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ʻO ka momona o nā manu e pili ana i ko lākou hiki ke mālama i ka sperm ola kūpono no kahi manawa lōʻihi i loko o nā tubules mālama sperm (SST). ʻO ke ʻano kikoʻī e komo ai nā spermatozoa, noho i loko, a haʻalele i ka SST e hoʻopaʻapaʻa mau nei. Ua hōʻike ka sperm o nā moa sharkasi i kahi ʻano kiʻekiʻe i ka agglutination, e hana ana i nā pūʻolo filamentous mobile e loaʻa ana nā cell he nui. Ma muli o ka paʻakikī o ka nānā ʻana i ka neʻe ʻana a me ke ʻano o ka spermatozoa i loko o kahi tube fallopian opaque, ua hoʻohana mākou i kahi mea microfluidic me kahi ʻāpana microchannel e like me ka spermatozoa e aʻo ai i ka agglutination a me ka neʻe ʻana o ka spermatozoa. Kūkākūkā kēia haʻawina pehea e hana ai nā pūʻolo sperm, pehea lākou e neʻe ai, a me kā lākou kuleana hiki i ka hoʻonui ʻana i ka noho ʻana o ka sperm ma ka SST. Ua noiʻi mākou i ka wikiwiki o ka sperm a me ke ʻano rheological i ka wā i hana ʻia ai ke kahe wai i loko o kahi kahawai microfluidic e ke kaomi hydrostatic (ka nui o ke kahe = 33 µm/s). ʻAuʻau pinepine ka spermatozoa i ke au (rheology maikaʻi) a ua emi nui ka wikiwiki o ka pūʻolo spermatozoon i hoʻohālikelike ʻia me ka spermatozoa hoʻokahi. Ua ʻike ʻia nā puʻupuʻu sperm e neʻe i loko o kahi ʻōwili a hoʻonui i ka lōʻihi a me ka mānoanoa i ka wā e hōʻiliʻili ʻia ai nā sperm hoʻokahi. Ua ʻike ʻia nā pūʻolo sperm e hoʻokokoke mai ana a pili ana i nā paia ʻaoʻao o nā kahawai microfluidic i ʻole e kaʻi ʻia me ka wikiwiki o ke kahe ʻana o ka wai > 33 µm/s. Ua ʻike ʻia nā pūʻolo sperm e hoʻokokoke mai ana a pili ana i nā paia ʻaoʻao o nā kahawai microfluidic i ʻole e kaʻi ʻia me ka wikiwiki o ke kahe ʻana o ka wai > 33 µm/s. Было замечено, что пучки сперматозоидов приближаются и прилипают к боковым стенкам микрофлюитодных каналботь сметания со скоростью потока жидкости> 33 мкм / с. Ua ʻike ʻia nā puʻupuʻu sperm e hoʻokokoke a pipili i nā paia ʻaoʻao o nā kahawai microfluidic i ʻole e lawe ʻia aku 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 микрожидкостногочять кабнала, потоком жидкости со скоростью > 33 мкм/с. Ua ʻike ʻia nā pūʻolo sperm e hoʻokokoke a pipili i nā paia ʻaoʻao o ke kahawai microfluidic i ʻole e lawe ʻia aku e ke kahe ʻana o ka wai ma >33 µm/s.Ua hōʻike ʻia e ka microscopy electron scanning a me ka hoʻoili ʻana ua kākoʻo ʻia nā pūʻolo sperm e nā mea paʻa he nui. Hōʻike ka ʻikepili i loaʻa i ka neʻe kūikawā o ka spermatozoa moa Sharkazi, a me ka hiki o ka spermatozoa ke hoʻohuihui a hana i nā pūʻolo mobile, kahi e hāʻawi ai i ka ʻike maikaʻi ʻana i ka mālama lōʻihi o ka spermatozoa ma SMT.
I mea e hoʻokō ai i ka hoʻoulu ʻana i loko o ke kanaka a me ka hapa nui o nā holoholona, ​​​​pono ka sperm a me nā hua manu e hōʻea i kahi o ka hoʻoulu ʻana i ka manawa kūpono. No laila, pono e hana ʻia ka male ʻana ma mua a i ʻole i ka manawa o ka ovulation. Ma ka ʻaoʻao ʻē aʻe, ʻo kekahi mau mammals, e like me nā ʻīlio, a me nā ʻano ʻaʻole mammalian, e like me nā pepeke, nā iʻa, nā mea kolo, a me nā manu, mālama i ka sperm i loko o kā lākou mau ʻōpū hānau no kahi manawa lōʻihi a hiki i ka mākaukau o kā lākou mau hua no ka hoʻoulu ʻana (fertilization asynchronous 1). Hiki i nā manu ke mālama i ke ola o ka spermatozoa hiki ke hoʻoulu i nā hua manu no 2-10 mau pule2.
He hiʻohiʻona kū hoʻokahi kēia e hoʻokaʻawale ai 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 i ka manawa like. ʻO ke kino mālama sperm nui, i kapa ʻia ʻo ka sperm storage tubule (SST), aia ma nā ʻāpana mucosal kūloko ma ka uterovaginal junction. A hiki i kēia lā, ʻaʻole maopopo loa nā ʻano hana e komo ai ka sperm, noho ai, a puka i waho o ka sperm bank. Ma muli o nā haʻawina i hala, ua waiho ʻia nā kuhiakau he nui, akā ʻaʻohe o lākou i hōʻoia ʻia.
Ua kuhi ʻo Forman4 e mālama nā spermatozoa i ko lākou noho ʻana ma ka lua SST ma o ka neʻe mau ʻana o ka oscillatory e kūʻē i ke kuhikuhi o ke kahe ʻana o ka wai ma o nā kahawai protein i loaʻa ma nā pūnaewele epithelial SST (rheology). Hoʻopau ʻia ka ATP ma muli o ka hana flagellar mau e pono ai e mālama i ka sperm i loko o ka lumen SST a emi hope ka neʻe ʻana a hiki i ka lawe ʻia ʻana o ka sperm mai ka waihona sperm e ke kahe wai a hoʻomaka i kahi huakaʻi hou i lalo o ka ʻōmole fallopian piʻi e hoʻoulu i ka sperm. Hua manu (Forman4). Kākoʻo ʻia kēia kumu hoʻohālike o ka mālama ʻana i ka sperm e ka ʻike ʻana e ka immunocytochemistry o nā aquaporins 2, 3 a me 9 i loaʻa i nā pūnaewele epithelial SST. A hiki i kēia lā, ua nele nā ​​​​haʻawina e pili ana i ka rheology semen moa a me kāna kuleana i ka mālama ʻana i ka SST, ke koho ʻana o ka sperm vaginal, a me ka hoʻokūkū sperm. I loko o nā moa, komo ka sperm i loko o ka vagina ma hope o ka male kūlohelohe, akā ʻoi aku ma mua o 80% o ka spermatozoa i kipaku ʻia mai ka vagina ma hope koke iho o ka male ʻana. Hōʻike kēia ʻo ka vagina ke kahua mua no ke koho ʻana o ka sperm i nā manu. Eia kekahi, ua hōʻike ʻia ʻaʻole i hiki i ka 1% o nā spermatozoa i hoʻoulu ʻia i loko o ka vagina e pau i nā SSTs2. I ka hoʻokomo ʻia ʻana o nā moa i loko o ka vagina, ʻoi aku ka nui o nā spermatozoa e hiki ana i ka SST i ka piʻi ʻana o 24 mau hola ma hope o ka hoʻokomo ʻia ʻana. I kēia manawa, ʻaʻole maopopo ke ʻano o ke koho ʻana o ka sperm i loko o kēia kaʻina hana, a he mea nui paha ka neʻe ʻana o ka sperm i ka lawe ʻana o ka sperm SST. Ma muli o nā paia mānoanoa a me ka opaque o nā ʻōmole fallopian, he paʻakikī ke nānā pololei i ka neʻe ʻana o ka sperm i loko o nā ʻōmole fallopian o nā manu. No laila, nele mākou i ka ʻike kumu pehea e neʻe ai ka spermatozoa i ka SST ma hope o ka hoʻokomo ʻana.
