생후 7일, 16일된 circling mouse 청각 뇌줄기에서 N-메틸-D 아스파르트산염 수용체(NMDA receptor)에 대한 면역염색학적 분포
Immunohistochemical Localization of NMDA Receptor in the Auditory Brain Stem of Postnatal 7, 16 Circling Mouse
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글루타메이트 수용체는 신경연접의 발달에서 신경연접과 정제 등에 중요한 역할을 한다. 포유류 청각뇌줄기의 가쪽위올리브핵-안쪽마름섬유체핵 신경연접에서의 흥분성 신경전달물질의 전달은 글루타메이트와 같은 많은 흥분성 신경전달물질의 전달을조절하므로 발생학적 연구에 매우 유용한 모델이다. 귀먹음은 흥분성 회로 또는 억제성 회로의 신경연접에서 신경전달물질들의변화뿐 아니라 형태학적 변화도 초래하게 된다. 이런 이유로 선천적으로 태어나면서 달팽이 기관이 퇴화되는 선회생쥐는 귀먹음의 병태생리를 연구하는데 가장 좋은 동물모델이다. 그러나,선회생쥐에서 이런 NMDA 수용체 각각의 아형들이 발달에 따라 어떻게 나타나는지에 관해서는 거의 알려진 사실이 없다. 따라서, 본 연구에서는 선천적으로 속귀에 돌연변이를 가진 선회생쥐를 이용하여 면역조직화학염색법으로 생후 7일과 16일의 쥐를 정상쥐와 비교해서 글루타메이트성의 흥분성 신호가 전달되는 청각뇌줄기의 가쪽위올리브핵(LSO)에서 N-메틸-D-아스파르트산염(NMDA) 수용체의 아형인 NR1, NR2A, NR2B의 분포를각각 조사하였다. 생후 7일째 선회생쥐의 NR1에 대한 면역반응성의 세기는 정상군에서 128.67±8.87로 나타났고 이형접합체에서는 111.06±8.04, 동형접합체에서는 108.09±5.94으로 나타났다. 그리고 생후 16일째 선회생쥐에서는 정상군에서 43.83±10.49, 이형접합체에서는 40±13.88, 동형접합체에서는 55.96±17.35으로 나타났다. 생후 7일째 선회생쥐의 NR2A에 대한 면역반응성의 세기는 정상군에서 97.97±9.71, 이형접합체에서 102.87±9.30, 동형접합체에서 106.85±5.79로 나타났다. 생후 16일 선회생쥐의 가쪽위올리브핵에서의 NR2A에 대한 면역반응성 세기는 정상군에서 47.4±20.6, 이형접합체에서 43.9±17.5, 동형접합체에서 49.2±20.1로 나타났다. 생후 7일의 선회생쥐의 NR2B의면역반응성 세기는 정상군에서 109.04±6.77, 이형접합체에서106.43±10.24, 동형접합체에서 105.98±4.10으로 나타났다. 생후 16일째 선회생쥐의 NR2B의 면역반응성의 세기는 정상군에서 101.47±11.50, 이형접합체에서 91.47±14.81, 동형접합체에서 93.93±15.71으로 나타났다. 이 결과들은 생후 7일과 16일의선회생쥐의 가쪽위올리브핵에서 NMDA 수용체의 면역반응성의변화를 나타낸 것으로 인간 귀먹음의 주요한 병태생리를 파악하는 데 기본적인 자료를 제시할 수 있을 것이다.
Abstract
Glutamate receptors may play a critical role in the refinement of developing synapses. The lateral superior olivary nucleus (LSO)-medial nucleus of trapezoid body (MNTB) synaptic transmission in the mammalian auditory brain stem mediate many excitatory transmitters such as glutamate, which is a useful model to study excitatory synaptic development. Hearing deficits are often accompanied by changes in the synaptic organization such as excitatory or inhibitory circuits as well as anatomical changes. Owing to this, circling mouse whose cochlea degenerates spontaneously after birth, is an excellent animal model to study deafness pathophysiology. However, little is known about the development regulation of the subunits composing these receptors in circling mouse. Thus, we used immunohistochemical method to compare the N-Methyl-D-aspartate receptor (NMDA receptor) NR1, NR2A, NR2B distribution in the LSO which project glutamergic excitatory input into the auditory brainstem, in circling mouse of postnatal (p) 7 and 16, which have spontaneous mutation in the inner ear, with wild-type mouse. The relative NMDAR1 immunoreactive density of the LSO in circling mouse p7 was 128.67±8.87 in wild-type,111.06±8.04 in heterozygote, and 108.09±5.94 in homozygote. The density of p16 circling mouse was 43.83±10.49 in wild-type, 40±13.88 in heterozygote, and 55.96±17.35 in homozygote. The relative NMDAR2A immunoreactive density of LSO in circling mouse p7 was 97.97±9.71 in wild-type, 102.87±9.30 in heterozygote, and 106.85±5.79 in homozygote. The density of LSO in p16 circling was 47.4±20.6 in wild-type, 43.9±17.5 in heterozygote, and 49.2±20.1 in homozygote. The relative NMDAR2B immunoreactive density of LSO in circling mouse p7 was 109.04±6.77 in wild-type, 106.43±10.24 in heterozygote, and 105.98±4.10 in homozygote. the density of LSO in p16 circling mouse was 101.47±11.5 in wild-type, 91.47±14.81 in heterozygote, and 93.93±15.71 in homozygote.
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