Identification of neuregulin as a factor required for formation of aligned spermatogonia

F. Kent Hamra, Karen M. Chapman, Derek Nguyen, David L. Garbers

Research output: Contribution to journalArticle

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Abstract

In the absence of somatic cells, medium conditioned by the SNL fibroblast line (SNL-CM) is able to stimulate primary cultures of rat type-A single spermatogonia to develop into chains of aligned spermatogonia at the 8-, 16-, and 32-cell stages. By comparison, medium conditioned by an MSC-1 Sertoli cell line is ineffective. Glial cell line-derived neurotrophic factor (GDNF)-like molecules were identified in SNL-CM and recombinant forms of GDNF, neurturin, and artemin were shown to stimulate formation of aligned spermatogonia, but principally to only the 4- and 8-cell stages. Because SNL-CM and GDNFlike molecules stimulated the formation of spermatogonial chain length differently, we purified components of SNL-CM to identify the additional contributing factor(s). A fraction was isolated that was dependent on GDNF, but required for effective formation of 16- and 32-cell chain lengths. Sequence analysis identified the factor as mouse neuregulin-1. At picomolar concentrations, recombinant neuregulin-1 in combination with GDNF effectively stimulated formation of aligned spermatogonia up to the 32-cell stage. Neuregulin in the absence of GDNF was relatively ineffective. Soluble receptors for neuregulins blocked the effects of GDNF and SNL-CM, suggesting that both neuregulin and GDNF are required for effective formation of long spermatogonial chains. Addition of neuregulin-1 to cultures on MSC-1 feeder layers resulted in spermatogonial behavior similar to that seen on feeder layers of SNL fibroblasts. In fact, SNL cells were found to express 100-fold higher levels of neuregulin-1 transcripts than MSC-1 cells. Thus, we identify neuregulin as a factor required for spermatogonial amplification and differentiation in culture.

Original languageEnglish (US)
Pages (from-to)721-730
Number of pages10
JournalJournal of Biological Chemistry
Volume282
Issue number1
DOIs
StatePublished - Jan 5 2007

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Neuregulins
Glial Cell Line-Derived Neurotrophic Factor
Spermatogonia
Neuregulin-1
Feeder Cells
Fibroblasts
Conditioned Culture Medium
Chain length
Neurturin
Molecules
Sertoli Cells
Amplification
Sequence Analysis
Rats
Cells
Cell Line

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of neuregulin as a factor required for formation of aligned spermatogonia. / Hamra, F. Kent; Chapman, Karen M.; Nguyen, Derek; Garbers, David L.

In: Journal of Biological Chemistry, Vol. 282, No. 1, 05.01.2007, p. 721-730.

Research output: Contribution to journalArticle

Hamra, F. Kent ; Chapman, Karen M. ; Nguyen, Derek ; Garbers, David L. / Identification of neuregulin as a factor required for formation of aligned spermatogonia. In: Journal of Biological Chemistry. 2007 ; Vol. 282, No. 1. pp. 721-730.
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