Ua ʻike ʻia nei ka rheology he mea nui e kaohi ana i ka lawe ʻana o ka sperm i loko o nā genitalia mammalian. Ma muli o ka hiki o ka spermatozoa motile ke neʻe i ka countercurrent, ua hoʻohana ʻo Zaferani et al8 i kahi ʻōnaehana corra microfluidic e hoʻokaʻawale i ka spermatozoa motile mai nā laʻana semen i hoʻopaʻa ʻia. He mea nui kēia ʻano hoʻokaʻawale semen no ka mālama ʻana i ka hānau ʻole lapaʻau a me ka noiʻi lapaʻau, a makemake ʻia ma mua o nā ʻano kuʻuna e nui ai ka manawa a me ka hana a hiki ke hoʻopilikia i ke ʻano o ka sperm a me ka pono o ke kūkulu ʻana. Eia nō naʻe, a hiki i kēia lā, ʻaʻohe haʻawina i hana ʻia ma ka hopena o nā mea huna mai nā ʻōpū genital o nā moa i ka neʻe ʻana o ka sperm.
Me ka nānā ʻole i ke ʻano hana e mālama ai i ka sperm i mālama ʻia i loko o ka SST, ua ʻike ka nui o nā mea noiʻi e hoʻohuihui ana nā spermatozoa noho poʻo i ke poʻo i loko o ka SST o nā moa 9, 10, nā manu quail 2, a me nā pelehū 11 e hana i nā pūʻolo sperm i hoʻohuihui ʻia. Manaʻo nā mea kākau aia kahi pilina ma waena o kēia agglutination a me ka mālama lōʻihi ʻana o ka spermatozoa i loko o ka SST.
Ua hōʻike ʻo Tingari lāua ʻo Lake12 i kahi pilina ikaika ma waena o ka spermatozoa i loko o ka gland sperm-receiving o ka moa a ua nīnau inā paha e agglutinate ka avian spermatozoa ma ke ʻano like me ka mammalian spermatozoa. Manaʻo lākou ʻo nā pilina hohonu ma waena o ka sperm i loko o ka vas deferens ma muli paha o ke kaumaha i hoʻokumu ʻia e ka nui o ka sperm i loko o kahi liʻiliʻi.
I ka loiloi ʻana i ke ʻano o ka spermatozoa ma nā paheʻe aniani hou, hiki ke ʻike ʻia nā hōʻailona pōkole o ka agglutination, ʻoiai 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, ka mea e wehewehe ai i ke ʻano pōkole o kēia hanana. Ua ʻike pū nā mea noiʻi i ka wā i hoʻohui ʻia ai ka mea hoʻoheheʻe i ka semen, ua ʻike ʻia nā hui cell "e like me ke kaula" elongated.
Ua hana ʻia nā hoʻāʻo mua e hoʻohālike i kahi spermatozoon ma ka wehe ʻana i kahi uea lahilahi mai kahi kulu kau, kahi i hopena ai i kahi vesicle e like me ka sperm e hoʻolōʻihi ʻia mai ka kulu o ka semen. Ua kū pololei ka spermatozoa ma ke ʻano like i loko o ka vesicle, akā ua nalowale 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 neʻe ʻana a me ke ʻano o ka spermatozoa i loko o nā tubules mālama sperm i hoʻokaʻawale ʻia, he mea paʻakikī ke hoʻokō. No laila, pono e hoʻomohala i kahi mea hana e hoʻohālike i ka spermatozoa e kākoʻo i nā haʻawina o ka neʻe ʻana o ka sperm a me ke ʻano agglutination. Ua hōʻike ʻo Brillard et al13 ʻo ka lōʻihi awelika o nā tubules mālama sperm i nā moa makua he 400-600 µm, akā hiki i kekahi mau SST ke lōʻihi a hiki i ka 2000 µm. Ua hoʻokaʻawale ʻo Mero lāua ʻo Ogasawara14 i nā kelepa seminiferous i nā tubules mālama sperm i hoʻonui ʻia a me nā tubules mālama sperm i hoʻonui ʻole ʻia, ʻo ia mau mea ʻelua he like ka lōʻihi (~500 µm) a me ka laulā o ka ʻāʻī (~38 µm), akā ʻo ke anawaena lumen awelika o nā tubules he 56.6 a me 56.6 µm. . , 11.2 μm kēlā me kēia. Ma ke aʻo ʻana o kēia manawa, ua hoʻohana mākou i kahi mea microfluidic me ka nui o ke kahawai o 200 µm × 20 µm (W × H), nona ka ʻāpana kea e kokoke iki ana i ko ka SST i hoʻonui ʻia. Eia kekahi, ua nānā mākou i ka neʻe ʻana o ka sperm a me ke ʻano agglutination i loko o ke kahe wai, kahi e kūlike me ka kuhiakau a 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 countercurrent (rheological).
ʻO ka pahuhopu o kēia haʻawina, ʻo ia ke lanakila i nā pilikia o ka nānā ʻana i ka neʻe ʻana o ka spermatozoa i loko o ka ʻōpū fallopian a me ka pale ʻana i nā pilikia o ke aʻo ʻana i ka rheology a me ke ʻano o ka spermatozoa i loko o kahi ʻano loli. Ua hoʻohana ʻia kahi mea microfluidic e hana ana i ke kaomi hydrostatic e hoʻohālike i ka neʻe ʻana o ka sperm i loko o nā ʻōpū o ka moa.
I ka wā i hoʻouka ʻia ai kahi kulu o kahi hāpana sperm i hoʻoheheʻe ʻia (1:40) i loko o ka hāmeʻa microchannel, hiki ke ʻike ʻia ʻelua ʻano o ka neʻe ʻana o ka sperm (sperm i hoʻokaʻawale ʻia a me ka sperm i hoʻopaʻa ʻia). Eia kekahi, ua ʻauʻau pinepine nā spermatozoa e kūʻē i ke au (rheology maikaʻi; wikiō 1, 2). ʻOiai ʻoi aku ka haʻahaʻa o ka wikiwiki o nā pūʻolo sperm ma mua o ka sperm lonesome (p < 0.001), ua hoʻonui lākou i ka pakeneka o ka sperm e hōʻike ana i ka rheotaxis maikaʻi (p < 0.001; Papa 2). ʻOiai ʻoi aku ka haʻahaʻa o ka wikiwiki o nā pūʻolo sperm ma mua o ka sperm lonesome (p < 0.001), ua hoʻonui lākou i ka pakeneka o ka sperm e hōʻike ana i ka rheotaxis maikaʻi (p < 0.001; Papa 2). Хотя пучки сперматозоидов имели более низкую скорость, чем у одиночных сперматозоидов (p < 0,001), оливалици сперматозоидов, демонстрирующих положительный реотаксис (p < 0,001; таблица 2). ʻOiai ʻoi aku ka haʻahaʻa o ka wikiwiki o nā pūʻolo 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流变性的精子百分比（p < 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 浐倧 流性百分比 （p <0.001 ； 2。。。。。））））） Хотя скорость пучков сперматозоидов была ниже, чем у одиночных сперматозоидов (p < 0,001), они увеличирдит с положительной реологией (p < 0,001; таблица 2). ʻOiai ʻoi aku ka haʻahaʻa o ka wikiwiki o nā pūʻolo 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).Ua manaʻo ʻia ka rheology maikaʻi no ka spermatozoa hoʻokahi a me nā tufts ma kahi o 53% a me 85%, kēlā me kēia.
