Ball B A, Dobrinski I, Fagnan M S, Thomas P G
Department of Clinical Sciences, Cornell University, Ithaca, NY 14853, USA.
Am J Vet Res. 1997 Aug;58(8):816-22.
To examine glycoconjugates in the isthmic and ampullar regions of the uterine tube (oviduct) of horses during estrus, diestrus, and pregnancy.
Oviductal samples from 17 mares.
Oviducts were collected during estrus (n = 3), diestrus (n = 3), or pregnancy (n = 3), embedded, and snap frozen in liquid nitrogen. Frozen sections (5 to 6 microns in thickness) were stained with 100 micrograms/ml of fluorescein-isothiocyanate-conjugated lectin (30 min at 38.5 C) and were evaluated by use of epifluorescence microscopy and video image analysis. Specificity of lectins was established by blocking with the corresponding carbohydrate. Dolichos biflorus agglutinin (DBA)-affinity studies on western blots of oviductal lavage fluid, oviductal explant conditioned media, and apical membrane proteins from isthmic and ampullar regions of oviducts were used to identify glycoproteins with galactosyl residues.
Use of 4 lectins resulted in differential labeling of the luminal surface of the oviductal epithelium. Both DBA and soybean agglutinin labeled the apical epithelium of the isthmus, but not the ampullar oviduct. Soybean agglutinin resulted in more-intensely labeled epithelium in the isthmic region of oviducts during estrus and pregnancy than during diestrus. The DBA labeled a number of glycoproteins in conditioned media from both regions of the oviduct. These glycoproteins ranged from 14 to 200 kd, with major glycoproteins identified at 31 and 57 kd.
The predominant glycoconjugates in the oviduct of horses are galactosyl residues. There are regional differences in the distribution of these galactosyl glycoconjugates in the isthmic and ampullar oviduct.
研究发情期、黄体期及妊娠期马输卵管峡部和壶腹部的糖缀合物。
17匹母马的输卵管样本。
在发情期(n = 3)、黄体期(n = 3)或妊娠期(n = 3)采集输卵管,进行包埋,并在液氮中速冻。将冷冻切片(厚度为5至6微米)用100微克/毫升的异硫氰酸荧光素结合凝集素染色(在38.5℃下孵育30分钟),并通过落射荧光显微镜和视频图像分析进行评估。通过用相应的碳水化合物封闭来确定凝集素的特异性。利用双花扁豆凝集素(DBA)对输卵管灌洗液、输卵管外植体条件培养基以及输卵管峡部和壶腹部顶端膜蛋白的western印迹进行亲和研究,以鉴定具有半乳糖基残基的糖蛋白。
使用4种凝集素导致输卵管上皮腔面的标记不同。DBA和大豆凝集素均标记峡部顶端上皮,但不标记壶腹部输卵管。在发情期和妊娠期,大豆凝集素使输卵管峡部区域的上皮标记比黄体期更强。DBA在输卵管两个区域的条件培养基中标记了许多糖蛋白。这些糖蛋白的分子量范围为14至200千道尔顿,主要糖蛋白的分子量为31和57千道尔顿。
马输卵管中主要的糖缀合物是半乳糖基残基。这些半乳糖基糖缀合物在输卵管峡部和壶腹部的分布存在区域差异。