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Ukuzala kwezinyoni kuncike ekhonweni lazo lokugcina isidoda esanele esisebenzayo isikhathi eside kumashubhu okugcina isidoda (SST).Indlela eqondile lapho i-spermatozoa ingena ngayo, ihlale, futhi ishiye i-SST ihlala iyimpikiswano.Isidoda sezinkukhu zika-sharkasi sabonisa ukuthambekela okuphezulu kokuhlangana, senza izinyanda ze-filamentous mobile eziqukethe amaseli amaningi.Ngenxa yobunzima bokubheka ukuhamba nokuziphatha kwe-spermatozoa eshubhuni le-fallopian opaque, sasebenzisa idivayisi ye-microfluidic ene-microchannel cross-section efana ne-spermatozoa ukuze sihlole i-spermatozoa agglutination kanye ne-motility.Lolu cwaningo ludingida ukuthi izinqwaba zesidoda zakha kanjani, zihamba kanjani, kanye neqhaza lazo ekwandiseni ukuhlala kwesidoda ku-SST.Siphenye isivinini sesidoda nokuziphatha kwe-rheological lapho ukugeleza koketshezi kukhiqizwa ngaphakathi kwesiteshi se-microfluidic nge-hydrostatic pressure (izinga lokugeleza = 33 µm/s).I-spermatozoa ijwayele ukubhukuda ngokumelene nesimanje (i-rheology enhle) futhi isivinini se-spermatozoon bundle sincipha kakhulu uma kuqhathaniswa ne-spermatozoa eyodwa.Izinqwaba zesidoda ziye zabonwa zihamba ngokuzungeza futhi zanda ngobude nokuqina njengoba kuqashwa isidoda esisodwa. Izigaxa zesidoda zibonwe zisondela futhi zibambelela emaceleni eziteshini ezincane ze-microfluidic ukugwema ukushanelwa ngesivinini sokugeleza koketshezi> 33 µm/s. Izigaxa zesidoda zibonwe zisondela futhi zibambelela emaceleni eziteshini ezincane ze-microfluidic ukugwema ukushanelwa ngesivinini sokugeleza koketshezi> 33 µm/s. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрофлюидных каналов, чтобы избения 3 смета mkm / p. Izigaxa zesidoda ziye zaqashelwa ukuze zisondele futhi zinamathele ezindongeni eziseceleni zamashaneli ama-microfluidic ukuze zigweme ukukhukhuleka ngezinga lokugeleza kwamanzi >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 扫过. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрожидкостного канала, чтобы избежать 3 приложение 3 км/с. Izinqwaba zesidoda ziye zaqashelwa ukuze zisondele futhi zinamathele ezindongeni eziseceleni zesiteshi esincane se-microfluidic ukugwema ukukhukhulwa ukugeleza koketshezi ku->33 µm/s.Ukuskena nokudluliswa kwe-electron microscopy yembula ukuthi izinyanda zesidoda zazisekelwa izinto eziningi eziminyene.Idatha etholiwe ibonisa ukuhamba okuyingqayizivele kwe-spermatozoa yenkukhu yase-Sharkazi, kanye nekhono le-spermatozoa ukuze ihlanganise futhi yakhe ama-bundle eselula, okunikela ekuqondeni kangcono ukugcinwa kwesikhathi eside kwesidoda ku-SMT.
Ukuze kutholakale ukuvundiswa kwabantu nasezilwaneni eziningi, isidoda namaqanda kufanele kufike endaweni okuvundiswa kuyo ngesikhathi esifanele.Ngakho-ke, ukuzalanisa kufanele kwenzeke ngaphambi noma ngesikhathi sokuvuthwa.Ngakolunye uhlangothi, ezinye izilwane ezincelisayo, njengezinja, kanye nezinhlobo ezingezona ezincelisayo, njengezinambuzane, izinhlanzi, izilwane ezihuquzelayo, nezinyoni, zigcina isidoda ezithweni zazo zokuzala isikhathi eside kuze kube yilapho amaqanda azo eselungele ukuvundiswa (ukuvundiswa kwe-asynchronous 1).Izinyoni ziyakwazi ukugcina ukusebenza kwe-spermatozoa ekwazi ukuvundisa amaqanda amaviki angu-2-102.
Lesi yisici esiyingqayizivele esihlukanisa izinyoni kwezinye izilwane, njengoba kunikeza amathuba aphezulu okukhulelwa ngemva kokuzala okukodwa amasonto ambalwa ngaphandle kokuhlangana kanyekanye kanye ne-ovulation.Isitho esiyinhloko sokugcina isidoda, esibizwa ngokuthi i-sperm storage tubule (SST), sitholakala kumafinyila angaphakathi lapho kuhlangana khona isibeletho.Kuze kube manje, izindlela lapho isidoda singena, sihlala futhi siphuma ngaso ebhange lesidoda aziqondakali ngokugcwele.Ngokusekelwe ezifundweni ezedlule, imibono eminingi iye yabekwa phambili, kodwa akukho neyodwa yazo eqinisekisiwe.
I-Forman4 icabange ukuthi isidoda sigcina ukuhlala kwaso emgodini we-SST ngokunyakaza okuqhubekayo kwe-oscillatory ngokumelene nokuqondisa kokugeleza koketshezi ngamashaneli amaprotheni atholakala kumaseli e-SST epithelial (rheology).I-ATP iyancipha ngenxa yomsebenzi oqhubekayo we-flagellar odingekayo ukuze kugcinwe isidoda ku-lumen ye-SST futhi i-motility igcina iyancipha kuze kube yilapho isidoda sikhishwa onqenqemeni lwesidoda ngokugeleza koketshezi futhi siqale uhambo olusha lwehla ngeshubhu le-fallopian elikhuphukayo ukuze livundise isidoda.Iqanda (Forman4).Le modeli yokugcina isidoda isekelwa ukutholwa yi-immunocytochemistry ye-aquaporins 2, 3 kanye ne-9 ekhona kumaseli e-SST epithelial.Kuze kube manje, izifundo ze-rheology yesidoda sezinkukhu kanye nendima yayo ekugcinweni kwe-SST, ukukhethwa kwesidoda sowesifazane, kanye nokuncintisana kwesidoda.Ezinkukhuni, isidoda singena emomozini ngemva kokuhlangana ngokwemvelo, kodwa ngaphezu kuka-80% wesidoda sikhishwa esithweni sangasese ngemva nje kokukhwelana.Lokhu kusikisela ukuthi isitho sangasese sowesifazane siyindawo eyinhloko yokukhetha isidoda ezinyonini.Ukwengeza, kubikwe ukuthi ngaphansi kwe-1% yesidoda esivundisiwe esithweni sangasese sigcina ku-SSTs2.Ekufakweni kokwenziwa kwamatshwele esithweni sangasese sowesifazane, inani le-spermatozoa elifinyelela ku-SST livame ukukhula emahoreni angama-24 ngemva kokuzala.Kuze kube manje, indlela yokukhethwa kwesidoda phakathi nale nqubo ayicacile, futhi ukuhamba kwesidoda kungase kudlale indima ebalulekile ekuthathweni kwesidoda se-SST.Ngenxa yokuqina kwezindonga zamashubhu e-fallopian, kunzima ukuqapha ngokuqondile ukuhamba kwesidoda kumashubhu ezinyoni.Ngakho-ke, asinalo ulwazi oluyisisekelo lokuthi i-spermatozoa ishintshela ku-SST ngemva kokukhulelwa.