Ua ʻike ʻia ua hana nā spermatozoa o nā moa sharkasi ma hope koke o ka ejaculation i nā pūʻolo linear, nona nā ʻumi o nā kānaka. Hoʻonui kēia mau puʻupuʻu i ka lōʻihi a me ka mānoanoa i ka hala ʻana o ka manawa a hiki ke noho i loko o ka vitro no kekahi mau hola ma mua o ka nalowale ʻana (wikiō 3). Ua like ke ʻano o kēia mau pūʻolo filamentous me ka echidna spermatozoa e hana ʻia ma ka hopena o ka epididymis. Ua ʻike ʻia he kiʻekiʻe ke ʻano o ka sperm hen Sharkashi e agglutinate a hana i kahi pūʻolo reticulate ma lalo o hoʻokahi minuke ma hope o ka hōʻiliʻili ʻana. He dynamic kēia mau kukuna a hiki ke hoʻopili i nā paia kokoke a i ʻole nā ​​​​mea static. ʻOiai e hōʻemi ana nā pūʻolo sperm i ka wikiwiki o nā cell sperm, ua maopopo he macroscopically e hoʻonui lākou i ko lākou linearity. ʻOkoʻa ka lōʻihi o nā pūʻolo ma muli o ka helu o ka sperm i hōʻiliʻili ʻia i loko o nā pūʻolo. Ua hoʻokaʻawale ʻia ʻelua mau ʻāpana o ka pūʻolo: ka ʻāpana mua, me ke poʻo manuahi o ka sperm agglutinated, a me ka ʻāpana hope, me ka huelo a me ka hopena distal holoʻokoʻa o ka sperm. Me ka hoʻohana ʻana i kahi kāmela wikiwiki (950 fps), ua ʻike ʻia nā poʻo manuahi o nā spermatozoa agglutinated ma ka ʻāpana mua o ka pūʻolo, kuleana no ka neʻe ʻana o ka pūʻolo ma muli o kā lākou neʻe oscillatory, e huki ana i nā mea i koe i loko o ka pūʻolo me ka neʻe helical (Wikiō 4). Eia nō naʻe, i loko o nā tufts lōʻihi, ua ʻike ʻia ua pili kekahi mau poʻo sperm manuahi i ke kino a ʻo ka ʻāpana hope o ka tuft e hana ma ke ʻano he vanes e kōkua i ka hoʻoneʻe ʻana i ka tuft.
ʻOiai i loko o kahi kahe lohi o ka wai, neʻe like nā pūʻolo sperm kekahi i kekahi, akā naʻe, hoʻomaka lākou e uhi a pili i nā mea āpau e noho mālie ana, i ʻole e holoi ʻia e ke kahe o ke au i ka piʻi ʻana o ka wikiwiki o ke kahe. Hoʻokumu ʻia nā pūʻolo i ka wā e hoʻokokoke aku ai kekahi mau cell sperm kekahi i kekahi, hoʻomaka lākou e neʻe like a puni kekahi i kekahi, a laila e pili i kahi mea pipili. Hōʻike nā Kiʻi 1 a me 2 pehea e hoʻokokoke aku ai ka sperm kekahi i kekahi, e hana ana i kahi hui ʻana i ka wā e hoʻopuni ai nā huelo kekahi i kekahi.
Ua hoʻopili nā mea noiʻi i ke kaomi hydrostatic e hana i ke 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 pahu me nā syringes i hoʻokomo ʻia ma nā wēlau. Ua hoʻohana ʻia ka waihoʻoluʻu meaʻai e ʻike nui ʻia ai nā kahawai.
E nakinaki i nā kaula hoʻohui a me nā mea pono i ka paia. Ua lawe ʻia ke wikiō me kahi microscope ʻokoʻa pae. Me kēlā me kēia kiʻi, hōʻike ʻia nā kiʻi microscopy ʻokoʻa pae a me nā kiʻi palapala ʻāina. (A) Kūʻē ka pilina ma waena o nā kahawai ʻelua i ke kahe ma muli o ka neʻe helical (pua ʻulaʻula). (B) ʻO ka pilina ma waena o ka pūʻolo paipu a me ka paia kahawai (nā pua ʻulaʻula), i ka manawa like ua hoʻopili ʻia lākou i ʻelua mau pūʻolo ʻē aʻe (nā pua melemele). (C) Hoʻomaka nā pūʻolo sperm ma ke kahawai microfluidic e hoʻopili kekahi me kekahi (nā pua ʻulaʻula), e hana ana i kahi ʻupena o nā pūʻolo sperm. (D) Hoʻokumu ʻia ʻana o kahi pūnaewele o nā pūʻolo sperm.
I ka wā i hoʻouka ʻia ai kahi kulu o ka sperm i hoʻoheheʻe ʻia i loko o ka mea hana microfluidic a hana ʻia ke kahe, ua ʻike ʻia ke kukuna sperm e neʻe kūʻē i ke kuhikuhi o ke kahe. Hoʻopili pono nā puʻupuʻu i nā paia o nā microchannels, a ʻo nā poʻo manuahi ma ka ʻāpana mua o nā puʻupuʻu e kūpono pono iā lākou (wikiō 5). Hoʻopili pū lākou i nā ʻāpana paʻa i ko lākou ala, e like me nā ʻōpala, e pale aku i ka lawe ʻia ʻana e ke au. I ka hala ʻana o ka manawa, lilo kēia mau puʻupuʻu i mau filaments lōʻihi e hoʻopaʻa ana i nā spermatozoa hoʻokahi ʻē aʻe a me nā puʻupuʻu pōkole (Wikiō 6). I ka hoʻomaka ʻana o ke kahe e lohi, hoʻomaka nā laina lōʻihi o ka sperm e hana i kahi pūnaewele o nā laina sperm (Wikiō 7; Kiʻi 2).
Ma ka wikiwiki o ke kahe ʻana (V > 33 µm/s), hoʻonui ʻia nā neʻe ʻōwili o nā kaula i mea e hopu ai i nā pūʻolo sperm pākahi e pale maikaʻi aku i ka ikaika neʻe o ke kahe. Ma ka wikiwiki o ke kahe ʻana (V > 33 µm/s), hoʻonui ʻia nā neʻe ʻōwili o nā kaula i mea e hopu ai i nā pūʻolo sperm pākahi e pale maikaʻi aku i ka ikaika neʻe o ke kahe. При высокой скорости потока (V > 33 мкм/с) спиралевидные движения нитей усиливаются, отдельных сперматозоидов, образующих пучки, которые лучше противостоят дрейфующей силе потока. Ma nā kahe kahe kiʻekiʻe (V > 33 µm/s), piʻi aʻe ka neʻe ʻana o nā kaula helical i ko lākou hoʻāʻo ʻana e hopu i nā spermatozoa pākahi e hana ana i nā puʻupuʻu i ʻoi aku ka maikaʻi o ke kūʻē ʻana i ka ikaika neʻe 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精子，从而更好地抵抗的单个精子，从而更好地抵抗的地抵抗的单个精子。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 ，地 抵抗 的 漂移力。。。。。。。。 При высоких скоростях потока (V > 33 мкм/с) спиральное движение нитей увеличивается в сперматозоидов, образующих пучки, чтобы лучше сопротивляться силам дрейфа потока. Ma nā helu kahe kiʻekiʻe (V > 33 µm/s), piʻi ka neʻe ʻana o nā filaments i ka hoʻāʻo ʻana e hopu i nā pūʻolo spermatozoa pākahi he nui e kūʻē maikaʻi aku i nā mana drift o ke kahe.Ua hoʻāʻo pū lākou e hoʻopili i nā microchannels i nā paia ʻaoʻao.
Ua ʻike ʻia nā pūʻolo sperm he mau hui o nā poʻo sperm a me nā huelo wili me ka hoʻohana ʻana i ka microscopy māmā (LM). Ua ʻike ʻia hoʻi nā pūʻolo sperm me nā hui like ʻole he mau poʻo wili a me nā hui flagellar, nā huelo sperm i hoʻohui ʻia he nui, nā poʻo sperm i hoʻopili ʻia i kahi huelo, a me nā poʻo sperm me nā nuclei kūlou e like me nā nuclei i hoʻohui ʻia he nui. microscopy electron hoʻoili (TEM). Ua hōʻike ka microscopy electron scanning (SEM) he mau hui o nā poʻo sperm i uhi ʻia nā pūʻolo sperm a ua hōʻike nā hui sperm i kahi pūnaewele pili o nā huelo i wahī ʻia.
