Mahony M C, Fan Q, Cartwright S, Hodgen G D
Jones Institute for Reproductive Medicine, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk 23507.
Am J Reprod Immunol. 1994 Mar-Apr;31(2-3):133-40. doi: 10.1111/j.1600-0897.1994.tb00858.x.
The present study was carried out to evaluate the changes in morphology and motility of spermatozoa retrieved from different regions of the epididymis of the cynomolgus monkey. The role of the epididymis in sperm maturation is assessed by assaying protein synthesis within different regions of the epididymis and by correlating these with changes in spermatozoal membrane surface components.
Spermatozoa retrieved from proximal caput (CP), midcorpus (CO), and distal cauda (CD) were assessed by morphological evaluation and computerized motion analysis. Membrane surface proteins of spermatozoa of different epididymal regions were extracted and separated on SDS-PAGE. Protein synthesis of different regions of the epididymis were assayed in vitro by [35S]-methionine incorporation.
Spermatozoa obtained from different regions of the epididymis differed morphologically only in the location of the cytoplasmic droplet. Specifically, from caput to corpus to cauda, sperm steadily exhibited a more distal cytoplasmic droplet. When the motion parameters of velocity and amplitude of the lateral head were examined, CP spermatozoa were not progressively motile, and poor duration of movement was most noticeable for CO spermatozoa compared with CD spermatozoa. Membrane extracts from CP, CO, and CD epididymal monkey spermatozoa differed in only several protein bands. Three major polypeptide bands (19, 30, and 60 kD) that were absent from CP sperm were present in CO and CD sperm, with the latter showing increased intensity. Several polypeptides were lost from the sperm during epididymal transit: a 25-kD band was lost in CD sperm; and bands at 27 kD, 50-52 kD, and 90 kD were only present for CP sperm. Additionally, regional differences exist for proteins secreted by the cynomolgus monkey epididymis. Proteins (15, 25 kD) were only secreted in the CP region; a 38-kD protein increased in intensity from the CP to CD regions, whereas a 21-kD protein was absent from CD-secreted medium.
These preliminary findings permitted the identification of several "maturational antigens" for cynomolgus monkey spermatozoa. Further characterization of these antigens that are modified during epididymal transit is warranted to determine their significance in the acquisition of progressive motility and fertilizing ability by epididymal spermatozoa.
本研究旨在评估从食蟹猴附睾不同区域获取的精子的形态和活力变化。通过检测附睾不同区域的蛋白质合成,并将其与精子细胞膜表面成分的变化相关联,来评估附睾在精子成熟过程中的作用。
通过形态学评估和计算机化运动分析,对从附睾近端头部(CP)、附睾体中部(CO)和附睾尾远端(CD)获取的精子进行评估。提取不同附睾区域精子的膜表面蛋白,并在SDS-PAGE上进行分离。通过[35S]-甲硫氨酸掺入法体外检测附睾不同区域的蛋白质合成。
从附睾不同区域获取的精子在形态上仅在细胞质滴的位置有所不同。具体而言,从头部到体部再到尾部,精子的细胞质滴逐渐向更远端移动。当检测头部横向运动的速度和幅度等运动参数时,CP精子不具有渐进性运动能力,与CD精子相比,CO精子的运动持续时间最短。CP、CO和CD附睾猴精子的膜提取物仅在几条蛋白条带上存在差异。CP精子中不存在的三条主要多肽条带(19、30和60kD)在CO和CD精子中存在,且后者强度增加。在附睾转运过程中,精子丢失了几种多肽:CD精子中丢失了一条25-kD条带;27kD、50-52kD和90kD的条带仅在CP精子中存在。此外,食蟹猴附睾分泌的蛋白质也存在区域差异。蛋白质(15、25kD)仅在CP区域分泌;一种38-kD蛋白质的强度从CP区域到CD区域增加,而CD分泌的培养基中不存在一种21-kD蛋白质。
这些初步发现有助于鉴定食蟹猴精子的几种“成熟抗原”。有必要进一步表征这些在附睾转运过程中发生修饰的抗原,以确定它们在附睾精子获得渐进性运动能力和受精能力中的意义。