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STUB1 directs FOXQ1-mediated transactivation of Ldha gene and facilitates lactate production in mouse Sertoli cells

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Abstract

Sertoli cells (SCs) preferentially use glucose to convert to lactate. As an energy source, lactate is essential for survival of developed germ cells (GCs) due to its anti-apoptotic effect. Failure to maintain lactate metabolism homeostasis leads to infertility or germ cell apoptosis. Several Sertoli cell-expressed genes, such as Foxq1 and Gata4, have been identified as critical regulators for lactate synthesis, but the pathways that potentially modulate their expression remain ill defined. Although recent work from our collaborators pointed to an involvement of STIP1 homology and U-box–containing protein 1 (STUB1) in the modulation of Sertoli cell response to GCs-derived IL-1α, a true physiological function of STUB1 signaling in SCs has not been demonstrated. We therefore conditionally ablated Stub1 in SCs using Amh-Cre. Stub1 knockout males exhibited impaired fertility due to oligozoospermia and asthenospermia, possibly caused by lactate deficiency. Furthermore, by means of chromatin immunoprecipitation, in vivo ubiquitination, and luciferase reporter assays, we showed that STUB1 directed forkhead box Q1 (FOXQ1)-mediated transactivation of the lactate dehydrogenase A (Ldha) gene via K63-linked non-proteolytic polyubiquitination, thus facilitating lactate production in follicle-stimulating hormone (FSH)-stimulated SCs. In agreement, overexpression of LDHA by lentivirus infection effectively rescued the lactate production in TM4Stub1−/− cells. Our results collectively identify STUB1-mediated transactivation of FOXQ1 signaling as a post-translationally modified transcriptional regulatory network underlying nursery function in SCs, which may nutritionally contribute to Sertoli cell dysfunction of male infertility.

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Data Availability statement

All data generated or analyzed during this study are included in this published article and its supplementary information files.

Abbreviations

SCs:

Sertoli cells

GCs:

Germ cells

BTB:

Blood–testis barrier

GLUT1:

Glucose transporter 1

LDHA:

Lactate dehydrogenase A

MCT1:

Monocarboxylate transporter 1

TCA:

Tricarboxylic acid

FSH:

Follicle-stimulating hormone

TH:

Thyroid hormone

TNF-α:

Tumor necrosis factor α

IL-1α:

Interleukin 1α

STUB1:

STIP1 homology and U-box–containing protein 1

CHIP:

C terminus of HSP70-interacting protein

ELK1:

ETS domain transcription factor Elk-1

FOXQ1:

Forkhead box Q1

CASA:

Computer-assisted semen analysis

DMEM/F-12:

Dulbecco’s Modified Eagle Medium/Nutrient Mixture F-12

FCS:

Fetal calf serum

qPCR:

Real-time quantitative polymerase chain reaction

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

H&E:

Hematoxylin and eosin

ChIP:

Chromatin immunoprecipitation

BSA:

Bovine serum albumin

ANOVA:

Analysis of variance

shRNA:

Short hairpin RNA

MDM2:

Murine double minute 2 homolog

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Acknowledgements

The authors thank Dr. Tao Li (Reproductive Center, Baoji Maternal and Child Health Hospital, China) for enthusiastic discussion concerning the potential role of STUB1 in Sertoli cells and excellent scientific support.

Funding

This work was supported by grants from National Natural Science Foundation of China (No. 81901510), and from Research Incubation Fund of Xi’an People’s Hospital (Xi’an NO.4 Hospital) (No. ZD-9).

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Authors and Affiliations

  1. Department of Gynecology and Obstetrics, the First Affiliated Hospital of Xi’an Jiaotong University, No. 277 West Yanta Road, Xi’an 710061, Shaanxi, People’s Republic of China

    Yang Yang & Ruifang An

  2. Reproductive Medicine Center, Xi’an People’s Hospital (Xi’an NO.4 Hospital), 710004, Shaanxi, People’s Republic of China

    Yang Yang, Hongli Zhu & Panpan Shi

  3. Reproductive Medicine Center, Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi’an 710038, Shaanxi, People’s Republic of China

    Yuan Ma & Mao Li

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  1. Yang Yang
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  2. Yuan Ma
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  3. Mao Li
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  4. Hongli Zhu
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  5. Panpan Shi
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Contributions

Study design: AR. Study conduct: YY, MY, LM, ZH, and SP. Data collection: YY, MY, LM, ZH, SP, and AR. Data analysis: YY and AR. Data interpretation: YY and AR. Contribution of reagents: AR. Drafting and submission of manuscript: YY and AR.

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Correspondence to Ruifang An.

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This study was approved by the Institutional Animal Care and Use Committee of First Affiliated Hospital of Xi’an Jiaotong University (permission number XJTU1AF2018LSK-224). All procedures in studies involving animals were performed in accordance with the ethical standards recommended by Guide for the Care and Use of Laboratory Animals (Eighth Edition).

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Yang, Y., Ma, Y., Li, M. et al. STUB1 directs FOXQ1-mediated transactivation of Ldha gene and facilitates lactate production in mouse Sertoli cells. Cell Tissue Res 392, 565–579 (2023). https://doi.org/10.1007/s00441-022-03705-x

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