Ua aʻo ʻia ke ʻano a me ka ultrastructure o ka spermatozoa, ke kūkulu ʻia ʻana o nā pūʻolo spermatozoa me ka hoʻohana ʻana i ka microscopy māmā (hapa ʻāpana), ka microscopy electron scanning (SEM) a me ka microscopy electron transmission (TEM), ua hoʻoluʻu ʻia nā smears sperm me ka acridine orange a ua nānā ʻia me ka hoʻohana ʻana i ka microscopy epifluorescence.
Ua hōʻike ʻia ka hoʻoluʻu ʻana o ka sperm smear me ka acridine ʻalani (Kiʻi 3B) ua paʻa pū nā poʻo sperm a uhi ʻia me nā mea huna, kahi i alakaʻi ai i ka hoʻokumu ʻia ʻana o nā puʻupuʻu nui (Kiʻi 3D). ʻO nā pūʻolo sperm i hana ʻia me nā hui sperm me kahi pūnaewele o nā huelo i hoʻopili ʻia (Kiʻi 4A-C). Hana ʻia nā pūʻolo sperm me nā huelo o nā spermatozoa he nui i paʻa pū (Kiʻi 4D). Ua uhi nā mea huna (Kiʻi 4E,F) i nā poʻo o nā pūʻolo spermatozoa.
Hoʻokumu ʻia ʻana o ka pūʻolo spermatozoa Ma ka hoʻohana ʻana i ka microscopy ʻokoʻa pae a me nā smears sperm i hoʻoluʻu ʻia me ka acridine ʻalani, ua hōʻike ʻia e pili pū ana nā poʻo o ka spermatozoa. (A) Hoʻomaka ka hoʻokumu mua ʻana o ka tuft sperm me kahi sperm (pōʻai keʻokeʻo) a me ʻekolu sperm (pōʻai melemele), me ka spiral e hoʻomaka ana ma ka huelo a hoʻopau i ke poʻo. (B) Photomicrograph o kahi smear sperm i hoʻoluʻu ʻia me ka acridine ʻalani e hōʻike ana i nā poʻo sperm pili (nā pua). Uhi ka hoʻokuʻu ʻana i ke poʻo (nā poʻo). Hoʻonui × 1000. (C) Hoʻomohala ʻana o kahi kukuna nui i lawe ʻia e ke kahe ʻana i loko o kahi kahawai microfluidic (me ka hoʻohana ʻana i kahi kāmela wikiwiki ma 950 fps). (D) Micrograph o kahi smear sperm i hoʻoluʻu ʻia me ka acridine ʻalani e hōʻike ana i nā tufts nui (nā pua). Hoʻonui: ×200.
ʻO ka micrograph electron scanning o kahi kukuna sperm a me kahi smear sperm i hoʻoluʻu ʻia me ka acridine ʻalani. ʻO (A, B, D, E) nā micrograph electron scanning kala kikohoʻe o ka spermatozoa, a ʻo C a me F nā micrograph o nā smears sperm i hoʻoluʻu ʻia me ka acridine ʻalani e hōʻike ana i ka hoʻopili ʻana o nā spermatozoa he nui e ʻōwili 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) Hoʻopili ʻia o kekahi mau spermatozoa (me ka mea hoʻopili, ka outline poni, ka pua) e ʻōwili ana a puni ka huelo. (E a ​​me F) Nā hōʻuluʻulu poʻo sperm (nā kuhikuhi) i uhi ʻia me nā mea hoʻopili (nā kuhikuhi). Ua hoʻokumu ka spermatozoa i nā puʻupuʻu me kekahi mau ʻano vortex-like (F). (C) ×400 a me (F) ×200 hoʻonui.
Ma ka hoʻohana ʻana i ka microscopy electron transmission, ua ʻike mākou he mau huelo i hoʻopili ʻia i loko o nā pūʻolo sperm (Kiʻi 6A, C), nā poʻo i hoʻopili ʻia i nā huelo (Kiʻi 6B), a i ʻole nā ​​poʻo i hoʻopili ʻia i nā huelo (Kiʻi 6D). He piʻo nā poʻo o ka spermatozoa i loko o ka pūʻolo, e hōʻike ana ma ka ʻāpana ʻelua o nā ʻāpana nuklea (Kiʻi 6D). I loko o ka pūʻolo ʻoki, he poʻo wili ko ka spermatozoa me ʻelua mau ʻāpana nuklea a me nā ʻāpana flagellar he nui (Kiʻi 5A).
He kiʻi uila kala kikohoʻe e hōʻike ana i nā huelo hoʻohui i loko o ka pūʻolo sperm a me ka mea hoʻohuihui e hoʻohui ana i nā poʻo sperm. (A) Huelo i hoʻopili ʻia o ka nui o nā spermatozoa. E nānā i ke ʻano o ka huelo ma nā kiʻi kiʻi (pua) a me nā kiʻi ʻāina (pua). (B) Ua hoʻopili ʻia ke poʻo (pua) o ka sperm i ka huelo (pua). (C) Ua hoʻopili ʻia kekahi mau huelo sperm (pua). (D) Hoʻohui ka mea hoʻohuihui (AS, polū) i ʻehā mau poʻo sperm (poni).
Ua hoʻohana ʻia ka microscopy electron scanning e ʻike i nā poʻo sperm i loko o nā puʻupuʻu sperm i uhi ʻia me nā mea huna a i ʻole nā ​​membrane (Kiʻi 6B), e hōʻike ana ua hoʻopaʻa ʻia nā puʻupuʻu sperm e nā mea extracellular. Ua hoʻopaʻa ʻia ka mea agglutinated i loko o ke poʻo sperm (hui e like me ke poʻo jellyfish; Kiʻi 5B) a ua hoʻonui ʻia ma ka distal, e hāʻawi ana i kahi ʻano melemele ʻālohilohi ma lalo o ka microscopy fluorescence i ka wā i hoʻoluʻu ʻia me ka acridine ʻalani (Kiʻi 6C). ʻIke maopopo ʻia kēia mea ma lalo o kahi microscope scanning a ua manaʻo ʻia he mea hoʻopaʻa. Ua hōʻike nā ʻāpana semi-thin (Kiʻi 5C) a me nā smears sperm i hoʻoluʻu ʻia me ka acridine ʻalani i nā puʻupuʻu sperm e loaʻa ana nā poʻo i hoʻopaʻa nui ʻia a me nā huelo ʻōwili (Kiʻi 5D).
Nā kiʻi photomicrograph 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 hana like ʻole. (A) Micrograph electron hoʻoili kala kikohoʻe cross-sectional o kahi pūʻulu sperm e hōʻike ana i kahi poʻo sperm i ʻōwili ʻia me kahi nucleus ʻelua ʻāpana (uliuli) a me kekahi mau ʻāpana flagellar (ʻōmaʻomaʻo). (B) Micrograph electron scanning kala kikohoʻe e hōʻike ana i kahi hui o nā poʻo sperm e like me ka jellyfish (nā pua) i ʻike ʻia ua uhi ʻia. (C) ʻĀpana semi-lahilahi e hōʻike ana i nā poʻo sperm i hōʻuluʻulu ʻia (nā pua) a me nā huelo ʻōwili (nā pua). (D) Micrograph o kahi smear sperm i hoʻoluʻu ʻia me ka acridine ʻalani e hōʻike ana i nā hōʻuluʻulu ʻana o nā poʻo sperm (nā pua) a me nā huelo pili ʻōwili (nā pua). E hoʻomaopopo he mea pipili (S) e uhi ana i ke poʻo o ka spermatozoon. (D) × 1000 hoʻonui.
Ma ka hoʻohana ʻana i ka microscopy electron transmission (Kiʻi 7A), ua ʻike ʻia hoʻi ua wili ʻia nā poʻo sperm a he ʻano wili ko nā nuclei, e like me ka hōʻoia ʻana e nā smears sperm i hoʻoluʻu ʻia me ka acridine ʻalani a nānā ʻia me ka hoʻohana ʻana i ka microscopy fluorescence (Kiʻi 7B).
(A) Micrograph electron hoʻoili kala kikohoʻe a me (B) ʻO ka palaki sperm i hoʻoluʻu ʻia me ka ʻalani Acridine e hōʻike ana i nā poʻo i ʻōwili ʻia a me ka hoʻopili ʻana o nā poʻo sperm a me nā huelo (nā pua). (B) Hoʻonui × 1000.
