Enhanced testosterone secretion in adult rams after establishment of a high-frequency, low-amplitude pattern of LH pulses in the nonbreeding season occurs without changes in the number or binding affinity of testicular LH receptors

1990 ◽  
Vol 122 (1) ◽  
pp. 55-61 ◽  
Author(s):  
Lee M. Sanford ◽  
Susan J. Baker

Abstract When the LH signal in the ram is changed from one of large and infrequent pulses to one of small and frequent pulses, the testes quickly become more responsive to LH and testosterone secretion is elevated, perhaps because the number and (or) binding affinity of testicular LH receptors have increased. An experiment was undertaken in the nonbreeding season (July) with 10 adult Dorset × Leicester × Suffolk rams that were about 3.5 years of age and 69 ± 2 kg in body weight. Rams were given injections into the jugular vein of either 5 μg NIH-LH-S24 (in 1 ml saline) or vehicle every 80 min for 6 days. LH treament produced a series of LH pulses that occurred three times more frequently and were 70% less in amplitude than pulses in the control rams, without causing mean LH concentration to increase. Endogenously produced LH pulses were not evident in the treated rams after LH injection began. The modified LH-pulse pattern elevated mean testosterone concentration by 150% (assessed on days 2 and 5), and caused the cumulative testosterone response to LH pulses, estimated by multiplying testosterone-pulse amplitude by frequency per 6 h, to increase progressively by 180% (days −2 through 5). Enhanced testicular steroidogenic activity, presumably due to greater enzymatic activity and cholesterol availability within Leydig cells, was not associated with increases in either the concentration or affinity of LH-binding sites in the testis (assessed on days 3 and 6).

1989 ◽  
Vol 120 (2) ◽  
pp. 180-186 ◽  
Author(s):  
Lee M. Sanford

Abstract. The testes of the ram become more responsive to LH stimulation following immunoneutralization of endogenous estradiol. The possibility that testosterone secretion is facilitated by increased LH-binding activity in the testes was investigated in the present study conducted with adult Dorset × Leicester × Suffolk rams during the time of testicular recrudescence. Patterns of episodic LH release and testosterone secretion (days –5, 10 and 24) and LH-binding activity in testicular biopsy samples (days –1, 14 and 28) were assessed on the days indicated relative to the onset of passive immunization and the establishment of relatively low titres (~1:200) of estradiol antiserum. During the experimental period, mean serum testosterone concentration increased by approximately 150% for the immunized rams as basal concentration and pulse amplitude increased, while all characteristics of testosterone secretion remained unchanged for the nonimmunized rams. Characteristics of LH release and the concentration of LH-binding sites in the testes, however, were always similar for both groups of rams. Further, group differences in FSH and PRL secretion and in the concentration of testicular FSH-binding sites did not occur. These results provide evidence for an estradiol direct (gonadotropin independent) negativefeedback component in the regulation of Leydig cell function in the ram.


1997 ◽  
Vol 136 (6) ◽  
pp. 633-639 ◽  
Author(s):  
Sandra Valenti ◽  
Massimo Giusti ◽  
Roberta Guido ◽  
Giulio Giordano

