Restart of steroidogenesis in dogs during recrudescence of testicular function following downregulation with a GnRH-agonist implant.
To date, no details are available concerning the restart of steroidogenesis following the downregulation of testicular endocrine and germinative function by gonadotrophin-releasing hormone (GnRH)-agonist implants. This restart was assessed by determining the expression of steroidogenic acute regulatory (StAR) protein, cytochrome P450 side-chain cleavage enzyme (P450scc) and cytochrome P450 17α-hydroxylase,17,20-lyase (P450c17). The re-establishment of steroidogenesis was initiated by the removal of the GnRH-agonist implant (18.5 mg azagly nafarelin, Gonazon) at 5 months after treatment. Testes were removed at 3-week intervals (weeks 0-24) and four groups were formed according to the stage of spermatogenesis as revealed by the most developed germ cells observed (developmental group [DG] spermatocytes to DG elongated spermatids). Five dogs served as untreated controls. Positive immunostaining for StAR, P450scc and P450c17 was restricted to Leydig cells. Western blot indicated the specifity of the respective antibodies with hints of a expression of canine-specific P450scc and P450c17 proteins. A significant effect of group was observed for a percentage of the immunopositive area (PIA) as an indicator of active Leydig cells for StAR (P<0.05), P450scc (P<0.001) and P450c17 (P<0.001), with PIA being lowest for the DG spermatocytes. With regard to the strength of the immunopositive signal, a significant effect of group was found for P450scc (P<0.01) and P450c17 (P<0.05), with the lowest intensity being observed in DG spermatocytes. At the mRNA level, the upregulation from DG spermatocytes to DG round spermatids was clearly evident but was only significant for P450scc (P<0.05). Thus, downregulation affects the whole cascade of steroidogenesis, whereas withdrawal of inhibition results in a rapid restart, in part indicating a rebound phenomenon.
Clinic for Obstetrics, Gynecology and Andrology of Large and Small Animals, Justus-Liebig-University Giessen, Germany. schulzmichaela@gmx.de
M
M
Gentil
B
B
Hoffmann
A
A
Spang
K
K
Failing
S
S
Goericke-Pesch
D000818:Animals; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000096:biosynthesis* / Q000235:genetics; D004285:Dogs; D015536:Down-Regulation / Q000187:drug effects; D004343:Drug Implants; D007987:Gonadotropin-Releasing Hormone / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D008297:Male; D017274:Nafarelin / Q000008:administration & dosage / Q000031:analogs & derivatives*; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D013254:Steroid 17-alpha-Hydroxylase / Q000096:biosynthesis* / Q000235:genetics; D013256:Steroids / Q000096:biosynthesis*; D013737:Testis / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D000098682:Steroidogenic Acute Regulatory Protein
D004343:Drug Implants; D010750:Phosphoproteins; D012333:RNA, Messenger; D013256:Steroids; C483115:azaglycylnafarelin; D000098682:Steroidogenic Acute Regulatory Protein; D017274:Nafarelin; D007987:Gonadotropin-Releasing Hormone; D013254:Steroid 17-alpha-Hydroxylase; D002786:Cholesterol Side-Chain Cleavage Enzyme