I-Rheology isanda kuqashelwa njengento ebalulekile elawula ukuthuthwa kwesidoda ezithweni zangasese ezincelisayo.Ngokusekelwe ekhonweni le-motile spermatozoa lokufuduka ngokuphambene, u-Zaferani et al8 usebenzise uhlelo lwe-corra microfluidic ukuze bahlukanise ngokuzenzakalelayo isidoda se-motile kumasampuli esidoda esibhaliwe.Lolu hlobo lokuhlungwa kwesidoda lubalulekile ekwelapheni ukungazali kanye nocwaningo lomtholampilo, futhi lukhethwa kunezindlela ezivamile ezithatha isikhathi kanye nomsebenzi onzima futhi ezingaphazamisa ukwakheka kwesidoda kanye nobuqotho besakhiwo.Kodwa-ke, kuze kube manje, azikho izifundo ezenziwe mayelana nomphumela wokuphuma kwezitho zangasese zezinkukhu ku-sperm motility.
Kungakhathaliseki ukuthi iyiphi indlela egcina isidoda esigcinwe ku-SST, abaphenyi abaningi baye baqaphela ukuthi i-permatozoa ehlala endaweni ihlanganisa ikhanda nekhanda ku-SST yezinkukhu ezingu-9, 10, izigwaca ezingu-2, nama-turkeys 11 ukuze zakhe izinqwaba zesidoda ezihlanganisiwe.Ababhali basikisela ukuthi kukhona ukuxhumana phakathi kwalokhu kuhlanganiswa nokugcinwa kwesikhathi eside kwe-spermatozoa ku-SST.
I-Tikiri kanye neLake12 babike ukuhlobana okuqinile phakathi kwe-spermatozoa endlaleni eyamukela isidoda yenkukhu futhi babuza ukuthi ingabe i-avian spermatozoa agglutinate ngendlela efanayo ne-mammalian spermatozoa.Bakholelwa ukuthi ukuxhumana okujulile phakathi kwesidoda ku-vas deferens kungase kube ngenxa yokucindezeleka okubangelwa ukuba khona kwenani elikhulu lesidoda endaweni encane.
Lapho kuhlolwa ukuziphatha kwe-spermatozoa kumaslayidi engilazi alengayo amasha, izimpawu zesikhashana ze-agglutination zingabonakala, ikakhulukazi emaphethelweni amaconsi wesidoda.Kodwa-ke, i-agglutination yayivame ukuphazanyiswa isenzo sokujikeleza esihambisana nokunyakaza okuqhubekayo, okuchaza isimo sesikhashana salesi senzakalo.Abacwaningi baphinde baqaphela ukuthi lapho i-diluent yengezwa esidodeni, amaseli amaseli “afana nochungechunge” avela.
Imizamo yokuqala yokulingisa i-spermatozoon yenziwa ngokukhipha ucingo oluncane ethonsini elilengayo, okuholele ekutheni kube ne-vesicle emide efana nesidoda ephuma ethonsini lesidoda.I-spermatozoa ngokushesha yahlanganiswa ngendlela efanayo ngaphakathi kwe-vesicle, kodwa yonke iyunithi yanyamalala ngokushesha ngenxa yomkhawulo we-3D.Ngakho-ke, ukutadisha i-spermatozoa agglutination, kuyadingeka ukubheka ukuhamba nokuziphatha kwe-spermatozoa ngokuqondile kuma-tubules wokugcina isidoda, okunzima ukukufeza.Ngakho-ke, kuyadingeka ukuthuthukisa ithuluzi elingisa i-spermatozoa ukusekela izifundo ze-sperm motility kanye nokuziphatha kwe-agglutination.U-Brillard et al13 babike ukuthi ubude obumaphakathi bamashubhu okugcina isidoda ematshwele amadala bungama-400–600 µm, kodwa amanye ama-SST angaba ubude obungu-2000 µm.I-Mero ne-Ogasawara14 ihlukanise izindlala zeseminiferous zibe amashubhu okugcina isidoda akhulisiwe futhi angakhulisiwe, womabili ayefana ngobude (~500 µm) nobubanzi bentamo (~38 µm), kodwa ububanzi be-lumen obumaphakathi kwamashubhu kwakungu-56.6 no-56.6 µm.., ngokulandelana 11.2 μm, ngokulandelana.Ocwaningweni lwamanje, sisebenzise idivayisi ye-microfluidic enosayizi wesiteshi ongu-200 µm × 20 µm (W × H), ingxenye yayo enqamulayo ithi isondele kuleyo ye-SST ekhulisiwe.Ngaphezu kwalokho, sihlole ukuhamba kwesidoda kanye nokuziphatha kwe-agglutination oketshezini olugelezayo, oluhambisana nenkolelo kaForeman yokuthi uketshezi olukhiqizwa amangqamuzana e-SST epithelial lugcina isidoda kulume ngendlela ephikisanayo (rheological).
Inhloso yalolu cwaningo kwakuwukunqoba izinkinga zokubheka ukuhamba kwe-spermatozoa epayipini le-fallopian nokugwema ubunzima bokutadisha i-rheology nokuziphatha kwe-spermatozoa endaweni eguquguqukayo.Kusetshenziswe i-microfluidic device eyenza i-hydrostatic pressure ilingise ukuhamba kwesidoda esithweni sangasese senkukhu.
Lapho iconsi lesampula lesidoda esihlanjululiwe (1:40) lilayishwa kudivayisi ye-microchannel, izinhlobo ezimbili ze-sperm motility zazingase zibonakale (isidoda esisodwa nesidoda esiboshwe).Ngaphezu kwalokho, i-spermatozoa yayivame ukubhukuda ngokumelene nesimanje (i-rheology enhle; ividiyo 1, 2). Nakuba izinqwaba zesidoda zinesivinini esiphansi kunesidoda esinesizungu (p <0.001), zandise iphesenti lesidoda esibonisa i-rheotaxis eqondile (p <0.001; Ithebula 2). Nakuba izinqwaba zesidoda zinesivinini esiphansi kunesidoda esinesizungu (p <0.001), zandise iphesenti lesidoda esibonisa i-rheotaxis eqondile (p <0.001; Ithebula 2). Хотя пучки сперматозоидов имели более низкую скорость, чем у одиночных сперматозоидов (p < 0,001), они увеличивали проезент прозент, проценд ложительный реотаксис (p < 0,001; таблица 2). Nakuba izinqwaba ze-spermatozoa zinejubane eliphansi kune-spermatozoa eyodwa (p <0.001), zandisa iphesenti ye-spermatozoa ebonisa i-rheotaxis enhle (p <0.001; Ithebula 2).尽管精子束的速度低于孤独精子的速度（p < 0.001），但它們增加了显示阳性流变性的爾0.表2）.尽管 精子束 的 速度 低于 孤独 的 速度 （p <0.001） , 但 增加 显示 阳性 流叾.001; 2......((())) Хотя скорость пучков сперматозоидов была ниже, чем у одиночных сперматозоидов (p <0,001), они увеличивали процент спелоий процент спедргий p <0,001; isiqephu 2). Nakuba ijubane lezinqwaba zesidoda laliphansi kunelodwa le-spermatozoa (p <0.001), bandise iphesenti le-spermatozoa nge-rheology enhle (p <0.001; Ithebula 2).I-rheology enhle ye-spermatozoa eyodwa kanye nama-tufts ilinganiselwa cishe ku-53% no-85%, ngokulandelana.