Abstract Previous studies have suggested that melatonin (MLT) acts directly on rat Leydig cells by modulating androgen production. In the present study, the site of action of MLT was investigated. The binding of 2-[125I]iodomelatonin (125I-MLT; 7–240 pmol/l) to Leydig cell membrane fragments was tested in the presence or absence of guanosine 5′-O-(3-thiotriphosphate) (GTP-γ-S; 50 μmol/l). Saturation studies and Scatchard analysis revealed the existence of a high-affinity binding site with a Bmax of 46·70± 3·50 fmol/mg protein and a Kd of 88·70±6·20 pmol/l; treatment with GTP-γ-S reduced the concentration of 125I-MLT binding sites (Bmax 34·03±4·50), while increasing the Kd to 106·5± 2·61 pmol/l. Pretreatment of the cells with pertussis toxin (PTX; 10 ng/ml for 16 h) resulted in a decreased binding of I-MLT and a lack of effect of GTP-γ-S. Moreover, the effect of MLT on testosterone secretion induced by LH (30 mIU/ml), forskolin (1 μmol/l) and LHRH (100 nmol/l) was studied after 3-h incubation of cells which had been precultured with or without PTX. The inhibition of testosterone secretion due to MLT administration was eliminated by PTX pretreatment during forskolin and LH, but not during LHRH administration. However, 17-hydroxyprogesterone levels were higher in all groups incubated in the presence of MLT, irrespective of PTX pretreatment. Our data suggest that: (a) MLT receptors are present on the membranes of adult rat Leydig cells; (b) they couple through PTX-sensitive G-protein-coupled binding sites; (c) the mechanism by which MLT blocks 17–20 desmolase enzymatic activity (thus leading to increased 17-hydroxyprogesterone levels), and testosterone secretion during LHRH stimulation is likely to depend on one or more different mechanism(s) of action. European Journal of Endocrinology 136 633–639


2001 ◽  
Vol 67 (3) ◽  
pp. 1085-1089 ◽  
Author(s):  
Salvador Herrero ◽  
Brenda Oppert ◽  
Juan Ferré

ABSTRACT Susceptibility to protoxin and toxin forms of Cry1Ab and the binding of 125I-labeled Cry1Ab and Cry1Ac has been examined in three Plodia interpunctella colonies, one susceptible (688s) and two resistant (198r and Dplr) to Bacillus thuringiensis. Toxicological studies showed that the 198r colony was 11-fold more resistant to Cry1Ab protoxin than to Cry1Ab activated toxin, whereas the Dplr colony was 4-fold more resistant to protoxin versus toxin. Binding results with 125I-labeled toxins indicated the occurrence of two different binding sites for Cry1Ab in the susceptible insects, one of them shared with Cry1Ac. Cry1Ab binding was found to be altered in insects from both resistant colonies, though in different ways. Compared with the susceptible colony, insects from the Dplr colony showed a drastic reduction in binding affinity (60-fold higher Kd ), although they had similar concentrations of binding sites. Insects from the 198r colony showed a slight reduction in both binding affinity and binding site concentration (five-fold-higherKd and ca. three-fold-lowerRt compared with the 688s colony). No major difference in Cry1Ac binding was found among the three colonies. The fact that the 198r colony also has a protease-mediated mechanism of resistance (B. Oppert, R. Hammel, J. E. Throne, and K. J. Kramer, J. Biol. Chem. 272:23473–23476, 1997) is in agreement with our toxicological data in which this colony has a different susceptibility to the protoxin and toxin forms of Cry1Ab. It is noteworthy that the three colonies used in this work derived originally from ca. 100 insects, which reflects the high variability and high frequency of B. thuringiensisresistance genes occurring in natural populations.


1993 ◽  
Vol 136 (2) ◽  
pp. R1-R4 ◽  
Author(s):  
D. Conte ◽  
P. Questino ◽  
S. Fillo ◽  
M. Nordio ◽  
A. Isidori ◽  
...  

ABSTRACT The present study was designed to evaluate the effects of endothelin (ET) on rat testicular steroidogenesis in vitro and the involvement of prostaglandins (PG) and extracellular calcium in its mechanism of action. To this purpose we examined the effects of ET-1 and ET-3 on basal testosterone secretion, the influence of ET-1 on PGE2, release, the interaction of ET-1 and ET-3 with human chorionic gonadotrophin (hCG) and the interference of indomethacin (an inhibitor of cycloxygenase) and nifedipine (a calcium-channel blocker) in purified rat Leydig cells. The data indicate that ET-1 and ET-3 stimulate basal and hCG-induced testosterone production although the effects of ET-3 were less marked. In addition, a concomitant release of PGE2, was observed after exposure to ET-1. A sinergistic interaction between ET-1 and hCG in stimulating testicular steroidogenesis was revealed. Indomethacin was ineffective in modifying ET-1 evoked testosterone output, while in the presence of nifedipine the stimulatory effect of ET-1 was completely abolished. Since it has been shown by others that ET-1 is produced by rat Sertoli cells and specific binding sites are present in Leydig cells, the results of our study indicate that such a peptide may be regarded as a new paracrine factor able to influence steroidogenesis in Leydig cells. The action of ET-1 requires the activity of voltage-operated Ca2+ channels, while PGE2 activation is not essential for its steroidogenic effect.


