The detection of thyrotropin-releasing hormone (TRH) and TRH receptor gene expression in Siberian hamster testes.
Thyrotropin-releasing hormone (TRH) from the hypothalamus is the major regulator of TSH synthesis and secretion. Most recently, TRH and TRH receptors (TRH-R), as well as their mRNAs, have been identified in rat testis. To expand our knowledge on the testicular TRH and TRH receptor gene expression in different species, in the present study the mRNA levels of testicular TRH and TRH-R were investigated in Siberian hamsters. To further localize the cellular sites of the gene expression, the animal model was treated with a single injection of ethylene dimethane sulfonate (EDS) (i.p., 80 mg/kg body weight), a compound known as to specifically eliminate testicular Leydig cells. The elimination of Leydig cells induced by EDS treatment was confirmed by histological studies of the testis sections and by serum hormonal analyses, which showed a dramatic reduction of serum testosterone (T) levels and significantly elevated serum LH concentrations. Messenger RNA levels of TRH and TRH-R in the testes were determined by Northern blot analyses quantitated with densitometry scanning. The results showed that specific TRH-R mRNA, 3.8 kb in size, was identified in Siberian hamster testes and the mRNA levels were significantly elevated in the EDS-treated testes compared to the controls (p < 0.01). Testicular TRH mRNA was also detected; however, no significant differences in TRH mRNA levels were found between EDS-treated and control groups. The size of TRH mRNA was characterized as about 1.2 kb in hamster testes, which was smaller than that observed in the rat hypothalamus (1.6 kb) and in the rat testis (2.0 kb). Further studies by RNase H digestion revealed the presence of smaller TRH transcripts in the hamster testes than those in the rat testis. No hybridization signal for TRH mRNA was detected by RNase protection assay, when a rat TRH riboprobe was applied to hamster testis RNA, suggesting the limited homology of TRH gene sequences between these two species. Our results demonstrate that both TRH and TRH-R genes are expressed in Siberian hamster testes, and a significant increase of TRH-R mRNA levels occurs in the Leydig cell eliminated hamster testes. Unlike the rat testicular TRH mRNA mainly detected in Leydig cells, in hamster TRH mRNA could also be detected in other testicular compartment.
Rao
J N
JN
Department of Medicine, University of Maryland, School of Medicine, Baltimore 21201, USA.
Debeljuk
L
L
Bartke
A
A
Gao
Y P
YP
Wilber
J F
JF
Feng
P
P
D000818:Animals; D000988:Antispermatogenic Agents / Q000633:toxicity; D015152:Blotting, Northern; D006224:Cricetinae; D015870:Gene Expression; D007985:Leydig Cells / Q000187:drug effects; D008297:Male; D008698:Mesylates / Q000633:toxicity; D017111:Phodopus; D012333:RNA, Messenger / Q000096:biosynthesis; D018025:Receptors, Thyrotropin-Releasing Hormone / Q000096:biosynthesis* / Q000235:genetics; D013737:Testis / Q000378:metabolism*; D013739:Testosterone / Q000097:blood; D013973:Thyrotropin-Releasing Hormone / Q000096:biosynthesis* / Q000235:genetics
D000988:Antispermatogenic Agents; D008698:Mesylates; D012333:RNA, Messenger; D018025:Receptors, Thyrotropin-Releasing Hormone; D013739:Testosterone; D013973:Thyrotropin-Releasing Hormone; C002994:ethylene dimethanesulfonate