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ʻO ka momona o nā manu e pili ana i ko lākou hiki ke mālama i ka sperm kūpono no ka manawa lōʻihi i loko o ka sperm storage tubules (SST).ʻO ka hana pololei e komo ai ka spermatozoa, noho i loko, a haʻalele i ka SST e hoʻopaʻapaʻa mau.ʻO ka sperm o nā moa sharkasi i hōʻike i kahi ʻano kiʻekiʻe i ka agglutination, e hana ana i nā puʻupuʻu filamentous mobile i loaʻa nā cell he nui.Ma muli o ka paʻakikī o ka nānā ʻana i ka motility a me ke ʻano o ka spermatozoa i loko o kahi opaque fallopian tube, ua hoʻohana mākou i kahi mea microfluidic me kahi microchannel cross-section e like me ka spermatozoa e aʻo ai i ka agglutination a me ka motility spermatozoa.Kūkākūkā kēia haʻawina i ke ʻano o ka hana ʻana o nā pūʻolo sperm, pehea lākou e neʻe ai, a me kā lākou kuleana kūpono i ka hoʻonui ʻana i ka noho sperm ma ka SST.Ua noiʻi mākou i ka wikiwiki o ka sperm a me ke ʻano rheological i ka wā i hoʻopuka ʻia ai ke kahe wai i loko o ke kahawai microfluidic e ke kaomi hydrostatic (ka kahe kahe = 33 µm/s).Hiki i ka spermatozoa keʻauʻau i ke au (positive rheology) a me ka wikiwiki o ka pūpū spermatozoon e hoʻemi nuiʻia i ka hoʻohālikelikeʻana i ka spermatozoa hoʻokahi.Ua ʻike ʻia ka neʻe ʻana o nā puʻupuʻu sperm i loko o kahi spiral a hoʻonui i ka lōʻihi a me ka mānoanoa i ka wā e hoʻonui ʻia ai ka sperm hoʻokahi. Ua ʻike ʻia nā pūʻolo sperm e hoʻokokoke a pili ana i nā ʻaoʻao ʻaoʻao o nā awāwa microfluidic e pale ʻia ai me ka wikiwiki o ka kahe wai > 33 µm/s. Ua ʻike ʻia nā pūʻolo sperm e hoʻokokoke a pili ana i nā ʻaoʻao ʻaoʻao o nā awāwa microfluidic e pale ʻia ai me ka wikiwiki o ka kahe wai > 33 µm/s. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрофлюитодных каналботься скоростью потока жидкости> 33 мкм / с. Ua ʻike ʻia nā pūʻolo sperm e hoʻokokoke a pili i nā paia ʻaoʻao o nā awāwa microfluidic e pale aku ai i ka ʻau ʻia ʻana i nā kahe kahe wai >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。33 µm/s 扫过. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрожидкостногочбеникостного кабнала, м жидкости со скоростью > 33 мкм/с. Ua ʻike ʻia nā pūʻolo sperm e hoʻokokoke a pili i nā paia ʻaoʻao o ke kahawai microfluidic i pale ʻia e ka kahe wai ma>33 µm/s.Ua hōʻike ʻia ka nānā ʻana a me ka hoʻouna ʻana i ka microscopy electron ua kākoʻo ʻia nā pūpū sperm e ka nui o nā mea paʻa.ʻO kaʻikepili i loaʻa e hōʻike ana i ka mobility kūikawā o Sharkazi chicken spermatozoa, a me ka hiki i ka spermatozoa ke agglutinate a me ka hanaʻana i nā puʻupuʻu kelepona, e kōkua ana i kaʻike maikaʻi o ka mālama lōʻihi o ka spermatozoa i SMT.
No ka hoʻokō ʻana i ka fertilization i loko o ke kanaka a me ka hapa nui o nā holoholona, ​​​​pono e hōʻea ka sperm a me nā hua i ke kahua o ka fertilization i ka manawa kūpono.No laila, pono ka male ma mua a i ka manawa o ka ovulation.Ma ka ʻaoʻao ʻē aʻe, mālama kekahi mau mea momona, e like me nā ʻīlio, a me nā ʻano manu ʻole, e like me nā iniseti, iʻa, nā mea kolo, a me nā manu, mālama i ka sperm i loko o kā lākou mau mea hānau no ka manawa lōʻihi a hiki i ka mākaukau ʻana o kā lākou mau hua no ka fertilization (asynchronous fertilization 1).Hiki i nā manu ke mālama i ke ola o ka spermatozoa hiki ke hoʻomomona i nā hua no 2-10 pule2.
He hiʻohiʻona kūʻokoʻa kēia e hoʻokaʻawale i nā manu mai nā holoholona ʻē aʻe, no ka mea, hāʻawi ia i kahi kiʻekiʻe o ka fertilization ma hope o hoʻokahi insemination no kekahi mau pule me ka ʻole o ka male ʻana a me ka ovulation.ʻO ka ʻōkana mālama sperm nui, i kapa ʻia ʻo ka sperm storage tubule (SST), aia i loko o nā ʻāpana mucosal kūloko ma ka hui uterovaginal.A hiki i kēia lā, ʻaʻole maopopo loa nā ʻano hana e komo ai ka sperm, noho, a puka i waho o ka panakō sperm.Ma muli o nā haʻawina mua, nui nā kuhiakau i waiho ʻia i mua, akā ʻaʻole i hōʻoia ʻia kekahi o lākou.
Ua manaʻo ʻo Forman4 e mālama ka spermatozoa i ko lākou noho ʻana i loko o ka lua SST ma o ka neʻe ʻana oscillatory mau e kūʻē i ke kuhikuhi o ke kahe ʻana o ka wai ma o nā ala protein e loaʻa ana ma nā cell epithelial SST (rheology).Hoʻopau ʻia ka ATP ma muli o ka hana hae hae mau e pono ai e mālama i ka sperm i loko o ka lumen SST a emi iho ka motility a hiki i ka lawe ʻia ʻana o ka sperm i waho o ka panakō sperm ma ke kahe wai a hoʻomaka i kahi huakaʻi hou i ka piʻi ʻana o ka fallopian tube e hoʻomomona i ka sperm.Huamoa (Forman4).Kākoʻo ʻia kēia ʻano hoʻohālike o ka mālama ʻana i nā sperm e ka ʻike ʻana e ka immunocytochemistry o nā aquaporins 2, 3 a me 9 i loaʻa i nā pūnaewele epithelial SST.I kēia lā, nele nā ​​haʻawina e pili ana i ka rheology semen moa a me kāna kuleana i ka mālama ʻana i ka SST, koho sperm vaginal, a me ka hoʻokūkū sperm.I loko o nā moa, komo ka sperm i loko o ka ʻōpū ma hope o ka male maoli, akā ʻoi aku ma mua o 80% o ka spermatozoa i hoʻokuʻu ʻia mai ka ʻōpū ma hope koke o ka male ʻana.Hōʻike kēia ʻo ka vagina ka pae mua no ke koho ʻana i ka sperm i nā manu.Eia kekahi, ua hōʻike ʻia ʻo ka liʻiliʻi ma mua o 1% o ka spermatozoa i hoʻomoʻa ʻia i loko o ka ʻōpū e pau i SSTs2.I ka hoʻokomo ʻia ʻana o nā moa i loko o ka ʻōpū, ʻoi aku ka nui o ka spermatozoa i hiki i ka SST ke piʻi aʻe i 24 mau hola ma hope o ka insemination.I kēia manawa, ʻaʻole maopopo ke ʻano o ke koho ʻana i ka sperm i kēia kaʻina hana, a he mea nui paha ka motility sperm i ka lawe ʻana i ka sperm SST.Ma muli o ka mānoanoa a me ka opaque paia o ka fallopian tubes, he paʻakikī ke nānā pono i ka sperm motility i loko o ka fallopian tubes o nā manu.No laila, nele mākou i ka ʻike kumu e pili ana i ka hoʻololi ʻana o ka spermatozoa i SST ma hope o ka fertilization.
