 txheej.
PDMS tau npaj los ntawm kev sib xyaw cov monomer thiab hardener hauv qhov sib piv ntawm qhov hnyav ntawm 10: 1, tom qab ntawd degassed hauv lub tshuab nqus tsev desiccator thiab nchuav mus rau SU-8 lub ntsiab ncej.PDMS tau kho nyob rau hauv qhov cub (120 ° C, 30 min), tom qab ntawd cov channel raug txiav tawm, sib cais los ntawm tus tswv, thiab perforated kom tso cov raj xa mus rau ntawm qhov nkag thiab qhov hluav taws xob ntawm microchannel.Thaum kawg, PDMS microchannels tau txuas mus tas li mus rau lub tshuab ntsuas hluav taws xob siv lub tshuab hluav taws xob corona (Electro-Technic Products, Chicago, IL) raws li tau piav qhia lwm qhov.Lub microchannel siv hauv qhov kev tshawb fawb no ntsuas 200 µm × 20 µm (W × H) thiab yog 3.6 cm ntev.
Cov kua dej ntws los ntawm hydrostatic siab nyob rau hauv lub microchannel yog ua tiav los ntawm kev tswj cov kua dej hauv lub inlet reservoir saum qhov siab sib txawv Δh39 hauv qhov hluav taws xob reservoir (Daim duab 1).
qhov twg f yog tus coefficient ntawm kev sib txhuam, txhais raws li f = C / Re rau laminar ntws nyob rau hauv ib lub voj voog channel, qhov C yog qhov tas li nyob ntawm qhov sib piv ntawm cov channel, L yog qhov ntev ntawm microchannel, Vav yog qhov nruab nrab tshaj tawm hauv microchannel, Dh yog hydraulic txoj kab uas hla ntawm channel, g - acceleration ntawm lub ntiajteb txawj nqus.Siv qhov sib npaug no, qhov nruab nrab channel tshaj tawm tuaj yeem suav nrog siv cov kab zauv hauv qab no: