Osmotic Tolerance Limits and Properties of Rhesus Monkey (Macaca mulatta) Spermatozoa

Abstract

Fundamental cryobiological characteristics of rhesus spermatozoa must be determined for successful cryopreservation techniques to be established. The main objectives of the present study were to determine the osmotic behavior and osmotic tolerance limits of rhesus macaque spermatozoa. Cell volume changes over anisotonic conditions were assessed using an electronic particle counter and sperm motility was evaluated with a computer-assisted sperm analysis system. Analysis of membrane integrity and mitochondrial membrane potential was performed using flow cytometry. Rhesus monkey spermatozoa behave as linear osmometers in the osmotic range tested (75-900 mOsmol kg Ϫ1 ), as shown by the Boyle van't Hoff plot (r 2 ϭ .99). Rhesus spermatozoa have a mean cell volume of 36.8 Ϯ 0.5 m 3 at 22ЊC, with 77.2% of the intracellular volume being osmotically inactive. Results regarding sperm tolerance to osmotic stress showed that sperm motility was more sensitive than membrane integrity to deviations from isotonicity and, in addition, that rhesus sperm motility and membrane integrity were more sensitive to hypertonic than hypotonic conditions. Mitochondrial membrane potential did not explain the lack of sperm motility observed under anisosmolal conditions in our study. Although most spermatozoa were able to recover initial volume after osmotic stress, they were not able to recover initial motility.

Reprogramming Huntington Monkey Skin Cells into Pluripotent Stem Cells

“…Although the mechanism of cell type-specific pathology may hold the key for understanding HD pathogenesis, it is important to investigate the impact of mutant htt in peripheral tissues and subsequent HD development (Martin et al, 2008;Sathasivam et al, 1999). Because stem cells retain multipotent differentiation capability, they are powerful tools for investigating the underlying mechanism of cell type-specific events and the influence of developmental events that lead to pathologic consequences (Byrne et al, 2007;Dimos et al, 2008;Hanna et al, 2007;Lakshmipathy et al, 2007aLakshmipathy et al, , 2007bLaurent et al, 2008;Muller et al, 2008;Park et al, 2008aPark et al, , 2008bPark et al, , 2008cPruszak et al, 2007;Rutllant et al, 2003). Thus, pluripotent stem cells that develop HD cellular features paralleling neural development is a useful model for studying HD pathogenesis.…”

The Macaque Sperm Actin Cytoskeleton Reorganizes in Response to Osmotic Stress and Contributes to Morphological Defects and Decreased Motility1

“…Sperm in low-osmolality medium (100 mmol/kg) showed a statistically significant increase in MCV (31.04 6 0.51 lm 3 ; P , 0.05), whereas the MCV for sperm in high-osmolality medium (600 mmol/kg) did not differ significantly (20.17 6 1.3 lm 3 ) when compared to control values. The increase in sperm cell volume following low-osmolality incubations was expected based on previous work on the osmotic tolerance limits for macaque sperm [34,37] and indicate that the sperm in our model were undergoing osmotic stress.…”

“…Sperm sample (5 ll) was placed on a prewarmed microscope slide and overlaid with a 22-mm 2 prewarmed cover slip, and the slide was maintained at 378C during analysis by a heated slide holder (Hamilton-Thorne Research). Instrument settings for motility analysis were as previously described [34]. Percent total motility (TM), percent progressive motility (PM), straight line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), linearity (LIN), amplitude of lateral head displacement (ALH), beat cross frequency (BCF), and straightness (STR) were determined.…”

Aging in male primates: reproductive decline, effects of calorie restriction and future research potential

“…Whereas histological techniques and 'live-dead' morphology stains only measure whether the membrane is morphologically intact, this assay evaluates the functional integrity of the sperm membrane and fertilizing potential (Jeyendran et al 1984). This assay has been used in research with nonhuman primates as well as applied clinically (Kholkute et al 2000;Jeyendran 2003;Rutllant et al 2003).…”