Ua ʻike ʻia ʻo Rheology ma ke ʻano he kumu koʻikoʻi e hoʻomalu ana i ka lawe ʻana i ka sperm i loko o ka ʻōpū mammalian.Ma muli o ka hiki ʻana o ka spermatozoa motile e neʻe kūʻē i ka manawa, ua hoʻohana ʻo Zaferani et al8 i kahi ʻōnaehana microfluidic corra e hoʻokaʻawale i ka spermatozoa motile mai nā mea hoʻohālike i kākau ʻia.He mea nui kēia ʻano o ka hoʻokaʻawale ʻana i ka hua no ka mālama ʻana i ka maʻi infertility a me ka noiʻi lapaʻau, a ʻoi aku ka maikaʻi ma mua o nā ʻano kuʻuna i ka manawa a me ka hana ikaika a hiki ke hoʻololi i ka sperm morphology a me ka hoʻonohonoho pono.Eia naʻe, a hiki i kēia lā, ʻaʻohe haʻawina i hana ʻia e pili ana i ka hopena o nā mea huna mai nā ʻōpū o nā moa i ka motility sperm.
Me ka manaʻo o ke ʻano o ka mālama ʻana i ka sperm i mālama ʻia i loko o ka SST, nui nā mea noiʻi i ʻike i ka spermatozoa kamaʻāina agglutinate poʻo-i-poʻo i ka SST o nā moa 9, 10, quails 2, a me nā turkeys 11 e hana i nā pūpū sperm agglutinated.Manaʻo nā mea kākau he pilina ma waena o kēia agglutination a me ka mālama lōʻihi o ka spermatozoa i ka SST.
Ua hōʻike ʻo Tingari a me Lake12 i kahi hui ikaika ma waena o ka spermatozoa i ka sperm loaʻa gland o ka moa a nīnau ʻia inā e hoʻohui ʻia ka spermatozoa avian ma ke ʻano like me ka spermatozoa mammalian.Manaʻo lākou ʻo ka pilina hohonu ma waena o ka sperm i loko o nā vas deferens ma muli paha o ke koʻikoʻi i hana ʻia e ka nui o ka sperm ma kahi wahi liʻiliʻi.
I ka loiloi ʻana i ke ʻano o ka spermatozoa ma nā kiʻi aniani hou e kau ana, hiki ke ʻike ʻia nā hōʻailona transient o ka agglutination, ʻoi aku ma nā kihi o nā kulu semen.Eia nō naʻe, ua hoʻopilikia pinepine ʻia ka agglutination e ka hana rotational e pili ana me ka neʻe mau ʻana, e wehewehe ana i ke ʻano transient o kēia ʻano.Ua ʻike pū ka poʻe noiʻi i ka wā i hoʻohui ʻia ai ka mea hoʻoheheʻe i ka semen, ua ʻike ʻia nā aggregates cell "like-thread".
ʻO nā hoʻāʻo mua e hoʻohālike i kahi spermatozoon i hana ʻia ma ka wehe ʻana i kahi uwea lahilahi mai kahi kulu e kau ana, kahi i puka mai ai kahi sperm-like vesicle e puka ana mai ka kulu o ka hua.Ua laina koke ka spermatozoa ma ke ʻano like i loko o ka vesicle, akā nalo koke ka ʻāpana holoʻokoʻa ma muli o ka palena 3D.No laila, no ke aʻo ʻana i ka agglutination spermatozoa, pono e nānā pono i ka motility a me ke ʻano o ka spermatozoa i loko o nā tubules mālama sperm kaʻawale, he paʻakikī ke hoʻokō.No laila, pono e hoʻomohala i kahi mea hana e hoʻohālike i ka spermatozoa e kākoʻo i nā noiʻi o ka motility sperm a me ka hana agglutination.Ua hōʻike ʻo Brillard et al13 he 400-600 µm ka lōʻihi o ka loaʻa ʻana o nā tubules mālama sperm i loko o nā moa makua, akā hiki i kekahi mau SST ke lōʻihi i ka 2000 µm.Ua hoʻokaʻawale ʻo Mero lāua ʻo Ogasawara14 i nā kelepa seminiferous i loko o ka nui a me ka hoʻonui ʻole ʻana i ka sperm storage tubules, ʻo ia mau mea ʻelua i like ka lōʻihi (~ 500 µm) a me ka laula ʻāʻī (~ 38 µm), akā ʻo ka mean lumen diameter o nā tubules he 56.6 a me 56.6 µm.., i kēlā me kēia 11.2 μm, kēlā me kēia.Ma ke aʻo ʻana i kēia manawa, ua hoʻohana mākou i kahi mea microfluidic me ka nui o ke kahawai ʻo 200 µm × 20 µm (W × H), nona ka pauku kea i kahi kokoke i ka SST i hoʻonui ʻia.Eia hou, ua nānā mākou i ka motility sperm a me ka ʻano agglutination i ka wai kahe, i kūlike me ke kuhiakau o Foreman e mālama ana ka wai i hana ʻia e nā cell epithelial SST i ka sperm i loko o ka lumen i kahi kuhikuhi kūʻē (rheological).
ʻO ka pahuhopu o kēia noiʻi ʻana ʻo ia ka lanakila ʻana i nā pilikia o ka nānā ʻana i ka motility spermatozoa i loko o ka ʻōpū fallopian a pale i nā pilikia o ke aʻo ʻana i ka rheology a me ke ʻano o ka spermatozoa i loko o kahi kaiapuni ikaika.Ua hoʻohana ʻia kahi mea microfluidic e hana ana i ke kaomi hydrostatic e hoʻohālikelike i ka motility sperm i loko o ke ʻano o ka moa.
Ke hoʻokomo ʻia kahi kulu o kahi laʻana sperm i hoʻoheheʻe ʻia (1:40) i loko o ka mīkini microchannel, hiki ke ʻike ʻia ʻelua ʻano o ka motility sperm (sperm kaʻawale a me ka sperm paʻa).Eia kekahi, ua maʻa ka spermatozoa e ʻauʻau kūʻē i kēia manawa (positive rheology; video 1, 2). ʻOiai he haʻahaʻa ka wikiwiki o nā pūpū sperm ma mua o ka sperm lonesome (p <0.001), ua hoʻonui lākou i ka pākēneka o ka sperm e hōʻike ana i ka rheotaxis maikaʻi (p <0.001; Papa 2). ʻOiai he haʻahaʻa ka wikiwiki o nā pūpū sperm ma mua o ka sperm lonesome (p <0.001), ua hoʻonui lākou i ka pākēneka o ka sperm e hōʻike ana i ka rheotaxis maikaʻi (p <0.001; Papa 2). Хотя пучки сперматозоидов имели более низкую скорость, чем у одиночныпх сперматозоидов (p < 0,001), олимицил дов, демонстрирующих положительный реотаксис (p < 0,001; таблица 2). ʻOiai he haʻahaʻa ka wikiwiki o nā pūpū spermatozoa ma mua o ka spermatozoa hoʻokahi (p <0.001), ua hoʻonui lākou i ka pakeneka o ka spermatozoa e hōʻike ana i ka rheotaxis maikaʻi (p <0.001; Papa 2).尽管精子束的速度低于孤独精子的速度（p < 0.001），但它们增加了显示阳性显示阳性流的示阳性流的示阳性流的示阳了< 0.001；表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 百浐倧 正（p <0.001 ； 2……..））））） Хотя скорость пучков сперматозоидов была ниже, чем у одиночных сперматозоидов (p < 0,001), они увеличирдит жительной реологией (p < 0,001; таблица 2). ʻOiai ua emi ka wikiwiki o nā pūpū sperm ma mua o ka spermatozoa hoʻokahi (p <0.001), ua hoʻonui lākou i ka pakeneka o ka spermatozoa me ka rheology maikaʻi (p <0.001; Papa 2).ʻO ka rheology maikaʻi no ka spermatozoa hoʻokahi a me nā tufts i manaʻo ʻia ma kahi o 53% a me 85%, kēlā me kēia.
Ua ʻike ʻia ʻo ka spermatozoa o nā moa sharkasi ma hope koke o ka ejaculation e hana i nā puʻupuʻu linear, ʻo ia ka nui o nā kānaka.Hoʻonui kēia mau puʻupuʻu i ka lōʻihi a me ka mānoanoa i ka manawa a hiki ke noho i loko o ka vitro no kekahi mau hola ma mua o ka pau ʻana (wikiō 3).Hoʻohālikelike ʻia kēia mau pūʻolo filamentous e like me ka echidna spermatozoa i hana ʻia ma ka hope o ka epididymis.Ua ʻike ʻia ʻo Sharkashi hen semen i ke ʻano kiʻekiʻe o ka agglutinate a hana i kahi puʻupuʻu reticulate ma lalo o hoʻokahi minuke ma hope o ka hōʻiliʻili ʻana.He ikaika kēia mau kaola a hiki ke hoʻopili i nā paia kokoke a i ʻole nā ​​mea paʻa.ʻOiai e hoʻemi ana nā pūʻolo sperm i ka wikiwiki o nā pūnaewele sperm, ua maopopo ka macroscopically hoʻonui lākou i kā lākou linearity.ʻOkoʻa ka lōʻihi o nā pūʻolo ma muli o ka nui o ka sperm i hōʻiliʻili ʻia i loko o nā pūʻolo.Ua hoʻokaʻawale ʻia ʻelua ʻāpana o ka pūʻolo: ʻo ka ʻāpana mua, me ke poʻo manuahi o ka sperm agglutinated, a me ka ʻaoʻao hope, me ka huelo a me ka hopena distal a pau o ka sperm.Ke hoʻohana nei i kahi kāmela kiʻekiʻe (950 fps), ua ʻike ʻia nā poʻo manuahi o ka spermatozoa agglutinated i ka ʻāpana mua o ka pūʻolo, ke kuleana o ka neʻe ʻana o ka puʻupuʻu ma muli o kā lākou neʻe oscillatory, e kauo ana i nā mea i koe i loko o ka pūpū me kahi neʻe helical (Video 4).Eia nō naʻe, i loko o nā puʻupuʻu lōʻihi, ua ʻike ʻia ua pili kekahi mau poʻo sperm manuahi i ke kino a ʻo ka ʻāpana hope o ka puʻupuʻu e hana ma ke ʻano he vane e kōkua ai i ka hoʻoulu ʻana i ka ʻōpala.
ʻOiai e kahe mālie ana ka wai, neʻe like nā pūʻolo sperm i kekahi i kekahi, akā naʻe, hoʻomaka lākou e uhi a pili i nā mea a pau e noho mālie, i ʻole e holoi ʻia e ke kahe o kēia manawa i ka piʻi ʻana o ka wikiwiki.Hoʻokumu ʻia nā pūʻolo i ka wā e hoʻokokoke aku ai ka lima o nā pūnaʻi sperm i kekahi i kekahi, hoʻomaka lākou e neʻe i ka synchrony a hoʻopuni a puni kekahi i kekahi, a laila pili i kahi mea paʻa.Hōʻike nā kiʻi 1 a me 2 i ke ʻano o ka hoʻokokoke ʻana o ka sperm i kekahi i kekahi, e hana ana i ka hui ʻana i ka wā e ʻōwili ai nā huelo i kekahi i kekahi.
Ua noi nā mea noiʻi i ke kaomi hydrostatic e hana i ka kahe wai i loko o kahi microchannel e aʻo ai i ka rheology sperm.Ua hoʻohana ʻia kahi microchannel me ka nui o 200 µm × 20 µm (W × H) a me ka lōʻihi o 3.6 µm.E hoʻohana i nā microchannels ma waena o nā ipu me nā syringes i hoʻopaʻa ʻia ma nā wēlau.Ua hoʻohana ʻia ke kala meaʻai i mea e ʻike nui ʻia ai nā kahawai.
E hoʻopaʻa i nā kaula pili a me nā mea pono i ka paia.Ua lawe ʻia ke wikiō me kahi microscope hoʻohālikelike.Me kēlā me kēia kiʻi, hōʻike ʻia nā microscopy māhele ʻokoʻa a me nā kiʻi palapala ʻāina.(A) ʻO ka pilina ma waena o nā kahawai ʻelua e pale i ke kahe ma muli o ka neʻe helical (pua ʻulaʻula).(B) ʻO ka pilina ma waena o ka puʻupuʻu paipu a me ka paia o ke kahawai (nā pua ʻulaʻula), i ka manawa like ua hoʻopili ʻia lākou i ʻelua pūʻolo ʻē aʻe (nā pua melemele).(C) Hoʻomaka nā pūʻolo hua manu i loko o ke kahawai microfluidic e hoʻopili kekahi me kekahi (nā pua ʻulaʻula), e hana ana i mau pūʻulu sperm.(D) Ka hoʻokumu ʻana i kahi pūnaewele o nā pūʻolo sperm.
Ke hoʻokomo ʻia kahi kulu o ka sperm diluted i loko o ka microfluidic device a hana ʻia kahi kahe, ʻike ʻia ka sperm beam e neʻe i ka ʻaoʻao o ke kahe.Hoʻopili pono nā pūʻolo i nā paia o nā microchannels, a ʻo nā poʻo manuahi i ka ʻāpana mua o nā pūʻolo pili pono iā lākou (wikiō 5).Hoʻopili pū lākou i nā ʻāpana paʻa i ko lākou ala, e like me ka ʻōpala, e pale aku i ka lawe ʻia ʻana e ke au.I ka wā lōʻihi, lilo kēia mau ʻōpala i mau filament lōʻihi e paʻa ana i nā spermatozoa hoʻokahi a me nā puʻupuʻu pōkole (Video 6).I ka hoʻomaka ʻana o ke kahe ʻana, hoʻomaka nā laina lōʻihi o ka sperm e hana i kahi pūnaewele o nā laina sperm (Video 7; Kiʻi 2).
Ma ka holo kiʻekiʻe (V > 33 µm/s), ua hoʻonui ʻia ka neʻe ʻana o nā kaula ma ke ʻano he hoʻāʻo e hopu i nā pūʻolo sperm hoʻokahi i ʻoi aku ka maikaʻi o ka pale ʻana i ka ikaika o ke kahe. Ma ka holo kiʻekiʻe (V > 33 µm/s), ua hoʻonui ʻia ka neʻe ʻana o nā kaula ma ke ʻano he hoʻāʻo e hopu i nā pūʻolo sperm hoʻokahi i ʻoi aku ka maikaʻi o ka pale ʻana i ka ikaika o ke kahe. При высокой скорости потока (V > 33 мкм/с) спиралевидные движения нитей усиливаются, ерматозоидов, образующих пучки, которые лучше противостоят дрейфующей силе потока. Ma nā kahe kahe kiʻekiʻe (V > 33 µm/s), piʻi ka neʻe ʻana o ka helical o nā kaula i ko lākou hoʻāʻo ʻana e hopu i nā pūʻolo spermatozoa pākahi i ʻoi aku ka maikaʻi o ka pale ʻana i ka ikaika o ke kahe.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，我們运动增加，以试图捕捉许多形成束的单个，更华个，从耶和流动的漂移力。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 运动 增加 ， 以 试图 许多 形成 束 单 个 ，抵抗 的 漂移力。。。。。。。。 При высоких скоростях потока (V > 33 мкм/с) бразующих пучки, чтобы лучше сопротивляться силам дрейфа потока. Ma nā kahe kahe kiʻekiʻe (V > 33 µm/s), piʻi ka neʻe ʻana o nā filament i ka hoʻāʻo ʻana e hopu i nā pūʻolo spermatozoa pākahi i mea e pale maikaʻi aku ai i nā ikaika neʻe o ke kahe.Ua ho'āʻo pū lākou e hoʻopili i nā microchannels i nā ʻaoʻao ʻaoʻao.
Ua ʻike ʻia nā pūʻolo sperm ma ke ʻano he pūʻulu o nā poʻo sperm a me nā huelo wili e hoʻohana ana i ka microscopy māmā (LM).Ua ʻike ʻia nā pūʻolo hua manu me nā ʻāpana like ʻole he poʻo wili a me nā hōʻiliʻili hae, nā huelo sperm i hoʻohui ʻia he nui, nā poʻo sperm i hoʻopili ʻia i kahi huelo, a me nā poʻo sperm me ka nuclei piʻo e like me nā nuclei fused lehulehu.ʻO ka microscopy electron transmission (TEM).Ua hōʻike ʻia ka nānā ʻana i ka microscopy electron (SEM) i nā pūʻolo sperm i uhi ʻia o nā poʻo sperm a ua hōʻike nā sperm aggregates i kahi pūnaewele pili o nā huelo i ʻōwili ʻia.
