?Hybrid? X-Y translocation chromosomes of Drosophila hydei and D. neohydei

Hennig, Ingrid

doi:10.1007/bf00330123

Cited by 7 publications

(1 citation statement)

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“…Based on differential Giemsa staining (16) and cytogenetic criteria (17,18), the "proximal loops" of D. neohydei and the pseudonucleolus of D. eohydei are considered as homologous to the pseudonucleolus of D. hydei. Both assignments are confirmed by the immunofluorescence staining with the antiserum.…”

Section: Resultsmentioning

confidence: 99%

Lampbrush loop-specific protein of Drosophila hydei.

Hulsebos¹,

Hackstein²,

Hennig³

1984

Proc. Natl. Acad. Sci. U.S.A.

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By immunofluorescence techniques and protein blotting experiments we have shown that an antiserum specifically reacts with a Mr 80,000 protein (the "Ps protein") in the lampbrush loop "pseudonucleolus" in spermatocyte nuclei of Drosophila hydei. Comparative studies of X/Y and X/0 testes indicate that the gene encoding the Ps protein is not located on the Y chromosome but on an autosome or the X chromosome. The Ps protein is tissue specific. It is likely to be a rather conserved protein since the antigenic determinant recognized by the antiserum could be detected in the spermatocyte nuclei of a number of other Drosophila species. For those species with prominent Y chromosomal lampbrush loops, it could be shown that the cross-reaction is, as in D. hydei, associated with a specific Y chromosomal loop.During the primary spermatocyte stage the Y chromosome of Drosophila hydei (and many other Drosophila species) develops characteristic lampbrush loops of distinct morphology (1). The loops (2) result from transcription (3) MATERIALS AND METHODS Drosophila Stocks. D. hydei, wild type, Drosophila neohydei, wild-type, and Drosophila melanogaster, wild-type, were from our laboratory collection. D. eohydei was obtained from H. Beck, Geneva. All other Drosophila species came from the Drosophila Stock Center (Austin, TX).The 74/0 containing a Y chromosomal fragment with the pseudonucleolus (complementation groups B and C); and -X/ms (Y)C4 male sterile mutation in complementation group C, which "lacks" the pseudonucleolus in phase contrast. Immunological Procedures. Preparation of the antiserum against sph155 and the immunoblotting procedure have been described (14).For immunofluorescence staining, spermatocyte regions were collected in a drop of testis preparation buffer (10 mM Tris'HCl, pH 6.8/47 mM NaCl/183 mM KCl) on a microscopic slide. Coverslips were applied, and the preparations were gently squashed and then frozen in liquid nitrogen for 15 s. The coverslips were removed with a surgical knife and the preparations were passed through a series of ethanol washes (90%, 60%, 30%, 10%, 2 min each step). They were washed twice with phosphate-buffered saline (P1/NaCl), fixed in 3.7% formaldehyde in P1/NaCl for 10 min, and then incubated for 10 min with 3.7% formaldehyde/45% acetic acid in Pi/NaCl and extensively washed with Pi/NaCl. For immunofluorescence staining of spermatids and spermatozoa, the preparations were permeabilized for 5 min with 1% (wt/vol) Triton X-100 in Pi/NaCl and then washed. This treatment was omitted in cases in which spermatocyte nuclei were examined. The preparations were incubated for 30 min with 50 ,ul of antiserum or with pre-immune serum (both diAbbreviation: Pi/NaCl, phosphate-buffered saline. 3404The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Section: Resultsmentioning

confidence: 99%