Kuye kwaqashelwa ukuthi i-spermatozoa yezinkukhu ze-sharkasi ngokushesha ngemva kokuphuma kwe-ejaculation yakha izinyanda eziqondile, ezihlanganisa inqwaba yabantu.Lawa ma-tufts akhula ngobude nokuqina ngokuhamba kwesikhathi futhi angase ahlale e-vitro amahora ambalwa ngaphambi kokuhlakazeka (ividiyo 3).Lezi zibopho ezinezintambo zimise okwe-echidna spermatozoa ezakheka ekugcineni kwe-epididymis.Isidoda sesikhukhukazi sika-Sharkashi sitholwe sinokuthambekela okuphezulu kokuhlanganisa kanye nokwenza inqwaba ye-reticulate ngaphansi komzuzu owodwa ngemuva kokuqoqwa.Le mishayo iyashintshashintsha futhi iyakwazi ukunamathela kunoma yiziphi izindonga eziseduze noma izinto ezimile.Nakuba izinqwaba zesidoda zinciphisa isivinini samangqamuzana esidoda, kuyacaca ukuthi ngokwe-macroscopically zandisa ukuhambisana kwazo.Ubude bezinqwaba buyahlukahluka kuye ngenani lesidoda esiqoqwe ngezinqwaba.Izingxenye ezimbili zenqwaba zahlukaniswa: ingxenye yokuqala, kuhlanganise nekhanda elikhululekile lesidoda esixubile, kanye nengxenye yokugcina, kuhlanganise nomsila kanye nayo yonke ingxenye ekude yesidoda.Kusetshenziswa ikhamera yejubane eliphezulu (i-950 fps), amakhanda amahhala e-spermatozoa agglutinated abonwa engxenyeni yokuqala yenqwaba, enesibopho sokunyakaza kwenqwaba ngenxa yokunyakaza kwawo kwe-oscillatory, ehudulela okusele emgodleni ngokunyakaza kwe-helical (Ividiyo 4).Nokho, emaphethelweni amade, kuye kwaphawulwa ukuthi amanye amakhanda esidoda akhululekile anamathela emzimbeni futhi ingxenye yokugcina ye-tuft isebenza njengama-vane ukusiza ukuqhubeza isidoda.
Ngesikhathi sokugeleza kancane koketshezi, izinqwaba zesidoda zihamba ngokufana komunye nomunye, nokho, ziqala ukunqwabelana futhi zinamathele kukho konke okusekhona, ukuze zingakhukhulwa ukugeleza kwamanje njengoba ijubane lokugeleza landa.Izibopho zakheka lapho idlanzana lamangqamuzana esidoda lisondela elinye nelinye, aqala ukunyakaza ngokuhambisana futhi agoqene, abese enamathela entweni enamathelayo.Umfanekiso 1 no-2 ubonisa indlela isidoda esisondelana ngayo, sakha ukuhlangana lapho imisila igoqana.
Abacwaningi basebenzise ingcindezi ye-hydrostatic ukuze benze ukugeleza koketshezi ku-microchannel ukuze batadishe i-rheology yesidoda.Isiteshi esincane esinosayizi ongu-200 µm × 20 µm (W × H) nobude obungu-3.6 µm sisetshenzisiwe.Sebenzisa iziteshi ezincane phakathi kweziqukathi ezinesirinji ezifakwe ekugcineni.Umbala wokudla wasetshenziselwa ukwenza iziteshi zibonakale kakhudlwana.
Bophela izintambo ze-interconnect kanye nezinsiza odongeni.Ividiyo ithathwe ngesibonakhulu sokungafani kwesigaba.Ngesithombe ngasinye, kuvezwa imakroskopu yesigaba sokuqhathanisa nezithombe zemephu.(A) Ukuxhumana phakathi kwemifudlana emibili imelana nokugeleza ngenxa yokunyakaza kwe-helical (umcibisholo obomvu).(B) Ukuxhumana phakathi kwenqwaba yeshubhu nodonga lwesiteshi (imicibisholo ebomvu), ngesikhathi esifanayo ixhunywe kwezinye izinqwaba ezimbili (imicibisholo ephuzi).(C) Izigaxa zesidoda kumzila we-microfluidic ziqala ukuhlangana (imicibisholo ebomvu), zenze uxhaxha lwenqwaba yesidoda.(D) Ukwakhiwa koxhaxha lwenqwaba yesidoda.
Lapho iconsi lesidoda esihlanjululwe lilayishwa emshinini we-microfluidic futhi kwakhiwa ukugeleza, uhlaka lwesidoda lwalubhekwa ukuthi luhamba ngokumelene nesiqondiso sokugeleza.Izinqwaba zilingana kahle ezindongeni zamashaneli amancane, futhi amakhanda amahhala engxenyeni yokuqala yenqwaba alingana kahle nawo (ividiyo 5).Aphinde anamathele kunoma yiziphi izinhlayiya ezimile endleleni yawo, njengemfucumfucu, ukuze amelane nokukhukhulwa umsinga.Ngokuhamba kwesikhathi, lezi zicucu ziba imicu ende ebambe esinye isidoda esisodwa kanye nezigaxa ezimfishane (Ividiyo 6).Njengoba ukugeleza kuqala ukwehla, imigqa emide yesidoda iqala ukwakha inethiwekhi yemigqa yesidoda (Ividiyo 7; Umfanekiso 2).
Ngesivinini sokugeleza okuphezulu (V > 33 µm/s), ukunyakaza okuzungezayo kwemicu kuyenyuka njengomzamo wokubamba izinqwaba zesidoda ngasinye zenza izinyanda ukumelana kangcono namandla akhukhulekayo okugeleza. Ngesivinini sokugeleza okuphezulu (V > 33 µm/s), ukunyakaza okuzungezayo kwemicu kuyenyuka njengomzamo wokubamba izinqwaba zesidoda ngasinye zenza izinyanda ukumelana kangcono namandla akhukhulekayo okugeleza. При высокой скорости потока (V > 33 мкм/с) спиралевидные движения нитей усиливаются, поскольку они пытаются поймать множествоы оздонгей х пучки, которые лучше противостоят дрейфующей силе потока. Emazingeni okugeleza aphezulu (V > 33 µm/s), ukunyakaza kwe-helical yezintambo kuyakhula njengoba bezama ukubamba inqwaba ye-spermatozoa eyenza izinyanda ezikwazi kangcono ukumelana namandla okukhukhuleka okugeleza.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旋旺旺子，漂移力.在 高 流速 (v> 33 µm/s) 时, 的 螺旋 运动 增加, 以漂移力。。。。。. При высоких скоростях потока (V > 33 мкм/с) спиральное движение нитей увеличивается в попытке захватить множество отделььное движение нитей увеличивается в попытке захватить множество отделььное зпихер, чтобы лучше сопротивляться силам дрейфа потока. Emazingeni okugeleza aphezulu (V > 33 µm/s), ukunyakaza kwe-helical kwemicu kuyenyuka ngomzamo wokubamba inqwaba yesidoda ngasinye senza izinyanda ukuze imelane kangcono namandla okukhukhuleka okugeleza.Baphinde bazama ukunamathisela ama-microchannel emaceleni.
Izigaxa zesidoda zikhonjwe njengamaqoqo amakhanda esidoda nemisila egoqekayo kusetshenziswa i-microscopy ekhanyayo (LM).Izigaxa zesidoda ezinamaqoqo ahlukahlukene nazo zikhonjwe njengamakhanda asontekile nama-flagellar aggregate, imisila yesidoda eminingi ehlangene, amakhanda esidoda axhunywe umsila, namakhanda esidoda anama-nuclei agobile njengama-nuclei ahlangene amaningi.i-electron microscopy (TEM) yokudlulisela.I-Scanning electron microscopy (SEM) ibonise ukuthi inqwaba yesidoda yayiyinhlanganisela yamakhanda esidoda futhi ama-sperm aggregate abonisa inethiwekhi enamathiselwe yemisila esongekile.
I-morphology kanye ne-ultrastructure ye-spermatozoa, ukwakheka kwenqwaba yesidoda kwahlolisiswa kusetshenziswa i-microscopy ekhanyayo (ingxenye yesigamu), i-scanning electron microscopy (SEM) kanye ne-transmission electron microscopy (TEM), ama-sperm smear ayengcoliswe nge-acridine orange futhi ahlolwa kusetshenziswa i-epifluorescence microscopy.