Ua aʻo ʻia ka morphology a me ka ultrastructure o ka spermatozoa, ka hoʻokumu ʻana i nā pūpū spermatozoa me ka microscopy māmā (hapa ʻāpana), scanning electron microscopy (SEM) a me transmission electron microscopy (TEM), sperm smears i hoʻopaʻa ʻia me ka ʻalani acridine a nānā ʻia me ka microscopy epifluorescence.
Ua hōʻike ʻia ka hoʻopaʻa ʻana o ka sperm smear me ka ʻalani acridine (Fig. 3B) i ka paʻa ʻana o nā poʻo sperm a uhi ʻia me nā mea huna, i alakaʻi ʻia i ke kūkulu ʻia ʻana o nā puʻupuʻu nui (Fig. 3D).ʻO nā pūʻolo sperm he mau sperm aggregates me kahi pūnaewele o nā huelo i hoʻopili ʻia (Fig. 4A-C).Hoʻokumu ʻia nā pūʻolo sperm me nā huelo o nā spermatozoa he nui i paʻa pū (Fig. 4D).Ua uhi nā mea huna (Fig. 4E,F) i nā poʻo o nā pūpū spermatozoa.
Hoʻokumu ʻia ka pūʻolo spermatozoa Me ka hoʻohana ʻana i ka microscopy ʻokoʻa a me nā sperm smears i hoʻopaʻa ʻia me ka ʻalani acridine, ua hōʻike ʻia e pili pū nā poʻo o ka spermatozoa.(A) Hoʻomaka ka hoʻokumu ʻana o ka sperm tuft me kahi sperm (poʻai keʻokeʻo) a me ʻekolu sperm (poʻai melemele), me ka hoʻomaka ʻana o ka spiral ma ka huelo a pau ma ke poʻo.(B) Photomicrograph o kahi sperm smear i hoʻopaʻa ʻia me ka ʻalani acridine e hōʻike ana i nā poʻo sperm pili (nā pua).Uhi ka hoʻokuʻu ʻana i ke poʻo.Hoʻonui × 1000. (C) Ka hoʻomohala ʻana i kahi kaola nui i lawe ʻia e ke kahe i loko o kahi kahawai microfluidic (e hoʻohana ana i kahi pahu pahu kiʻekiʻe ma 950 fps).(D) Micrograph o kahi sperm smear i hoʻopaʻa ʻia me ka ʻalani acridine e hōʻike ana i nā puʻu nui (nā pua).Hoʻonui: ×200.
E nānā ana i ka micrograph electron o ka sperm beam a me ka sperm smear i hoʻopaʻa ʻia me ka ʻalani acridine.(A, B, D, E) he mau micrographs electron scanning kala kikohoʻe o ka spermatozoa, a ʻo C a me F he micrographs o nā sperm smears ʻalani ʻalani e hōʻike ana i ka hoʻopili ʻana o nā spermatozoa lehulehu e kāʻei ana i ka pūnaewele caudal.(AC) Hōʻike ʻia nā hōʻuluʻulu sperm ma ke ʻano he pūnaewele o nā huelo i hoʻopili ʻia (nā pua).(D) Ka hoʻopili ʻana o kekahi mau spermatozoa (me ka mea hoʻopili, ka ʻulaʻula ʻulaʻula, ka pua) e hoʻopuni ana i ka huelo.(E a me F) ʻO nā hui poʻo sperm (pointers) i uhi ʻia me nā mea hoʻopili (pointers).Hoʻokumu ka spermatozoa i nā pūʻolo me nā ʻano like me ka vortex (F).(C) ×400 a me (F) ×200 hoʻonui.
I ka hoʻohana ʻana i ka microscopy electron transmission, ua ʻike mākou ua pili nā pūpū sperm i nā huelo (Fig. 6A, C), nā poʻo i pili i nā huelo (Fig. 6B), a i ʻole nā ​​​​poʻo i pili i nā huelo (Fig. 6D).ʻO nā poʻo o ka spermatozoa i loko o ka puʻupuʻu he curved, e hōʻike ana ma ka ʻāpana ʻelua mau ʻāpana nuklea (Fig. 6D).I loko o ka pūʻolo incision, he poʻo wili ka spermatozoa me ʻelua mau ʻāpana nuklea a me nā ʻāpana flagellar he nui (Fig. 5A).
ʻO ka micrograph electron kala kikohoʻe e hōʻike ana i nā huelo hoʻohui i ka pūʻolo sperm a me ka mea hoʻohui e hoʻopili ana i nā poʻo sperm.(A) Hoʻopili ʻia ka huelo o ka nui o ka spermatozoa.E nānā i ke ʻano o ka huelo ma ke kiʻi (arrow) a me ka ʻāina (arrow).(B) Ua pili ke poo (arrow) o ka sperm i ka huelo (arrow).(C) Hoʻopili ʻia kekahi mau huelo sperm (nā pua).(D) Hoʻohui ʻia nā mea hoʻohuihui (AS, uliuli) i ʻehā poʻo sperm (ʻulaʻula).
Ua hoʻohana ʻia ka microscopy electron scanning no ka ʻike ʻana i nā poʻo sperm i loko o nā pūʻolo sperm i uhi ʻia me nā mea huna a i ʻole nā ​​membranes (Figure 6B), e hōʻike ana ua hoʻopaʻa ʻia nā pūpū sperm e nā mea extracellular.Hoʻopili ʻia ka mea i hoʻohui ʻia i loko o ke poʻo sperm (e like me ke poʻo o ka jellyfish; Fig. 5B) a hoʻonui ʻia ma kahi mamao loa, e hāʻawi ana i kahi hiʻohiʻona melemele nani ma lalo o ka microscopy fluorescence ke hoʻopaʻa ʻia me ka ʻalani acridine (Fig. 6C).ʻIke maopopo ʻia kēia mea ma lalo o kahi microscope scanning a manaʻo ʻia he mea paʻa.Ua hōʻike ʻia nā ʻāpana semi-maniʻi (Fig. 5C) a me nā sperm smears i hoʻopaʻa ʻia me ka ʻalani acridine i nā pūʻolo sperm i loaʻa nā poʻo paʻa paʻa a me nā huelo wili (Fig. 5D).
Nā kiʻi kiʻi like ʻole e hōʻike ana i ka hōʻuluʻulu ʻana o nā poʻo sperm a me nā huelo i pelu ʻia me ka hoʻohana ʻana i nā ʻano like ʻole.(A) E hōʻike ana i ke poʻo sperm i wili ʻia me kahi nucleus ʻelua ʻāpana (uliuli) a me kekahi mau ʻāpana hae ('ōmaʻomaʻo).(B) E hōʻike ana i kahi puʻupuʻu o nā poʻo sperm like me ka jellyfish (nā pua) i uhi ʻia.(C) ʻāpana ʻāpana ʻāpana ʻāpana e hōʻike ana i nā poʻo sperm aggregated (nā pua) a me nā huelo wili (nā pua).(D) Micrograph o kahi sperm smear i hoʻomaʻemaʻe ʻia me ka ʻalani acridine e hōʻike ana i nā hōʻuluʻulu o nā poʻo sperm (nā pua) a me nā huelo pili wili ʻia (nā pua).E hoʻomaopopo i ka uhi ʻana o kahi mea lāʻau (S) i ke poʻo o ka spermatozoon.(D) × 1000 hoʻonui.
Me ka hoʻohana ʻana i ka microscopy electron transmission (Fig. 7A), ua ʻike pū ʻia ua wili ʻia nā poʻo sperm a he ʻano spiral ka nuclei, e like me ka mea i hōʻoia ʻia e nā sperm smears i hoʻopaʻa ʻia me ka ʻalani acridine a nānā ʻia me ka microscopy fluorescence (Fig. 7B).
(A) ʻO ka micrograph electron hoʻouna kala kikohoʻe a me (B) ʻO ka sperm smear stained acridine e hōʻike ana i nā poʻo wili a me ka hoʻopili ʻana o nā poʻo sperm a me nā huelo (nā pua).(B) × 1000 hoʻonui.
ʻO kahi ʻike hoihoi, ʻo ka hui ʻana o ka sperm a Sharkazi e hana i nā puʻupuʻu filamentous mobile.ʻO nā waiwai o kēia mau pūʻolo e hiki ai iā mākou ke hoʻomaopopo i ko lākou kuleana kūpono i ka absorption a me ka mālama ʻana o ka spermatozoa i ka SST.
