title: The development of rat Leydig cell progenitors in vitro: how essential is luteinising hormone?
abstract: Luteinising hormone (LH) appears to be important for the establishment of the adult-type Leydig cell population. The role of LH in the initial steps of stem Leydig cell/precursor cell differentiation is less clear. The aim of the present study was to elucidate the role of LH in the differentiation of spindle-shaped mesenchymal-like cells into Leydig cell progenitors. Interstitial cells were isolated from rat testes at three different ages reflecting different phases in development, and cultured in the presence of increasing concentrations of LH (ranging from 0.01 to 10 ng/ml). Cells were isolated from 10-day-old rats when stem Leydig cells/precursor cells are abundant; 13-day-old rats when the first 3beta-hydroxysteroid dehydrogenase (3beta-HSD)-positive Leydig cell progenitors have developed in the strain of rats used in this study; and 18-day-old rats just prior to the wave of progenitor proliferation. Immunohistochemistry revealed that before stem Leydig cells differentiate into progenitor cells, they acquire functional LH receptors and become precursor cells. This was confirmed by an in vivo immunohistochemical double-labelling experiment. Addition of LH to the cultures increased the percentage of LH/3beta-HSD-labelled Leydig cell progenitors, while the percentage of cells solely expressing the LH receptor decreased. Cell proliferation was negligible, suggesting that the increase in 3beta-HSD-positive cells is the result of precursor cell differentiation. When interstitial cells were isolated from 13-day-old rats and to a lesser extent from 10-day-old rats, a small proportion of the precursors could develop into progenitor cells independent of the presence of LH.
  
  
  CONCLUSIONS
  before Leydig stem cells differentiate into 3beta-HSD-positive progenitor cells, they acquire LH receptors and become precursor cells. LH appears to be essential, even at very low doses for the differentiation of these precursor cells into 3beta-HSD-positive progenitors, although a subpopulation of precursor cells can develop into progenitors independently of LH.
authors.affiliation: Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. katja.teerds@wur.nl
authors.forename: Katja J
authors.initials: KJ
authors.lastname: Teerds
authors.forename: Eddy
authors.initials: E
authors.lastname: Rijntjes
authors.forename: Margarita B
authors.initials: MB
authors.lastname: Veldhuizen-Tsoerkan
authors.forename: Focko F G
authors.initials: FF
authors.lastname: Rommerts
authors.forename: Mieke
authors.initials: M
authors.lastname: de Boer-Brouwer
mesh_terms: D015096:3-Hydroxysteroid Dehydrogenases / Q000032:analysis / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001973:Bromodeoxyuridine; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006651:Histocytochemistry; D017403:In Situ Hybridization; D007985:Leydig Cells / Q000166:cytology*; D007986:Luteinizing Hormone / Q000378:metabolism / Q000494:pharmacology*; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D011974:Receptors, LH / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*
chemical_list: D015415:Biomarkers; D011974:Receptors, LH; D007986:Luteinizing Hormone; D000242:Cyclic AMP; D015096:3-Hydroxysteroid Dehydrogenases; D001973:Bromodeoxyuridine