ʻO kahi mea hoihoi i ʻike ʻia, ʻo ia ka hōʻuluʻulu ʻana o ka sperm a Sharkazi e hana i nā puʻupuʻu filamentous mobile. ʻO nā ʻano o kēia mau puʻupuʻu e hiki ai iā mākou ke hoʻomaopopo i kā lākou kuleana hiki i ka omo ʻana a me ka mālama ʻana o ka spermatozoa i loko o ka SST.
Ma hope o ka male ʻana, komo ka sperm i loko o ka vagina a hana i kahi kaʻina koho ikaika, e hopena ana i kahi helu palena wale nō o ka sperm e komo ana i ka SST15,16. A hiki i kēia lā, ʻaʻole maopopo nā ʻano hana e komo ai ka sperm a puka i waho o ka SST. I loko o ka manu, mālama ʻia nā spermatozoa i loko o ka SST no kahi manawa lōʻihi o 2 a 10 mau pule, ma muli o ke ʻano6. Ke mau nei ka hoʻopaʻapaʻa e pili ana i ke kūlana o ka semen i ka wā e mālama ʻia ai i loko o ka SST. Ke neʻe nei lākou a i ʻole e hoʻomaha? I nā huaʻōlelo ʻē aʻe, pehea e mālama ai nā cell sperm i ko lākou kūlana ma ka SST no ka lōʻihi?
Ua hōʻike ʻo Forman4 e hiki ke wehewehe ʻia ka noho ʻana o SST a me ka hoʻokuʻu ʻana ma ke ʻano o ka neʻe ʻana o ka sperm. Manaʻo nā mea kākau e mālama ka sperm i ko lākou kūlana ma ka ʻauʻau ʻana e kūʻē i ke kahe wai i hana ʻia e ka epithelium SST a ua hoʻokuʻu ʻia ka sperm mai ka SST ke hāʻule ko lākou wikiwiki ma lalo o ke kiko e hoʻomaka ai lākou e neʻe i hope ma muli o ka nele o ka ikehu. Ua hōʻoia ʻo Zaniboni5 i ke alo o nā aquaporins 2, 3, a me 9 ma ka ʻāpana apical o nā cell epithelial SST, kahi e kākoʻo pololei ʻole ai i ke kumu hoʻohālike mālama sperm a Foreman. Ma ke aʻo ʻana o kēia manawa, ua ʻike mākou ua kokoke ka hapalua o ka spermatozoa a Sharkashi e hōʻike ana i ka rheology maikaʻi i loko o ke kahe wai, a ʻo nā pūʻolo sperm agglutinated e hoʻonui i ka helu o nā spermatozoa e hōʻike ana i ka rheology maikaʻi, ʻoiai ʻo ka agglutination e hoʻolohi iā lākou. ʻAʻole maopopo loa pehea e hele ai nā cell sperm i luna o ka fallopian tube o ka manu i kahi o ka fertilization. I loko o nā mammals, huki ka chemoattractants i ka spermatozoa. Eia nō naʻe, ua manaʻo ʻia nā chemoattractants e kuhikuhi i ka spermatozoa e hoʻokokoke i nā mamao lōʻihi7. No laila, ʻo nā ʻano hana ʻē aʻe ke kuleana no ka lawe ʻana o ka sperm. ʻO ka hiki o ka sperm ke kuhikuhi a kahe e kūʻē i ka wai o ka fallopian tube i hoʻokuʻu ʻia ma hope o ka male ʻana ua hōʻike ʻia he kumu nui i ka hoʻopaʻa ʻana i ka sperm i nā ʻiole. Ua hōʻike ʻo Parker 17 e hele ka spermatozoa i nā oviducts ma ka ʻauʻau ʻana e kūʻē i ke kahe ciliary i nā manu a me nā reptile. ʻOiai ʻaʻole i hōʻike ʻia ma ke ʻano hoʻokolohua i nā manu, ʻo Adolphi18 ka mea mua i ʻike e hāʻawi ka sperm avian i nā hopena maikaʻi ke hana ʻia kahi papa lahilahi o ka wai ma waena o kahi uhi a me ka slide me kahi ʻāpana pepa kānana. Rheology. Ua kau ʻo Hino lāua ʻo Yanagimachi [19] i kahi complex ovary-tubal-uterine mouse i loko o kahi apo perfusion a hoʻokomo i 1 µl o ka ʻīnika i loko o ka isthmus e nānā i ke kahe ʻana o ka wai i loko o nā fallopian tubes. Ua ʻike lākou i kahi neʻe ikaika loa o ka ʻaelike a me ka hoʻomaha ʻana i loko o ka fallopian tube, kahi e neʻe mau ana nā pōpō ʻīnika a pau i ka ampulla o ka fallopian tube. Hoʻoikaika nā mea kākau i ke koʻikoʻi o ke kahe ʻana o ka wai tubal mai nā fallopian tubes haʻahaʻa a i luna no ka hoʻokiʻekiʻe ʻana o ka sperm a me ka fertilization. Ua hōʻike ʻo Brillard20 i loko o nā moa a me nā pelehū, neʻe nā spermatozoa ma o ka neʻe ikaika mai ka puka komo o ka vagina, kahi e mālama ʻia ai lākou, a i ka hui utero-vaginal, kahi e mālama ʻia ai lākou. Eia naʻe, ʻaʻole pono kēia neʻe ʻana ma waena o ka hui uterovaginal a me ka infundibulum no ka mea ua lawe ʻia nā spermatozoa e ka neʻe ʻana o ka passive. Ma ka ʻike ʻana i kēia mau ʻōlelo aʻoaʻo mua a me nā hopena i loaʻa i ke aʻo ʻana 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 vagina a me ko lākou komo ʻana i loko o ka CCT no ka mālama ʻana. E like me ka manaʻo o Forman4, 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 waho i ka wā e hoʻomaka ai ko lākou wikiwiki e lohi.
Ma ka ʻaoʻao ʻē aʻe, ua hōʻike ʻo Matsuzaki lāua ʻo Sasanami 21 e loli ana ka neʻe ʻana o ka spermatozoa manu mai ka dormancy a i ka neʻe ʻana i loko o nā ala hānau kāne a me ka wahine. Ua hāpai ʻia ka pale ʻana i ka neʻe ʻana o ka sperm noho ma ka SST e wehewehe i ka manawa mālama lōʻihi o ka sperm a laila hoʻoulu hou ʻia ma hope o ka haʻalele ʻana i ka SST. Ma lalo o nā kūlana hypoxic, ua hōʻike ʻo Matsuzaki et al. 1 i ka hana kiʻekiʻe a me ka hoʻokuʻu ʻana o ka lactate i ka SST, kahi e alakaʻi ai i ka pale ʻana i ka neʻe ʻana o ka sperm noho. I kēia hihia, ʻike ʻia ke koʻikoʻi o ka rheology sperm i ke koho ʻana a me ka omo ʻana o ka spermatozoa, ʻaʻole i ko lākou mālama ʻana.