Ma hope o ka male ʻana, komo ka sperm i loko o ka puʻupaʻa a loaʻa i kahi kaʻina koho koʻikoʻi, e hopena i kahi helu liʻiliʻi o ka sperm i komo i ka SST15,16.I kēia lā, ʻaʻole maopopo nā hana e komo ai ka sperm a puka i waho o ka SST.I loko o ka moa, mālama ʻia ka spermatozoa i loko o ka SST no ka manawa lōʻihi o 2 a 10 mau pule, ma muli o ka ʻano6.Ke mau nei ka hoʻopaʻapaʻa e pili ana i ke ʻano o ka pua i ka wā e waiho ai i ka SST.Ke holo nei a hoomaha paha?I nā huaʻōlelo ʻē aʻe, pehea e mālama ai nā sperm cell i ko lākou kūlana i ka SST no ka lōʻihi?
Ua manaʻo ʻo Forman4 e hiki ke wehewehe ʻia ka noho SST a me ka hoʻokuke ʻana ma ke ʻano o ka motility sperm.Manaʻo nā mea kākau e mālama ka sperm i ko lākou kūlana ma ka ʻauʻau ʻana i ke kahe wai i hana ʻia e ka epithelium SST a ua hoʻokuʻu ʻia ka sperm mai ka SST i ka wā e hāʻule ai ko lākou wikiwiki ma lalo o kahi e hoʻomaka ai lākou e neʻe i hope ma muli o ka nele o ka ikehu.Ua hōʻoia ʻo Zaniboni5 i ka loaʻa ʻana o nā aquaporins 2, 3, a me 9 ma ka ʻāpana apical o nā pūnaewele epithelial SST, hiki ke kākoʻo ʻole i ke kumu hoʻohālike sperm storage o Foreman.Ma ka haʻawina o kēia manawa, ua ʻike mākou ua kokoke ka hapalua o ka spermatozoa o Sharkashi e hōʻike i ka rheology maikaʻi i loko o ka wai kahe, a ʻo nā pūpū sperm agglutinated e hoʻonui i ka helu o ka spermatozoa e hōʻike ana i ka rheology maikaʻi, ʻoiai ke hoʻolōʻihi nei ka agglutination iā lākou.ʻAʻole maopopo loa ke ʻano o ka holo ʻana o nā pūnana sperm i ka ʻōpū fallopian o ka manu i kahi o ka hoʻomomona ʻana.I loko o nā mammals, hoʻokaʻawale ka wai follicular i ka spermatozoa.Eia naʻe, ua manaʻo ʻia nā chemoattractants e kuhikuhi i ka spermatozoa e hoʻokokoke i kahi mamao7.No laila, ʻo nā hana ʻē aʻe ke kuleana no ka lawe ʻana i ka sperm.Ua hōʻike ʻia ka hiki o ka sperm e kuhikuhi a kahe i ka wai o ka fallopian tube i hoʻokuʻu ʻia ma hope o ka male ʻana he kumu nui ia i ka huli ʻana i ka sperm i nā ʻiole.Manaʻo ʻo Parker 17 e hele ka spermatozoa i nā oviducts ma ka ʻauʻau ʻana i ke au ciliary i nā manu a me nā mea kolo.ʻOiai ʻaʻole i hoʻāʻo ʻia i nā manu, ʻo Adolphi18 ka mea mua i ʻike i ka hāʻawi ʻana o ka sperm avian i nā hopena maikaʻi ke hana ʻia kahi ʻāpana wai ma waena o ka uhi uhi a me ka paheʻe me kahi pepa kānana.Rheology.Hoʻokomo ʻo Hino lāua ʻo Yanagimachi [19] i kahi paʻakikī ovary-tubal-uterine ʻiole i loko o ke apo perfusion a hoʻokomo i 1 µl o ka inika i loko o ka isthmus e ʻike ai i ke kahe ʻana o ka wai i loko o nā ʻōpū fallopian.Ua ʻike lākou i ka neʻe ikaika loa o ka ʻoki ʻana a me ka hoʻomaha ʻana i loko o ka puʻu fallopian, kahi e neʻe mau ai nā pōpō inika a pau i ka ampulla o ka ʻōpū fallopian.Hoʻoikaika nā mea kākau i ke koʻikoʻi o ke kahe ʻana o ka wai tubal mai ka lalo a i ka ʻōpū fallopian luna no ka hāpai ʻana o ka sperm a me ka hoʻoulu ʻana.Ua hōʻike ʻo Brillard20 i loko o nā moa a me nā turkey, neʻe ka spermatozoa ma o ka neʻe ikaika ʻana mai ka puka komo, kahi i mālama ʻia ai, a i ka hui utero-vaginal, kahi i mālama ʻia ai.Eia naʻe, ʻaʻole koi ʻia kēia neʻe ma waena o ka hui uterovaginal a me ka infundibulum no ka mea, lawe ʻia ka spermatozoa e ka neʻe ʻana.I ka ʻike ʻana i kēia mau ʻōlelo aʻoaʻo mua a me nā hopena i loaʻa i ka haʻawina o kēia manawa, hiki ke manaʻo ʻia ʻo ka hiki o ka spermatozoa ke neʻe i luna (rheology) kekahi o nā waiwai i hoʻokumu ʻia ai ke kaʻina koho.Hoʻoholo kēia i ka hele ʻana o ka spermatozoa ma o ka waha a me ko lākou komo ʻana i ka CCT no ka mālama ʻana.E like me ka Forman4 i ʻōlelo ai, hiki i kēia ke hoʻomaʻamaʻa i ke kaʻina hana o ka sperm e komo ana i ka SST a me kona wahi noho no kekahi manawa a laila puka i ka wā e hoʻomaka ai ka lohi.
Ma ka ʻaoʻao ʻē aʻe, ua manaʻo ʻo Matsuzaki a me Sasanami 21 e loaʻa i ka spermatozoa avian i nā loli motility mai ka dormancy i ka motility i loko o nā ʻāpana hānau kāne a me ka wahine.ʻO ka pale ʻana i ka motility sperm noho ma ka SST ua manaʻo ʻia e wehewehe i ka lōʻihi o ka mālama ʻana o ka sperm a laila rejuvenation ma hope o ka haʻalele ʻana i ka SST.Ma lalo o nā kūlana hypoxic, Matsuzaki et al.Ua hōʻike ʻo 1 i ka hana kiʻekiʻe a me ka hoʻokuʻu ʻana o ka lactate i ka SST, hiki ke alakaʻi i ka pale ʻana i ka motility sperm noho.I kēia hihia, ʻike ʻia ke koʻikoʻi o ka rheology sperm i ke koho ʻana a me ka absorption o ka spermatozoa, ʻaʻole i kā lākou mālama.
Ua manaʻo ʻia ke ʻano hoʻohālikelike o ka sperm agglutination he wehewehe kūpono no ka lōʻihi o ka mālama ʻana o ka sperm i ka SST, no ka mea he ʻano maʻamau kēia o ka paʻa ʻana o ka sperm i ka moa2,22,23.ʻO Bakst et al.Ua ʻike ʻia ʻo ka hapa nui o ka spermatozoa i pili kekahi i kekahi, e hana ana i nā ʻāpana fascicular, a ʻaʻole loaʻa ka spermatozoa hoʻokahi i loko o ka quail CCM.Ma kekahi ʻaoʻao, ʻo Wen et al.Ua ʻike ʻia ʻo 24 i ʻoi aku ka nui o nā spermatozoa i hoʻopuehu ʻia a ʻoi aku ka liʻiliʻi o nā ʻōpala spermatozoa i ka lumen SST i loko o nā moa.Ma muli o kēia mau ʻike, hiki ke manaʻo ʻia he ʻokoʻa ka propensity no ka sperm agglutination ma waena o nā manu a ma waena o ka spermatozoa i ka ejaculate hoʻokahi.Eia kekahi, ʻo Van Krey et al.Manaʻo ʻo 9 ʻo ka wehe ʻole ʻana o ka spermatozoa agglutinated ke kuleana no ke komo mālie ʻana o ka spermatozoa i loko o ka lumen o ka ʻōpū fallopian.Wahi a kēia kuhiakau, pono e kipaku mua ʻia ka spermatozoa me ka mana agglutination haʻahaʻa mai ka SST.Ma kēia pōʻaiapili, hiki i ka hiki o ka spermatozoa ke agglutinate ke kumu e hoʻololi ai i ka hopena o ka hoʻokūkū sperm i nā manu lepo.Eia kekahi, ʻoi aku ka lōʻihi o ka wehe ʻana o ka sperm agglutinated, ʻoi aku ka lōʻihi o ka ulu ʻana.