1985 ◽  
Vol 21 (2) ◽  
pp. 199-205 ◽  
Author(s):  
Bunzo Sato ◽  
Yoshiaki Maeda ◽  
Makoto Nakao ◽  
Keizo Noma ◽  
Susumu Kishimoto ◽  
...  

2009 ◽  
Author(s):  
James P. Ryle ◽  
Mohammed Al-Kalbani ◽  
Unnikrishnan Gopinathan ◽  
Gerard Boyle ◽  
Davis Coakley ◽  
...  

2013 ◽  
Vol 368 (1632) ◽  
pp. 20130018 ◽  
Author(s):  
Andrea I. Ramos ◽  
Scott Barolo

In the era of functional genomics, the role of transcription factor (TF)–DNA binding affinity is of increasing interest: for example, it has recently been proposed that low-affinity genomic binding events, though frequent, are functionally irrelevant. Here, we investigate the role of binding site affinity in the transcriptional interpretation of Hedgehog (Hh) morphogen gradients . We noted that enhancers of several Hh-responsive Drosophila genes have low predicted affinity for Ci, the Gli family TF that transduces Hh signalling in the fly. Contrary to our initial hypothesis, improving the affinity of Ci/Gli sites in enhancers of dpp , wingless and stripe , by transplanting optimal sites from the patched gene, did not result in ectopic responses to Hh signalling. Instead, we found that these enhancers require low-affinity binding sites for normal activation in regions of relatively low signalling. When Ci/Gli sites in these enhancers were altered to improve their binding affinity, we observed patterning defects in the transcriptional response that are consistent with a switch from Ci-mediated activation to Ci-mediated repression. Synthetic transgenic reporters containing isolated Ci/Gli sites confirmed this finding in imaginal discs. We propose that the requirement for gene activation by Ci in the regions of low-to-moderate Hh signalling results in evolutionary pressure favouring weak binding sites in enhancers of certain Hh target genes.


1990 ◽  
Vol 70 (1) ◽  
pp. 121-128 ◽  
Author(s):  
V. L. TRUDEAU ◽  
L. M. SANFORD

Seasonal variations in LH, FSH, and testosterone secretion were investigated for adult Landrace boars housed in different social environments for 1 yr. Socially nonrestricted boars (n = 4) were penned adjacent to ovariectomized gilts that were hormonally brought into estrus every 2 wk, while socially restricted boars (n = 4) were kept in pens with solid walls. Mean hormone concentrations were determined from the assay of single AM and PM blood samples collected from the jugular vein by venipuncture once a month. In November, February, May and August, blood samples were collected serially over 12 h from jugular catheters for assessment of pulsatile LH and testosterone secretion, and the LH response to a GnRH injection (1 μg kg−1 body weight). Mean LH and testosterone concentrations were relatively high in all boars during the late summer and fall, and often were greater for the socially nonrestricted versus the restricted boars (group × month), P < 0.05) in the winter (December and January). Mean FSH concentration also varied with month (P < 0.05). Pulse analysis indicated that higher mean testosterone concentrations in November and August were the result of increases (month, P < 0.05) in testosterone-pulse frequency and basal concentration. Maximal mean LH concentration in August was associated with maximal (month, P < 0.05) LH-pulse amplitude and basal concentration. The amplitude of the LH peak following GnRH injection increased (P < 0.05) between November and May, and remained high in August. Key words: Gonadotropins, testosterone, blood, season, social environment, boar


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