Ua manaʻo ʻia ke ʻano o ka hoʻohuihui ʻana o ka sperm he wehewehe kūpono no ka manawa mālama lōʻihi o ka sperm ma ka SST, ʻoiai he ʻano maʻamau kēia o ka paʻa ʻana o ka sperm i loko o ka manu2,22,23. Ua ʻike ʻo Bakst et al. 2 ua pili ka hapa nui o nā spermatozoa kekahi i kekahi, e hana ana i nā hui fascicular, a ʻaʻole i loaʻa pinepine nā spermatozoa hoʻokahi ma ka quail CCM. Ma ka ʻaoʻao ʻē aʻe, ua ʻike ʻo Wen et al. 24 i nā spermatozoa i hoʻopuehu ʻia a me nā ʻāpana spermatozoa liʻiliʻi i loko o ka lumen SST i nā moa. Ma muli o kēia mau ʻike ʻana, hiki ke manaʻo ʻia he ʻokoʻa ka makemake no ka hoʻohuihui ʻana o ka sperm ma waena o nā manu a ma waena o nā spermatozoa i loko o ka ejaculate like. Eia kekahi, ua hōʻike ʻo Van Krey et al. 9 ʻo ka dissociation random o ka spermatozoa agglutinated ke kuleana no ke komo mālie ʻana o ka spermatozoa i loko o ka lumen o ka fallopian tube. Wahi a kēia kuhiakau, pono e kipaku mua ʻia nā spermatozoa me ka hiki ke hoʻohuihui haʻahaʻa mai ka SST. Ma kēia ʻano, ʻo ka hiki o ka spermatozoa ke hoʻopili i ka hui ʻana he mea ia e hoʻopilikia ai i ka hopena o ka hoʻokūkū sperm i nā manu lepo. Eia kekahi, ʻo ka lōʻihi o ka hoʻokaʻawale ʻana o ka sperm i hoʻopili ʻia, ʻo ka lōʻihi o ka mālama ʻia ʻana o ka momona.
ʻOiai ua ʻike ʻia ka hōʻuluʻulu ʻana o ka spermatozoa a me ka hōʻuluʻulu ʻana i loko o nā puʻupuʻu i kekahi mau haʻawina2,22,24, ʻaʻole i wehewehe kikoʻī ʻia lākou 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 agglutination sperm in vitro. Ua ʻike ʻia ka hōʻuluʻulu nui akā transient i ka wā i wehe ʻia ai ke kaula lahilahi mai ka hāʻule hua e kau ana. ʻO kēia ke alakaʻi i ka ʻoiaʻiʻo e puka mai ana kahi pehu elongated mai ka hāʻule ʻana, e hoʻohālike ana i ka seminal gland. Ma muli o nā palena 3D a me nā manawa maloʻo pōkole, ua hāʻule koke ka poloka holoʻokoʻa i ka hōʻino9. I ke aʻo ʻana o kēia manawa, me ka hoʻohana ʻana i nā moa Sharkashi a me nā ʻāpana microfluidic, ua hiki iā mākou ke wehewehe pehea e hana ai kēia mau tufts a pehea lākou e neʻe ai. Ua hana koke ʻia nā puʻupuʻu sperm ma hope o ka hōʻiliʻili ʻana o ka semen a ua ʻike ʻia e neʻe i loko o kahi spiral, e hōʻike ana i ka rheology maikaʻi i ka wā e kū nei i ke kahe. Eia kekahi, i ka nānā ʻana ma ke ʻano macroscopic, ua ʻike ʻia nā puʻupuʻu sperm e hoʻonui i ka linearity o ka motility i hoʻohālikelike ʻia me ka spermatozoa i hoʻokaʻawale ʻia. Hōʻike kēia e hiki ke hana ʻia ka agglutination sperm ma mua o ke komo ʻana o SST a ʻaʻole i kaupalena ʻia ka hana ʻana o ka sperm i kahi wahi liʻiliʻi ma muli o ke kaumaha e like me ka mea i hōʻike mua ʻia (Tingari lāua ʻo Lake12). I ka wā o ka hoʻokumu ʻana o ka tuft, ʻauʻau ka spermatozoa ma ka synchrony a hiki i ka wā e hana ai lākou i kahi hui, a laila ʻōwili ko lākou huelo a puni kekahi i kekahi a noho manuahi ke poʻo o ka spermatozoon, akā ʻo ka huelo a me ka ʻāpana distal o ka spermatozoon e pili pū me kahi mea pipili. No laila, ʻo ke poʻo manuahi o ka ligament ke kuleana no ka neʻe ʻana, e huki ana i ke koena o ka ligament. Ua hōʻike ka microscopy electron scanning o nā pūʻolo sperm i nā poʻo sperm i hoʻopili ʻia i uhi ʻia me ka nui o nā mea pipili, e hōʻike ana ua hoʻopili ʻia nā poʻo sperm i nā pūʻolo hoʻomaha, kahi i hana ʻia ma hope o ka hiki ʻana i kahi mālama (SST).
Ke hoʻoluʻu ʻia kahi ʻōmole sperm me ka acridine ʻalani, hiki ke ʻike ʻia nā mea hoʻopili extracellular a puni nā cell sperm ma lalo o kahi microscope fluorescent. ʻAe kēia mea i nā puʻupuʻu sperm e pipili a hoʻopili i nā ʻili a puni a i ʻole nā ​​​​​​mea liʻiliʻi i ʻole lākou e holo pū me ke kahe a puni. No laila, hōʻike kā mākou mau nānā ʻana i ke kuleana o ka hoʻopili ʻana o ka spermatozoa ma ke ʻano o nā puʻupuʻu mobile. ʻO ko lākou hiki ke ʻauʻau kūʻē i ke au a pili i nā ʻili kokoke e hiki ai i ka sperm ke noho lōʻihi ma ka SST.
Ua hoʻohana ʻo Rothschild25 i kahi kāmela hemocytometry e aʻo i ka hoʻolaha lana ʻana o ka semen bipi i loko o kahi kulu o ka hoʻokuʻu ʻana, e lawe ana i nā photomicrographs ma o kahi kāmela me ke axis optical vertical a me ka horizontal o ka microscope. Ua hōʻike nā hopena ua huki ʻia nā spermatozoa i ka ʻili o ke keʻena. Manaʻo nā mea kākau he mau pilina hydrodynamic paha ma waena o ka sperm a me ka ʻili. Ke noʻonoʻo nei i kēia, me ka hiki o ka semen moa Sharkashi ke hana i nā pufts sticky, hiki ke hoʻonui i ka likelika e pili ka semen i ka paia SST a mālama ʻia no ka manawa lōʻihi.
Ua hōʻike ʻo Bccetti lāua ʻo Afzeliu26 e pono ana ka glycocalyx sperm no ka ʻike ʻana i nā gamete a me ka agglutination. Ua ʻike ʻo Forman10 ʻo ka hydrolysis o nā pilina α-glycosidic i loko o nā uhi glycoprotein-glycolipid ma ka mālama ʻana i ka semen manu me neuraminidase i hopena i ka emi ʻana o ka momona me ka ʻole o ka hoʻopilikia ʻana i ka neʻe ʻana o ka sperm. Manaʻo nā mea kākau e hoʻopilikia ka hopena o ka neuraminidase ma ka glycocalyx i ka sequestration sperm ma ka utero-vaginal junction, a laila e hōʻemi ana i ka momona. ʻAʻole hiki i kā lākou mau nānā ke hoʻowahāwahā i ka hiki ke hōʻemi ka mālama ʻana i ka neuraminidase i ka ʻike ʻana i ka sperm a me ka oocyte. Ua ʻike ʻo Forman lāua ʻo Engel10 ua emi ka momona i ka wā i hoʻokomo ʻia ai nā moa i loko o ka vagina me ka semen i mālama ʻia me neuraminidase. Eia nō naʻe, ʻaʻole i hoʻopilikia ka IVF me ka sperm i mālama ʻia me neuraminidase i ka momona i hoʻohālikelike ʻia me nā moa kaohi. Ua hoʻoholo nā mea kākau ua hōʻemi nā loli i ka uhi glycoprotein-glycolipid a puni ka membrane sperm i ka hiki o ka sperm ke hoʻoulu ma o ka hoʻopilikia 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 utero-vaginal junction, akā ʻaʻole ia e hoʻopilikia i ka ʻike ʻana o ka sperm a me ka hua manu.
I loko o nā turkeys, ua loaʻa iā Bakst lāua ʻo Bauchan 11 nā vesicles liʻiliʻi a me nā ʻāpana membrane i loko o ka lumen o ka SST a ua 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 ma SST. Eia naʻe, ʻaʻole i kuhikuhi nā mea noiʻi i ke kumu o kēia mau ʻāpana, inā paha 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 hana a hūnā nā cell epididymal epithelial i kahi protein kikoʻī e pono ai no ka hoʻokumu ʻana o nā ala seminal hoʻokahi-pore. Hōʻike pū nā mea kākau e pili ana ka hoʻopuehu ʻana o kēia mau puʻupuʻu i ka launa pū ʻana o nā protein epididymal. Ua ʻike ʻo Nixon et al28 e hūnā ka adnexa i kahi protein, ʻo ia ka osteonectin waiwai cysteine ​​acidic; Pili ʻo SPARC i ka hoʻokumu ʻana o nā tufts sperm i nā echidnas pōkole a me nā platypuses. ʻO ka hoʻopuehu ʻana o kēia mau kaola e pili ana i ka nalowale ʻana o kēia protein.