ʻOiai uaʻikeʻia ka hōʻuluʻuluʻana o ka spermatozoa a me ka hōʻiliʻiliʻana i loko o nā pūpū i loko o nā haʻawina2,22,24,ʻaʻole lākou i wehewehe ponoʻia ma muli o ka paʻakikī o kā lākou nānā kinematic i loko o ka SST.Ua hana ʻia kekahi mau hoʻāʻo e aʻo i ka sperm agglutination in vitro.Ua ʻike ʻia ka hōʻuluʻulu lōʻihi akā i ka wā i wehe ʻia ai ka uwea lahilahi mai ka hāʻule ʻana o ka ʻanoʻano.Ke alakaʻi nei kēia i ka ʻoiaʻiʻo e puka mai kahi ʻōpū elongated mai ka hāʻule, e hoʻohālike ana i ka seminal gland.Ma muli o nā palena 3D a me nā manawa hoʻomaloʻo kulu pōkole, ua hāʻule koke ka poloka holoʻokoʻa.Ma ka haʻawina o kēia manawa, me ka hoʻohana ʻana i nā moa Sharkashi a me nā ʻāpana microfluidic, ua hiki iā mākou ke wehewehe i ke ʻano o kēia mau puʻupuʻu a me ka neʻe ʻana.Hoʻokumu koke ʻia nā pūʻolo sperm ma hope o ka hōʻiliʻili ʻana i ka hua manu a ua ʻike ʻia e neʻe i loko o kahi spiral, e hōʻike ana i ka rheology maikaʻi ke loaʻa i ke kahe.Eia kekahi, ke nānā ʻia ma ka macroscopically, ua ʻike ʻia nā pūpū sperm e hoʻonui i ka linearity o ka motility i hoʻohālikelike ʻia me ka spermatozoa kaʻawale.Hōʻike kēia e hiki ke hoʻohui ʻia ka sperm agglutination ma mua o ke komo ʻana o SST a ʻaʻole i kaupalena ʻia ka hana sperm i kahi wahi liʻiliʻi ma muli o ke koʻikoʻi e like me ka manaʻo mua (Tingari a me Lake12).I ka wā o ka hoʻokumu ʻana o ka sperm, ʻauʻau ka spermatozoa i ka synchrony a hiki i ka hana ʻana i kahi hui, a laila ʻōwili ko lākou huelo i kekahi i kekahi a noho manuahi ke poʻo o ka spermatozoon, akā pili ka huelo a me ka ʻāpana o ka spermatozoon me kahi mea paʻa.No laila, ʻo ke poʻo manuahi o ka ligament ke kuleana no ka neʻe ʻana, e huki ana i ke koena o ka ligament.ʻO ka nānā ʻana i ka microscopy electron o nā pūʻolo sperm i hōʻike ʻia i nā poʻo sperm i hoʻopili ʻia i uhi ʻia me ka nui o nā mea paʻa, e manaʻo ana ua hoʻopili ʻia nā poʻo sperm i nā pūʻolo hoʻomaha, aia paha ma hope o ka hiki ʻana i ke kahua mālama (SST).
Ke hoʻopaʻa ʻia ka sperm smear me ka ʻalani acridine, hiki ke ʻike ʻia nā mea hoʻopili extracellular a puni nā pūnana sperm ma lalo o kahi microscope fluorescent.ʻO kēia mea e hiki ai i nā pūʻolo sperm ke hoʻopili a pili i nā ʻili a i ʻole nā ​​ʻāpana a puni i ʻole lākou e ʻauheʻe me ke kahe a puni.No laila, hōʻike kā mākou mau ʻike i ke kuleana o ka adhesion spermatozoa ma ke ʻano o nā pūpū mobile.ʻO ko lākou hiki ke ʻauʻau i ke au a pili i nā wahi kokoke e hiki ai i ka sperm ke noho lōʻihi i ka SST.
Ua hoʻohana ʻo Rothschild25 i kahi pahupaʻi hemocytometry no ke aʻo ʻana i ka puʻunaue lana ʻana o ka pua puaʻa i kahi kulu o ka hoʻokuʻu ʻana, e lawe ana i nā kiʻi kiʻi kiʻi ma o ka pahupaʻikiʻi me ka axis optical a me ke koʻikoʻi o ka microscope.Ua hōʻike ʻia nā hopena ua huki ʻia ka spermatozoa i ka ʻili o ke keʻena.Manaʻo nā mea kākau e loaʻa paha nā pilina hydrodynamic ma waena o ka sperm a me ka ʻili.Ma ka noʻonoʻo ʻana i kēia, a me ka hiki o Sharkashi semen moa ke hana i nā puʻupuʻu paʻa, hiki ke hoʻonui i ka likelike e pili ai ka semen i ka pā SST a mālama ʻia no ka manawa lōʻihi.
Ua hōʻike ʻo Bccetti a me Afzeliu26 e koi ʻia ka sperm glycocalyx no ka ʻike a me ka agglutination gamete.Ua ʻike ʻo Forman10 i ka hydrolysis o nā paʻa α-glycosidic i loko o ka glycoprotein-glycolipid coatings ma o ka mālama ʻana i ka pua manu me ka neuraminidase i hoʻemi ʻia ka momona me ka ʻole o ka hoʻopili ʻana i ka motility sperm.Manaʻo nā mea kākau i ka hopena o ka neuraminidase ma ka glycocalyx e hōʻemi i ka sperm sequestration ma ka utero-vaginal junction, a laila e hōʻemi ana i ka momona.ʻAʻole hiki i kā lākou ʻike ke haʻalele i ka hiki ke hōʻemi i ka mālama ʻana i ka neuraminidase i ka ʻike sperm a me ka oocyte.Ua ʻike ʻo Forman lāua ʻo Engel10 ua hoʻemi ʻia ka ulu ʻana i ka wā i hoʻokomo ʻia ai nā moa i loko o ka ʻōpū me ka pua i mālama ʻia me ka neuraminidase.Eia naʻe, ʻaʻole i pili ka IVF me ka neuraminidase i ka sperm mālama ʻia i ka ulu ʻana i ka hoʻohālikelike ʻana i nā moa mālama.Ua hoʻoholo nā mea kākau i ka hoʻololi ʻana i ka uhi glycoprotein-glycolipid a puni ka membrane sperm i hoʻemi i ka hiki o ka sperm e hoʻomoʻa ʻia e ka hōʻino ʻana i ka sequestration o ka sperm ma ka utero-vaginal junction, ka mea i hoʻonui i ka nalowale o ka sperm ma muli o ka wikiwiki o ka hui utero-vaginal, akā ʻaʻole pili i ka sperm a me ka ʻike hua manu.
I loko o nā turkeys ʻo Bakst a me Bauchan 11 i loaʻa nā vesicles liʻiliʻi a me nā ʻāpana membrane i ka lumen o ka SST a ʻike ʻia ua hui pū kekahi o kēia mau granules me ka membrane sperm.Manaʻo nā mea kākau e hiki i kēia mau pilina ke kōkua i ka mālama lōʻihi o ka spermatozoa i SST.Eia naʻe, ʻaʻole i wehewehe ka poʻe noiʻi i ke kumu o kēia mau ʻāpana, inā i hūnā ʻia e nā cell epithelial CCT, i hana ʻia a hūnā ʻia e ka ʻōnaehana hānau kāne, a i hana ʻia e ka sperm ponoʻī.Eia kekahi, ʻo kēia mau ʻāpana ke kuleana no ka agglutination.Ua hōʻike ʻo Grützner et al27 e hoʻopuka ana nā epididymal epithelial cell a hūnā i kahi protein kikoʻī e pono ai no ka hoʻokumu ʻana i nā ʻāpana seminal single-pore.Hōʻike pū nā mea kākau e pili ana ka hoʻopuehu ʻana o kēia mau pūʻolo i ka pilina o nā protein epididymal.Ua ʻike ʻo Nixon et al28 ua huna ka adnexa i kahi protein, ka osteonectin waiwai nui o ka cysteine ​​acid;Hoʻokomo ʻia ʻo SPARC i ka hoʻokumu ʻana o nā sperm tufts i nā echidnas pōkole a me nā platypuses.Hoʻopili ʻia ka hoʻopuehu ʻana o kēia mau kaola me ka nalowale o kēia pūmua.
