A genetic selection for isolating cDNAs encoding secreted proteins

Jacobs, Kenneth; Collins-Racie, Lisa A.; Colbert, Maureen; Duckett, McKeough; Golden-Fleet, Margaret; Kelleher, Kerry; Kriz, Ronald; LaVallie, Edward R.; Merberg, David; Spaulding, Vikki; Stover, Julia; Williamson, Mark J.; McCoy, John

doi:10.1016/s0378-1119(97)00330-2

Cited by 277 publications

(281 citation statements)

References 33 publications

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“…We have initiated a systematic analysis of membrane and secreted proteins that are expressed by sperm cells, particularly those expressed exclusively by the germ cells. The approach is to screen an enriched spermatid cDNA library by using a yeast-based signal peptide selection method (16,17). With this method, a functional bias is not introduced and the cell population can remain heterogeneous with respect to physiological status.…”

Section: Resultsmentioning

confidence: 99%

“…To alleviate the problem of asynchrony, we have applied a method known as signal peptide trapping (16,17) to selectively define membrane and secreted proteins of spermatozoa. This yeast-based method has distinct advantages in that cDNA libraries can be both subtracted from other tissue cDNA and normalized to aid in identifying rare, sperm-specific surface proteins.…”

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confidence: 99%

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A voltage-gated ion channel expressed specifically in spermatozoa

Quill

Ren

Clapham

et al. 2001

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

281

276

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Calcium ions play a primary role in the regulation of sperm cell behavior. We report finding a voltage-gated ion channel (CatSper2) that is expressed in male germ cells but not in other cells. The putative channel contains 6 transmembrane segments, making it more similar to the voltage-gated potassium channels, but the ion selectivity pore domain sequence resembles that of a Ca v channel. The mRNA is expressed during the meiotic or postmeiotic stages of spermatogenesis, and the protein is localized to the sperm flagellum, suggesting a role in the regulation of sperm motility. The mRNA for the channel is present in mouse, rat, and human sperm cells, and the gene is found on chromosome 2 E5-F1 in the mouse and 15q13 in the human. Recently, another voltagegated channel (CatSper) that has features similar to the one reported here was discovered. It also is expressed within the flagellum and is required for normal fertility of mice. However, expression of CatSper2 alone or coexpression with CatSper in cultured cells, or attempts to coimmunoprecipitate the two proteins from germ cells failed to demonstrate that these two unique but similar ␣-like subunits form either a homo-or heterotetramer. It is possible, therefore, that two independent ␣ subunits, different from other known voltage-gated channels, regulate sperm motility.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

A voltage-gated ion channel expressed specifically in spermatozoa

Quill

Ren

Clapham

et al. 2001

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

281

276

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“…Yeast transformants from the otocyst cDNA library were screened, and 440 positive clones were obtained by the signal sequence trap method as described previously (Jacobs et al, 1997). Nucleotide sequences of some positive clones were determined, and redundant clones among the 440 positive clones were identified by dot-blot hybridization.…”

Section: Isolation Of Oc29 From Rat Otocystmentioning

confidence: 99%

“…Poly (A) ϩ RNA was extracted from otocysts with TRIzol reagent (GIBCO) and Oligotex-MAG mRNA Purification Kit (TaKaRa). The construction of the cDNA library and the screening by the signal sequence trap method was carried out as described previously (Jacobs et al, 1997). The sequence of the partial cDNA fragment of OC29 was determined.…”

Section: Signal Sequence Trap Screening and Cloning Of Rat Oc29mentioning

confidence: 99%

“…For this purpose, we used the signal sequence trap (SST) method (Tashiro et al, 1993Jacobs et al, 1997), which allowed us to effectively isolate many secreted and membrane proteins with a wide variety of functions (Tashiro et al, 1993;Yabe et al, 1997;Nakamura et al, 1998Nakamura et al, , 1999Kato et al, 2000;Nakashiba et al, 2000;Kobuke et al, 2001;Toda et al, 2003), including a tissue-specific molecule (Michishita et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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OC29 is preferentially expressed in the presumptive sensory organ region of the otocyst

Nishida

Kobuke

Kojima

et al. 2004

Developmental Dynamics

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The mammalian inner ear derives from the otocyst. Molecular mechanisms underlying inner ear development are largely unknown. We have isolated a secreted molecule, OC29, from a rat otocyst cDNA library by the signal sequence trap method. OC29 was revealed to be a rat homologue of human WFIKKN. OC29 is preferentially expressed in the developing inner ear and the dorsal neural tube. In the inner ear, the expression of OC29 is first detectable at embryonic day 11.5 (E11.5), broadly in the dorsolateral region of the otocyst, which gives rise to the vestibular organ. At E12.5, the expression of OC29 becomes restricted to the presumptive sensory region, mainly to the BMP4-positive presumptive cristae, and expression becomes reduced at later stages. These results suggest that OC29 may have a role in the early development of the inner ear sensory organ, particularly in the formation of the cristae of the semicircular canals. Developmental Dynamics 231:766 -774, 2004.

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