I-sperm smear staining nge-acridine orange (Fig. 3B) yabonisa ukuthi amakhanda e-sperm ayenamathele ndawonye futhi embozwe ngezinto eziyimfihlo, okwaholela ekwakhekeni kwama-tufts amakhulu (Fig. 3D).Izinqwaba zesidoda zazihlanganisa izilinganiso zesidoda ezinoxhaxha lwemisila enamathiselwe (Fig. 4A-C).Izigaxa zesidoda zakhiwe ngemisila yama-spermatozoa amaningi anamathelene (Fig. 4D).Izimfihlo (Fig. 4E,F) zimboze amakhanda enqwaba ye-spermatozoa.
Ukwakhiwa kwenqwaba ye-spermatozoa Ukusebenzisa i-microscope yesigaba sokuqhathanisa kanye ne-sperm smear egcotshwe nge-acridine orange, kwabonisa ukuthi amakhanda e-spermatozoa ayanamathelana.(A) Ukwakheka kwasekuqaleni kwesidoda kuqala ngesidoda (isiyingi esimhlophe) nesidoda ezintathu (isiyingi esiphuzi), okuzungezayo kuqale emsileni kugcine ekhanda.(B) I-Photomicrograph ye-sperm smear egcotshwe nge-acridine orange ebonisa amakhanda esidoda anamathelayo (imicibisholo).Ukukhishwa kuvala ikhanda(ama)khanda.Ukukhulisa × 1000. (C) Ukuthuthukiswa kwe-beam enkulu ethuthwa ngokugeleza kushaneli ye-microfluidic (kusetshenziswa ikhamera yejubane eliphezulu ku-950 fps).(D) I-Micrograph ye-sperm smear egcotshwe nge-acridine orange ebonisa izigaxa ezinkulu (imicibisholo).Ukukhulisa: × 200.
Iskena i-electron micrograph ye-sperm beam kanye ne-sperm smear egcotshwe nge-acridine orange.(A, B, D, E) ama-electron micrographs e-electron micrographs ethwebula imibala ye-spermatozoa, kanti u-C no-F ama-micrographs we-acridine orange stained sperm smear abonisa ukunamathiselwe kwe-spermatozoa eminingi esonga iwebhu ye-caudal.(AC) Izilinganiso zesidoda ziboniswa njengenethiwekhi yemisila enamathiselwe (imicibisholo).(D) Ukunamathela kwe-spermatozoa (enezinto ezinamathelayo, uhlaka oluphinki, umcibisholo) ezisonga umsila.(E kanye no-F) Amaqoqo ekhanda lesidoda (izinkomba) ambozwe ngezinto ezinamathelayo (izikhombi).I-spermatozoa yakha izinyanda ezinezakhiwo eziningana ezifana ne-vortex (F).(C) × 400 kanye (F) × 200 ukukhuliswa.
Ngokusebenzisa i-microscopy ye-electron yokudlulisela, sithole ukuthi izinqwaba zesidoda zinemisila enamathiselwe (Fig. 6A, C), amakhanda axhunywe emisileni (Fig. 6B), noma amakhanda axhunywe emisileni (Fig. 6D).Amakhanda e-spermatozoa ku-bundle agobile, ethula esigabeni sezifunda ezimbili zenuzi (Fig. 6D).Ku-incision bundle, i-spermatozoa yayinekhanda elisontekile elinezifunda ezimbili zenuzi kanye nezifunda eziningi ze-flagellar (Fig. 5A).
I-electron micrograph yombala wedijithali ebonisa imisila exhumayo kunqwaba yesidoda kanye nezinto ezihlanganisayo ezixhuma amakhanda esidoda.(A) Umsila onamathiselwe wenombolo enkulu yesidoda.Qaphela ukuthi umsila ubukeka kanjani kukho kokubili ukuma ngobude (umcibisholo) kanye nokwakheka kwezwe (umcibisholo).(B) Inhloko (umcibisholo) yesidoda ixhunywe emsileni (umcibisholo).(C) Imisila yesidoda eminingana (imicibisholo) inamathiselwe.(D) I-agglutination material (AS, blue) ixhuma amakhanda amane esidoda (onsomi).
I-scanning electron microscopy yasetshenziselwa ukuthola amakhanda esidoda kunqwaba yesidoda embozwe uketshezi noma ulwelwesi (Umfanekiso 6B), okubonisa ukuthi inqwaba yesidoda yayigxilwe ngezinto ezingaphandle kwamangqamuzana.I-agglutinated material yayigxiliswe ekhanda lesidoda (i-jellyfish head-like assembly; Fig. 5B) futhi yanwetshwa ngokuqhelelana, inika ukubukeka okuphuzi okukhazimulayo ngaphansi kwe-fluorescence microscopy lapho ingcoliswe nge-acridine orange (Fig. 6C).Le nto ibonakala ngokucacile ngaphansi kwe-microscope yokuskena futhi ithathwa njenge-binder.Izingxenye ezincane (Fig. 5C) kanye nama-sperm smear agcotshwe nge-acridine orange abonisa izinyanda zesidoda ezinamakhanda aminyene kanye nemisila egoqene (Fig. 5D).
Ama-photomicrographs ahlukahlukene abonisa ukuhlangana kwamakhanda esidoda nemisila esongekile kusetshenziswa izindlela ezahlukahlukene.(A) I-electron micrograph ye-electron micrograph yombala wedijithali ehlukene yenqwaba yesidoda ebonisa ikhanda lesidoda eligoqiwe eline-nucleus enezingxenye ezimbili (oluhlaza okwesibhakabhaka) kanye nezingxenye ezimbalwa ze-flagellar (eziluhlaza).(B) I-electron micrograph yokuskena umbala wedijithali ebonisa iqoqo lamakhanda esidoda afana ne-jellyfish (imicibisholo) abonakala emboziwe.(C) Isigaba esincanyana kancane esibonisa amakhanda esidoda ahlanganisiwe (imicibisholo) nemisila egoqekile (imicibisholo).(D) I-Micrograph ye-sperm smear egcotshwe nge-acridine orange ebonisa inhlanganisela yamakhanda esidoda (imicibisholo) kanye nemisila enamathelayo egoqekile (imicibisholo).Qaphela ukuthi into enamathelayo (S) ivala ikhanda le-spermatozoon.(D) × 1000 ukukhulisa.
Kusetshenziswa i-microscopy ye-electron yokudlulisela (Fig. 7A), kwaphawulwa nokuthi amakhanda esidoda asontekile futhi i-nuclei inomumo ovunguzayo, njengoba kuqinisekiswa ama-sperm smear agcotshwe nge-acridine orange futhi ahlolwe kusetshenziswa i-fluorescence microscopy (Fig. 7B).
(A) I-Digital color transmission electron micrograph kanye (B) ne-Acridine orange stained sperm smear ebonisa amakhanda abophene nokunamathiselwe kwamakhanda nemisila yesidoda (imicibisholo).(B) × 1000 ukukhulisa.
Okuthakazelisayo okutholakele ukuthi isidoda sika-Sharkazi siyahlangana ukuze kwakheka izinyanda ezinama-filamentous.Izakhiwo zalezi zinqwaba zisivumela ukuba siqonde indima yazo engenzeka ekumunceni nasekugcinweni kwe-spermatozoa ku-SST.