Ma ka noiʻi o kēia manawa, hōʻike ka ultrastructural analysis me ka microscopy electron ua pili ka spermatozoa i ka nui o nā mea paʻa.Ua manaʻo ʻia ʻo kēia mau mea ke kuleana no ka agglutination e condenses ma waena a a puni nā poʻo pili, akā ma nā haʻahaʻa haʻahaʻa ma ka ʻāpana huelo.Manaʻo mākou ua hoʻokuʻu ʻia kēia mea agglutinating mai ka ʻōnaehana hānau kāne (epididymis a i ʻole vas deferens) me ka semen, no ka mea, ʻike pinepine mākou i ka semen e hoʻokaʻawale ana mai ka lymph a me ka plasma seminal i ka wā ejaculation.Ua hōʻike ʻia i ka hele ʻana o ka spermatozoa avian i loko o ka epididymis a me nā vas deferens, e hana ana lākou i nā loli pili i ka maturation e kākoʻo ana i ko lākou hiki ke hoʻopaʻa i nā protein a loaʻa i ka plasma lemma-pili glycoproteins.ʻO ka hoʻomau ʻana o kēia mau protein i nā membrane sperm noho ma ka SST e hōʻike ana e hiki i kēia mau protein ke hoʻoikaika i ka loaʻa ʻana o ka paʻa sperm membrane 30 a hoʻoholo i ko lākou momona 31.Ua hōʻike ʻo Ahammad et al32 i ka spermatozoa i loaʻa mai nā ʻāpana like ʻole o ka ʻōnaehana hānau kāne (mai nā testes a hiki i ka distal vas deferens) i hōʻike i ka piʻi holomua o ka viability ma lalo o nā kūlana mālama wai, me ka nānā ʻole i ka mahana o ka mālama ʻana, a piʻi aʻe ka viability i nā moa i loko o ka fallopian tubes ma hope o ka insemination artificial.
ʻOkoʻa nā hiʻohiʻona a me nā hana o ka Sharkashi chicken sperm tufts ma mua o nā ʻano ʻē aʻe e like me ka echidnas, platypuses, nā ʻiole lāʻau, nā ʻiole dia, a me nā puaʻa guinea.I loko o nā moa sharkasi, ua hoʻemi ka hoʻokumu ʻana o nā pūpū spermatozoa i ko lākou holo ʻauʻau ma mua o ka spermatozoa hoʻokahi.Eia naʻe, ua hoʻonui kēia mau puʻupuʻu i ka pakeneka o ka spermatozoa rheologically positive a hoʻonui i ka hiki o ka spermatozoa e hoʻopaʻa iā lākou iho i kahi kaiapuni ikaika.No laila, ua hōʻoia kā mākou hopena i ka manaʻo mua e pili ana ka sperm agglutination i SST me ka mālama sperm lōʻihi.Manaʻo nō hoʻi mākou e hiki ke hoʻololi i ka hopena o ka hoʻokūkū sperm i ka nui o ka nalowale o ka sperm ma SST.Wahi a kēia manaʻo, ʻo ka spermatozoa me ka haʻahaʻa haʻahaʻa haʻahaʻa e hoʻokuʻu i ka SST mua, ʻoiai ʻo ka spermatozoa me ka mana agglutination kiʻekiʻe e hana i ka hapa nui o nā hua.ʻO ka hoʻokumuʻiaʻana o nā pūpū sperm pore hoʻokahi he mea maikaʻi a pili i ka ratio makua-keiki, akā hoʻohana i kahi hana'ē aʻe.I loko o nā echidnas a me nā platypuses, ua hoʻonohonoho like ka spermatozoa i kekahi i kekahi e hoʻonui i ka wikiwiki o mua o ke kukui.ʻO nā pūʻolo echidnas e neʻe ʻekolu manawa ʻoi aku ka wikiwiki ma mua o ka spermatozoa hoʻokahi.Manaʻo ʻia ʻo ka hoʻokumu ʻia ʻana o ia mau sperm tufts i loko o ka echidnas he hoʻololi evolutionary e mālama ai i ka noho aliʻi ʻana, no ka mea he moe kolohe nā wahine a maʻa mau me kekahi mau kāne.No laila, hoʻokūkū ikaika ka spermatozoa mai nā ejaculates like ʻole no ka fertilization o ka hua.
He maʻalahi ka nānā ʻana i ka spermatozoa agglutinated o nā moa sharkasi me ka hoʻohana ʻana i ka microscopy phase contrast, i manaʻo ʻia he mea maikaʻi no ka mea hiki ke aʻo maʻalahi i ke ʻano o ka spermatozoa in vitro.He ʻokoʻa nō hoʻi ke ʻano o ka hoʻokumu ʻana o ka sperm tuft i ka hoʻohua ʻana i nā moa sharkasi me ka mea i ʻike ʻia ma kekahi mau mea momona placental e hōʻike ana i ka hana sperm hui like e like me nā ʻiole lāʻau, kahi e hiki ai kekahi spermatozoa i nā hua, e kōkua ana i nā poʻe ʻē aʻe e pili a hōʻino i kā lākou mau hua.e hooiaio ia oe iho.hana kūʻokoʻa.ʻO ka hoʻomaʻamaʻa ponoʻī 34. Ua loaʻa kekahi laʻana o ka hana hui i loko o ka spermatozoa i nā mice deer, kahi i hiki ai i ka spermatozoa ke ʻike a hui pū me ka spermatozoa pili loa i ka genetically a hoʻokumu i nā hui hui e hoʻonui i ko lākou wikiwiki e hoʻohālikelike ʻia me ka spermatozoa pili ʻole35.
ʻAʻole kūʻē nā hopena i loaʻa i kēia haʻawina i ka manaʻo o Foman no ka mālama lōʻihi o ka spermatozoa ma SWS.Hōʻike nā mea noiʻi e hoʻomau ka neʻe ʻana o nā sperm cell i ke kahe o nā epithelial cell e hoʻopaʻa ana i ka SST no kahi manawa lōʻihi, a ma hope o kekahi manawa, ua pau nā hale kūʻai ikehu o nā sperm cell, ka hopena i ka emi ʻana o ka wikiwiki, e hiki ai ke kipaku ʻia nā mea kaumaha molekala liʻiliʻi.ka ikehu o ka spermatozoa me ke kahe ana o ka wai mai ka lumen o ka SST Ka lua o ka opu fallopian.Ma ka haʻawina o kēia manawa, ua ʻike mākou ua hōʻike ka hapalua o ka sperm hoʻokahi i ka hiki ke ʻauʻau i nā wai kahe, a ʻo kā lākou adhesion i loko o ka pūpū i hoʻonui i ko lākou hiki ke hōʻike i ka rheology maikaʻi.Eia kekahi, ua kūlike kā mākou ʻikepili me nā mea o Matsuzaki et al.1 ka mea i hōʻike i ka piʻi ʻana o ka lactate huna ma SST hiki ke kāohi i ka motility sperm noho.Eia nō naʻe, wehewehe kā mākou hopena i ka hoʻokumu ʻana o nā ligaments motile sperm a me kā lākou ʻano rheological i ke alo o kahi ʻano ikaika i loko o kahi microchannel i kahi hoʻāʻo e wehewehe i kā lākou ʻano ma SST.Hiki ke nānā aku ka noiʻi e hiki mai ana i ka hoʻoholo ʻana i ka haku mele a me ke kumu o ka agglutinating agent, ʻo ia ka mea e kōkua i ka poʻe noiʻi e hoʻomohala i nā ala hou e mālama ai i ka wai puna a hoʻonui i ka lōʻihi o ka momona.