Ma ke aʻo ʻana o kēia manawa, ua hōʻike ka loiloi ultrastructural e hoʻohana ana i ka microscopy electron ua pili ka spermatozoa i kahi nui o nā mea paʻa. Manaʻo ʻia kēia mau mea ke kuleana no ka agglutination e hoʻopili ana ma waena a puni nā poʻo pili, akā ma nā ʻano 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, ʻoiai 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 wā e hele ai ka spermatozoa manu ma o ka epididymis a me vas deferens, loaʻa iā lākou nā loli e pili ana i ka maturation e kākoʻo ana i ko lākou hiki ke hoʻopaʻa i nā protein a loaʻa nā glycoproteins e pili ana i ka lemma plasma. ʻO ka hoʻomau ʻana o kēia mau protein ma nā membrane sperm noho ma ka SST e hōʻike ana e hiki i kēia mau protein ke hoʻopilikia i ka loaʻa ʻana o ka paʻa o ka membrane sperm 30 a hoʻoholo i ko lākou momona 31. Ua hōʻike ʻo Ahammad et al32 ua hōʻike nā spermatozoa i loaʻa mai nā ʻāpana like ʻole o ka ʻōnaehana hānau kāne (mai nā testes a i nā vas deferens distal) i ka piʻi mau ʻana o ke ola ma lalo o nā kūlana mālama wai, me ka nānā ʻole i ka mahana mālama, a piʻi pū ke ola i nā moa i loko o nā ʻōpū fallopian ma hope o ka insemination artificial.
He ʻokoʻa nā ʻano a me nā hana o nā ʻōpuʻu sperm moa Sharkashi ma mua o nā ʻano ʻē aʻe e like me nā echidnas, platypuses, ʻiole lāʻau, ʻiole dia, a me nā puaʻa guinea. I loko o nā moa sharkasi, ua hoʻemi ka hoʻokumu ʻia ʻana o nā pūʻolo spermatozoa i ko lākou wikiwiki ʻauʻau i hoʻohālikelike ʻia me nā spermatozoa hoʻokahi. Eia nō naʻe, ua hoʻonui kēia mau pūʻolo i ka pakeneka o nā spermatozoa rheologically maikaʻi a hoʻonui i ka hiki o ka spermatozoa ke hoʻopaʻa iā lākou iho i kahi ʻano loli. No laila, ua hōʻoia kā mākou mau hopena i ka manaʻo mua e pili ana ka agglutination sperm ma SST me ka mālama ʻana i ka sperm no ka manawa lōʻihi. Ke kuhi nei hoʻi mākou ʻo ka hiki o ka sperm ke hana i nā tufts e kāohi i ka wikiwiki o ka nalowale o ka sperm ma SST, kahi e hoʻololi ai i ka hopena o ka hoʻokūkū sperm. Wahi a kēia kuhiakau, ʻo ka spermatozoa me ka hiki ke agglutination haʻahaʻa e hoʻokuʻu mua iā SST, ʻoiai ʻo ka spermatozoa me ka hiki ke agglutination kiʻekiʻe e hana i ka hapa nui o nā keiki. He mea maikaʻi ka hoʻokumu ʻia ʻana o nā pūʻolo sperm hoʻokahi-pore a hoʻopilikia i ka ratio makua-keiki, akā hoʻohana i kahi ʻano hana ʻokoʻa. I loko o nā echidnas a me nā platypuses, ua hoʻonohonoho like ʻia nā spermatozoa kekahi i kekahi e hoʻonui i ka wikiwiki o ke kukuna i mua. Neʻe nā puʻupuʻu o nā echidnas ma kahi o ʻekolu manawa wikiwiki ma mua o nā spermatozoa hoʻokahi. Ua manaʻoʻiʻo ʻia ʻo ka hoʻokumu ʻia ʻana o ia mau puʻupuʻu sperm i loko o nā echidnas he hoʻololi evolutionary e mālama i ka mana, ʻoiai he moekolohe nā wahine a pili pinepine me kekahi mau kāne. No laila, hoʻokūkū ikaika nā spermatozoa mai nā ejaculates like ʻole no ka hoʻoulu ʻana o ka hua manu.
He maʻalahi ke ʻike ʻia nā spermatozoa agglutinated o nā moa sharkasi me ka hoʻohana ʻana i ka microscopy ʻokoʻa pae, kahi i manaʻo ʻia he pono no ka mea e ʻae maʻalahi i ke aʻo ʻana i ke ʻano o ka spermatozoa in vitro. ʻOkoʻa nō hoʻi ke ʻano hana e hoʻolaha ai ka hoʻokumu ʻana o ka sperm tuft i ka hana hou ʻana i nā moa sharkasi mai ka mea i ʻike ʻia i kekahi mau mammals placental e hōʻike ana i ke ʻano sperm cooperative e like me nā ʻiole lāʻau, kahi e hiki ai kekahi spermatozoa i nā hua, e kōkua ana i nā poʻe pili ʻē aʻe e hiki a hōʻino i kā lākou mau hua. e hōʻoia iā ʻoe iho. ʻano altruistic. Hoʻoulu ponoʻī 34. Ua loaʻa kekahi laʻana o ke ʻano hana like i loko o ka spermatozoa i loko o nā ʻiole dia, kahi i hiki ai i nā spermatozoa ke ʻike a hui pū me nā spermatozoa pili genetically loa a hana i nā hui hana like e hoʻonui i ko lākou wikiwiki i hoʻohālikelike ʻia me nā spermatozoa pili ʻole35.
ʻAʻole kūʻē nā hopena i loaʻa ma kēia haʻawina i ke kumumanaʻo o Foman no ka mālama lōʻihi ʻana o ka spermatozoa ma SWS. Hōʻike nā mea noiʻi e hoʻomau ka neʻe ʻana o nā cell sperm i ke kahe o nā cell epithelial e hoʻopili ana i ka SST no kahi manawa lōʻihi, a ma hope o kekahi manawa, ua pau nā waihona ikehu o nā cell sperm, e hopena ana i ka emi ʻana o ka wikiwiki, kahi e hiki ai ke kipaku ʻia nā mea kaumaha molekala liʻiliʻi. ikehu o ka spermatozoa me ke kahe o ka wai mai ka lumen o ka SST ʻO ka lua o ka fallopian tube. Ma ke aʻo ʻana o kēia manawa, ua ʻike mākou ua hōʻike ka hapalua o ka sperm hoʻokahi i ka hiki ke ʻauʻau kūʻē i nā wai e kahe ana, a ʻo ko lākou hoʻopili ʻana i loko o ka pūʻolo i hoʻonui i ko lākou hiki ke hōʻike i ka rheology maikaʻi. Eia kekahi, kūlike kā mākou ʻikepili me nā mea o Matsuzaki et al. 1 nāna i hōʻike e hiki i ka hoʻonui ʻia ʻana o ka lactate secretion ma SST ke kāohi i ka neʻe ʻana o ka sperm noho. Eia naʻe, wehewehe kā mākou mau hopena i ka hoʻokumu ʻia ʻana o nā ligaments motile sperm a me kā lākou ʻano rheological i ke alo o kahi kaiapuni ikaika i loko o kahi microchannel i kahi hoʻāʻo e wehewehe i kā lākou ʻano ma SST. Hiki i nā noiʻi e hiki mai ana ke kālele i ka hoʻoholo ʻana i ka haku mele kemika a me ke kumu o ka mea hoʻohuihui, ka mea e kōkua maoli i nā mea noiʻi e hoʻomohala i nā ala hou e mālama ai i ka sperm wai a hoʻonui i ka lōʻihi o ka momona.
