A Guide to the Use of Spermatogenesis Markers

A Guide to the use of Spermatogenesis Markers: which ones should you use?

DDX4/VASA/MVH

Present in both male and female germ cells’ cytoplasms is germ cell-specific ATP-dependent RNA helicase.

P6 CD-1 testis stained with anti-DDX4/MVH (ab13840) primary antibody and AlexaFluor^® 488 secondary antibody (green). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

ZBTB16/PLZF

Functioning as a putative transcriptional repressor, this is a nuclear protein.

P6 CD-1 testis stained with anti-PLZF (ab189849) primary antibody and AlexaFluor^® 555 secondary antibody (green). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

POU5F1/OCT4

Functioning as a transcription factor, this is a homeodomain-containing protein.

P6 mouse testis stained with anti-OCT4 (ab181557) antibody and AlexaFluor^®488 secondary antibody (green). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

STRA8

With an unknown function, this nuclear protein is necessary for meiotic entry in both sexes of mice to occur.

Adult CD-1 testis stained with anti-STRA8 antibody (ab49602) and AlexaFluor^®488 secondary antibody (green). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

KIT/c-KIT

Expressed in primordial germ cells (PGCs), receptor tyrosine kinase is necessary to their survival and also in differentiating spermatogonia.

P6 CD-1 testis stained with anti-KIT primary antibody (ab65525) and goat anti-rabbit AlexaFluor^® 488 secondary antibody (green, ab150081). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

SYCP3

A structural element of the synaptonemal complex, SYCP3 is formed during meiosis at the zygotene stage.

P6 CD-1 testis stained with anti-SCP3 primary antibody (ab15093), using AlexaFluor^® 488 secondary antibody (green). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

DAZL

Normally located in the cytoplasm, this is an autosomally-encoded RNA binding protein which is expressed solely in male and female germ cells.

P6 CD-1 testis stained with anti-DAZL primary antibody (ab34139) and ab150081 goat anti-rabbit AlexaFluor^® 488 secondary antibody (green). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

SALL4

During embryonic development, the transcription factor SALL4 regulates axis formation and can generally be detected in populations of stem and progenitor cells.

P6 CD-1 testis stained with anti-SALL4 primary antibody (ab 29112) and goat anti-rabbit AlexaFluor^® 488 secondary antibody (green, ab150081). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

TRA98

Without a known function, TRA98 is a testis-specific nuclear protein which can be found in all post-natal male germ cells.

Shown above: P6 CD-1 testis stained with anti-TRA98 rat monoclonal (ab82527) primary antibody and ab150165 goat anti-rat AlexaFluor^® 488 secondary antibody (green). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

P6 CD-1 testis stained with anti-TRA98 rat monoclonal (ab82527) primary antibody and goat anti-rat AlexaFluor^® 488 secondary antibody (green, ab150165). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

DPPA3/STELLA

This nuclear protein is part of epigenetic regulation in the fetal gonad, but is not necessary for fertility and is dispensable during male germ cell development.

Adult testis stained with anti-STELLA primary antibody (ab19878) and goat anti-rabbit AlexaFluor^® 488 secondary antibody (green, ab150081). Phalloidin-594 was used to label F-actin (red), and DAPI was used to label nuclei (blue).

References

• Kehler J, Tolkunova E, Koschorz B, Pesce M, Gentile L, Boiani M, Lomelí H, Nagy A, McLaughlin KJ, Schöler HR, Tomilin A. Oct4 is required for primordial germ cell survival. EMBO Rep. 5, 1078–83 (2004).
• Mithraprabhu S, Loveland KL. Control of KIT signalling in male germ cells: what can we learn from other systems? Reproduction 138:743–57 (2009).
• Orr-Urtreger A, Avivi A, Zimmer Y, Givol D, Yarden Y, Lonai P. Developmental expression of c-kit, a proto-oncogene encoded by the W locus. Development 109,911–23 (1990).
• Sato M, Kimura T, Kurokawa K, Fujita Y, Abe K, Masuhara M, Yasunaga T, Ryo A, Yamamoto M, Nakano T. Identification of PGC7, a new gene expressed specifically in preimplantation embryos and germ cells. Mech Dev. 113, 91–4 (2002).

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