| OPTIMIZATION OF A FILTER-LYSIS PROTOCOL TO PURIFY RAT TESTICULAR HOMOGENATES FOR AUTOMATED SPERMATID COUNTING. | |
| | |
MedLine Citation:
|
PMID: 22240558 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
|
Objectives: Quantifying testicular homogenization resistant spermatid heads (HRSH) is a powerful indicator of spermatogenesis. These counts have traditionally been performed manually using a hemocytometer, but this method can be time consuming and biased. We aimed to develop a protocol to reduce debris for the application of automated counting, which would allow for efficient and unbiased quantification of rat HRSH. Findings: We developed a filter-lysis protocol that effectively removes debris from rat testicular homogenates. After filtering and lysing the homogenates, we found no statistical differences between manual (classic and filter-lysis) and automated (filter-lysis) counts using one-way ANOVA with Bonferroni's multiple comparison test. In addition, Pearson's correlation coefficients were calculated to compare the counting methods and there was a strong correlation between the classic manual counts and the filter-lysis manual (r = 0.85, p = 0.002) and the filter-lysis automated (r = 0.89, p = 0.0005) counts. We also tested the utility of the automated method in a low dose exposure model known to decrease HRSH. Adult Fischer 344 rats exposed to 0.33% 2,5-hexanedione (HD) in the drinking water for 12 weeks demonstrated decreased body (p = 0.02) and testes (p = 0.002) weights. In addition, there was a significant reduction in the number of HRSH per testis (p = 0.002) when compared to control. Conclusions: A filter-lysis protocol was optimized to purify rat testicular homogenates for automated HRSH counts. Automated counting systems yield unbiased data and can be applied to detect changes in the testis after low dose toxicant exposure. |
| | |
Authors:
|
Sara E Pacheco; Linnea M Anderson; Kim Boekelheide |
Related Documents
:
|
6521818 - Turnover of ethanolamine phosphoglycerides in different brain areas of adult and aged r... 18589308 - [18f]fbau 3',5'-dibenzoate, a lipophilic prodrug, enhances brain uptake of the cell pro... 12913878 - Locoregional opening of the rodent blood-brain barrier for paclitaxel using nd:yag lase... 19625028 - Maintenance of white matter integrity in a rat model of radiation-induced cognitive imp... 7480138 - Effect of hematoporphyrin derivative on hematological parameters in rats. 10220048 - The effect of age and injury on the expression of inducible nitric oxide synthase in fa... |
Publication Detail:
|
Type: JOURNAL ARTICLE Date: 2012-1-12 |
Journal Detail:
|
Title: Journal of andrology Volume: - ISSN: 1939-4640 ISO Abbreviation: - Publication Date: 2012 Jan |
Date Detail:
|
Created Date: 2012-1-13 Completed Date: - Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 8106453 Medline TA: J Androl Country: - |
Other Details:
|
Languages: ENG Pagination: - Citation Subset: - |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
|
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Secretory Phospholipase A2 Group IID is Involved in Progesterone-induced Acrosomal Exocytosis of Hum...
Next Document: The Comparison of the Aging Male Symptoms (AMS) Scale and Androgen Deficiency in the Aging Male (ADA...