Category Archives: Adult Rat Testis - Part 2

Local Regulation of Macrophage Subsets in the Adult Rat Testis: RESULTS(4)

Total testicular macrophage numbers (i.e., all ED1+ cells and ED2+ cells) declined by approximately 40-50% in T3-implanted and in T24-implanted rat testes (Fig. 6A). This decline was primarily the result of a 40% reduction in the number of resident macrophages (ED2+ cells only) (Fig. 6B). There was a more pronounced decline (75%) in the less numerous ED1+ED2~ monocyte/macrophage subset in the T3-implanted animals, while the … Continue reading

Local Regulation of Macrophage Subsets in the Adult Rat Testis: RESULTS(3)

Testicular MIF Concentrations Immunoreactive MIF levels in testicular IF measured by ELISA (Fig. 4) and by Western blot analysis (Fig. 5) were unaffected in all groups with scrotal testes but were reduced in the abdominal testis. Both T3- and T24-implants at least partially restored immunoreactive MIF levels in the abdominal testes. Accurate quantification of MIF protein levels from the Western data was not considered because … Continue reading

Local Regulation of Macrophage Subsets in the Adult Rat Testis: RESULTS(2)

Hormonal and Accessory Organ Observations Serum LH and FSH levels were increased 2-fold in cryptorchid rats (Fig. 2, A and B). Serum LH was reduced to 30% of normal control levels by either length of T-im-plant in rats with either scrotal or abdominal testes (Fig. 2A), while serum FSH was reduced to a lesser degree (5080% of normal) under the same conditions (Fig. 2B).

Local Regulation of Macrophage Subsets in the Adult Rat Testis: RESULTS(1)

General Morphological Observations Testis weights were reduced by 22% in T3-implanted rats compared with normal controls, but weights were fully maintained in T24-implanted rats (Fig. 1A). In bilaterally cryptorchid rats, testis weights declined by 67% compared with those in controls, but neither the T3- nor T24-implants had any additional effect. These weight changes were attributable to a loss of some late-stage spermatogenic cells in the … Continue reading

Local Regulation of Macrophage Subsets in the Adult Rat Testis: MATERIALS AND METHODS(6)

Western BlotAnalysis Residual samples of testicular IF from each experimental group were pooled and were subjected to Western blot analysis as previous described, with some modifications. The pooled samples were diluted 1:1 with sample buffer (0.125 M Tris, 4% SDS, 20% glycerol, 2.5% (3-mercapto-ethanol, 0.02% bromophenol blue, pH 6.8). The equivalent of 6-10 ^l of IF from each experimental group was run on a 15% … Continue reading

Local Regulation of Macrophage Subsets in the Adult Rat Testis: MATERIALS AND METHODS(5)

For counting and subsequent analysis, the macrophages were separated into total monocytes and macrophages (labeled by combined ED1 and ED2 immunohistochemistry) and total resident macrophages (all ED2+ cells). In the previous study, the ED2+ resident macrophages represented 80% of the total testicular mononuclear phagocyte population. Intratesticular ED1+ED2- monocyte/macrophages and putative ‘‘long-term resident’’ macrophages (ED2+ED1- cells) were calculated by subtraction, as previously described. birth control pills Hormone RIAs Serum … Continue reading

Local Regulation of Macrophage Subsets in the Adult Rat Testis: MATERIALS AND METHODS(4)

Immunohistochemistry and Stereological Analysis Sections were stained with the mouse anti-rat monoclonal antibodies ED1, which labels a lysosomal antigen specific to monocytes, dendritic cells, and some macrophages (1:4 dilution), or ED2, which labels resident macrophages specifically (1:800 dilution), using immunoperoxi-dase detection and subsequent hematoxylin counterstaining as previously described. In order to label all cells of the mononuclear phagocyte and dendritic cell lineages, sections were double-stained … Continue reading

Local Regulation of Macrophage Subsets in the Adult Rat Testis: MATERIALS AND METHODS(3)

The remaining testis was fixed by lower body perfusion via the descending aorta, with 2% paraformaldehyde, 75 mM lysine, 10 mM periodate, containing 1.0% glutaralde-hyde, and then weighed and processed for collection of 6-^m frozen sections, as previously described. Animals were not pretreated with any anti-clotting agents. buy levaquin online After collection of the testes, several androgen-responsive tissues, i.e., the thymus, seminal vesicles (including coagulating … Continue reading

Local Regulation of Macrophage Subsets in the Adult Rat Testis: MATERIALS AND METHODS(2)

Groups of untreated rats and rats that had been made cryptorchid 7 days previously received low-dose (3 cm) or high-dose (3 X 8-cm implants: 24-cm total) T-implants (s.c.) and were maintained for a further 21 days. Consequently, all cryptorchid rats were maintained for a total of 28 days prior to assessment. Each treatment group consisted of 6 animals, as follows: 1) scrotal testes and no … Continue reading

Local Regulation of Macrophage Subsets in the Adult Rat Testis: MATERIALS AND METHODS(1)

Animals Adult male Sprague-Dawley rats (110-120 days old) were obtained from the Monash University Central Animal House and were housed at Monash Medical Centre for the duration of the experimental procedure under conditions of controlled day length (12L:12D) with unlimited access to food and water. This study was conducted in accordance with the Guiding Principles for the Care and Use of Research Animals of the … Continue reading