Ngemva kokuhlangana, isidoda singena emomozini bese singenela inqubo eqinile yokukhetha, okuholela enanini elilinganiselwe lesidoda esingena ku-SST15,16.Kuze kube manje, izindlela lapho isidoda singena futhi siphuma ngaso ku-SST azikacaci.Ezinkukhu, i-spermatozoa igcinwa ku-SST isikhathi eside samaviki angu-2 kuya kwangu-10, kuye ngezinhlobo6.Ukungqubuzana kusasele mayelana nesimo sesidoda ngesikhathi sokugcinwa ku-SST.Ingabe ziyanyakaza noma ziphumule?Ngamanye amazwi, amangqamuzana esidoda agcina kanjani isikhundla sawo ku-SST isikhathi eside kangaka?
I-Forman4 iphakamise ukuthi ukuhlala kwe-SST nokukhishwa kungachazwa ngokuya ngokuhamba kwesidoda.Ababhali bacabanga ukuthi isidoda sigcina isikhundla saso ngokubhukuda ngokumelene nokugeleza koketshezi okudalwe yi-SST epithelium nokuthi isidoda sikhishwa ku-SST lapho isivinini saso siwela ngaphansi kwephuzu lapho siqala khona ukubuyela emuva ngenxa yokuntuleka kwamandla.I-Zaniboni5 iqinisekise ukuba khona kwama-aquaporin 2, 3, no-9 engxenyeni ye-apical yamaseli e-SST epithelial, angase asekele ngokungaqondile imodeli yokugcina isidoda sika-Foreman.Ocwaningweni lwamanje, sithole ukuthi cishe ingxenye yesidoda sika-Sharkashi ibonisa i-rheology enhle oketshezini olugelezayo, nokuthi izinyanda zesidoda ezihlanganisiwe zandisa inani lesidoda esibonisa i-rheology enhle, nakuba i-agglutination iwanciphisa.Ukuthi amangqamuzana esidoda ahamba kanjani ngeshubhu lenyoni lenyoni aye lapho ikhulela khona akuqondakali ngokugcwele.Ezilwaneni ezincelisayo, uketshezi lwe-follicular lukhanga i-spermatozoa.Nokho, ama-chemoattractants kukholakala ukuthi aqondisa i-spermatozoa ukuze isondele amabanga amade7.Ngakho-ke, ezinye izindlela zinesibopho sokuthutha isidoda.Ikhono lesidoda ukuze liqondise futhi ligeleze ngokumelene noketshezi lweshubhu ye-fallopian olukhishwe ngemva kokuhlangana kubikwe njengembangela enkulu ekuqondiseni isidoda kumagundane.U-Parker 17 uphakamise ukuthi isidoda siwele ama-oviducts ngokubhukuda ngokumelene nomoya we-ciliary wezinyoni nezilwane ezihuquzelayo.Nakuba kungazange kuboniswe ngokuhlolwa ezinyonini, i-Adolphi18 yaba ngowokuqala ukuthola ukuthi isidoda sezinyoni sinikeza imiphumela emihle lapho ungqimba oluncane loketshezi phakathi kwe-coverlip ne-slide kwakhiwa ngocu lwephepha lokuhlunga.I-Rheology.U-Hino no-Yanagimachi [19] babeke inkimbinkimbi yegundane ye-ovary-tubal-uterine eringini yokuxutshwa futhi bajova u-1 µl kayinki ku-isthmus ukuze babone ukugeleza koketshezi kumashubhu e-fallopian.Baqaphela ukunyakaza okusebenzayo kokufinyela nokuphumula eshubhuni yesibeletho, lapho wonke amabhola kayinki ayehamba kancane eya ku-ampula yeshubhu le-fallopian.Ababhali bagcizelela ukubaluleka kokugeleza kwe-tubal fluid kusuka phansi kuya kumashubhu e-fallopian aphezulu ukuze kukhushulwe isidoda nokuvundiswa.I-Brillard20 ibike ukuthi ezinkukhwini kanye nama-turkeys, i-spermatozoa ifuduka ngokunyakaza okusebenzayo isuka emnyango wesitho sangasese sowesifazane, lapho igcinwa khona, iye endaweni ye-uterine-vaginal, lapho igcinwa khona.Kodwa-ke, lokhu kunyakaza akudingekile phakathi kokuhlangana kwe-uterovaginal kanye ne-infundibulum ngoba i-spermatozoa ithuthwa ngokususwa kwe-passive.Ukwazi lezi zincomo zangaphambilini kanye nemiphumela etholwe esifundweni samanje, kungacatshangwa ukuthi ikhono le-spermatozoa lokukhuphuka (rheology) lingenye yezakhiwo lapho inqubo yokukhetha isekelwe khona.Lokhu kunquma ukudlula kwe-spermatozoa ngokusebenzisa isitho sangasese sowesifazane kanye nokungena kwabo ku-CCT ukuze kugcinwe.Njengoba i-Forman4 yasikisela, lokhu kungase futhi kusize inqubo yokungena kwesidoda ku-SST nendawo yayo isikhathi esithile bese iphuma lapho ijubane lazo liqala ukwehla.
Ngakolunye uhlangothi, i-Matsuzaki ne-Sasanami 21 iphakamise ukuthi isidoda se-avian sibhekane nezinguquko ze-motility ukusuka ku-dormancy kuya ku-motility emapheshaneni okuzala abesilisa nabesifazane.Ukuvinjelwa kokuhamba kwesidoda esihlala endaweni ku-SST kuhlongoziwe ukuze kuchazwe isikhathi eside sokugcinwa kwesidoda kanye nokuvuselelwa ngemva kokushiya i-SST.Ngaphansi kwezimo ze-hypoxic, uMatsuzaki et al.1 ibike ukukhiqizwa okuphezulu nokukhishwa kwe-lactate ku-SST, okungase kuholele ekuvinjweni kokuhamba kwesidoda esihlala khona.Kulokhu, ukubaluleka kwe-rheology yesidoda kubonakala ekukhethweni nasekumunyweni kwe-spermatozoa, hhayi ekugcinweni kwabo.
Iphethini ye-sperm agglutination ibhekwa njengencazelo ezwakalayo yesikhathi eside sokugcinwa kwesidoda ku-SST, njengoba lena kuyiphethini evamile yokugcinwa kwesidoda enkukhu2,22,23.Bakst et al.2 yabona ukuthi iningi le-spermatozoa linamathela komunye nomunye, lakha ama-fascicular aggregates, futhi i-spermatozoa eyodwa yayingavamile ukutholakala ku-CCM yezigwaca.Ngakolunye uhlangothi, u-Wen et al.24 ibone isidoda esihlakazekile kakhulu kanye nezigaxa ze-spermatozoa ezimbalwa ku-lumen ye-SST ezinkukhuni.Ngokusekelwe kulokhu okubonwayo, kungacatshangwa ukuthi ukuthambekela kwe-sperm agglutination kuyahluka phakathi kwezinyoni naphakathi kwe-spermatozoa ku-ejaculate efanayo.Ngaphezu kwalokho, uVan Krey et al.9 iphakamise ukuthi ukuhlukaniswa okungahleliwe kwesidoda esihlanganisiwe kunesibopho sokungena kancane kancane kwe-spermatozoa ku-lumen yeshubhu le-fallopian.Ngokwalokhu kucatshangelwa, i-spermatozoa enamandla aphansi we-agglutination kufanele ikhishwe ku-SST kuqala.Kulo mongo, ikhono le-spermatozoa lokuhlanganisa i-agglutinate lingase libe yisici esithonya umphumela wokuncintisana kwesidoda ezinyoni ezingcolile.Ngaphezu kwalokho, uma isidoda esihlanganisiwe sihlukana isikhathi eside, ukuzala okude kuyagcinwa.
