Wistar Sıçan Sperminin Elektron Mikroskobik ve Morfometrik Analizi

Yıl 2024, , 157 - 161, 30.06.2024

Emre Demirci

https://doi.org/10.35229/jaes.1440801

Öz

Wistar sıçanlarda epididimal spermin taramalı elektron mikroskobu (SEM) ile
görüntülenmesi ve morfolojik incelenmesi çalışmanın amacını oluşturmaktadır. Çalışmanın
materyalini üç Wistar Sıçanından alınan epididimal spermalar oluşturdu. Elde edilen epididimal
spermalara % 2,5 glutaraldehit solüsyonu kullanılarak rutin fiksasyon işlemi yapıldı. Spermaların
morfometrik ölçümleri Taramalı elektron mikroskobu kullanılarak gerçekleştirildi. Çalışma, baş,
orta parça ve flagellar bölümün kalan kısmı dahil olmak üzere spermin çeşitli bölgelerinin
incelenmesine odaklanmıştır. Farklı sahalarda bütünlüğünü korumuş spermalarda ölçümler
dikkate alındı. Bu bölgelerin ölçümleri İmageJ programı ile yapıldı. Yapılan ölçümler Sonucunda
sıçan spermasının baş uzunluğunun ortalama 7,821 ± 0,844 μm total flagellum uzunluğunun
ortalama 168,549 ± 9,19 μm ve ortalama flagellum/baş oranının 21,550 olduğu ve flagellumun
oldukça uzun olduğu belirlendi. Araştırmada kullanılan sıçan sperminin ölçümlerinin diğer sıçan
spermlerinin ölçümleriyle tutarlı olduğu görüldü. Yapılan çalışmalar incelendiğinde, sıçan
spermasının vücut boyutuna göre son derece uzun bir kamçıya sahip olduğu sonucuna varıldı.
Ortaya konulan spermaların morfolojik farklılıkları yapılacak deneysel çalışmalara bir rehber
olacaktır.

Anahtar Kelimeler

Elektron Mikroskop, Sıçan Sperma, Morfoloji

Electron Microscopic and Morphometric Analysis of Wistar Rat Sperm

Öz

The aim of the study is to image and morphologically examine epididymal sperm in
Wistar rats with scanning electron microscopy (SEM). The material of the study was epididymal
sperm taken from three Wistar Rats. Routine fixation was performed on the obtained epididymal
sperm using 2.5% glutaraldehyde solution. Morphometric measurements of sperm were carried
out using scanning electron microscopy. The study focused on examining various regions of the
sperm, including the head, midpiece, and the remainder of the flagellar segment. Measurements
were taken into account in semen that preserved its integrity in different fields. Measurements of
these regions were made with the ImageJ program. As a result of the measurements, it was
determined that the average head length of rat semen was 7.821 ± 0.844 μm, the average total
flagellum length was 168.549 ± 9.19 μm and the average flagellum/head ratio was 21.550 and the
flagellum was quite long. It was observed that the measurements of the rat sperm used in the
study were consistent with the measurements of other rat sperm. When the studies were examined,
it was concluded that rat sperm has an extremely long flagellum compared to its body size. The
morphological differences of the sperms revealed will be a guide for experimental studies.

Anahtar Kelimeler

Electron Microscope, Morphology, Rat sperm

Etik Beyan

Etik Kurul Onayına İhtiyacı Yoktur Yazısı Yüklenmiştir.

