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Aging has the opposite effect on cAMP and cGMP circadian variations in rat Leydig cells

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Abstract

The Leydig cell physiology displays a circadian rhythm driven by a complex interaction of the reproductive axis hormones and circadian system. The final output of this regulatory process is circadian pattern of steroidogenic genes expression and testosterone production. Aging gradually decreases robustness of rhythmic testosterone secretion without change in pattern of LH secretion. Here, we analyzed effect of aging on circadian variation of cAMP and cGMP signaling in Leydig cells. Results showed opposite effect of aging on cAMP and cGMP daily variation. Reduced amplitude of cAMP circadian oscillation was probably associated with changed expression of genes involved in cAMP production (increased circadian pattern of Adcy7, Adcy9, Adcy10 and decreased Adcy3); cAMP degradation (increased Pde4a, decreased Pde8b, canceled rhythm of Pde4d, completely reversed circadian pattern of Pde7b and Pde8a); and circadian expression of protein kinase A subunits (Prkac/PRKAC and Prkar2a). Aging stimulates expression of genes responsible for cGMP production (Nos2, Gucy1a3 and Gucy1b3/GUCYB3) and degradation (Pde5a, Pde6a and Pde6h) but the overall net effect is elevation of cGMP circadian oscillations in Leydig cells. In addition, the expression of cGMP-dependent kinase, Prkg1/PRKG1 is up-regulated. It seems that aging potentiate cGMP- and reduce cAMP-signaling in Leydig cells. Since both signaling pathways affect testosterone production and clockwork in the cells, further insights into these signaling pathways will help to unravel disorders linked to the circadian timing system, aging and reproduction.

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Abbreviations

5-AMP:

5′ Adenosine monophosphate

Adcy/ADCY:

Gene/protein for adenylyl cyclase

BSA:

Bovine serum albumin

cAMP:

Cyclic adenosine monophosphate

Car2 :

Gene for carbonic anhydrase 2

cGMP:

Cyclic guanosine monophosphate

CREB:

Protein for cAMP response element-binding protein

Cyp11a :

Gene for cytochrome P450 side chain cleavage enzyme

Cyp17a :

Gene for 17α-hydroxylase/C17–20 lyase

DHT:

Dihydrotestosterone

Gapdh :

Gene for glyceraldehyde 3-phosphate dehydrogenase

GATA4:

Protein for GATA4

Gucy/GUCY:

Gene/protein for guanylyl cyclases

Hsd17b :

Gene for hydroxysteroid dehydrogenase 17β

Hsd3b/HSD3B:

Gene/protein for hydroxysteroid dehydrogenase 3β

LH:

Luteinizing hormone

Lhr/LHR:

Gene/protein for luteinizing hormone receptor

LIPE:

Protein for hormone-sensitive lipase

Nos/NOS:

Gene/protein for nitric oxide synthase

Pde/PDE:

Gene/protein for phosphodiesterase

Prka/PRKA:

Gene/protein for cAMP-dependent protein kinase

Prkac :

Gene for cAMP-dependent protein kinase, catalytic subunit

Prkar :

Gene for cAMP-dependent protein kinase, regulatory subunit

Prkg/PRKG:

Gene/protein for cGMP-dependent protein kinase

RIA:

Radioimmunoassay

ROS:

Reactive oxygen species

RQ-PCR:

Relative quantification polymerase chain reaction

SCN:

Suprachiasmatic nucleus

SF1:

Protein for steroidogenic factor 1

Star/StAR:

Gene/protein for steroidogenic acute regulatory protein

T:

Testosterone

ZT:

Zeitgeber (German: “time giver”) time

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Acknowledgements

We are very grateful to Professor Gordon Niswender (Colorado State University) for supplying antibodies for radioimmunoassay analysis. Also, we are thankful to Ms Marica Jovic for technical assistance. Figure 2a is reused from Experimental Gerontology 73 (2016) 5–13 (license number 3895230854733).

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Authors and Affiliations

  1. LaRES, Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Dositeja Obradovica Sq 2, Novi Sad, 21000, Serbia

    Aleksandar Z. Baburski, Srdjan J. Sokanovic, Silvana A. Andric & Tatjana S. Kostic

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  1. Aleksandar Z. Baburski
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  2. Srdjan J. Sokanovic
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  3. Silvana A. Andric
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  4. Tatjana S. Kostic
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Correspondence to Tatjana S. Kostic.

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Supported by the Serbian Ministry of Science grant No. 173057 and the Autonomic Province of Vojvodina Grant No. 2856.

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The authors have nothing to disclose.

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Communicated by G. Heldmaier.

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Baburski, A.Z., Sokanovic, S.J., Andric, S.A. et al. Aging has the opposite effect on cAMP and cGMP circadian variations in rat Leydig cells. J Comp Physiol B 187, 613–623 (2017). https://doi.org/10.1007/s00360-016-1052-7

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  • DOI: https://doi.org/10.1007/s00360-016-1052-7

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