Nakuba ukuhlanganiswa kwe-spermatozoa nokuhlanganiswa kube yizinqwaba kuye kwabonwa ezifundweni eziningana ze-2,22,24, azizange zichazwe ngokuningiliziwe ngenxa yobunzima bokubuka kwabo kwe-kinematic ngaphakathi kwe-SST.Imizamo eminingana yenziwe yokutadisha i-sperm agglutination in vitro.Ukuhlanganisa okunabile kodwa okudlulayo kwabonwa lapho ucingo oluncane lususwa ethonsi lembewu elengayo.Lokhu kuholela eqinisweni lokuthi ibhamuza elide liphuma ethonsini, lilingisa indlala yesidoda.Ngenxa yemikhawulo ye-3D kanye nezikhathi ezimfushane zokumisa ama-drip, yonke ibhulokhi yawela ngokushesha ekumeni9.Ocwaningweni lwamanje, sisebenzisa izinkukhu ze-Sharkashi nama-microfluidic chips, sikwazile ukuchaza ukuthi lawa ma-tuft akheka kanjani nokuthi ahamba kanjani.Izinqwaba zesidoda zakhiwe ngokushesha ngemva kokuqoqwa kwesidoda futhi zatholakala zihamba ngokuzungeza, zibonisa i-rheology enhle lapho ikhona ekugelezeni.Ngaphezu kwalokho, lapho kubukwa nge-macroscopically, ama-sperm bundles aye aqashelwa ukwandisa umugqa we-motility uma kuqhathaniswa ne-spermatozoa eyedwa.Lokhu kuphakamisa ukuthi ukuhlanganiswa kwesidoda kungase kwenzeke ngaphambi kokungena kwe-SST nokuthi ukukhiqizwa kwesidoda akukhawulelwe endaweni encane ngenxa yengcindezi njengoba kwakusikiselwe ngaphambilini (iTingari neLake12).Ngesikhathi sokwakheka kwe-tuft, i-spermatozoa ibhukuda nge-synchrony ize yakha ukuhlangana, khona-ke imisila yaso igoqana futhi ikhanda le-spermatozoon lihlala likhululekile, kodwa umsila nengxenye ekude ye-spermatozoon kunamathelana nento enamathelayo.Ngakho-ke, inhloko yamahhala ye-ligament inesibopho sokunyakaza, idonsa yonke i-ligament.Ukuskena i-electron microscopy yenqwaba yesidoda kubonise amakhanda esidoda anamathiselwe embozwe izinto eziningi ezinamathelayo, okusikisela ukuthi amakhanda esidoda axhunywe kunqwaba yokuphumula, okungenzeka kwenzeke ngemva kokufika endaweni yokugcina (SST).
Uma i-sperm smear ingcoliswe i-acridine orange, izinto zokunamathisela ezingaphandle kwamangqamuzana esidoda zingabonakala ngaphansi kwesibonakhulu se-fluorescent.Le nto ivumela inqwaba yesidoda ukuthi inamathele futhi ibambelele kunoma yiziphi izindawo ezizungezile noma izinhlayiya ukuze zingakhukhuli nokugeleza okuzungezile.Ngakho-ke, ukubuka kwethu kubonisa indima ye-spermatozoa adhesion ngendlela yezibopho ezihambayo.Ikhono labo lokubhukuda ngokumelene ne-current futhi linamathele ezindaweni eziseduze livumela isidoda ukuthi sihlale isikhathi eside ku-SST.
I-Rothschild25 isebenzise ikhamera ye-hemocytometry ukutadisha ukusatshalaliswa okuntantayo kwesidoda sezinkomo endaweni yokumiswa, ithatha ama-photomicrographs ngekhamera ene-eksisi yokubona eqondile nevundlile yesibonakhulu.Imiphumela yabonisa ukuthi i-spermatozoa yakhangwa ebusweni bekamelo.Ababhali basikisela ukuthi kungase kube nokusebenzisana kwe-hydrodynamic phakathi kwesidoda nendawo engaphezulu.Ukucabangela lokhu, kanye nekhono lesidoda sephuphu lika-Sharkashi ukwenza izigaxa ezinamathelayo, kungase kwandise amathuba okuthi isidoda sinamathele odongeni lwe-SST futhi sigcinwe isikhathi eside.
I-Bccetti ne-Afzeliu26 babike ukuthi i-sperm glycocalyx iyadingeka ukuze kubonwe i-gamete nokuhlanganisa.I-Forman10 yaphawula ukuthi i-hydrolysis yamabhondi e-α-glycosidic embotsheni ye-glycoprotein-glycolipid ngokwelapha isidoda sezinyoni nge-neuraminidase kuphumele ekunciphiseni ukuzala ngaphandle kokuphazamisa ukuhamba kwesidoda.Ababhali basikisela ukuthi umthelela we-neuraminidase ku-glycocalyx ukhinyabeza ukuhlukaniswa kwesidoda lapho kuhlangana khona isibeletho nesitho sangasese sowesifazane, ngaleyo ndlela kunciphise inzalo.Ukuqaphela kwabo akukwazi ukuziba ukuthi kungenzeka ukuthi ukwelashwa kwe-neuraminidase kunganciphisa ukuqashelwa kwesidoda kanye ne-oocyte.I-Forman ne-Engel10 bathola ukuthi inzalo yehla lapho izinkukhu zifakwa esithweni sangasese ngesidoda esiphathwe nge-neuraminidase.Kodwa-ke, i-IVF ne-neuraminidase isidoda esilashwayo ayizange ithinte ukuzala uma kuqhathaniswa nokulawula izinkukhu.Ababhali baphethe ngokuthi izinguquko ezimbotsheni ze-glycoprotein-glycolipid ezizungeze ulwelwesi lwesidoda zanciphisa ikhono lesidoda lokuvundisa ngokukhinyabeza ukuqoqwa kwesidoda lapho kuhlangana khona isibeletho, okubuye kwandise ukulahlekelwa kwesidoda ngenxa yejubane lokuhlangana kwesibeletho, kodwa akuthinti isidoda nokubonwa kweqanda.
Kumaturkeys u-Bakst no-Bauchan 11 bathola ama-vesicle amancane nezingcezu zolwelwesi kulume lwe-SST futhi baqaphela ukuthi amanye alawa ma-granules ayehlangene nolwelwesi lwesidoda.Ababhali basikisela ukuthi lobu budlelwano bungaba nomthelela ekugcinweni kwesikhathi eside kwe-spermatozoa ku-SST.Kodwa-ke, abacwaningi abazange bacacise umthombo walezi zinhlayiya, kungakhathaliseki ukuthi zifihliwe ngamaseli e-CCT epithelial, akhiqizwa futhi afihliwe uhlelo lokuzala wesilisa, noma akhiqizwa yisidoda ngokwaso.Futhi, lezi zinhlayiya zinesibopho se-agglutination.U-Grützner et al27 ubike ukuthi amangqamuzana e-epididymal epithelial akhiqiza futhi akhiphe iphrotheni ethile edingekayo ukuze kwakhiwe amapheshana e-seminal e-pore eyodwa.Ababhali baphinde babike ukuthi ukuhlakazeka kwalezi zinqwaba kuncike ekusebenzisaneni kwamaprotheni e-epididymal.I-Nixon et al28 ithole ukuthi i-adnexa ikhiqiza iphrotheni, i-osteonectin ene-acidic cysteine-rich;I-SPARC ibandakanyeka ekwakhekeni kwama-sperm tufts kuma-echidna anoqhwaku olufushane nama-platypus.Ukuhlakazeka kwale mishayo kuhlotshaniswa nokulahlekelwa yile phrotheni.
