1987
DOI: 10.1095/biolreprod36.3.769
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Effect of Luteinizing Hormone Deprivation In Situ on Steroidogenesis of Rat Leydig Cells Purified by a Multistep Procedure12

Abstract: Depriving rats of luteinizing hormone (LH) causes Leydig cells to lose smooth endoplasmic reticulum and diminishes their P450 C17-hydroxylase/C17,20-lyase activity (Wing et al., 1984). LH administration to hypophysectomized rats prevents these changes in Leydig cell structure and function (Ewing and Zirkin, 1983). We adopted a multistep procedure of rat Leydig cell isolation to study the trophic effects of LH on steroidogenesis in the Leydig cell. Our method employs vascular perfusion, enzymatic dissociation, … Show more

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Cited by 282 publications
(165 citation statements)
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“…Primary Rat Leydig Cell Isolation and Lentiviral InfectionLeydig cells were isolated from 60-to 64-day-old Sprague-Dawley rats (Charles River Laboratories) as described previously (67). In brief, the testis were decapsulated, digested in M-199 medium containing 0.05% collagenase/dispase, 0.005% soy trypsin inhibitor, and 0.001% deoxyribonuclease I (all from Sigma) and shaken at 90 cycles/min for 45 min at 34°C.…”
Section: Methodsmentioning
confidence: 99%
“…Primary Rat Leydig Cell Isolation and Lentiviral InfectionLeydig cells were isolated from 60-to 64-day-old Sprague-Dawley rats (Charles River Laboratories) as described previously (67). In brief, the testis were decapsulated, digested in M-199 medium containing 0.05% collagenase/dispase, 0.005% soy trypsin inhibitor, and 0.001% deoxyribonuclease I (all from Sigma) and shaken at 90 cycles/min for 45 min at 34°C.…”
Section: Methodsmentioning
confidence: 99%
“…This suspensions of interstitial cells (Klinefelter et al, 1987) was used to prepare primary cultures of purified Leydig cells (Andric et al, 2007(Andric et al, , 2010aKostic et al, 2008Kostic et al, , 2010Kostic et al, , 2011 by centrifugation on a Percoll gradient consisting of four 2 mL layers of Percoll with densities of 1.090, 1.080, 1.065 and 1.045 g/mL (formed by mixing isotonic Percoll consisting of 109 concentrated DMEM/F12 enriched with 3% of BSA and the corresponding amount of Percoll and distilled water). A crude suspension of interstitial cells (approximately 35-40 9 10 6 cells), containing besides Leydig cells macrophages and endothelial cells (Klinefelter et al, 1987), was applied to each Percoll gradient and centrifuged at 500 g for 28 min at room temperature. Fractions containing Leydig cells were collected from the 1.080/1.065 g/mL and 1.065/1.045 g/mL interfaces, washed in 50 mL M199-0.1% BSA and centrifuged at 200 g for 5 min at room temperature.…”
Section: Extraction Of Steroids From Testicular Tissuementioning
confidence: 99%
“…It is important to emphasize that the structural organization of the testis may account for the integration of autocrine and paracrine factors to regulate testicular steroidogenesis and TIF characteristics and/or content. Although testosterone is an exclusive product of Leydig cells, secretory products originated from seminiferous tubule and/or Sertoli cells and/or non-steroidogenic cells in the testicular interstitium (Klinefelter et al, 1987;Saez, 1994;Hutson, 2006) could take place in regulation of TIF characteristics. Accordingly, reported changes in the TIF and testosterone content in testis could be the consequence of the complex effects of secretory contribution of the different cell types including, besides HSD3B-positive Leydig cells, also macrophages, endothelial cells, fibroblasts, peritubular cells, Sertoli cells etc.…”
Section: (A) (B) (D) (C)mentioning
confidence: 99%
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“…Cells were centrifuged again at 900×g for 5 min, resuspended and seeded on coverslips. The efficiency of this process was confirmed by cytochemical detection of the enzyme 3β-HSD as described elsewhere [28].…”
Section: Osmotic Shockmentioning
confidence: 99%