Kaynakça

• Avidor-Reiss, T., Khire, A., Fishman, E. L. & Jo, K. H. (2015). Atypical centrioles during sexual reproduction. Frontiers in cell and developmental biology, 3, 21.
• Cummins, J.M. & Woodall, P.F. (1985). On mammalian sperm dimensions. Journal of Reproduction and Fertility, 75, 153-175.
• Davis, R.O., Gravance, C.G., Thal, D.M. & Miller, M.G. (1994). Automated analysis of toxicantinduced changes in rat sperm head morphometry. Reproductive Toxicity, 8(6), 521-529.
• Demirci, B., Kandil, B., Yüksel, S. & Gültiken, M. E. (2023). Morphological structure of rat tongue using light and scanning electron microscopy. Microscopy Research and Technique, 86(1), 75- 83.
• Devireddy, R.V., Swanlund, D.J., Roberts, K.P. & Bischof, J.C. (1999). Subzero water permeability parameters of mouse spermatozoa in the presence of extracellular ice and cryoprotective agents. Biology of Reproduction, 61, 764-775.
• Dibal, N.I., Garba, S. H. & Jacks, T.W. (2020). Morphological assessment of epididymal sperm in Wistar rats using different histological stains. Acta Veterinaria Eurasia, 46(3), 132-137.
• Eddy, E. M., Toshimori, K., & O'Brien, D. A. (2003). Fibrous sheath of mammalian spermatozoa. Microscopy research and technique, 61(1), 103- 115.
• Firman, R.C. & Simmons, L.W. (2010). Experimental evolution of sperm quality via postcopulatory sexual selection in house mice. Evolution, 64, 1245-1256.
• Gage, M.J.G. (1998). Mammalian sperm morphometry. Proceedings of the Royal Society of London. Series B. Biological Sciences, 265, 97-103.
• Gomendio, M. & Roldan, E.R.S. (1991). Sperm size and sperm competition in mammals. Proceeding of the Royal Society London Series B, 243, 181-185.
• Gu, N.H., Zhao, W.L., Wang, G.S. & Sun, F. (2019). Comparative analysis of mammalian sperm ultrastructure reveals relationships between sperm morphology, mitochondrial functions and motility. Reproductive Biology and Endocrinology, 17(1), 1-12.
• Holt, W.V. (2000). Fundamental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology, 53, 47- 58.
• Lazar, J., Moreno, C., Jacob, H.J. & Kwitek, A.E. (2005). Impact of genomics on research in the rat. Genome Research, 15, 1717-1728.
• Lindemann, C. B., & Lesich, K. A. (2016). Functional anatomy of the mammalian sperm flagellum. Cytoskeleton, 73(11), 652-669.
• Mortimer, D. (2018). The functional anatomy of the human spermatozoon: relating ultrastructure and function. Molecular Human Reproduction, 24, 567-592.
• Nakatsukasa, E., Inomata, T., Ikeda, T., Shino, M. & Kashiwazaki, N. (2001). Generation of live rat offspring by intrauterine insemination with epididymal spermatozoa cryopreserved at -196°C. Reproduction, 122, 463-467.
• Nakatsukasa, E., Kashiwazaki, N., Takizawa, A., Shino, M., Kitada, K., Serikawa, T., Hakamata, Y., Kobayashi, E., Takahashi, R., Ueda, M., Nakashima, T. & Nakagata, N. (2003). Cryopreservation of spermatozoa from closed colonies, and inbred, spontaneous mutant, and transgenic strains of rats. Comparative Medicine, 53, 639-641.
• Oh, S.H., Miyoshi, K. & Funahashi, H. (1998). Rat oocytes fertilized in modified rat 1 cell embryo culture medium containing a high sodium chloride concentration and bovine serum albumin maintain developmental ability to the blastocyst stage. Biology of Reproduction, 59, 884-889.
• Tesson, L., Cozzi, J., Menoret, S., Remy, S., Usal, C., Fraichard, A. & Anegon, I. (2005). Transgenic modifications of the rat genome. Transgenic Research, 14, 531-546.
• Tourmente, M., Gomendio, M. & Rolando, A. (2011). Sperm competition and the evolution of sperm design in mammals. BMC Evolutionary Biology, 11, 12.
• Varisli, O., Uguz, C., Agca, C. & Agca, Y. (2009). Effect of chilling on the motility and acrosomal integrity of rat sperm in the presence of various extenders. Journal of American Association for Laboratory Animal, 48, 1-7.
• Varisli, O., Scott, H., Agca, C. & Agca, Y. (2013). The effects of cooling rates and type of freezing extenders on cryosurvival of rat sperm. Cryobiology, 67, 109-116.
• Vashishat, N., Dhanju, C.K. & Cheema, R.S. (2012). Morphology and morphometry of cauda epididymal spermatozoa in Rattus rattus L. Proceeding of the National Academy of Sciences, India, Section B Biological Sciences, 82, 285-290.
• Woolley, D.M., Carter, D.A. & Tilly, G.N. (2008). Compliance in the neck structures of the guinea pig spermatozoon, as indicated by rapid freezing and electron microscopy. Journal of Anatomy, 213(3), 336-341.
• Yamashiro, H., Han, Y.J., Sugawara, A., Tomioka, I., Hoshino, Y. & Sato, E. (2007). Freezability of rat epididymal sperm induced by raffinose in modified Krebs-Ringer bicarbonate (mKRB) based extender solution. Cryobiology, 55, 285- 294.
• Ward, W.S. & Coffey, D.S. (1991). DNA packaging and organization in mammalian spermatozoa: comparison with somatic cells. Biology of Reproduction, 44, 569-574.