He ʻumikūmālima 30-wiki-ʻāʻī kāne sharkasi kāne (homozygous dominant; Na Na) i koho ʻia e like me nā mea hāʻawi sperm ma ke aʻo ʻana.Ua hānai ʻia nā manu ma ka Research Poultry Farm o ka Faculty of Agriculture, Ashit University, Ashit Governorate, Egypt.Hoʻokomo ʻia nā manu i loko o nā hīnaʻi pākahi (30 x 40 x 40 cm), i hoʻopaʻa ʻia i kahi papahana māmā (16 mau hola o ka mālamalama a me 8 mau hola o ka pōʻeleʻele) a hānai ʻia i kahi meaʻai e loaʻa ana he 160 g o ka protein crude, 2800 kcal o ka ikehu metabolizable, 35 g o ka calcium kēlā me kēia.5 grams o ka phosphorus i loaʻa no ka kilo o ka meaʻai.
Wahi a ka ʻikepili 36, 37, ua hōʻiliʻili ʻia ka semen mai nā kāne e ka lomilomi ʻōpū.Ua hōʻiliʻili ʻia he 45 mau mea hoʻohālike mai nā kāne 15 i nā lā 3.Semen (n = 15 / lā) ua hoʻoheheʻe koke ʻia 1: 1 (v: v) me Belsville Poultry Semen Diluent, aia i loko o ka potassium 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) a me H2O (100 ml), pH = 7, 5, osmolarity 333 mOsmolarityUa nānā mua ʻia nā laʻana semen i hoʻoheheʻe ʻia ma lalo o kahi microscope māmā e hōʻoia i ka maikaʻi o ka semen maikaʻi (moisture) a laila mālama ʻia i loko o ka wai ʻauʻau ma 37 ° C a hiki i ka hoʻohana ʻana i loko o ka hapalua hola ma hope o ka hōʻiliʻili ʻana.
Hōʻike ʻia nā kinematics a me ka rheology o spermatozoa me ka hoʻohana ʻana i kahi ʻōnaehana microfluidic.Ua hoʻoheheʻe hou ʻia nā mea hoʻohālike i ka 1:40 ma Beltsville Avian Semen Diluent, i hoʻouka ʻia i loko o kahi mea microfluidic (e ʻike i lalo), a ua hoʻoholo ʻia nā ʻāpana kinetic me ka hoʻohana ʻana i kahi ʻōnaehana Computerized Semen Analysis (CASA) i kūkulu mua ʻia no ka microfluidics characterization.e pili ana i ka neʻe ʻana o ka spermatozoa i ka media wai (Department of Mechanical Engineering, Faculty of Engineering, Assiut University, Egypt).Hiki ke hoʻoiho ʻia ka plugin ma: http://www.assiutmicrofluidics.com/research/casa39.Ua anaʻia ka wikiwiki o ka curve (VCL, μm / s), ka laina laina (VSL, μm / s) a me ka wikiwiki trajectory (VAP, μm / s).Ua lawe ʻia nā wikiō o ka spermatozoa me ka hoʻohana ʻana i kahi microscope hoʻohālikelike Optika XDS-3 i hoʻohuli ʻia (me ka pahuhopu 40x) i hoʻopili ʻia i kahi pahu pahu Tucson ISH1000 ma 30 fps no 3 mau kekona.E hoʻohana i ka polokalamu CASA e aʻo i ʻekolu mau wahi a me 500 sperm trajectories no kēlā me kēia laʻana.Ua hana ʻia ka wikiō i hoʻopaʻa ʻia me ka CASA homemade.ʻO ka wehewehe ʻana o ka motility i ka CASA plug-in e pili ana i ka wikiwiki ʻauʻau o ka sperm i hoʻohālikelike ʻia me ka kahe kahe, a ʻaʻole i hoʻokomo i nā ʻāpana ʻē aʻe e like me ka neʻe ʻana o ka ʻaoʻao a me ka ʻaoʻao, no ka mea, ua ʻike ʻia kēia i ʻoi aku ka hilinaʻi o ka kahe wai.Hōʻike ʻia ka neʻe Rheological e like me ka neʻe ʻana o nā pūpū sperm e kūʻē i ke ala o ka kahe wai.ʻO Spermatozoa me nā waiwai rheological i māheleʻia e ka helu o ka spermatozoa motile;ʻO ka spermatozoa e hoʻomaha ana a me ka sperm neʻe convectively ua kāpae ʻia mai ka helu.
Ua kiʻi ʻia nā kemika āpau i hoʻohana ʻia mai Elgomhoria Pharmaceuticals (Cairo, Egypt) ke ʻole i ʻike ʻia.Ua hana ʻia ka mea hana e like me ka wehewehe ʻana e El-sherry et al.40 me kekahi mau hoʻololi.ʻO nā mea i hoʻohana ʻia no ka hana ʻana i nā microchannels ʻo ia nā pā aniani (Howard Glass, Worcester, MA), SU-8-25 kūʻē kūʻē (MicroChem, Newton, CA), diacetone alcohol (Sigma Aldrich, Steinheim, Germany), a me polyacetone.-184, Dow Corning, Midland, Michigan).Hana ʻia nā microchannels me ka lithography palupalu.ʻO ka mea mua, ua paʻi ʻia kahi pale maka pale me ka hoʻolālā microchannel i makemake ʻia ma kahi paʻi kiʻekiʻe (Prismatic, Cairo, Egypt a me Pacific Arts and Design, Markham, ON).Ua hana ʻia nā haku me ka hoʻohana ʻana i nā pā aniani ma ke ʻano he substrates.Ua hoʻomaʻemaʻe ʻia nā papa i ka acetone, isopropanol a me ka wai deionized a laila uhi ʻia me kahi papa 20 µm o SU8-25 e ka uhi ʻana (3000 rpm, 1 min).Hoʻomaloʻo mālie ʻia nā ʻāpana SU-8 (65 ° C, 2 min a me 95 ° C, 10 min) a ʻike ʻia i ka radiation UV no 50 s.ʻO ka bake ma hope o ka wehe ʻana ma 65 ° C a me 95 ° C no 1 min a me 4 min e crosslink i ʻike ʻia i nā papa SU-8, a ukali ʻia e ka hoʻomohala ʻana i ka waiʻona diacetone no 6.5 min.E hoʻomoʻa ikaika i nā waffles (200 ° C no 15 min) e hoʻopaʻa hou i ka papa SU-8.
Ua hoʻomākaukau ʻia ʻo PDMS ma ka hui ʻana i ka monomer a me ka paʻakikī i ka ratio paona o 10: 1, a laila hoʻoheheʻe ʻia i loko o kahi desiccator vacuum a ninini ʻia ma ke kiʻi nui SU-8.Hoʻōla ʻia ka PDMS i loko o ka umu (120 ° C, 30 min), a laila ua ʻoki ʻia nā kahawai, hoʻokaʻawale ʻia mai ka haku, a ʻae ʻia e hoʻopili ʻia nā paipu ma ka inlet a me ka puka o ka microchannel.ʻO ka mea hope loa, ua hoʻopili mau ʻia nā microchannels PDMS i nā paheʻe microscope me ka hoʻohana ʻana i kahi kaʻina corona portable (Electro-Technic Products, Chicago, IL) e like me ka wehewehe ʻana ma nā wahi ʻē aʻe.ʻO ka microchannel i hoʻohana ʻia ma kēia noiʻi ʻana he 200 µm × 20 µm (W × H) a he 3.6 knm ka lōʻihi.
ʻO ke kahe wai i hoʻoulu ʻia e ka hydrostatic pressure i loko o ka microchannel e hoʻokō ʻia ma ka mālama ʻana i ka pae wai i loko o ka waihona inlet ma luna o ka ʻokoʻa kiʻekiʻe Δh39 i loko o ka waihona puka (Fig. 1).
kahi f ka coefficient o ka friction, i hoakakaia e like me f = C/Re no ke kahe lamina i ke kanal rectangular, kahi o C he mau ma muli o ka ratio hiohiona o ke kahawai, L ka loa o ka microchannel, Vav ka awelika velocity i loko o ka microchannel, Dh ke anawaena hydraulic o ke kahawai, g - ka wikiwiki o ka umekaumaha.Me ka hoʻohana ʻana i kēia hoohalike, hiki ke helu ʻia ka wikiwiki o ka awelika me ka hoʻohana ʻana i kēia hoohalike: