| Hyperosmotic tolerance of human spermatozoa: separate effects of glycerol, sodium chloride, and sucrose on spermolysis. | |
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MedLine Citation:
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PMID: 8353176 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Hyperosmotic stress, which cells experience during the freezing process, and its release during the warming process are both related to cryoinjury. To define optimal cooling or warming rates and prevent osmotic injury to human sperm, information is required regarding the osmotic tolerance of the cells as a function of 1) time, 2) temperature, 3) type of solute, and 4) solute concentration. Human sperm samples were divided into three aliquots. The aliquots were equilibrated at 0, 8, and 22 degrees C, respectively. Different hyperosmotic solutions were prepared by addition of either a permeating cryoprotective agent (glycerol) or nonpermeating solutes (sucrose, non-ionic; or NaCl, ionic) to isotonic Mann's Ringer solution. Aliquots of the prepared solutions were equilibrated at 0, 8, and 22 degrees C, respectively. A small volume (2.5 microliters) of each sperm aliquot was quickly mixed with 50 microliters of each hyperosmotic solution at the corresponding temperature. After times ranging from 5 s to 5 min, 10 microliters of each hyperosmotic cell suspension was abruptly returned to an isosmotic environment by mixing with 500 microliters of Mann's Ringer solution at the corresponding temperature. The plasma membrane integrity of cells after exposure to hyperosmotic stress and after return to isosmotic conditions was measured by a dual staining (carboxyfluoroscein diacetate and propidium iodide) technique and flow cytometry. The morphology of the treated cells was observed by scanning electron microscopy of freeze-substituted sperm. The results indicate that human spermatozoa exhibited a significant posthypertonic lysis/injury, i.e., loss of membrane integrity, when returned to isosmotic conditions after exposure to hyperosmotic solutions of NaCl or sucrose. The higher the hyperosmolality, the more serious the cell injury. The majority of the cells (> 50%) lost membrane integrity when the osmolality was > or = 2000 mOsm. In contrast, if the sperm were not returned to isosmotic conditions, the majority of the sperm in the hyperosmotic solutions appeared to maintain membrane integrity. For a given higher hyperosmolality (> 1000 mOsm), posthypertonic spermolysis was reduced with a decrease of temperature. Cell survival was also affected by time of cell exposure to hyperosmotic environments before cells were returned to the isotonic condition. The shorter the time, the higher the cell survival. When exposed to hyperosmotic glycerol solutions that were isotonic with respect to electrolytes, few cells lost their membrane integrity if the osmolality of glycerol was < 3000 mOsm. For a fixed high osmolality (> 3000 mOsm), the lower the temperature, the higher the percentage spermolysis.(ABSTRACT TRUNCATED AT 400 WORDS) |
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Authors:
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D Y Gao; E Ashworth; P F Watson; F W Kleinhans; P Mazur; J K Critser |
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Publication Detail:
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Type: In Vitro; Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Biology of reproduction Volume: 49 ISSN: 0006-3363 ISO Abbreviation: Biol. Reprod. Publication Date: 1993 Jul |
Date Detail:
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Created Date: 1993-09-22 Completed Date: 1993-09-22 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0207224 Medline TA: Biol Reprod Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 112-23 Citation Subset: IM |
Affiliation:
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Center for Reproduction and Transplantation Immunology, Methodist Hospital of Indiana, Indianapolis 46202. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Body Water
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metabolism Cell Survival Cryopreservation / methods Cryoprotective Agents* / pharmacology Flow Cytometry Glycerol / pharmacology Humans Hypertonic Solutions Male Microscopy, Electron, Scanning Osmotic Pressure Saline Solution, Hypertonic Spermatozoa* / cytology, drug effects, metabolism Sucrose / pharmacology |
| Grant Support | |
ID/Acronym/Agency:
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HD00980/HD/NICHD NIH HHS; HD25949/HD/NICHD NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Cryoprotective Agents; 0/Hypertonic Solutions; 0/Saline Solution, Hypertonic; 56-81-5/Glycerol; 57-50-1/Sucrose |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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