Ocwaningweni lwamanje, ukuhlaziywa kwe-ultrastructural kusetshenziswa i-electron microscopy kubonise ukuthi i-spermatozoa inamathela kumthamo omkhulu wezinto eziminyene.Lezi zinto kucatshangwa ukuthi zinesibopho sokuhlangana okuhlangana phakathi noma eduze kwamakhanda abambelelayo, kodwa ezindaweni eziphansi endaweni yomsila.Sicabanga ukuthi le nto ehlanganisayo ikhishwa ohlelweni lokuzala wesilisa (epididymis noma vas deferens) kanye nesidoda, njengoba sivame ukubona isidoda sihlukana ne-lymph kanye ne-seminal plasma ngesikhathi sokuphuma.Kuye kwabikwa ukuthi njengoba i-spermatozoa yezinyoni idlula ku-epididymis kanye ne-vas deferens, zithola izinguquko ezihlobene nokuvuthwa ezisekela ikhono lazo lokubopha amaprotheni futhi zithole i-plasma lemma-associated glycoprotein.Ukuphikelela kwalawa maprotheni kulwelwesi lwesidoda esihlala khona ku-SST kuphakamisa ukuthi lawa maprotheni angase abe nomthelela ekutholweni kokuqina kolwelwesi lwesidoda 30 futhi anqume ukuzala kwawo 31.U-Ahammad et al32 ubike ukuthi i-spermatozoa etholakala ezingxenyeni ezihlukahlukene zesistimu yokuzala yabesilisa (kusuka kumasende kuya ku-distal vas deferens) ibonise ukwanda okuqhubekayo kokusebenza ngaphansi kwezimo zokugcina uketshezi, kungakhathaliseki izinga lokushisa lokugcina, nokusebenza kwezinkukhu nakho kuyanda kumashubhu e-fallopian ngemva kokufakelwa kokufakelwa.
Izigaxa zesidoda senkukhu ye-Sharkashi zinezici nemisebenzi ehlukile kunezinye izinhlobo ezifana nama-echidna, ama-platypus, amagundane okhuni, amagundane enyamazane, nezingulube ze-Guinea.Ezinkukhu ze-sharkasi, ukwakheka kwenqwaba ye-spermatozoa kunciphisa isivinini sabo sokubhukuda uma kuqhathaniswa ne-spermatozoa eyodwa.Kodwa-ke, lezi zinqwaba zandisa iphesenti le-rheologically positive spermatozoa futhi zandisa ikhono le-spermatozoa lokuzinza endaweni eguquguqukayo.Ngakho-ke, imiphumela yethu iqinisekisa isiphakamiso sangaphambili sokuthi i-sperm agglutination ku-SST ihlotshaniswa nokugcinwa kwesidoda isikhathi eside.Siphinde sicabange ukuthi ukuthambekela kwesidoda ukwenza ama-tuft kungase kulawule izinga lokulahlekelwa kwesidoda ku-SST, okungase kuguqule umphumela wokuncintisana kwesidoda.Ngokwalokhu kucatshangelwa, i-spermatozoa ene-low agglutination capacity ikhulula i-SST kuqala, kuyilapho i-spermatozoa enamandla amakhulu e-agglutination ikhiqiza iningi lenzalo.Ukwakhiwa kwenqwaba yesidoda esinembotshana eyodwa kuyazuzisa futhi kuthinta isilinganiso somzali nengane, kodwa kusebenzisa indlela ehlukile.Ku-echidnas kanye ne-platypus, i-spermatozoa ihlelwe ngokufana ukuze kwandiswe ijubane elihamba phambili le-beam.Inqwaba ye-echidnas ihamba ngokushesha ngokuphindwe kathathu kune-spermatozoa eyodwa.Kukholelwa ukuthi ukwakheka kwalezi zingqimba zesidoda kuma-echidna kuwukuziphendukela kwemvelo ukuze kugcinwe ukubusa, njengoba abesifazane baziphatha kabi futhi ngokuvamile bahlangana nabesilisa abambalwa.Ngakho-ke, i-spermatozoa evela ku-ejaculate ehlukene incintisana kakhulu ngokuvundiswa kweqanda.
I-spermatozoa ene-agglutinated yezinkukhu zika-sharkasi kulula ukuyibona ngeso lengqondo kusetshenziswa i-microscope yesigaba sokuqhathanisa, okubhekwa njengenzuzo ngoba ivumela ukufunda okulula kokuziphatha kwe-spermatozoa in vitro.Indlela ukwakheka kwe-sperm tuft okugqugquzela ngayo ukuzala kwezinkukhu ze-sharkasi nayo ihlukile kuleyo ebonakala kwezinye izilwane ezincelisayo ezimele ukubambisana kwesidoda njengamagundane okhuni, lapho isidoda esithile sifinyelela amaqanda, sisize abanye abantu abahlobene ukuthi bafinyelele futhi balimaze amaqanda abo.ukuze uzibonakalise.ukuziphatha okungafanele.Ukuzilungiselela 34. Esinye isibonelo sokuziphatha kokubambisana ku-spermatozoa satholakala kumagundane enyamazane, lapho i-spermatozoa ikwazi ukukhomba futhi ihlanganise ne-spermatozoa ehlobene kakhulu nezakhi zofuzo futhi yakha amaqembu okubambisana ukuze kwandiswe isivinini sawo uma kuqhathaniswa ne-spermatozoa35 engahlobene.
Imiphumela etholwe kulolu cwaningo ayiphikisani nombono kaFoman wokugcinwa kwe-spermatozoa isikhathi eside ku-SWS.Abacwaningi babika ukuthi amangqamuzana esidoda ayaqhubeka nokuhamba ekugelezeni kwamangqamuzana e-epithelial ahlanganisa i-SST isikhathi eside, futhi ngemva kwesikhathi esithile, izitolo zamandla zamangqamuzana esidoda ziyancipha, okuholela ekunciphiseni kwejubane, okuvumela ukuxoshwa kwezinto ezincane zesisindo samangqamuzana.amandla e-spermatozoa ngokugeleza koketshezi oluvela kulume lwe-SST Umgodi we-fallopian tube.Ocwaningweni lwamanje, sibonile ukuthi ingxenye yesidoda esisodwa yabonisa ikhono lokubhukuda ngokumelene noketshezi olugelezayo, futhi ukunamathela kwabo emgodleni kwandisa ikhono labo lokukhombisa i-rheology enhle.Ngaphezu kwalokho, idatha yethu iyahambisana naleyo kaMatuzaki et al.1 obike ukuthi ukwanda kokuphuma kwe-lactate ku-SST kungase kuvimbele ukuhamba kwesidoda sabahlali.Kodwa-ke, imiphumela yethu ichaza ukwakheka kwemigqa ye-sperm motile kanye nokuziphatha kwabo kwe-rheological phambi kwendawo eguquguqukayo ngaphakathi kwe-microchannel emzamweni wokucacisa ukuziphatha kwabo ku-SST.Ucwaningo lwesikhathi esizayo lungase lugxile ekunqumeni ukwakheka kwamakhemikhali kanye nemvelaphi ye-agglutinating agent, okuzosiza ngokungangabazeki abacwaningi ukuba bathuthukise izindlela ezintsha zokugcina isidoda esiwuketshezi futhi andise ubude besikhathi sokuzala.
Ushaka wesilisa oneminyaka eyishumi nanhlanu onamasonto angu-30 ubudala ongenalutho (ophethe i-homozygous; Na Na) ukhethwe njengabanikeli besidoda ocwaningweni.Lezi zinyoni zikhuliswe e-Research Poultry Farm ye-Faculty of Agriculture, e-Ashit University, e-Ashit Governorate, e-Egypt.Izinyoni zazihlaliswa emakhejini ngamanye (30 x 40 x 40 cm), ngaphansi kohlelo lokukhanya (amahora angu-16 okukhanya namahora angu-8 obumnyama) futhi zondliwa ukudla okuqukethe i-160 g yamaprotheni aluhlaza, 2800 kcal wamandla we-metabolizable, 35 g we-calcium lilinye.5 amagremu e-phosphorus etholakalayo ngekhilogremu ngayinye yokudla.
