Targeted disruption of genes in the
            <i>Bombyx mori</i>
            sex pheromone biosynthetic pathway

Ohnishi, Atsushi; Hull, J. Joe; Matsumoto, Shogo

doi:10.1073/pnas.0511270103

Cited by 109 publications

(81 citation statements)

References 37 publications

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“…The knockdown efficacy, however, was not complete, as bombykol production was not abolished. This partial reduction may have been the result of inefficient uptake of the dsRNAs by the pheromone producing cells, which were injected on the day of emergence and not during PG development as previously described (40). Alternatively, the partial reduction may be an indication that more than one type of channel is opened following PBAN binding.…”

Section: Discussionmentioning

confidence: 91%

“…These data provide a clear role for the STIM1-Orai1 pathway in ROC Ca 2ϩ influx and suggest that under physiological conditions the PBAN-PBANR complex in the PG triggers the opening of SOCs via translocation of endogenous ER-bound BmSTIM1 to regions of the plasma membrane replete with BmOrai1B molecules. In Vivo Knockdown of BmSTIM1 and BmOrai1-We previously demonstrated the effectiveness of dsRNA-mediated RNA interference in elucidating the downstream cellular targets of PBAN binding (40). We expected that a similar strategy would allow us to determine whether BmSTIM1 and BmOrai1B function within the biological context of sex pheromone produc- tion.…”

Section: Journal Of Biological Chemistry 31207mentioning

confidence: 99%

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Bombyx mori Homologs of STIM1 and Orai1 Are Essential Components of the Signal Transduction Cascade That Regulates Sex Pheromone Production

Hull

Lee

Kajigaya

et al. 2009

Journal of Biological Chemistry

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Sex pheromone production in the pheromone gland (PG) of the silkmoth, Bombyx mori, is mediated by store-operated channels (SOCs) acting downstream of pheromone biosynthesis activating neuropeptide (PBAN) binding. Although recent studies have implicated STIM1 and Orai1 as essential components of SOCs, little is known about the molecular nature of the SOCs involved in sex pheromone production. In this study we cloned silkmoth homologs of STIM1 and Orai1 and sought to determine whether they comprise the PG SOC pathway. BmSTIM1 is expressed in multiple tissues and, in the PG, is encoded by two transcripts of differing size. BmOrai1A and BmOrai1B, which are identical except for a 37-residue N-terminal truncation in BmOrai1B, arise from alternative splicing of the bmorai1 locus and are expressed as independent transcripts in various tissues. In the PG, only BmOrai1B is actively transcribed. Fluorescent chimeras demonstrated that BmSTIM1 expression is restricted to the endoplasmic reticulum, whereas both BmOrai1A and BmOrai1B localize to the cell surface. In Ca 2؉ -free medium, thapsigargin-mediated depletion of endoplasmic reticulum Ca 2؉ stores resulted in redistribution of BmSTIM1 to the plasma membrane, but only when the BmOrai1 homologs were also overexpressed. Translocation was dependent on the BmSTIM1 C terminus "CRAC activation domain." Ala mutation of Lys 380 , Lys 383 , Lys 384 , Arg 382 , and Arg 385 suggests that translocation involves electrostatic interactions. Translocation was also seen following PBAN stimulation in cells co-expressing BmSTIM1, BmOrai1B, and the PBAN receptor. In vivo RNA interference-mediated knockdown of BmSTIM1 and BmOrai1 significantly reduced sex pheromone production without affecting cell viability.

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Section: Discussionmentioning

confidence: 91%

Section: Journal Of Biological Chemistry 31207mentioning

confidence: 99%

Bombyx mori Homologs of STIM1 and Orai1 Are Essential Components of the Signal Transduction Cascade That Regulates Sex Pheromone Production

Hull

Lee

Kajigaya

et al. 2009

Journal of Biological Chemistry

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“…This result supports the notion that expression of these genes is associated with pheromone production and regulation. However, further investigations are needed, including establishment of a primary culture of sebaceous gland cells [17,18] and functional analysis of these genes using cultured cells [19], in order to provide conclusive evidence that the pheromone is produced in this gland and that these genes play important roles in pheromone synthesis and regulation. These in vitro approaches, in conjunction with in vivo studies, will also help to elucidate the mechanism underlying production of the 'male effect' pheromone.…”

Section: Discussionmentioning

confidence: 99%

Localization of the Candidate Genes ELOVL5 and SCD1 for Male Effect' Pheromone Synthesis in Goats (Capra hircus)

et al. 2007

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Abstract. The 'male effect' is a well-known phenomenon in female sheep and goats, whereby pheromone-induced activation of reproductive function occurs. In a previous study, we showed that the genes for elongation of long-chain fatty acids family member 5 (ELOVL5) and stearoyl-CoA desaturase 1 (SCD1) increased their expression significantly, concomitant with induction of pheromone synthesis. Therefore, these genes were considered to be prime candidate genes for pheromone synthesis. In the present study, we performed in situ hybridization to investigate where these two genes are expressed in goat skin. Strong positive signals were detected for both genes in the head skin of the male goat, which is the main site of pheromone production, and were mainly in the basal layer of the sebaceous gland cells, with the remaining cells showing negligible signals. None of the cells in the rump skin of the male goat or the head skin of the orchidectomized goat, neither of which produce pheromone, exhibited strong positive signals. The present study demonstrates that expression of these two candidate genes for pheromone synthesis is primarily localized in the sebaceous glands of the pheromone-producing skin region. , the 'male effect' is well known as a pheromone-induced phenomenon in which seasonally anovulatory females resume ovarian cyclicity in response to introduction of a male. In goats, we have previously shown that the 'male effect' pheromone is produced in a testosterone (T)-dependent manner in the skin of the head region, but not in the skin of the rump region [4,5], using a bioassay for pheromone activity that we had developed previously [6,7]. Dihydrotestosterone (DHT) treatment induces pheromone production in the skin of the rump region, where pheromones are not produced constitutively [8]. Interestingly, 'male effect' pheromone activity is also induced in female skin by androgens [9]. We found that expression of the genes for elongation of long-chain fatty acids family member 5 (ELOVL5) and stearoyl-CoA desaturase 1 (SCD1) increased significantly, concomitant with induction of pheromone synthesis, in three types of skin models, i.e., the head skin of Ttreated orchidectomized (ODX) male goats, rump skin of DHT-treated ODX male goats and head skin of DHT-treated ovariectomized female goats, thereby implicating these two genes as prime can-

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“…These PSG cells are responsible for the active intracellular transport during secretion [24], which may require a broad participation of kinesins and other transport-specific proteins. In addition, RNAi technology recently established in silkworm [25] has enabled researchers to knockdown target gene transcripts by injection of dsRNA into adult or juvenile insects. Thus, RNAi technology, in combination with the BmNPV baculoviruses transfection system [26], transgenic silkworm technique [27], and micromanipulation tools in silkworm cells [28], offers a great opportunity to assess the physiological function of individual protein in certain cellular processes and developmental stages.…”

Section: Introductionmentioning

confidence: 99%

Characterization of kinesin-like proteins in silkworm posterior silkgland cells

et al. 2010

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Targeted disruption of genes in the Bombyx mori sex pheromone biosynthetic pathway

Cited by 109 publications

References 37 publications

Bombyx mori Homologs of STIM1 and Orai1 Are Essential Components of the Signal Transduction Cascade That Regulates Sex Pheromone Production

Bombyx mori Homologs of STIM1 and Orai1 Are Essential Components of the Signal Transduction Cascade That Regulates Sex Pheromone Production

Localization of the Candidate Genes ELOVL5 and SCD1 for Male Effect' Pheromone Synthesis in Goats (Capra hircus)

Characterization of kinesin-like proteins in silkworm posterior silkgland cells

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