Ua koho ʻia he ʻumikūmālima kāne sharkasi ʻāʻī ʻōlohelohe he 30 pule (homozygous dominant; Na Na) i mau mea hāʻawi sperm ma ke aʻo ʻana. Ua hānai ʻia nā manu ma ka Research Poultry Farm o ke Faculty of Agriculture, Ashit University, Ashit Governorate, Egypt. Ua hoʻokomo ʻia nā manu i loko o nā hale manu pākahi (30 x 40 x 40 cm), i hoʻomālamalama ʻia i kahi papahana kukui (16 hola o ka mālamalama a me 8 hola o ka pouli) a hānai ʻia i kahi meaʻai e loaʻa ana he 160 g o ka protein maka, 2800 kcal o ka ikehu metabolizable, 35 g o ka calcium pākahi. 5 grams o ka phosphorus i loaʻa no kēlā me kēia kilokani o ka meaʻai.
Wahi a ka ʻikepili 36, ​​37, ua hōʻiliʻili ʻia ka sperm mai nā kāne ma o ka lomilomi ʻōpū. Ua hōʻiliʻili ʻia he 45 mau laʻana sperm mai 15 mau kāne ma luna o 3 mau lā. Ua hoʻoheheʻe koke ʻia ka Semen (n = 15/lā) 1:1 (v:v) me ka Belsville Poultry Semen Diluent, nona 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 mOsm/kg38. Ua nānā mua ʻia nā hāpana sperm i hoʻoheheʻe ʻia ma lalo o kahi microscope māmā e hōʻoia i ka maikaʻi o ka sperm (makū) 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.
Ua wehewehe ʻia ke kinematics a me ka rheology o ka spermatozoa me ka hoʻohana ʻana i kahi ʻōnaehana o nā mea hana microfluidic. Ua hoʻoheheʻe hou ʻia nā laʻana semen i 1:40 ma Beltsville Avian Semen Diluent, i hoʻouka ʻia i loko o kahi mea hana microfluidic (e nānā i lalo), a ua hoʻoholo ʻia nā palena kinetic me ka hoʻohana ʻana i kahi ʻōnaehana Computerized Semen Analysis (CASA) i hoʻomohala mua ʻia no ke ʻano microfluidics. ma ka neʻe ʻana o ka spermatozoa i loko o nā 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 ana ʻia ka wikiwiki o ke kuʻekuʻe (VCL, μm/s), ka wikiwiki linear (VSL, μm/s) a me ka wikiwiki o ke alapine awelika (VAP, μm/s). Ua lawe ʻia nā wikiō o ka spermatozoa me ka hoʻohana ʻana i kahi microscope ʻokoʻa Optika XDS-3 phase inverted (me ka pahuhopu 40x) i hoʻopili ʻia i kahi kāmela Tucson ISH1000 ma 30 fps no 3 s. E hoʻohana i ka polokalamu CASA e aʻo ai ma ka liʻiliʻi he ʻekolu mau wahi a me 500 mau ala sperm no kēlā me kēia laʻana. Ua hana ʻia ke wikiō i hoʻopaʻa ʻia me ka hoʻohana ʻana i kahi CASA hana home. Hoʻokumu ʻia ka wehewehe ʻana o ka neʻe ʻana i loko o ka plug-in CASA ma ka wikiwiki ʻauʻau o ka sperm i hoʻohālikelike ʻia me ka nui o ke kahe ʻana, a ʻaʻole hoʻokomo i nā palena ʻē aʻe e like me ka neʻe ʻana o ka ʻaoʻao a i ka ʻaoʻao, no ka mea, ua ʻike ʻia he ʻoi aku ka hilinaʻi o kēia i ke kahe ʻana o ka wai. Ua wehewehe ʻia ka neʻe ʻana o ka rheological ʻo ia ka neʻe ʻana o nā cell sperm e kūʻē i ke kuhikuhi o ke kahe ʻana o ka wai. Ua puʻunaue ʻia nā spermatozoa me nā waiwai rheological e ka helu o nā spermatozoa motile; ua kāpae ʻia nā spermatozoa i hoʻomaha a me nā spermatozoa e neʻe ana me ka convective mai ka helu ʻana.
Ua loaʻa nā kemika a pau i hoʻohana ʻia mai Elgomhoria Pharmaceuticals (Kairo, ʻAigupita) ke ʻole i ʻōlelo ʻia. Ua hana ʻia ka hāmeʻa e like me ka mea i wehewehe ʻia 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 papa aniani (Howard Glass, Worcester, MA), SU-8-25 negative resist (MicroChem, Newton, CA), diacetone alcohol (Sigma Aldrich, Steinheim, Kelemānia), a me polyacetone. -184, Dow Corning, Midland, Michigan). Hana ʻia nā Microchannels me ka hoʻohana ʻana i ka lithography palupalu. ʻO ka mea mua, ua paʻi ʻia kahi pale maka pale maopopo me ka hoʻolālā microchannel i makemake ʻia ma kahi mīkini paʻi kiʻekiʻe (Prismatic, Cairo, ʻAigupita a me Pacific Arts and Design, Markham, ON). Ua hana ʻia nā haku me ka hoʻohana ʻana i nā papa aniani ma ke ʻano he substrates. Ua hoʻomaʻemaʻe ʻia nā papa i ka acetone, isopropanol a me ka wai deionized a laila ua uhi ʻia me kahi papa 20 µm o SU8-25 e ka uhi wili (3000 rpm, 1 min). A laila ua hoʻomaloʻo mālie ʻia nā papa SU-8 (65°C, 2 min a me 95°C, 10 min) a ua hoʻokuʻu ʻia i ka radiation UV no 50 kekona. Ma hope o ka hoʻokuʻu ʻana, e hoʻomoʻa ma 65°C a me 95°C no 1 min a me 4 min e hoʻopili i nā papa SU-8 i hōʻike ʻia, a ukali ʻia e ka hoʻomohala ʻana i loko o ka waiʻona diacetone no 6.5 min. E hoʻomoʻa paʻa i nā waffles (200°C no 15 min) e hoʻopaʻa hou aku i ka papa SU-8.
Ua hoʻomākaukau ʻia ka PDMS ma ke kāwili ʻana i ka monomer a me ka hardener ma kahi lakio kaumaha o 10:1, a laila hoʻohemo ʻia ke kinoea i loko o kahi desiccator vacuum a ninini ʻia ma luna o ka ʻōnaehana nui SU-8. Ua 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 ua puka ʻia i mea e hiki ai ke hoʻopili ʻia nā paipu ma ke komo ʻana a me ka puka ʻana o ka microchannel. ʻO ka hope loa, ua hoʻopili mau ʻia nā microchannels PDMS i nā slide microscope me ka hoʻohana ʻana i kahi mea hana corona lawe lima (Electro-Technic Products, Chicago, IL) e like me ka mea i wehewehe ʻia ma kahi ʻē. ʻO ke ana o ka microchannel i hoʻohana ʻia ma kēia haʻawina he 200 µm × 20 µm (W × H) a he 3.6 cm ka lōʻihi.
Hoʻokō ʻia ke kahe ʻana o ka wai i hoʻokumu ʻia e ke kaomi hydrostatic i loko o ka microchannel ma ka mālama ʻana i ka pae wai i loko o ka waihona komo ma luna o ka ʻokoʻa kiʻekiʻe Δh39 i loko o ka waihona puka (Kiʻi 1).
kahi ʻo f ke koina o ka friction, i wehewehe ʻia ʻo f = C/Re no ke kahe laminar i loko o kahi kahawai rectangular, kahi ʻo C kahi mau ma muli o ka lakio hiʻohiʻona o ke kahawai, ʻo L ka lōʻihi o ka microchannel, ʻo Vav ka wikiwiki awelika i loko o ka microchannel, ʻo Dh ke anawaena hydraulic o ke kahawai, g - ka wikiwiki o ka umekaumaha. Ma ka hoʻohana ʻana i kēia kaulike, hiki ke helu ʻia ka wikiwiki awelika o ke kahawai me ka hoʻohana ʻana i kēia kaulike: