Difference between revisions of "Radovic 2019b MiP2019"

Revision as of 14:48, 26 September 2019

Conditional deletion of insulin and IGF1 receptors disturbs transcription of the main markers of mitochondrial biogenesis and architecture/fusion in mouse prepubertal steroidogenic cells of testes but not ovaries.

Link: MiP2019

Radovic SM, Starovlah IM, Capo I, Miljkovic D, Nef S, Kostic TS, Andric SA (2019)

Event: MiP2019

Controlled changes in mitochondrial biogenesis and morphology are required for cell survival and homeostasis, but the molecular mechanisms are largely unknown. Here, the male and female knock out prepubertal mice (P21), with insulin and IGF1 receptors deletion in steroidogenic tissues (Insr/Igf1r-DKO), were used to investigate transcription of key regulators of mitochondrial biogenesis (Ppargc1a, Ppargc1b, Pparg, Nrf1, Tfam) in Leydig cells, ovaries and adrenals as well as mitochondrial architecture (Mfn1, Mfn2, Opa1) in Leydig cells.

Accordingly, gene expression was followed employing polymerase chain reaction in real time (RQ-PCR), proteins expression detected by western blot, while hormones level was measured by radioimmuno assay.

Results showed that expression of PGC1, the master regulator of mitochondrial biogenesis and integrator of environmental signals, as well as its downstream target Tfam, significantly decreased in testosterone-producing Leydig cells from Insr/Igf1r-DKO animals. This was followed with reduced transcription of Mtnd1, the core subunit of NADH dehydrogenase belonging to the minimal assembly required for catalysis. Transcription of mitochondrial biogenesis markers remained unchanged in ovaries. Differently, in adrenals, the pattern of mitochondrial markers transcripts was the same in both sexes and, besides Pparg and Tfam, opposite from Leydig cells. Number of mitochondria was decreased but their volume and diameter were increased in Leydig cells from Insr/Igf1r-DKO animals. The transcription level of mitochondrial architecture markers (Mfn1, Mfn2) was significantly increased in Leydig cells from Insr/Igf1r-DKO mice suggesting that the mitochondrial architecture and mitochondrial phase of steroidogenesis was affected in males.

Our results are the first to show that insulin and IGF1 receptors are important for mitochondrial biogenesis in gonadal steroidogenic cells of prepubertal males, but not females, as important regulators of both mitochondrial biogenesis and architecture markers transcription.

• Bioblast editor: Plangger M, Tindle-Solomon L

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Affiliations and support

Radovic SM(1), Starovlah IM(1), Capo I(2), Miljkovic D(2), Nef S(3), Kostic TS(1), Andric SA(1)

Lab Reproductive Endocrinology Signaling, Lab Chronobiology Aging, Centre Excellence CeRES, Fac Sciences
Dept Histology Embryology, Fac Medicine, Univ Novi Sad, Novi Sad, Serbia
Lab Molecular Developmental Biology Sexual Development, Univ Geneva, Geneva, Switzerland. – [email protected]

This work was supported by the grant no. 173057 and grant no. 451-0302807 Centre of Excellence CeRES from the Ministry of Education, Science and Technological Development, Republic of Serbia, grant no. 2130 from the Autonomous Province of Vojvodina as well as grant no. IZ73Z0-128070 from Swiss National Science Foundation.

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 {{Abstract
-|title=[[Image:Photo Sava Radovic.jpg|left|90px|Sava Radovic]] Conditional deletion of insulin and IGF1 receptors in steroidogenic cells changes profile of molecular markers of mitochondrial dynamics in myocard of prepubertal male, but not female mice.
+|title=[[Image:Photo Sava Radovic.jpg|left|90px|Sava Radovic]] Conditional deletion of insulin and IGF1 receptors disturbs transcription of the main markers of mitochondrial biogenesis and architecture/fusion in mouse prepubertal steroidogenic cells of testes but not ovaries.
 |info=[[MiP2019]]
-|authors=Radovic SM, Starovlah IM, Nef S, Kostic TS, Andric SA
+|authors=Radovic SM, Starovlah IM, Capo I, Miljkovic D, Nef S, Kostic TS, Andric SA
 |year=2019
 |event=MiP2019
 |abstract=[[Image:MITOEAGLE-logo.jpg|left|100px|link=http://www.mitoglobal.org/index.php/MITOEAGLE|COST Action MitoEAGLE]]
-Mitochondria are essential for cardiac function since they produce ATP and other essential cellular mitocrine signals, store and buffer calcium, initiate cell death signals, etc. Not surprisingly, mitochondrial dysfunction has been linked to several severe cardiovascular disorders. The contribution of cellular signaling is required for heart homeostasis, but link between them and mitochondrial dynamic is not described well. We used prepubertal male and female mice with insulin/IGF1 receptors deletion in steroidogenic cells (Insr/Igf1r-DKO) to determine the transcription profile of markers of mitochondrial dynamics in myocard, since these mice have decreased level of circulating-sex-steroid-hormones.
+Controlled changes in mitochondrial biogenesis and morphology are required for cell survival and homeostasis, but the molecular mechanisms are largely unknown. Here, the male and female knock out prepubertal mice (P21), with insulin and IGF1 receptors deletion in steroidogenic tissues (Insr/Igf1r-DKO), were used to investigate transcription of key regulators of mitochondrial biogenesis (Ppargc1a, Ppargc1b, Pparg, Nrf1, Tfam) in Leydig cells, ovaries and adrenals as well as mitochondrial architecture (Mfn1, Mfn2, Opa1) in Leydig cells.
-In order to determine relative expression of the genes coding mitochondrial dynamics markers, total RNA was isolated from mouse myocard, first strand cDNA was synthesized and polymerase chain reaction in real time (RQ-PCR) was preformed.
+Accordingly, gene expression was followed employing polymerase chain reaction in real time (RQ-PCR), proteins expression detected by western blot, while hormones level was measured by radioimmuno assay.
-RQ-PCR-results revealed increased transcription of Ppargc1a, the master regulator of mitochondrial dynamics and integrator of environmental signals, as well as his downstream targets Pparg and Nrf1 in hearts of Insr/Igf1r-DKO males, but not females, comparing to controls. The marker of mitochondrial fusion/architecture, Opa1, decreased in hearts from Insr/Igf1r-DKO males, but not females, while Mfn1/Mfn2 remained unchanged in mice of both sexes. The transcription of the main markers of mitophagy (Pink, Prkn) remained unchanged in mice of both sexes, while transcript for Tfeb (master gene involved in autophagy) declined in hearts of both sexes. This was followed by decreased mtDNA copy numbers in hearts of Insr/Igf1r-DKO males, but increased in females, suggesting the importance of sex-steroid- hormones in regulation of mtDNA copy numbers.
+Results showed that expression of PGC1, the master regulator of mitochondrial biogenesis and integrator of environmental signals, as well as its downstream target Tfam, significantly decreased in testosterone-producing Leydig cells from Insr/Igf1r-DKO animals. This was followed with reduced transcription of Mtnd1, the core subunit of NADH dehydrogenase belonging to the minimal assembly required for catalysis. Transcription of mitochondrial biogenesis markers remained unchanged in ovaries. Differently, in adrenals, the pattern of mitochondrial markers transcripts was the same in both sexes and, besides Pparg and Tfam, opposite from Leydig cells. Number of mitochondria was decreased but their volume and diameter were increased in Leydig cells from Insr/Igf1r-DKO animals. The transcription level of mitochondrial architecture markers (Mfn1, Mfn2) was significantly increased in Leydig cells from Insr/Igf1r-DKO mice suggesting that the mitochondrial architecture and mitochondrial phase of steroidogenesis was affected in males.
-According to our best knowledge, these results are the first data showing the transcriptional profile of mitochondrial dynamics markers in myocard of prepubertal mice and can be used as a solid base for further research.
+Our results are the first to show that insulin and IGF1 receptors are important for mitochondrial biogenesis in gonadal steroidogenic cells of prepubertal males, but not females, as important regulators of both mitochondrial biogenesis and architecture markers transcription.
 |editor=[[Plangger M]], [[Tindle-Solomon L]]
 }}
-{{Labeling
+{{Labeling}}
-|area=nDNA;cell genetics, Genetic knockout;overexpression, Gender
-|organism=Mouse
-|tissues=Heart
-}}
 == Affiliations and support ==
-::::Radovic SM(1), Starovlah IM(1), Nef S(2), Kostic TS(1), Andric SA(1)
+::::Radovic SM(1), Starovlah IM(1), Capo I(2), Miljkovic D(2), Nef S(3), Kostic TS(1), Andric SA(1)
+::::#Lab Reproductive Endocrinology Signaling, Lab Chronobiology Aging, Centre Excellence CeRES, Fac Sciences
-::::#Lab Reproductive Endocrinology Signaling, Lab Chronobiology Aging, Centre Excellence CeRES, Fac Sciences, Univ Novi Sad, Novi Sad, Serbia
+::::#Dept Histology Embryology, Fac Medicine, Univ Novi Sad, Novi Sad, Serbia
 ::::#Lab Molecular Developmental Biology Sexual Development, Univ Geneva, Geneva, Switzerland. – [email protected]
 ::::This work was supported by the grant no. 173057 and grant no. 451-0302807 Centre of Excellence CeRES from the Ministry of Education, Science and Technological Development, Republic of Serbia, grant no. 2130 from the Autonomous Province of Vojvodina as well as grant no. IZ73Z0-128070 from Swiss National Science Foundation.