We had previously identified a new mammalian A-type cyclin, cyclin A1, in addition to the already known cyclin A2. Both A-type cyclins are expressed in the mouse testis. Expression of the cyclin A1 gene, Ccna1, is germ cell-specific and restricted to late meiotic prophase spermatocytes while Ccna2 is expressed more broadly, in both somatic cells and in spermatogonia and pre-leptotene spermatocytes. Targeted mutagenesis of Ccna1 resulted in viable progeny but male sterility, while females were fully fertile. Spermatocytes in cyclin A1-deficient mice cannot enter into the first meiotic division, the activation of MPF is deficient and the cells undergo apoptosis. In contrast, cyclin A2-deficient mice are embryonic lethal. We now wish to understand the unique regulation and function of cyclin A1 in spermatocytes, to explore the function of cyclin A2 in the male germ line, and to dissect the regulatory pathways that are driven by cyclin A1 and its catalytic Cdk partners during male meiosis. We will also investigate the extraordinarily robust induction of apoptosis that occurs in the cyclin A1- and Cdk2-deficient models. We will identify regulatory elements and corresponding transcription factors required for the proper in vivo expression of Ccna1. This will involve DNase 1 hypersensitivity mapping, electrophoretic mobility shift assays (EMSA), chromatin immunoprecipitation (ChIP), expression of reporter constructs in transgenic mice, sequence analysis, and a novel transcription factor screen to identify the regulatory network responsible for both activating and repressing its expression. We will also examine the changes in the events in the cell cycle, chromosome dynamics, and phosphorylation status of putative substrates that result in the arrest at diplotene and apoptosis in cyclin A1 and Cdk2-defident spermatocytes. This will also involve a novel phosphorylation screen to identify substrates of cyclin A1-complexes. Finally, we will determine the affect of targeted conditional mutagenesis of cyclin A2 in spermatogonia. These studies will provide new and novel insight into the function of the mammalian A-type cyclins, information that cannot be obtained from simpler model systems such as yeast, which lack this class of cyclins, or Drosophila, which contain only a single A-type cyclin. They will further enhance our understanding of the control of meiosis as it relates to male infertility and possible new targets for contraception.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD034915-12
Application #
7816867
Study Section
Cellular, Molecular and Integrative Reproduction Study Section (CMIR)
Program Officer
Taymans, Susan
Project Start
1997-09-01
Project End
2012-04-30
Budget Start
2010-05-01
Budget End
2011-04-30
Support Year
12
Fiscal Year
2010
Total Cost
$308,885
Indirect Cost
Name
Columbia University (N.Y.)
Department
Genetics
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
Panigrahi, Sunil K; Manterola, Marcia; Wolgemuth, Debra J (2017) Meiotic failure in cyclin A1-deficient mouse spermatocytes triggers apoptosis through intrinsic and extrinsic signaling pathways and 14-3-3 proteins. PLoS One 12:e0173926
Manterola, Marcia; Sicinski, Piotr; Wolgemuth, Debra J (2016) E-type cyclins modulate telomere integrity in mammalian male meiosis. Chromosoma 125:253-64
Martinerie, Laetitia; Manterola, Marcia; Chung, Sanny S W et al. (2014) Mammalian E-type cyclins control chromosome pairing, telomere stability and CDK2 localization in male meiosis. PLoS Genet 10:e1004165
Wolgemuth, Debra J; Manterola, Marcia; Vasileva, Ana (2013) Role of cyclins in controlling progression of mammalian spermatogenesis. Int J Dev Biol 57:159-68
Panigrahi, Sunil K; Vasileva, Ana; Wolgemuth, Debra J (2012) Sp1 transcription factor and GATA1 cis-acting elements modulate testis-specific expression of mouse cyclin A1. PLoS One 7:e47862
Wolgemuth, Debra J (2011) Function of the A-type cyclins during gametogenesis and early embryogenesis. Results Probl Cell Differ 53:391-413
Wolgemuth, Debra J; Roberts, Shelby S (2010) Regulating mitosis and meiosis in the male germ line: critical functions for cyclins. Philos Trans R Soc Lond B Biol Sci 365:1653-62
Joshi, Ayesha R; Jobanputra, Vaidehi; Lele, Karen M et al. (2009) Distinct properties of cyclin-dependent kinase complexes containing cyclin A1 and cyclin A2. Biochem Biophys Res Commun 378:595-9
Kalaszczynska, Ilona; Geng, Yan; Iino, Tadafumi et al. (2009) Cyclin A is redundant in fibroblasts but essential in hematopoietic and embryonic stem cells. Cell 138:352-65
Wolgemuth, Debra J (2008) Function of cyclins in regulating the mitotic and meiotic cell cycles in male germ cells. Cell Cycle 7:3509-13

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