Effects of Trehalose Supplementation on Lipid Composition of Rooster Spermatozoa Membranes in a Freeze/Thaw Protocol

Abstract

The plasma membrane of spermatozoa plays an important role in the formation and maintenance of many functions of spermatozoa, including during cryopreservation. As a result of chromatographic analysis, the content of lipids and fatty acids in the membranes of spermatozoa of roosters of two breeds was determined under the influence of cryoprotective media containing trehalose LCM-control (0 mM), Treh20 (9.5 mM), and Treh30 (13.4 mM). The use of the cryoprotective diluent Treh20 made it possible to maintain a dynamic balance between the synthesis and degradation of phospholipids and sterols in the plasma membranes of frozen/thawed spermatozoa, close to that of native spermatozoa. This contributed to an increase in the preservation of frozen/thawed spermatozoa membranes from 48.3% to 52.2% in the egg breed and from 30.0% to 35.1% in the meat- and-egg breed. It was also noted that their kinetic apparatus (mobility indicators) remained at the level of 45.6% (egg breed) and 52.4% (meat-and-egg breed). An increase in the concentration of trehalose to 13.4 mM in a cryoprotective diluent for rooster sperm resulted in a decrease in the morphofunctional parameters of frozen/thawed spermatozoa.

Lipid composition of plasma membranes of rooster sperm (Gallus gallus domesticus) and its dynamics during cryopreservation

“…Россия; Motic BA410E, Китай). Полученные эякуляты объединяли и делили на 2 части: добавляли криозащитную среду 1 часть -ЛКСконтроль [8], 2 часть -ЛКС-Т20 [9] в соотношении 1:1. Замораживали в гранулах по протоколу, описанному Станишевской и др.…”

“…Замораживали в гранулах по протоколу, описанному Станишевской и др. (2023) [9]. Хранили замороженное семя в сосудах Дьюара в течение 30 дней.…”

“…Анализ растворимых углеводов цитозоля и мембранных липидов сперматозоидов. Липиды и углеводы анализировали, как описано ранее [9,12] Ответом на стресс, вызванный протоколом замораживания/оттаивания, является перестройка мембранных связей сперматозоидов за счет изменения соотношения не только основных (ФЭ, ФС, ФХ, СМ и СТ), но и изменения минорных липидов (ГЛ, КЛ, СГЛ). Известно, что изменения в соотношении глицеринфосфолипид-холестерин играют существенную роль в саморегуляции мембран сперматозоидов, связанной с криотолерантностью [13,14].…”

Hydrolysis of Phospholipids in Presence of Phospholipase D:Thrmodynamic and Kinetic Studies of Hydrolysis in Water and Alcoholic Environments

“…It is normally the most abundant phospholipid in animals and plants, often constituting nearly 50% of total complex lipids, and as such appears to be a key component of membrane bilayers [12]. Phosphatidylserine (PS), which makes up around 5-10% of all phospholipids, and phosphatedylethanolamine (PE), which accounts for 20-30% of all phospholipids, are the two main types in the inner layer of cell membranes [13][14][15]. An analogy to other phospholipids, PS has a polar head group neutral serine group attached to the phosphate and thus has an overall negative charged linked to position sn-3 and two acyl chains at locations sn-1 and sn-2 of the glycerol moiety [16][17][18].…”

Trehalose as a Stabilizer of the Lipid Composition of Membranes and the Composition of the Cytosol of Frozen/Thawed Rooster Spermatozoa

“…The lipid ratio of the plasma membranes can be postulated as a general basis for the predicted cryodamage of spermatozoa. Spermatozoa of both domestic fowl and other animal species have plasma membranes highly specialized in their lipid composition, with a high content of polyunsaturated fatty acids (PUFAs), sterols, phosphatidylcholines (PC), and sphingomyelins (SM), which play an important role in the interactions between spermatozoa and oocyte and fertilization ability [15,16].…”