| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2000: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Research Abstract |
It is well known about the idea that antidunesis mav be contmlled hv cells in the, hypothalamus acting as 'vesicular osmometer'. There has been considerable debate on the identity and sensitivity of such cells, focusing mainly on the neurons located in osmosensitive nuclei. The aniannrin l(AQR) family of membrane bound water transporting channels is expressed in a variety of origanisms. Channel- forming integral protein of 28 kDa was the first aquaporinto be jdentified and characterized AQP4 is a newly discoberedmembrane of this family. AQP4 iswidely distri but expression of brain cells through AQP4 cDNA did not yet identified. The purpose of this study was the cloning of AQP4 cDNA from the birds and understood the physiological function of AQP4 in related with osmoreception in birds,. The total RNA was extracted from one-day old chick brain and was reverse translated into cDNA was used for PCR with the primenrs designed from mammmalian AQP4 cDNA scquence. The PCR product was subcloned
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sith pGEM-T easy vector, and was sequenced. Five one-day old chicks were dehydrated for on day and their brain was removed. IN control chicks, five brains were removed without the dehydration treatment. The total RNA was extracted from the chick brain and was reverse-translated into cDNA. PCR was performed with chicken AQP4 primers. PCR products was 376bp. Using BLAST program, this PCR products was searchedthe homologous gene. The results showed that this PCR product was highoy similar to AQP4 cDNA sequence of mammalian. For example, between human AQP4 and this PCR product, identity was 312/368(86%), and Score was 333 bits, and between mouse AQP4 and this PCR product, identity was 312/368(86%), and Score was 303 bits. These results highly suggeste that this PCR product might be chiken AQP4. But, this is still partial sequence, and then we are working on the cloning of full sequence of this gene. With RT-PCR, the AQP4 mRNA expression in the dehydrated chicken brain was slightly higher than those in control brains. This result suggest that AQP4 may be relate with osmoregulation. Less
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