Ngokwedatha engu-36, 37, isidoda saqoqwa kwabesilisa ngokubhucungwa kwesisu.Kwaqoqwa amasampula esidoda angama-45 emadodeni ayi-15 ezinsukwini ezi-3.Isidoda (n = 15/day) sihlanjululwe ngokushesha ngo-1: 1 (v:v) nge-Belsville Poultry Semen Diluent, equkethe i-potassium diphosphate (1.27 g), i-monosodium glutamate monohydrate (0.867 g), i-fructose (0.5 d) i-anhydrous sodium.i-acetate (0.43 g), i-tris(hydroxymethyl)aminomethane (0.195 g), i-potassium citrate monohydrate (0.064 g), i-potassium monophosphate (0.065 g), i-magnesium chloride (0.034 g) ne-H2O (100 ml), i-pH = 7, 5, i-osmolarity 3/33 m33 m8 m.Amasampula esidoda ahlanjululiwe aqale ahlolwa ngaphansi kwesibonakhulu esikhanyayo ukuze kuqinisekiswe ikhwalithi enhle yesidoda (umswakama) bese egcinwa endaweni yokugeza yamanzi engu-37°C kuze kusetshenziswe phakathi nengxenye yehora ngemva kokuqoqwa.
I-kinematics kanye ne-rheology ye-spermatozoa ichazwa ngokusebenzisa uhlelo lwamadivayisi we-microfluidic.Amasampula wesidoda aphinde ahlanjululwa ukuze abe ngu-1:40 e-Beltsville Avian Semen Diluent, alayishwe kudivayisi encane ye-microfluidic (bona ngezansi), futhi imingcele ye-kinetic yanqunywa kusetshenziswa uhlelo lwe-Computerized Semen Analysis (CASA) olwakhelwe ngaphambilini ukuze luveze ama-microfluidics.ekuhambeni kwe-spermatozoa emithonjeni ye-liquid (uMnyango Wezobunjiniyela Bemishini, i-Faculty of Engineering, i-Assiut University, e-Egypt).I-plugin ingalandwa ku-: http://www.assiutmicrofluidics.com/research/casa39.Isivinini sejika (VCL, μm/s), isivinini somugqa (VSL, μm/s) kanye nesivinini se-trajectory esimaphakathi (VAP, μm/s) kukalwa.Amavidiyo e-spermatozoa athathwe kusetshenziswa isibonakhulu sesigaba sokuqhathanisa se-Optika XDS-3 (enenjongo engu-40x) exhunywe kukhamera ye-Tucson ISH1000 ngo-30 fps ngemizuzwana emi-3.Sebenzisa isofthiwe ye-CASA ukufunda okungenani izindawo ezintathu kanye nama-trajectories wesidoda angu-500 isampula ngayinye.Ividiyo erekhodiwe icutshungulwe kusetshenziswa i-CASA eyenziwe ekhaya.Incazelo ye-motility ku-plug-in ye-CASA isekelwe esivinini sokubhukuda sesidoda uma kuqhathaniswa nesilinganiso sokugeleza, futhi ayifaki ezinye imingcele ezifana nokunyakaza okuhlangene, njengoba lokhu kutholakale ukuthi kunokwethenjelwa kakhulu ekugelezeni kwamanzi.Ukunyakaza kwe-Rheological kuchazwa njengokuhamba kwamangqamuzana esidoda ngokumelene nesiqondiso sokugeleza koketshezi.I-Spermatozoa enezici ze-rheological yahlukaniswa ngenani le-spermatozoa ehamba phambili;i-spermatozoa eyayiphumule futhi isidoda esinyakazayo asifakiwe ekubaleni.
Wonke amakhemikhali asetshenzisiwe atholwe kwa-Elgomhoria Pharmaceuticals (Cairo, Egypt) ngaphandle uma kuboniswe ngenye indlela.Idivayisi yenziwe njengoba kuchazwe ngu-El-sherry et al.40 nezinye izinguquko.Izinto ezisetshenziswa ukwenza iziteshi ezincane zihlanganisa amapuleti engilazi (Howard Glass, Worcester, MA), SU-8-25 resistant negative (MicroChem, Newton, CA), diacetone alcohol (Sigma Aldrich, Steinheim, Germany), kanye ne-polyacetone.-184, eDow Corning, eMidland, eMichigan).Amashaneli amancane akhiwa kusetshenziswa i-lithography ethambile.Okokuqala, imaskhi yobuso evikelayo ecacile enomklamo we-microchannel oyifunayo yanyatheliswa kuphrinta enesinqumo esiphezulu (Prismatic, Cairo, Egypt and Pacific Arts and Design, Markham, ON).Izingcweti zenziwa kusetshenziswa amapuleti engilazi njengama-substrates.Amapuleti ahlanzwa nge-acetone, i-isopropanol namanzi enziwe i-deionized abese embozwa ngesendlalelo esingu-20 µm se-SU8-25 nge-spin coating (3000 rpm, 1 min).Izendlalelo ze-SU-8 zase zomiswa kahle (65°C, 2 min kanye no-95°C, 10 min) futhi zavezwa emisebeni ye-UV imizuzwana engu-50.Ngemuva kokuchayeka bhaka ku-65°C naku-95°C iminithi elingu-1 nemizuzu emi-4 ukuze uphambanise izendlalelo eziveziwe ze-SU-8, okulandelwa ukuthuthukiswa kotshwala be-diacetone imizuzu engu-6.5.Bhaka ama-waffles kanzima (200 ° C imizuzu engu-15) ukuze uqhubeke uqinisa ungqimba lwe-SU-8.
I-PDMS yalungiswa ngokuxuba i-monomer ne-hardener ngesilinganiso sesisindo esingu-10:1, yase ikhishwa ku-vacuum desiccator futhi ithululelwa kuhlaka oluyinhloko lwe-SU-8.I-PDMS yelashwa kuhhavini (i-120 ° C, i-30 min), bese iziteshi zisikwa, zihlukaniswe ne-master, futhi zenziwe nge-perforated ukuvumela amashubhu ukuba afakwe endaweni yokungena nokuphuma kwe-microchannel.Ekugcineni, iziteshi ezincane ze-PDMS zanamathiselwa unomphela kumaslayidi e-microscope kusetshenziswa iphrosesa ephathekayo ye-corona (Imikhiqizo ye-Electro-Technic, Chicago, IL) njengoba kuchazwe kwenye indawo.I-microchannel esetshenziswe kulolu cwaningo ikala u-200 µm × 20 µm (W × H) futhi inobude obungu-3.6 cm.
Ukugeleza koketshezi okubangelwa ukucindezela kwe-hydrostatic ngaphakathi kwe-microchannel kufinyelelwa ngokugcina izinga loketshezi endaweni yokugcina amanzi ngaphezu komehluko wobude Δh39 ku-outlet reservoir (Fig. 1).
lapho u-f eyi-coefficient of friction, echazwa ngokuthi f = C/Re yokugeleza kwe-laminar esiteshini esingunxande, lapho u-C ehlala njalo kuye ngokuthi isici seshaneli, L ubude besiteshi esincane, i-Vav isilinganiso sesivinini ngaphakathi kwe-microchannel, i-Dh ububanzi be-hydraulic yesiteshi, g – ukusheshisa amandla adonsela phansi.Ngokusebenzisa lesi sibalo, isivinini sesiteshi esimaphakathi singabalwa kusetshenziswa isibalo esilandelayo: