2010 Fiscal Year Final Research Report
Dynamic functional morphology of pheromone signaling-Identification of estrous pheromone and its receptive vomeronasal neurons-
| Project/Area Number |
21790188
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | Iwate Medical University |
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Principal Investigator |
AKUTSU Hitomi Iwate Medical University, 医学部, 助教 (30398482)
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| Project Period (FY) |
2009 – 2010
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| Keywords | フェロモン / カルシウムイメージング / 鋤鼻器 / 生理活性物質 / バイオアッセイ / ラット / 尿 |
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| Research Abstract |
We found out in earlier study that female rats' urine excreted on proestrous stage (proestrous urine) had some bioactivity which induced a specific increase of intracellular Ca^<2+> concentration ([Ca^<2+>]_i) in vomeronasal sensory neurons of male rat. By set such specific [Ca^<2+>]_i increase as a indicator of bioactivity, in this study, we aimed to identify a bioactive substance, a pheromone, which was specific contained in proestrous urine with bioassay systems of calcium imaging of vomeronasal sensory neuron (VSN). Female rat's urine sample, treated with ultrafiltration membrane or boiled, were tested bioactivity with the bioassay system. It became clear that urinary substances which seemed to induce a specific [Ca^<2+>]_i increase have properties as a low molecular and hard to denature by heat. By purification of female rats' urine with HPLC using hydrophobic column, most components of urine were eluted in low concentration acetonitril (0-15%). When HPLC fractions were examined its bioactivity by "a bioassay system based on fluorescent microscope" which was built up in this study, bioactivity was observed as a specific [Ca^<2+>]_i increase in some HPLC fractions of proestrous urine. We have applied the bioassay system using real time laser scanning microscope to measure the responses in individual VSNs, the new bioassay system based on fluorescent microscope was developed to measure the responses of the whole vomeronasal sensory epithelium and the mass of VSN. The purification of urinary component and the bioactivity examination by two ways of bioassay raised a possibility to identify a proestrous urine specific pheromone.
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