The spermatid manchette is both a source of fascination and neglect for reproductive and cell biologists. In recent years, there have been severable notable developments in our laboratory that have rekindled attention to the molecular organization and function of the manchette in sperm nuclear shaping and tail morphogenesis. Examples include the isolation of intact manchettes from rodents, the findings that the manchette's perinuclear ring contains keratin 9 (K9) and that tubulin isoforms of the manchette and sperm tail axoneme are associated to Sak 57 (for spermatogenic cell/sperm-associated keratin, Mr 57 kDa). K9 and Sak 57 will prove to be of profound developmental significance in spermatogenesis. K9 is of particular interest because its gene expression occurs in two sites only: the footpad epidermis and testis. In addition, point mutations in a subdomain of the helical rod of human K9 cause epidermolytic palmoplantar keratoderma, a skin disease linked to familial internal malignancies (esophagus, bronchi, breast and ovary). The working hypotheses in this proposal are: 1. K9 gene knockout male mouse will be """"""""manchette-less"""""""". 2. Spermatogenesis in a Sak 57 knockout male mouse will not advance beyond meiotic prophase. The following aims are pursued:
Aim 1 will test that K9 is essential for the assembly of the perinuclear ring of the manchette.
Aim 2 will complete the cDNA cloning of Sak 57, an essential step for further knockout studies to determine the role of this keratin in meiotic prophase and spermiogenesis.
Aim 3 will test that the temporal K9 gene expression correlates with the development of the manchette and that actin stabilizes the manchette at a postacrosomal location.
Aim 4 will test that abnormal tubulin isoforms in the manchette and tail account for nuclear shaping and tail abnormalities in the azh/azh mutant (for abnormal spermatozoon headshape).
Aim 5 will test that the disruption of the Sak 57 subcortical framework in pachytene spermatocytes will result in the cessation of gene expression and arrest of meiosis. Engineering mouse models in which the expression of K9 is null, is important for both reproductive biology and cancer research.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD037282-04
Application #
6521142
Study Section
Reproductive Biology Study Section (REB)
Program Officer
Rankin, Tracy L
Project Start
1999-07-01
Project End
2005-06-30
Budget Start
2002-07-01
Budget End
2005-06-30
Support Year
4
Fiscal Year
2002
Total Cost
$237,570
Indirect Cost
Name
City College of New York
Department
Anatomy/Cell Biology
Type
Other Domestic Higher Education
DUNS #
603503991
City
New York
State
NY
Country
United States
Zip Code
10031
Liska, Frantisek; Gosele, Claudia; Popova, Elena et al. (2013) Overexpression of full-length centrobin rescues limb malformation but not male fertility of the hypodactylous (hd) rats. PLoS One 8:e60859
Kierszenbaum, Abraham L; Rivkin, Eugene; Tres, Laura L et al. (2011) GMAP210 and IFT88 are present in the spermatid golgi apparatus and participate in the development of the acrosome-acroplaxome complex, head-tail coupling apparatus and tail. Dev Dyn 240:723-36
Kierszenbaum, Abraham L; Rivkin, Eugene; Tres, Laura L (2011) Cytoskeletal track selection during cargo transport in spermatids is relevant to male fertility. Spermatogenesis 1:221-230
Kierszenbaum, Abraham L; Rivkin, Eugene; Talmor-Cohen, Anat et al. (2009) Expression of full-length and truncated Fyn tyrosine kinase transcripts and encoded proteins during spermatogenesis and localization during acrosome biogenesis and fertilization. Mol Reprod Dev 76:832-43
Rivkin, Eugene; Kierszenbaum, Abraham L; Gil, Mara et al. (2009) Rnf19a, a ubiquitin protein ligase, and Psmc3, a component of the 26S proteasome, tether to the acrosome membranes and the head-tail coupling apparatus during rat spermatid development. Dev Dyn 238:1851-61
Behnen, Martina; Murk, Kai; Kursula, Petri et al. (2009) Testis-expressed profilins 3 and 4 show distinct functional characteristics and localize in the acroplaxome-manchette complex in spermatids. BMC Cell Biol 10:34
Kierszenbaum, Abraham L; Rivkin, Eugene; Tres, Laura L (2008) Expression of Fer testis (FerT) tyrosine kinase transcript variants and distribution sites of FerT during the development of the acrosome-acroplaxome-manchette complex in rat spermatids. Dev Dyn 237:3882-91
Rivkin, Eugene; Eddy, Edward M; Willis, William D et al. (2005) Sperm tail abnormalities in mutant mice with neo(r) gene insertion into an intron of the keratin 9 gene. Mol Reprod Dev 72:259-71
Kierszenbaum, Abraham L (2004) Polycystins: what polycystic kidney disease tells us about sperm. Mol Reprod Dev 67:385-8
Kierszenbaum, Abraham L; Tres, Laura L; Rivkin, Eugene et al. (2004) The acroplaxome is the docking site of Golgi-derived myosin Va/Rab27a/b- containing proacrosomal vesicles in wild-type and Hrb mutant mouse spermatids. Biol Reprod 70:1400-10

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